Note: Descriptions are shown in the official language in which they were submitted.
CA 02795952 2012-10-09
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TREATMENT OF CANCERS HAVING K-RAS MUTATIONS
RELATED APPLICATION
This application claims the benefit of U.S. Provisional Application No.
61/324,912,
filed on April 16, 2010. The entire teachings of the above application are
incorporated
herein by reference.
BACKGROUND OF THE INVENTION
The members of the ras gene family encode membrane-bound proteins of about 21
kD which are referred to as p21-ras. These proteins are believed to relay
growth and
differentiation signals from activated receptors to intracellular protein
kinases. The ras
gene family includes K-ras, H-ras and N-ras. The ras genes are proto-
oncogenes; mutated
ras genes are found in a variety of cancer types. These mutants typically
exhibit a single
base pair substitution, usually in codon 12, 13, 146 or 161, which leads to an
expressed
p21-ras product with a point mutation at the corresponding amino acid
position. In wild
type K-ras, codon 12 codes for glycine, but in the most common mutations this
codon
codes for aspartic acid, valine, arginine or cysteine (Mu et at., World J.
Gastroenterol.
2004, 10: 471-475). While wild-type p21-ras is activated only by an activated
receptor and
deactivated after downstream signaling, the p21-ras mutants are constitutively
active, and
thus stimulate growth and differentiation continuously, in the absence of a
signal (Bos, J.L.,
Cancer Res. 1989, 49: 4682-4689).
Mutated ras family genes occur in variety of cancers. In particular, K-ras
mutations
are prevalent in pancreatic, colorectal, thyroid, lung, cervical and
endometrial cancer. K-
ras mutations have also been identified in certain precancerous conditions,
such as
myelodysplastic syndrome and adenomas of the thyroid and the colon.
Given the prevalence of cancers associated with mutated K-ras, there is a need
for
new therapeutic approaches particularly suitable for cancers of this type.
SUMMARY OF THE INVENTION
The present invention provides methods and compositions for treating cancers
and
precancerous conditions which are characterized by proliferating cells having
a K-ras
mutation.
In one embodiment, the invention provides a method of inhibiting the
proliferation
of cells having a K-ras mutation, such as pre-cancerous cells or cancer cells.
The method
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CA 02795952 2012-10-09
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comprises the steps of (1) identifying cells which have a K-ras mutation; (2)
inhibiting the
P13 kinase signaling pathway in the cells; and (3) inhibiting HDAC activity in
the cells. In
one embodiment, the P13 kinase signaling pathway and HDAC activity are
inhibited by
contacting the cells with a first compound which inhibits P13 kinase activity
and a second
compound which inhibits HDAC activity. In a second embodiment, the P13 kinase
signaling pathway and HDAC activity are inhibited by contacting the cells with
a
compound which inhibits both P13 kinase activity and HDAC activity.
In another embodiment, the invention provides a method of treating cancer or a
precancerous condition associated with a K-ras mutation in a subject in need
thereof. The
method comprises the steps of (1) identifying a subject having a cancer or a
precancerous
condition associated with a K-ras mutation; (2) administering to the subject
(i) an amount
of a P13 kinase inhibitor and (ii) an amount of an HDAC inhibitor, where the
amount of a
compound which inhibits P13 kinase and the amount of a compound which inhibits
HDAC
together are therapeutically effective. The P13 kinase inhibitor and the HDAC
inhibitor can
be administered simultaneously or sequentially and as separate compositions or
combined
in a single composition.
In yet another embodiment, the invention provides a method of treating a
cancer or
a precancerous condition associated with mutated K-ras in a subject in need
thereof. The
method comprises the steps of (1) identifying a subject having a cancer or
precancerous
condition associated with a K-ras mutation; (2) administering to the subject
an effective
amount of a compound is a P13 kinase inhibitor and an HDAC inhibitor.
The invention additionally includes compounds and compositions for use in the
disclosed methods.
DETAILED DESCRIPTION OF THE INVENTION
Phosphoinositides (PIs), which are phosphorylated derivatives of
phosphatidylinositol, are essential in eukaryotic cells, regulating nuclear
processes,
cytoskeletal dynamics, signalling and membrane trafficking. Among the enzymes
involved
in PI metabolism, P13-kinases (P13K) have attracted special attention because
of their
oncogenic properties and potential as drug targets. P13-kinases phosphorylate
phosphatidylinositols or PIs at the 3-position of the inositol ring. (Lindmo
et. al. Journal of
Cell Science 119, 605-614, 2006). The 3-phosphorylated phospholipids generated
by P13K
activity bind to the pleckstrin homology (PH) domain of protein kinase B (PKB
or AKT),
causing translocation of PKB to the cell membrane and subsequent
phosphorylation of
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PKB. Phosphorylated PKB inhibits apoptosis-inducing proteins such as FKHR,
Bad, and
caspases, and is thought to play an important role in cancer progression. The
PI3Ks are
divided into classes I-III, and class I is further subclassified into classes
la and lb. Among
these isoforms, class la enzymes are thought to play the most important role
in cell
proliferation in response to growth factor-tyrosine kinase pathway activation
(Hayakawa et.
al., Bioorganic & Medicinal Chemistry 14 6847-6858, 2006). Three frequent
mutations in
cancer constitutively activate PI3Ka and, when expressed in cells, they drive
the oncogenic
transformation and chronic activation of downstream signalling by molecules
such as PKB,
S6K and 4E bpl that is commonly seen in cancer cells. (Stephens et. al.,
Current Opinion
in Pharmacology, 5(4) 357-365, 2005). As such, P13-kinases are attractive
targets for the
treatment of proliferative diseases.
There are several known P13-kinase inhibitors including Wortmannin and
LY294002. Although wortmannin is a potent P13K inhibitor with a low nanomolar
IC50
value, it has low in vivo anti-tumor activity. (Hayakawa et al, Bioorg Med
Chem 14(20),
6847-6858 (2006)). Recently, a group of morpholine substituted quinazoline,
pyridopyrimidine and thienopyrimidine compounds have been reported to be
effective in
inhibiting Pl3kinase pl l0a. (Hayakawa, 6847-6858). Oral dosage of a
morpholine
substituted thienopyrimidine compound (GDC-0941) has shown tumor suppression
in
glioblastoma xenografts in vivo. (Folkes et. al., Journal of Medicinal
Chemistry, 51, 5522-
5532, 2008). The following publications disclose a series of thienopyrimidine,
pyridopyrimidine and quinazoline based P13-Kinase inhibitors: WO 2008/073785;
WO
2008/070740; WO 2007/127183; U.S. Patent Publication 20080242665.
Histone acetylation is a reversible modification, with deacetylation being
catalyzed
by a family of enzymes termed histone deacetylases (HDACs). HDAC's are
represented by
18 genes in humans and are divided into four distinct classes (JMo1 Biol,
2004, 338:1, 17-
31). In mammalians class I HDAC's (HDAC 1-3, and HDAC8) are related to yeast
RPD3
HDAC, class 2 (HDAC4-7, HDAC9 and HDAC10) related to yeast HDA1, class 4
(HDAC11), and class 3 (a distinct class encompassing the sirtuins which are
related to
yeast Sir2).
Csordas, Biochem. J., 1990, 286: 23-38 teaches that histones are subject to
post-
translational acetylation of the, r,-amino groups of N-terminal lysine
residues, a reaction
that is catalyzed by histone acetyl transferase (HAT 1). Acetylation
neutralizes the positive
charge of the lysine side chain, and is thought to impact chromatin structure.
Indeed,
access of transcription factors to chromatin templates is enhanced by histone
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WO 2011/130628 PCT/US2011/032683
hyperacetylation, and enrichment in underacetylated histone H4 has been found
in
transcriptionally silent regions of the genome (Taunton et at., Science, 1996,
272:408-411).
In the case of tumor suppressor genes, transcriptional silencing due to
histone modification
can lead to oncogenic transformation and cancer.
Several classes of HDAC inhibitors currently are being evaluated by clinical
investigators. Examples include hydroxamic acid derivatives, Suberoylanilide
hydroxamic
acid (SAHA), PXD101 and LAQ824, are currently in the clinical development. In
the
benzamide class of HDAC inhibitors, MS-275, MGCDO103 and CI-994 have reached
clinical trials. Mourne et at. (Abstract #4725, AACR 2005), demonstrate that
thiophenyl
modification of benzamides significantly enhance HDAC inhibitory activity
against
HDAC 1.
Although combined use of inhibitors of P13 kinase inhibitors and HDAC
inhibitors
for treating cancers has been described (Denlinger, C.E. et at., J. Thorac.
Cardiovasc. Surg.
2005: 1422-1429; WO 2009/058895; WO 2009/155659), the particular
susceptibility of
cancers associated with mutant K-ras to such therapy has not been previously
recognized.
The present invention provides methods for treating cancers and precancerous
conditions associated with mutated K-ras. The invention relates to the
discovery that such
cancers are particularly susceptible to the effects of concomitant inhibition
of P13 kinase
and HDAC.
In one embodiment, the invention provides a method of treating a cancer
comprising cancer cells having a K-ras mutation comprising the step of
concurrently
inhibiting P13 kinase activity and inhibiting HDAC activity within said the
cancer cells.
As set forth herein, a cancer cell or cells associated with a precancerous
condition
are said to have a mutated K-ras gene if the gene for K-ras within the cell
has been mutated
relative to the corresponding wild type gene in such a way as to produce a
gene product
having an amino acid sequence which differs from that of the wild-type K-ras
protein.
Preferably, the mutated gene has one or more nucleotide sunbstitutions in one
or more
codons and encodes a protein having one or more point mutations relative to
the wild type
protein. More preferably, the mutation is in a single codon of the K-ras gene,
such as
codon 12, 13 or 16. Most preferably the mutation is in codon 12 or 13.
As set forth herein, a subject has a cancer or a precancerous condition is
"associated
with mutated K-ras" if cells obtained from the subject have a mutated K-ras
gene. The
identification of a K-ras mutation is within the skill in the art, and in one
embodiment is
done using, for example, methods disclosed herein or other methods as are
known in the
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WO 2011/130628 PCT/US2011/032683
art. In another embodiment, the subject is identified as having a cancer
associated with
mutated K-ras if the subject is diagnosed with a cancer type which with a high
probability
of mutated K-ras, including pancreatic cancer, such as pancreatic
adenocarcinoma, or
adenocarcinoma of the colon or lung.
A K-ras mutation can be identified using methods known in the art. For
example,
cells from a tumor biopsy, such as a surgical biopsy or a needle aspirate, can
be analyzed
for the presence of a mutation. The cells can also be obtained from a body
fluid, such as
blood or urine, or from a stool sample. Methods for detecting K-ras mutations
are known
in the art and include, but are not limited to, those set forth in Poehlmann,
A. et at., Pathol.
Res. Pract. 2007, 203: 489-497; van Heck, N.T. et at., J. Clin. Pathol. 2005,
58, 1315-
1320; Taback, B. et at., Int. J. Cancer 2004, 111: 409-414; Lleonart, M.E. et
at., Nucleic
Acids Res. 2006, 34: e12; Bjornheim J. et at. Mutat. Res. 1998, 403: 103-112;
Chen, C.Y. et
at., Clin. Chem. 2004, 50: 481-489; Maekawa, M. et at., Clin. Chem. 2004, 50:
1322-1327;
Bos, J.L. et at., Nucleic Acids Res. 1984, 12: 9155-9163; Corominas, M. et
at., Environ.
Health Perspect. 1991, 93: 19-25; Su, Y.-H. et at., Ann. NYAcad. Sci. 2008,
1137: 197-
206.
Cancers which can be associated with mutated K-ras include pancreatic cancer,
such as adenocarcinoma of the pancreas, including ductal adenocarcinoma; lung
cancer,
such as non-small cell lung cancer and lung adenocarcinoma; colorectal cancer,
such as
adenocarcinoma of the colon; thyroid cancer, such as follicular carcinoma,
undifferentiated
carcinoma, and papillary carcinoma; testicular cancer, such as seminoma;
leukemia, such as
acute myologenous leukemia, chronic myologenous leukemia, multiple myeloma and
acute
lymphocytic leukemia; multiple myeloma, liver cancer, breast cancer, prostate
cancer,
ovarian cancer and endometrial cancer.
Non-cancerous conditions which can be associated with mutated K-ras include
adenomas, such as colon adenomas and thyroid adenomas, and myelodysplastic
syndrome.
In one embodiment, a subject is determined to have a cancer associated with a
K-ras
mutation if the subject is diagnosed as having a cancer in which the
prevalence of K-ras
mutation is greater than about 50%. Such cancers include pancreatic cancer,
such as ductal
adenocarcinoma, which is the cancer type with the highest rate of K-ras
mutation. Thus,
such a cancer is significantly more likely than not to be associated with
mutated K-ras, and
a clinician may determine that treatment with an inhibitor of P13 kinase and
an inhibitor of
HDAC according to the invention is warranted on the basis of the diagnosis of
pancreatic
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cancer, such as ductal adenocarcinoma, without the need for additional testing
to confirm
the presence of a K-ras mutation.
The term "P13 kinase inhibitor", as used herein, is a compound which inhibits
cellular signaling through the PI3K/AKT signaling pathway. The P13 kinase
inhibitor can
be, for example, an inhibitor of one or more proteins involved in this
signaling pathway,
including P13 kinase, such as PI3Ka, PI3K(3, PI3Ky, and PI3k6, PKB (AKT),
mammmalian
Target of Rapamycin ("mTOR") and/or another protein in the pathway. Methods
for
assessing the ability of a compound to inhibit such proteins include those set
forth herein
and others known in the art. In one embodiment, the P13 kinase inhibitor is
selected from
LY294002 (Eli Lily), Wortmannin, Wortmannin analogues, pegylated Wortmannin,
pegylated 17-hydroxy-Wortmannin, PX-866, SF1124 (Semafore Pharmaceuticals),
SF1126
(Semafore Pharmaceuticals), BEZ235 (Novartis), BGT226 (Novartis), BKM120
(Novartis),
XL-765 (Exelexis), XL-147 (Exelexis), GDC-0941 (Genentech), AS-252424, ONC-201
(Oncalis), CAL-101 (Calistoga), CAL-263 (Calistoga), Atu-027, PF-4691502, PBI-
05204
(Phoenix Biotechnology), GSK-2126458, PIK-90, PIK-75, PI-103, ZSTK-474,
TGX115,
TGX-221 and TGX126.
In one embodiment, the P13 kinase inhibitor is selected from the compounds set
forth below:
N
O O
O O NO
PN/
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O)_CH, N-S
o N
Mew 01"
O
O
N NH
o I \ o
OH aN NH
H N
0
N
CN
/ \ \ N N"
O~-N N NH
\ I
(0)
N N
S N .N I
N
\ NH
N N \ \O N~NH
NJ / O ON
/\O
N NN
rN-
S
NO2 bN /N F
S
OZ
off
7
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(0)
N
N
N \N
O
N
N r N N N \
OH
eN \ O` J F \
~O I O~ O
/ N
aN NNH N N N
NH II
O~ N N
O
O NH2
N O
N N
H2N N N O
OH
N N
O N
O
N
H
F F N
2-morpholin-4-yl-8-phenylchromen-4-one; acetic acid (1 S,4E, l OR,11 R,13
S,14R)-[4-
diallylaminomethylene-6-hydroxy- l -methoxymethyl- 10, 1 3-dimethyl-3,7,17-
trioxo-
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1,3,4,7,10,11,12,13,14,15,16,17-dodecahydro-2-oxa-cyclopenta[a]phenanthren-1l-
yl ester;
9-(3-pyridinylmethyl)oxy-2-morpholinyl-4H-pyrido[1,2-a]pyrimidin-4-one (TGX-
140); 7-
methyl-9-phenylaminomethyl-2-morpholinyl-4H-pyrido[1,2-a]pyrimidin-4-one (TGX-
183); 8-(4-methylphenl)2-)4-morpholinyl)-4(1H)-quinolinone (TGX- 113); 8-(4-
fluorophenoxy)-2-(4-morpholinyl)-4(1H)-quinolinone (TGX-121); 2-morpholinyl-8-
(phenylmethyl)-4H-1-benzopyran-4-one (TGX-90); 2-(4-morpholinyl)-8-(4-fluoro-2-
methylphenyl)oxy-4H-1-benzopyran-4-one (TGX-184); 9-[[(2-chlorophenyl)-
methyl] amino -7-methyl-2-(4-morpholinyl)-4H-pyrido [ 1,2-a]pyrimidin-4-one
(TGX-167);
9- [[(2-methoxyphenyl)-methyl] amino] -7-methyl-2-(4-morpholinyl)-4H-pyrido [
1,2-
a]pyrimidin-4-one (TGX-137); 7-methyl-2-(4-morpholinyl)-9-
[(phenylmethyl)amino]-4H-
pyrido[1,2-a]pyrimidin-4-one (TGX-126); 9-[[(4-fluoro-2-methylphenyl)amino]-7-
methyl-
2-(4-morpholinyl)-4H-pyrido[l- ,2-a]pyrimidin-4-one (TGX-170); 7-methyl-2-(4-
morpholinyl)-9-[[(1R)-1-phenylethyl]amino] -4H-pyrido[1,2-a]pyrimidin-4-one
(TGX-123);
7-methyl-2-(4-morpholinyl)-9-[(2-pyridinylmethyl)amino] -4H-pyrido [ l ,2-
a]pyrimidin-4-
one (TGX-161); 9-[ [(4-chlorophenyl)methyl] amino] -7-methyl-2-(4-morpholinyl)-
4H-
pyrido[1,2-a]pyrimidin-4-one (TGX-108); 2-(4-morpholinyl)-9-(phenylmethyl)-4H-
pyrido[1,2-a]pyrimidin-4-one (TGX-040); 7-methyl-9-(N-Methyl-N-
phenyl)aminomethyl-
2-(4-morpholinyl)-4H-pyrido[1,2- a]pyrimidin-4-one (TGX-195); 2-(4-
morpholinyl)-8-
(phenylmethyl)oxy-4H-1-benzopyran-4-one (TGX-102); 2-(4-morpholinyl)-8-
(phenylmethyl)amino-4H-1-benzopyran-4-one (TGX-204); 2-(4-morpholinyl)-8-
phenylamino-4H- 1-benzopyran-4-one (TGX-324); 8-(3-chlorophenyl)oxy-2-(4-
morpholinyl)-4H- 1-benzopyran-4-one (TGX-259); 8-(3-methylphenyl)-2-(4-
morpholinyl)-
4(1 H)-quinolinone (TGX-127); 8-(2-fluorophenyl)-2-(4-morpholinyl)-4(1H)-
quinolinone
(TGX-143); (+/-)-7-methyl-2-morpholin-4-yl-9-[1-(3-pyridinylamino)ethyl]-
pyrido[1,2--
a]pyrimidin-4-one (KN-304); (+/-)-7-methyl-9-{[methyl(phenyl)amino]methyl}-2-
morpholin-4-yl-pyrido[l,- 2-a]pyrimidin4-one (TGX-195); (+/-)-7-methyl-2-
morpholin-4-
yl-9-(1-phenylaminoethyl)-pyrido[1,2-a]pyrimidin-4-one (TGX-221); (+/-)-7-
methyl-2-
morpholin-4-yl-9-[1-(4-fluorophenylamino)ethyl]-pyrido[1,2-a]pyrimidin-4-one
(TGX-
224); (+/-)-9-[1-(3,4-difluorophenylamino)ethyl]-7-methyl-2-morpholin-4-yl-
pyrido[1,2-
a]pyrimidin4-one (TGX-237); (+/-)-9-[1-(2,5-difluorophenylamino)ethyl]-7-
methyl-2-
morpholin-4-yl-pyrido[l,2-a]pyrimidin-4-one (TGX-238); (+/-)-9-[1-(3,5-
difluorophenylamino)ethyl] -7-methyl-2-morpholin-4-yl-pyrido [ l ,2-
a]pyrimidin-4-one
(TGX-239); (+/-)-9-[1-(4-fluoro-2-methylphenylamino)ethyl]-7-methyl-2-
morpholin-4-yl-
pyrido[1,2-a]pyrimidin-4-one (TGX-240); (+/-)-9-[1-(4-chlorophenylamino)ethyl]-
7-
9
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methyl-2-morpholin-4-yl-pyrido[1,2-a]pyrimidin-4-one (TGX-243); (+/-)-9-[1-
(3,4-
dichlorophenylamino)ethyl]-7-methyl-2-morpholin-4-yl-pyrido [ l ,2-
a]pyrimidin4-one
(TGX-244); (+/-)-9-[1-(3 fluorophenylamino)ethyl]-7-methyl-2-morpholin-4-yl-
pyrido[1,2-
a]pyrimidin-4-one (TGX-247); (+/-)-9-[1-(3-chlorophenylamino)ethyl]-7-methyl-2-
morpholin-4-yl-pyrido[1,2-a]pyrimidin-4-one (TGX-248); (+/-)-7-methyl-2-
morpholin-4-
yl-9-[l-(2-thiazolylamino)ethyl]-pyrido[1,2-a]pyrimidin-4-one (TGX-261);
(+/-)-7-methyl-9-[ 1-(3-methylphenylamino)ethyl]-2-morpholin-4-yl-pyrido [ 1,2-
a]pyrimidin-4-one (TGX-262); (+/-)-7-methyl-2-morpholin-4-yl-9-[1-(3-
trifluoromethylphenylamino)ethyl]- -pyrido[1,2-a]pyrimidin-4-one (TGX-264);
(+/-)-7-
methyl-2-morpholin-4-yl-9-[1-(2-pyridinylamino)ethyl]-pyrido[1,2-a]pyrimidin-4-
one
(TGX-295); (+/-)-2-({1-[7-methyl-2-(morpholin4-yl)-4-oxo-pyrido[1,2-
a]pyrimidin-9-yl-
]ethyl}amino)benzoic acid (KN-309);
(+/-) methyl 2-({1-[7-methyl-2-(morpholin-4-yl)-4-oxo-pyrido[1,2-a]pyrimidin-9-
yl]ethy-
1}amino)benzoate (KN-321); (+/-)-2-({1-[7-methyl-2-(morpholi-4-yl)-4-oxo-
pyrido[1,2-
a]pyrimidin-9-yl- ]ethyl} amino)benzonitrile (KN-320); (+/-)-7-methyl-2-
(morpholin-4-yl)-
9-(l - {[2-(2H-tetrazol-5 -yl)phenyl] amino- }ethyl)-pyrido[1,2-a]pyrimid-4-
one (KN-325);
(+/-)-2-(4-morpholinyl)-8[1-(phenylamino)ethyl]-4H-1-benzopyran-4-one (TGX-
280); (3-
(4-(4-morpholinyl)pyrido[3',': 4,5]furo[3,2-d]pyrimidin-2-yl)phenol); 2-methyl-
2-(4-(3-
methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydro-1 H-imidazo [4,5-c] quinolin-l -
yl)phenyl)propanenitrile (BEZ-235); 5-[[5-(4-Fluoro-2-hydroxyphenyl)-2-
furanyl]methylene]-2,4-thiazolidinedione (AS252424); 3-(4-(4-
Morpholinyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidin-2-yl)phenol (PI-103); 2-(2-
Difluoromethylbenzimidazol- 1-yl)-4,6-dimorpholino-1,3,5-triazine (ZSTK474).
Additional P13 kinase inhibitors of use herein include 42-(3-hydroxy-2-
(hydroxymethyl)-rapamycin 2-methylpropanoate (temsirolimus, Wyeth); 42-0-(2-
hydroxyethyl)-rapamycin (Novartis), and 42-(dimethylphopsinoyl)-rapamycin
(Ariad).
Additional P13 kinase inhibitors which can be used in the methods of the
invention
include those set forth in US 7,598,377; 7,662,977; 7,544,677; 7,524,850;
7,511,041; and
US published applications 2009/0098135; 2009/0029997; 2009/0029998 and
2010/0061982, WO 2006/065601, WO 2008/144463, WO 2009/071888, WO 2009/03265 1,
WO 2009/032652, WO 2009/032653, WO 2009/017822, WO 2009/053716; WO
2009/143317; W02008/152387, W02010/005558, W02010/001126, WO/2009/143313,
WO 2009/021083, W02008/127226, W02009/155052 and W02009/155121, each of
which is incorporated herein by reference in its entirety.
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P13 kinase inhibitors of use in the present methods further include
pharmaceutically
acceptable salts, esters and prodrugs of any of the P13 kinase inhibitors
described herein.
The term "HDAC inhibitor", as used herein, refers to a compound which inhibits
the enzymatic activity of one or more isoforms of histone deacetylase. The
ability of a
compound to inhibit HDAC activity can beassessed using methods known in the
art,
including, but not limited to, the method described herein.
Suitable HDAC inhibitors include, but are not limited to, hydroxamic acids,
cyclic
peptides, benzamides and aliphatic acids. In one embodiment, the HDAC
inhibitor is
selected from the group consisting of suberoylanilide hydroxamic acid (SAHA),
butyric
acid, valproic acid, and Romidepsin. In another embodiment, the HDAC inhibitor
is
belinostat, mocetinostat, LAQ824, LBH589, C1994, BML-210, M344, MS275, JNJ-
26481585 or MGCDO103. In other embodiments, the HDAC inhibitor is selected
from:
OH
HN
O NH
ct NH2
OH
O
O
O H
NH NH
N \ I N
O N I / / NH2 O NH2
O
O O NI~OH
11OH H I H
I / H H I N /
N N
H
11
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0
NH
N,', N NH2
NCO /
N~OH
O O N
O
N \ N1OH
0 ~--~` / I H
N~OH N DO
jN/~N I H \N
'~r
H
iN LN
0
C \ / \ 1OH N
N~~ H N H I H NH2
N N
NI O 0
;
((E)-N-hydroxy-3-[4-[ [2-hydroxyethyl-[2-(1 H-indol-3-yl)ethyl]
amino]methyl]phenyl]prop-
2-enamide;
4-(acetylamino)-N-(2-aminophenyl)benzamide;
Pyridin-3-ylmethyl 4-(2-aminophenylcarbamoyl)benzylcarbamate;
N-(2-aminophenyl)-N'-phenyloctanediamide;
4-(dimethylamino)-N-[7-(hydroxyamino)-7-oxoheptyl]benzamide;
(2E)-3-[3-(anilinosulfonyl)phenyl]-N-hydroxyacrylamide;
N-(2-aminophenyl)-4-[[(4-pyridin-3-ylpyrimidin-2-yl)amino]methyl]benzamide;
and
2-(6- {[(6-fluoroquinolin-2-yl)methyl] amino }bicyclo [3. 1. 0] hex-3 -yl)-N-
hydroxypyrimidine-5-carboxamide.
In another embodiment, the HDAC inhibitor is EVP-0334, MGCD-290, CHR-3996
or DAC-0060.
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Additional HDAC inhibitors which can be used in the methods of the invention
include those set forth in WO 2008/019025, WO 2006/094068, WO 2007/054719, WO
2005/007091, WO 2010/028193, WO 2008/082646, WO 2007/045962, WO 2005/087724,
WO 2005/065681, WO 2004/113366, WO 2004/054999, WO 2003/01185 1, WO
2009/026446, WO 2009/006403, W02009/150129, WO 2007/113644, WO 2009/045440,
WO 2008/087514 and WO 2008/068176, each of which is incorporated herein by
reference
in its entirety.
HDAC inhibitors of use in the present methods further include pharmaceutically
acceptable salts, esters and prodrugs of any of the HDAC inhibitors described
herein.
In one embodiment, the inhibitor of P13 kinase and the HDAC inhibitor are
separate
compounds. In another embodiment, the inhibitor of P13 kinase and the HDAC
inhibitor
are the same compound, that is, a single bifunctional compound which functions
as both an
inhibitor of P13 kinase and an HDAC inhibitor. In one embodiment, the
bifunctional
molecule comprises a moiety which inhibits P13 kinase and a moiety which
inhibits histone
deacetylase. Suitable bifunctional molecules include those having the general
formula A-
B-C, where A is a moiety that inhibitors P13 kinase, C is a moiety which
inhibis HDAC and
B is a linker.
In a preferred embodiment, the bifunctional molecule is selected from the
compounds set forth in US Patent Publication No. 2010/0222343 and US Patent
Application No. 13/078,769, the contents of which are incorporated herein by
reference in
their entirety.
In one embodiment, the bifunctional compound has the general formula (I):
(R1)n (C-B gRaNb
G~1 N -G3 X N
1C-B S N G2-(B-C
)r
(R2)p
Formula I
or a geometric isomer, enantiomer, diastereomer, racemate, pharmaceutically
acceptable
salt or prodrug thereof,
wherein represents a single or double bond;
q, r and s are independently 0 or 1, wherein at least one of q, r and s is 1;
preferably, one
of q, r and s is 1 and the rest are 0;
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n is 0, 1, 2, 3 or 4;
p is 0, 1 or 2, preferably 0 or 1;
t is 0 or 1; preferably, when s is 1, t is 0;
X and Y are independently CR1, N(R8), S or 0; wherein when one of X and Y is
CR1,
the other is N(R8), S or 0; preferably X is S and Y is CR1;
Gi is CR1, S, 0, NRio or NS(O)2R10;
G2 is substituted or unsubstituted aryl, substituted or unsubstituted
heteroaryl or,
substituted or unsubstituted heterocyclic;
G3 is substituted or unsubstituted C1-Cg alkyl, substituted or unsubstituted
C2-Cg alkenyl
or substituted or unsubstituted C2-Cg alkynyl;
each Rs is independently hydrogen, acyl, aliphatic or substituted aliphatic;
each R1 and R2 is absent or is independently selected from hydrogen, hydroxy,
amino,
halogen, alkoxy, alkylamino, dialkylamino, CF3, CN, NO2, sulfonyl, acyl,
aliphatic,
substituted aliphatic, aryl, substituted aryl, heteroaryl, substituted
heteroaryl,
heterocyclic, and substituted heterocyclic;
Ra is optionally substituted alkyl, optionally substituted aryl or optionally
substituted
heteroaryl;
Rb is hydrogen, optionally substituted alkyl, optionally substituted aryl or
optionally
substituted heteroaryl;
or Ra and Rb, together with the nitrogen atom to which they are attached, form
an
optionally substituted heterocyclic group;
Rio is selected from hydrogen, hydroxy, amino, alkoxy, alkylamino,
dialkylamino,
sulfonyl, acyl, aliphatic, substituted aliphatic, aryl, substituted aryl,
heteroaryl,
substituted heteroaryl, heterocyclic, and substituted heterocyclic; preferably
Rio is
hydrogen, acyl, aliphatic or substituted aliphatic;
B is a linker; and
C is selected from:
W
HO "
(a) R31 III 1
; where W is 0 or S; J is 0, NH or NCH3; and R31 is
hydrogen or lower alkyl;
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W
R34)
Z
(b) 133 R32 ; where W is O or S; Y2 is absent, N, or CH; Z is N or CH; R32
and R34 are independently hydrogen, hydroxy, aliphatic group, provided that if
R32 and R34 are both present, one of R32 or R34 must be hydroxy and if Y2 is
absent, R34 must be hydroxy; and R33 is hydrogen or aliphatic group;
W
H0,,z1 lj~ Y1-
(c) ; where W is 0 or S; Y1 and Z1 are independently N, C or CH;
and
R21 NH2
1 W
R22
Z Y2
Rz3 I I
(d) R12 R11 ; where Z, Y2, and W are as previously defined; R11 and
R12 are independently selected from hydrogen or aliphatic; R21, R22 and R23
are independently selected from hydrogen, hydroxy, amino, halogen, alkoxy,
alkylamino, dialkylamino, CF3, CN, NO2, sulfonyl, acyl, aliphatic, substituted
aliphatic, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic,
and substituted heterocyclic.
In another embodiment, each R1 and R2 is independently absent, hydrogen,
hydroxy,
amino, halogen, alkoxy, alkylamino, dialkylamino, CF3, CN, NO2, sulfonyl,
acyl, aliphatic,
substituted aliphatic, aryl, substituted aryl, heteroaryl, substituted
heteroaryl, heterocyclic,
and substituted heterocyclic; arylalkyl, substituted arylalkyl,
heteroarylalkyl, substituted
heteroarylalkyl, heterocyclylalkyl or substituted heterocyclylalkyl.
In one embodiment, the bifunctional compound is represented by formula (II) or
formula (III), or a geometric isomer, enantiomer, diastereomer, racemate,
pharmaceutically
acceptable salt, or prodrug thereof:
( G
(R1)n (B C)4
KN
1)n N
N
X -
G, G3 Y G
(C-Br N////~\G2-4B-C) r
(R2)P
(R2)P
(II) (III),
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wherein represents a single or double bond and G1, G2, G3, R1, R2, X, Y, n, p,
q, r, s,
B and C are as defined above. Preferably, in Formula (III), q is 1 and r is 0.
In another embodiment of the bifunctional compound is represented by formula
(IV) or (V) as illustrated below, or a geometric isomer, enantiomer,
diastereomer, racemate,
pharmaceutically acceptable salt or prodrug thereof:
1
(C-t~ r 1') (Rj )n (C-BI-7,--r G, I (Rj )n
q ~NJ q /
~G4 N \N
(C-B)s N'G2 (B-C)1 (C-4 s 'a N' ~G2 (B-C),
(R2)p (R2)P
Formula IV Formula V,
wherein ------------represents a single or double bond; G1, G2, R1, R2, Rs, n,
p, q, r, s, B and C
are as defined above; and G4 is NRg, S or 0, preferably S. Preferably Gi is O.
In another embodiment, the bifunctional compound is represented by formula
(VI)
and (VII) as illustrated below, or a geometric isomer, enantiomer,
diastereomer, racemate,
pharmaceutically acceptable salt, or prodrug thereof:
Gl> (R1)n c) q R8 G11 (R1)n (B C)q
R
N~ `Ga N I=NNBC~S N 8 BC)s
N ~G4 N
o N
2)P (R3)m (R2)P (R3)m
Formula VI Formula VII,
wherein represents a single or double bond; G1, G4, R1, R2, R3, Rg, n, m, p,
q, , s, B and
C are as defined above; and o is 1, 2, 3 or 4.
In preferred embodiments of the compounds of Formulas Ito V, q and r are 0 and
s
is 1. In another preferred embodiment q is 1 while r and s are 0. In another
preferred
embodiment q and s are 0 and r is 1.
In preferred embodiments of the compounds of Formula III, r is 0 and q is 1.
In preferred embodiments of the compounds of Formulas VI and VII, q is 1 and s
is
0.
In a preferred embodiment, B is a CI-Cg alkyl where one or more CH2 can be
optionally replaced by O, S, SO2, NRg or -CONH-, C is -C(O)N(H)OH, and Gi is -
0. In
another preferred embodiment, B is a CI-Cs alkyl where one or more CHz can be
optionally
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replaced by 0, S, SO2, NRg or -CONH-, C is -C(O)N(H)OH, and Gi is NS(O)2CH3.
In
another preferred embodiment, B is an aryl, heteroaryl, Ci-Cio-alkylaryl, Ci-
Cio-
alkylheteroaryl group, Ci-Cio-alkylheterocyclylaryl, Ci-Cio-
alkylheterocyclylheteroaryl,
Ci-Cio-alkylheterocyclylaryl-Ci-Cio-alkyl, or Ci-C io-
alkylheterocyclylheteroaryl-Ci-Cio-
alkyl group where one or more CH2 can be optionally replaced by 0, S, S02, NRg
or -
CONH-, and Gi is -O or -NS(O)2CH3.
In another embodiment, the bifunctional compound is represented by formula
(VIII) or (IX) as illustrated below, or a geometric isomer, enantiomer,
diastereomer,
racemate, pharmaceutically acceptable salt or prodrug thereof:
(C B)q Ra\ ,Rb (C-B)q Ra\N /Rb
G~1 (R~)n N Gj\~(Rj)n
G4 N CN) N
N 1
V0~ %\G2 (B-C)S Ga i G2 (B-C)S
N
N o N
(R2)p (R2)P
Formula VIII Formula IX,
wherein represents a single or double bond; G1, G2, G4, Ra, Rb, Ri, R2, n, p,
q, s, B and
C are as defined above; and o is 1, 2, 3 or 4. In preferred embodiments, G2 is
optionally
substituted phenyl, pyridyl, pyrimidyl, indolyl, indazolyl, pyridopyrrolyl,
pyrrolyl,
imidazolyl, pyrazolyl or benzimidazolyl.
In a preferred embodiment of the compounds of Formulas VIII and IX, q is 0 and
s
is 1. In another preferred embodiment, q is 1 and s is 0. In a preferred
embodiment, B is a
CI-Cg alkyl where one or more CH2 can be optionally replaced by O, S, S02, NRg
or -
CONH-, C is -C(O)N(H)OH, and Gi is -0. In another preferred embodiment, B is a
CI-Cs
alkyl where one or more CH2 can be optionally replaced by O, S, S02, NRg or -
CONH-, C
is -C(O)N(H)OH, and Gi is -NS(O)2CH3.
In another embodiment, the bifunctional compound is represented by formula X
or
XI as illustrated below, or a geometric isomer, enantiomer, diastereomer,
racemate,
pharmaceutically acceptable salt or prodrug thereof:
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G~\
(R1)n
N'1-j /B J (Rj )n
C N/
S
\ ~ S N
C B N G2
I N G2
(R2)P (R2)p
Formula X Formula XI,
wherein G1, G2, n, p, B, C, Ri and R2 are as defined above. In preferred
embodiments, G2
is optionally substituted phenyl, pyridyl, pyrimidyl, indolyl, indazolyl,
pyridopyrrolyl,
pyrrolyl, imidazolyl, pyrazolyl or benzimidazolyl.
In preferred embodiment of the compounds of Formulas X and XI, B is a CI-CS
alkyl where one or more CH2 can be optionally replaced by 0, S, SO2, N(R8), -
CONH-,
aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic or
substituted
heterocyclic groups, C is -C(O)N(H)OH, and Gi is O. In another preferred
embodiment, B
is a CI-Cs alkyl where one or more CH2 can be optionally replaced by 0, S,
SO2, NRg, -
CONH-, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic or
substituted heterocyclic groups, C is -C(O)N(H)OH, and Gi is -NS(O)2CH3.
In another embodiment, the bifunctional compound is represented by formula
XII,
as illustrated below, or a geometric isomer, enantiomer, diastereomer,
racemate,
pharmaceutically acceptable salt or prodrug thereof:
C
B
NC
C) (R' )n
N
S N
%\
I N G2
(R2)p
XII
wherein n, p, B, C, R1, G2 and R2 are as defined above.
In another embodiment, the bifunctional compound is represented by formula
XIII
as illustrated below, or a geometric isomer, enantiomer, diastereomer,
racemate,
pharmaceutically acceptable salt or prodrug thereof:
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G1~
J (R,)n
N
R33 R32 G4 N
R34 M5 M4 M3 M2 Ml N G2
W (R2)P
Formula XIII,
wherein n, p, Y2, W, Z, G1, G4, G2, R1, R2, R32, R33 and R34 are as defined
above; Mi is
absent, 0, S, NRg, Ci-C6 alkyl, , C2-C6 alkenyl, C2-C6 alkynyl, aryl,
heteroaryl,
heterocyclic, SO, SO2 or C=O; M2 is absent, C1-C6 alkyl, 0, NRgõ heterocyclic,
aryl,
heteroaryl, or C=O; M3 is absent, 0, NRg, S, SO, SO2, CO, C1-C6 alkyl, C2-C6
alkenyl, C2-
C6 alkynyl, aryl, heteroaryl, or heterocyclic; M4 is absent, 0, NRg,
heteroaryl, heterocyclic
or aryl; and M5 is absent, CI-Cs alkyl, C2-Cg alkenyl, C2-Cgalkynyl,
heteroaryl,
heterocyclic or aryl. In preferred embodiments, G2 is optionally substituted
phenyl,
pyridyl, pyrimidyl, indolyl, indazolyl, pyridopyrrolyl, pyrrolyl, imidazolyl,
pyrazolyl or
benzoimidazolyl. In more preferred embodiments, G2 is optionally substituted
phenyl,
pyridyl, pyrimidyl, indazolyl, pyrrolyl or benzimidazolyl.
In a another embodiment, the bifunctional compound is represented by formula
(XIV) as illustrated below, or a geometric isomer, enantiomer, diastereomer,
racemate,
pharmaceutically acceptable salt or prodrug thereof:
G1,)
G7~, (R')n
(R5)v N
W(R6) N )t G4
N
H0- G6
NH C I \ / %
N G5 N G
2
)u (R2)P
0
Formula XIV,
wherein G1, G2, G4, n, p, Ri and R2 are as defined above; t, v and w are
independently 0, 1,
2 or 3; u is 0, 1, 2, 3, 4, 5, 6, 7 or 8; G5 is absent, CI-Cg alkyl or a CI-Cg
alkyl interrupted by
one or more 0, S, S(O), SO2, N(R8), or C(O); preferably G5 is -N(R8)-Ci-C4-
alkyl, t is 1
and u is 0;
G6 is selected from CRi or NRg, wherein Ri and Rs are as defined above;
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G7 is selected from -CR1, -NR8, S or 0 wherein Ri and Rs are as defined above;
or
G7 is selected from -C(R1)2 , and -N; R5 and R6 are independently selected
from absent,
hydrogen, hydroxy, amino, halogen, alkoxy, alkylamino, dialkylamino, CF3, CN,
NO2,
sulfonyl, acyl, aliphatic, substituted aliphatic, aryl, substituted aryl,
heteroaryl, substituted
heteroaryl, heterocyclic, and substituted heterocyclic. In preferred
embodiments, G2 is
optionally substituted phenyl, pyridyl, pyrimidyl, indolyl, indazolyl,
pyridopyrrolyl,
pyrrolyl, imidazolyl, pyrazolyl or benzimidazolyl. In more preferred
embodiments, G2 is
optionally substituted phenyl, pyridyl, pyrimidyl, indazolyl, pyrrolyl or
benzimidazolyl.
Preferably, when G7 is CRi or N, the pyrimidine ring is directly bonded to G7.
In a another embodiment, the bifunctional compound is represented by formula
(XV) as illustrated below, or a geometric isomer, enantiomer, diastereomer,
racemate,
pharmaceutically acceptable salt, or prodrug thereof-
(G
J (Rj)n
N
/G N
W(R6) 4
HO--NH --GN fiG2
--- N (R2)P
)u
O
Formula XV,
wherein G1, G2, G4, G5, n, p, w, u, R1, R2 and R6 are as defined above. In
preferred
embodiments, G2 is optionally substituted phenyl, pyridyl, pyrimidyl, indolyl,
indazolyl,
pyridopyrrolyl, pyrrolyl, imidazolyl, pyrazolyl or benzimidazolyl. In more
preferred
embodiments, Gi is 0, G2 is optionally substituted phenyl, pyridyl, pyrimidyl,
indazolyl,
pyrrolyl or benzimidazolyl, G5 is -N(R8)-Ci-C4-alkyl and u is 0.
In more preferred embodiments of the compounds of any of the foregoing
formulas,
G2 is optionally substituted phenyl, pyridyl, pyrimidyl or pyrrolyl selected
from the group:
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~R3)m (R3)m (R3)m 3)
-- ri N
N\ ~\ N R$ N:
3)m 3)m R8 (R3)m (R3)m
Preferably in these groups, m is 1 and R3 is hydroxyl, hydroxymethyl, amino,
acylamino,
such as acetylamino, or methylamino. In another preferred embodiment,
G2 is an optionally substituted indazolyl or benzimidazolyl group selected
from the groups
shown below:
R8 R
N
N N
(R3)m (R3)m
where R3, Rs and m are as defined above.
In certain preferred embodiments of the compounds of the foregoing formulas,
G2
is an optionally substituted monoaryl or monoheteroaryl group. In more
preferred
embodiments, G2 is phenyl, pyridyl, pyrimidyl or pyrrolyl with one or more
substituents
include but not limited to hydroxyl, hydroxymethyl, amino and substituted
amino; Gi is 0,
G5 is -N(R8)-Ci-C4-alkyl and u is 0. For example, G2 can be phenyl, pyridyl,
pyrimidyl or
pyrrolyl substituted by a hydroxyl, hydroxymethyl, acetylamino, amino or
methylamino
group. Such compounds have significant inhibitory activity toward mTOR, as
well as P13
kinase and HDAC.
O
HO1N~ `
I ~"
The most preferred embodiment for C is: R33 ; where R33 is selected from
hydrogen and lower alkyl.
In a preferred embodiment, the bivalent B is a direct bond or straight- or
branched-,
substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl,
substituted or
unsubstituted alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl,
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heteroarylalkenyl, heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl,
heterocyclylalkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl,
alkylarylalkyl,
alkylarylalkenyl, alkylarylalkynyl, alkenylarylalkyl, alkenylarylalkenyl,
alkenylarylalkynyl,
alkynylarylalkyl, alkynylarylalkenyl, alkynylarylalkynyl,
alkylheteroarylalkyl,
alkylheteroarylalkenyl, alkylheteroarylalkynyl, alkenylheteroarylalkyl,
alkenylheteroarylalkenyl, alkenylheteroarylalkynyl, alkynylheteroarylalkyl,
alkynylheteroarylalkenyl, alkynylheteroarylalkynyl, alkylheterocyclylalkyl,
alkylheterocyclylalkenyl, alkylhererocyclylalkynyl, alkenylheterocyclylalkyl,
alkenylheterocyclylalkenyl, alkenylheterocyclylalkynyl,
alkynylheterocyclylalkyl,
alkynylheterocyclylalkenyl, alkynylheterocyclylalkynyl, alkylaryl,
alkenylaryl, alkynylaryl,
alkylheteroaryl, alkenylheteroaryl, or alkynylhereroaryl, which one or more
methylenes can
be interrupted or terminated by 0, S, S(O), SO2, N(R8), C(O), substituted or
unsubstituted
aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted
heterocyclic; such
divalent B linkers include but are not limited to alkyl, alkenyl, alkynyl,
alkylaryl,
alkenylaryl, alkynylaryl, alkylheterocyclylaryl, alkylheterocyclylarylalkyl,
alkylheterocyclylheteroaryl, alkylheterocyclylheteroarylalkyl, alkoxyaryl,
alkylaminoaryl,
alkoxyalkyl, alkylaminoalkyl, alkylheterocycloalkyl, alkylheteroarylalkyl,
alkylamino,
N(R8)alkenyl, N(R8)alkynyl, N(R8)alkoxyalkyl, N(R8)alkylaminoalkyl,
N(R8)alkylaminocarbonyl, N(R8)alkylaryl, N(R8)alkenylaryl, N(R8)alkynylaryl,
N(R8)alkoxyaryl, N(R8)alkylaminoaryl, N(R8)cycloalkyl, N(R8)aryl,
N(R8)heteroaryl,
N(R8)heterocycloalkyl, N(R8)alkylheterocycloalkyl, alkoxy, 0-alkenyl, 0-
alkynyl, 0-
alkoxyalkyl, 0-alkylaminoalkyl, O-alkylaminocarbonyl, O-alkylaryl, 0-
alkenylaryl, 0-
alkynylaryl, 0-alkoxyaryl, O-alkylaminoaryl, O-cycloalkyl, O-aryl, 0-
heteroaryl, 0-
heterocycloalkyl, O-alkylheterocycloalkyl, C(O)alkyl, C(O)-alkenyl,
C(O)alkynyl,
C(O)alkylaryl, C(O)alkenylaryl, C(O)alkynylaryl, C(O)alkoxyalkyl,
C(O)alkylaminoalkyl,
C(O)alkylaminocarbonyl, C(O)cycloalkyl, C(O)aryl, C(O)heteroaryl,
C(O)heterocycloalkyl, CON(R8), CON(R8)alkyl, CON(R8)alkenyl, CON(R8)alkynyl,
CON(R8)alkylaryl, CON(R8)alkenylaryl, CON(R8)alkynylaryl, CON(R8)alkoxyalkyl,
CON(R8)alkylaminoalkyl, CON(R8)alkylaminocarbonyl, CON(R8)alkoxyaryl,
CON(R8)alkylaminoaryl, CON(R8)cycloalkyl, CON(R8)aryl, CON(R8)heteroaryl,
CON(R8)heterocycloalkyl, CON(R8)alkylheterocycloalkyl, N(R8)C(O)alkyl,
N(R8)C(O)alkenyl, N(R8)C(O)- alkynyl, N(R8)C(O)alkylaryl,
N(R8)C(O)alkenylaryl,
N(R8)C(O)alkynylaryl, N(R8)C(O)alkoxyalkyl, N(R8)C(O)alkylaminoalkyl,
N(R8)C(O)alkylaminocarbonyl, N(R8)C(O)alkoxyaryl, N(R8)C(O)alkylaminoaryl,
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N(R8)C(O)cycloalkyl, N(R8)C(O)aryl, N(R8)C(O)heteroaryl,
N(R8)C(O)heterocycloalkyl,
N(R8)C(O)alkylheterocycloalkyl, NHC(O)NH, NHC(O)NH-alkyl, NHC(O)NH-alkenyl,
NHC(O)NH-alkynyl, NHC(O)NH-alkylaryl, NHC(O)NH-alkenylaryl, NHC(O)NH-
alkynylaryl, NHC(O)NH-alkoxyaryl, NHC(O)NH-alkylaminoaryl, NHC(O)NH-
cycloalkyl,
NHC(O)NH-aryl, NHC(O)NH-heteroaryl, NHC(O)NH-heterocycloalkyl, NHC(O)NH-
alkylheterocycloalkyl, S-alkyl, S-alkenyl, S-alkynyl, S-alkoxyalkyl, S-
alkylaminoalkyl, S-
alkylaryl, S-alkylaminocarbonyl, S-alkylaryl, S-alkynylaryl, S-alkoxyaryl, S-
alkylaminoaryl, S-cycloalkyl, S-aryl, S-heteroaryl, S-heterocycloalkyl, S-
alkylheterocycloalkyl, S(O)alkyl, S(O)alkenyl, S(O)alkynyl, S(O)alkoxyalkyl,
S(O)alkylaminoalkyl, S(O)alkylaminocarbonyl, S(O)alkylaryl, S(O)alkenylaryl,
S(O)alkynylaryl, S(O)alkoxyaryl, S(O)alkylaminoaryl, S(O)cycloalkyl, S(O)aryl,
S(O)heteroaryl, S(O)heterocycloalkyl, S(O)alkylheterocycloalkyl, S(O)2alkyl,
S(O)2alkenyl, S(O)2alkynyl, S(O)2alkoxyalkyl, S(O)2alkylaminoalkyl,
S(O)2alkylaminocarbonyl, S(O)2alkylaryl, S(O)2alkenylaryl, S(O)2alkynylaryl,
S(O)2alkoxyaryl, S(O)2alkylaminoaryl, S(O)2yycloalkyl, S(O)2aryl,
S(O)2heteroaryl,
S(O)2heterocycloalkyl, S(O)2alkylheterocycloalkyl, S(O)2heterocyclylalkyl,
S(O)2heterocyclylalkenyl, S(O)2heterocyclylalkynyl, SO2NH, SO2NH-alkyl, SO2NH-
alkenyl, SO2NH-alkynyl, SO2NH-alkylaryl, SO2NH-alkenylaryl, SO2NH-alkynylaryl,
SO2NH-cycloalkyl, SO2NH-aryl, SO2NH-heteroaryl, SO2NH-heterocycloalkyl, SO2NH-
alkylheterocycloalkyl, alkylaryloxyalkoxy, alkylaryloxyalkylamino,
alkylarylaminoalkoxy,
alkylarylaminoalkylamino, alkylarylalkylaminoalkoxy,
alkylarylalkylaminoalkoxy,
alkenylaryloxyalkoxy, alkenylaryloxyalkylamino, alkenylarylaminoalkoxy,
alkenylarylaminoalkylamino, alkenylarylalkylaminoalkoxy,
alkenylarylalkylaminoalkylamino.
In a more preferred embodiment, B is a straight chain alkyl, alkenyl, alkynyl,
arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl,
heteroarylalkynyl,
heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, aryl, heteroaryl,
heterocyclyl,
cycloalkyl, cycloalkenyl, alkylarylalkyl, alkylarylalkenyl, alkylarylalkynyl,
alkenylarylalkyl, alkenylarylalkenyl, alkenylarylalkynyl, alkynylarylalkyl,
alkynylarylalkenyl, alkynylarylalkynyl, alkylheteroarylalkyl,
alkylheteroarylalkenyl,
alkylheteroarylalkynyl, alkenylheteroarylalkyl, alkenylheteroarylalkenyl,
alkenylheteroarylalkynyl, alkynylheteroarylalkyl, alkynylheteroarylalkenyl,
alkynylheteroarylalkynyl, alkylheterocyclylalkyl, alkylheterocyclylalkenyl,
alkylhererocyclylalkynyl, alkenylheterocyclylalkyl,
alkenylheterocyclylalkenyl,
23
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl,
alkynylheterocyclylalkenyl,
alkynylheterocyclylalkynyl, alkylaryl, alkenylaryl, alkynylaryl,
alkylheteroaryl,
alkenylheteroaryl, or alkynylhereroaryl,. One or more methylenes can be
interrupted or
terminated by -0-, -N(R8)-, -C(O)-, -C(O)N(R8)-, or -C(O)O-. Preferably, the C
group is
attached to B via an aliphatic moiety within B.
In one embodiment, the linker B is between 1-24 atoms, preferably 4-24 atoms,
preferably 4-18 atoms, more preferably 4-12 atoms, and most preferably about 4-
10 atoms.
In a preferred embodiment, B is selected from straight chain Ci-Cio alkyl, Ci-
Cio
alkenyl, Ci-Cio alkynyl, Ci-Cio alkoxy, alkoxyCi-Cioalkoxy, Ci-Cio alkylamino,
alkoxyCi-
Cioalkylamino, Ci-Cio alkylcarbonylamino, Ci-Cio alkylaminocarbonyl, aryloxyCi-
Cioalkoxy, aryloxyCi-Cioalkylamino, aryloxyCi-Cioalkylamino carbonyl, Ci-Cio-
alkylaminoalkylaminocarbonyl, Ci-Cio alkyl(N-alkyl)aminoalkyl-aminocarbonyl,
alkylaminoalkylamino, alkylcarbonylaminoalkylamino, alkyl(N-
alkyl)aminoalkylamino,
(N-alkyl)alkylcarbonylaminoalkylamino, alkylaminoalkyl,
alkylaminoalkylaminoalkyl,
alkylpiperazinoalkyl, piperazinoalkyl, alkylpiperazino, alkenylaryloxyCi-
Cioalkoxy,
alkenylarylaminoCi-Cioalkoxy, alkenylaryllalkylaminoCi-Cioalkoxy,
alkenylaryloxyCi-
Cioalkylamino, alkenylaryloxyCi-C ioalkylaminocarbonyl, piperazinoalkylaryl,
heteroarylCi-Cioalkyl, heteroarylC2-Cioalkenyl, heteroarylC2-Cioalkynyl,
heteroarylCi-
Cioalkylamino, heteroarylCi-Cioalkoxy, heteroaryloxyCi-Cioalkyl,
heteroaryloxyC2-
Cioalkenyl, heteroaryloxyC2-Cioalkynyl, heteroaryloxyCi-Cioalkylamino,
heteroaryloxyCi-
Cioalkoxy. In the most preferred embodiments, the C group is attached to B via
an aliphatic
moiety carbon chain, an aryl group or a heteroaryl group within B.
In a particularly preferred embodiment, B is an aryl, heteroaryl, Ci-Cio-
alkylaryl,
Ci-Cio-alkylheteroaryl group, Ci-Cio-alkylheterocyclylaryl, Ci-Cio-
alkylheterocyclylheteroaryl, Ci-Cio-alkylheterocyclylaryl-Ci-Cio-alkyl, or Ci-
Cio-
alkylheterocyclylheteroaryl-Ci-Cio-alkyl group.
It is understood that alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl,
heterocyclyl and the like can be further substituted.
In certain embodiments of the compounds of Formulas I-XIX, B is selected from
the group:
R100 R100
(CH2 I (CH2 I (CH2)
~ ~ e
24
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
100 11100
(CH2)x-C N (CH2-N C
(CH2)cS (CH2)(rO
(CH2fS (CH2)e (CH2 O (CH2)e
11100 II II 100
I (CH2fN C-(CH2)e (CH2f( N-(CH2)e
(CH2)CT--- N N-(CH2)e (CH2 N (CH2)e
(CH2 N-(CH2),
N
(/ N N-(CH2) e
N `~
N
(CH2 / N (CH2)e
N
In another embodiment, B is
N 8100
((CH2)e
In the foregoing formulas, d and e are independently 0, 1, 2, 3, 4, 5, 6, 7 or
8; and
R100 is hydrogen or a group selected from CI-Cg alkyl, C2-Cg alkenyl, C2-Cg
alkynyl, and
C3-Cs cycloalkyl. Preferred alkyl groups are -CH3, -CH2CH3, -CH2CH2CH3, -
CH(CH3)CH3, -C(CH3)2CH3, -C(CH3)3. Preferably, R100 is hydrogen or methyl.
CA 02795952 2012-10-09
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Representative bifunctional compounds of use herein are those selected from
Table
A below or the geometric isomers, enantiomers, diastereomers, racemates,
pharmaceutically acceptable salts, and prodrugs thereof. It is to be
understood that in any
structure in Table A in which a nitrogen atom is represented with an open
valence, that
valence is occupied by a hydrogen atom.
Table A
Compound No. Structure
1 H3C.O S
% N (0)
O ~
N
N S N _N H
\ I ` N- OH
N I O
2
H3C O
S 0
O N~ J
CN1
O OH
S N .N H
%
N
N
3 H3C O (0) sN H% O
o N
N S
N N
`
N
N
4 H3C O'S
. (O) O
ON N O H
S N _N N % N-/-~
N I \
26
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
FiOis~O N.OH % N EN)
O< O
`N S N N
% N
N
6 H3C. = 0 O O
S O Nci~-<xaN
IEN) % 7 H3C, .O
0
' (0)
N N
N
N
0N O
S N N OH
% N
N
g O
HO
.N
H3C'..O 0
S N N
O ON S N N
N
9 H3% o O's %
N (0)
D N
LS ~ I " N N O % N N
N OH
27
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
( )
S 1N \ N N
N N I \
N
O,
HO-N\/\Y
H
11 CO/
S N _N
N
N N I \
N
N
HO 0
12 c0
N
iN NN
N N I \
N
0
~_J~
HO-N
13 C /
S \N N
\N
N N I \
N
N
HO O
14 HO 0
N-
0 N I`J
N N
~N S Nz N N
i N
N
28
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
15 CD O N,
N -N N
OH N S / / \
N
0
16 C
O
N,
N rN N N
OH N
N
17 E0
N
H S I N
O~N i NH N
HO-N
18 (0)
N
N S N N
NH
H N
Hd O
19 (0)
N
H S --- N _N
N NH
HO- N
H
29
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
20 EN)
pN e'c,'-Z N
\ NH
N
H
IN
HO p
21 0
c)
N
S N _N
O\ O NH
7'~~
HO-N
22 (0)
N
S N _N
O NH
H N
HO p
23 EN)
pS N _N
O NH
p~ N I \
HO-N
H
24 (0)
N
S N _N
O I , \ NH
H
,N
HO p
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
25 (0)
N
H S I N N
N \ NH
N _O N
HO O
26 (0)
N
H S ~N _N
I
\ N NH
0
0
HO-N
H
27 (0)
NN S ~N N
NH
O
H I/
,N
HO O
28 (0)
N
H S I N" N .N
N A\ \ NH
O N
HO-N
H
29
0
~N~OH
r -' H
McSO2
,ON Jl
S N N _N
NH
N I \
31
CA 02795952 2012-10-09
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30 H
N, OH
McS02, N` O
CN r`Jl
S N N -N
NH
N
31 0
OH
N-
H
McSOZ` (N
NJ
N S NZ N -N
NH
N
32 NH
, OH
McS02 (N)
CN S N NN _N
NH
N I \
33 McS02
H
NOH
HN
~-N ,
S N O
N
N-NH
34 McS02
~ 0
N H O H
C HN '
N S N
N
N-NH
32
CA 02795952 2012-10-09
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35 McS02
H
,
HN N 'OH
N S Nz N O
N
N-NH
36 McS02
, 0
N
C HN H 'OH
N S NZ N
N
N-NH
37 \ ,O 0
o- s% I N)
N \
`N S S N
N
Q N
N-NH
HO-N
H
38 \ o (0)
0,S`N
N
N S Nz N
N
H
N
\
N N-NH
HO
O
39 \ 'o (0)
O %N
C 0 N
N S N
N
H
N
o
N-NH
HON
H
33
CA 02795952 2012-10-09
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40 \ ,o
o % (0)
N N S N S NN
H
N
H N-NH
HO
0
41 (0)
N
H S ~N -N
HO'NN , I i NH
0 H N
42
CO
)
N
O S N N
HO,H H \ NH N N
43 CND
oH S N _N %
HON N , I i NH
0 H N
Jl
f`
44 (0)
0 S N _N
HO,N N \ \ N%
H
H H N
34
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
45 0
C~
N
H S N _N
HON NH
0 N
46 (0)
N
S N _N
O
' NH
N
HO-N
47 HO 0
N
N ~ `O
N
N
N S N
I
N
O N
48 HO 0
%
N
H 0-
N-( N NH
N
N
N
N
H
49 HO% 0
N
H 0
~N \ NH
N-(
-N
S N
N
CA 02795952 2012-10-09
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50 CNJ
OH
HO-N H S N _N
N
o sot \ N I c N
51 HO 0
N
N~ rO
N `NJ
c -
`nl S s `N N
sot.
N
52 HOB 0
N
H _ 0
N / N,
N -
qo,,s NN / NH
S N \
53 HO 0
N 0
iN
N CNJ
NH S N -N
~N.
54 HQ 0
N N (0)
N ~
N S N N
N N
36
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
55 HO 0 0
N - C 1
N
N S N -N
N H N. [ N
56 HO 0 O
N
N a\N J N -N
/ N .N
57 HO 0 0
N - ~ 1
N
N S `N -N
O N N
58 C)
O\L// \ N N
HO-N N S N _N
H 0 NH
N
)
59 CN
N HO-N N S N _N
H 0 NH
N
37
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
H
(N)
N
OH S N N
HN/ N NH
0 N\ N
N I /
61
N
C~
N
OH S N N
HN N NH
N N/
O \ //N
62 O I
CN)
N
OH S N
HN/ N
~ NH
N/ I \
N
O \ N
63 Ho~
(N)
N
OH S N N
HN/ N \ NH
O /\ N\ N/
N
64
LN
S N N
-ONN N
~> N
HON N
H
38
CA 02795952 2012-10-09
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65 "~(O)"**'
N N -O~ / S ~ N ~N
HO NH N ~- N \ I NH
N
U
66
N
S N N
/ \ NH
N/ \
N
HO-NH
67
ON
~N
/ \ S NH
N\ N HN N I /
OH
68 (0)
N
-O, ~N/ S \ N N
HO N/H}~--(\ -N I NH
U
69 (O)
N
S N N
/ \ NH
~N\ N~
HO-NH -N
F
39
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
70 (0)
N
-C N \
-O~
S N -N
HO NH N \ I / NH
U
71 (0)
N
O~ -NH S N N
H -
NH N \ I / NH
N I \
U
72 (0)
N
O S N _N
N
N N
N
HO H N I /
73 (0)
N
S --N -N
~-CNH
NH HO-NH N
74 (0)
N
S --N -N
~-C N I NH
~N\
H O - N H N
CA 02795952 2012-10-09
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75 co)
N
S N N
N \ NH
'-
HO-NH N N
N
U
76 co)
0 N
/ NH S
N
HO-NH N NH
N
77 (0)
0 N
/ N S
N -N
HO-NH N NH
N
78 (0)
O N
Y-CD~-NH S
N N
HO-NH NH
N/
79 (0)
N
N S N N
\N HO-NH \ / H
N
41
CA 02795952 2012-10-09
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80 (0)
N
O NH S
/ \ I \N NNH
HO-NH N
U
81
co)
N
O S ~ N -N
NH
HO-NH \ / \ N N
U
82 (0)
O N
N N
Y-O-NH g
HO-NH NH
N
83 (0)
N
S N N
/ N
I NH
/
HO-NH -N
84 co)
N
S ~N .N
HO-NH N C/~ NH
~-N N
O N
loo
42
CA 02795952 2012-10-09
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85 (0)
N
S N N
HO-NH / \ NH
~-N N
O -N
U
86
C
)
N
O N S N N
NH
~-('//~~
N
HO NH N
U
OH
87 (0)
N
OH S N ~N
HN/ / \ I NH
N N/
o N I /
O
88 (0)
N
S ~N .N
0 _C N I N
NH
HO-NH N
0
OH
43
CA 02795952 2012-10-09
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89 (0)
N
S I ~N N
_H ON N N
N
HO N
Y-C
O=~
0
90 (0)
N
/ OH
N N
N S
HN
NH
N N
O N /
0
91 (0)
N
S N N
HO-NH N
NH
-N N
O N
U
OH
92 (0)
N
S N N
/ \ NH
~N N/ \
HO- NH N
93 (0)
N
/ \ \ I NH
HO _NH -C
~N N/ \
U
44
CA 02795952 2012-10-09
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94 (0)
N
9 N .N
NH
\ N
HO-NH N
U
NH2
95 COD
N
S N .N
/ \ \ NH
~-N N HO NH N
U
N -
96 COD
N
S N N
/ \ NH
~-N N
HO-NH -N
U
NH
O-/
97 (0)
N
S N .N
/ \ \ NH
~~ N N/
HO-NH N
U
0
NH2
CA 02795952 2012-10-09
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98 COD
N
9 N N
/ \ \ I H
N
~-N N/
HO-NH -N
99 co)
N
S N N
/ \ NH
- ; ~-N N/ \
HO NH N
N
100 (0)
N
S I ~N .N
O N N \ N
r N
_
HO N N O
Y-C
H
101 0
C~
HO 0 N
N
-jl S I N _N
N N
CH3
102 0
N
S
N ~N _N
0
CH3 N N
HO-N
46
CA 02795952 2012-10-09
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O
103 CN
S
III .N _N %
HON N N
0 CH3
104 (O)
N
O S --N _N
HO,N \ I N N
CH3
105 (0)
N
S
N \N H
N N N/
HOB
0
106 (0)
N
N
S N N
HN
1 \ NH
N H0
N 0
H
107 0
N
O / ~N/ S
N
HNH
OH N)
N
47
CA 02795952 2012-10-09
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108 (0)
N
S
0 N
/ _N
HO NH N
109
N
S N
~N\ N N
HO-NH N
110
(0)
N
0 Y-C N S N
HN N~-- \ I /
OH N N
NH2
111
(O)
N
S XtN
Y-C HO-N N
H NH
112 (o)
N
S N
o
N N-0 N 0
HO-N
H N
H
48
CA 02795952 2012-10-09
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O
113 CNI
S I ~N
N Ni NH2
HO-N N N
H
114 (0)
N
N S N
HO N N
H N I N
N
115 (0)
N
0 ~-C / NS N
HN N \ I /
OH N N
N NH2
116
(0)
N
S N
O> N \ \ i
N\ N \ N
HO NH N
N N
H
117
co)
N
S N
0
N
N\ N N
HO NH N
N N
49
CA 02795952 2012-10-09
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118
CNJ
OS LN Y-C \\- N N I ~N O
HON N
H
N~N
H
119 (0)
N
% H S
N
HN \ 1
N N N
O N
N
120
N
O / \~ N/ (Cll N
HO-NH -N N/ N
I
N
121 o
EN)
S LN
O N
N N N
HO H N Ni CH3
122 OEN)
S LN
o
N N N
HO_N N I N NH2
H 0
CA 02795952 2012-10-09
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123 O)
CN
S N
O N
N N N
H0_N N H
H N~l
N II
0
124 0
EN)
S N
O N
N\ N I N
HO-N ~- H N N NH2
125 (0)
N
S N
-OK ~-N \ N/ N
HO NH
N~NHZ
126 o
EN)
S N
O N
N N ~
HO N N
H NN CI
127 O
EN)
S N
NN N ~ N
O ~~N
HO-H N l
C
51
CA 02795952 2012-10-09
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128 O
EN)
S N
O N
\N N N
HO -N N
H NCN
129 (0)
N
~ N S ~N
HO-N N
N
130 (0)
N
0 ~-c ~-N N
/
HN N O
\
OH N
131 (0)
N
N S N
HO H N N
N I \
O
132 co)
N
0 ~-c N-N/ S N
HN N \ I /
OH
OH N
52
CA 02795952 2012-10-09
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133 (0)
O N
N S ~ N
HO H N
N
OH
134 (0)
N
O NL(, N
HN\ -N N
HZ
OH N
135
C
)
N
S \ N
HO NH
NH2
136 0
O N
-Q- N S N
HO-H
N
H
Ny
0
137 co)
N
S N
O / \ N
N NHZ
HO NH
53
CA 02795952 2012-10-09
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138 (0)
N
S N
O / \ ~-N\ N OH
HO-NH -N
139 (0)
N
O/ N
N
N S
HN\
OH N/ O
140 (0)
N
6 N
O / ~N \ N
HO-N N
OH
141 (0)
N
0 Y-C \\ N/ OH
/ N
HN N
OH N
142 (o)
N
S N O
~N N I \ NH2
_O
HO -N N
Y-C
H
54
CA 02795952 2012-10-09
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143 (O)
N
N S ~N
HO H N
N I \
NH2
0
144 (o)
N
S N O
O- \ N \ Nll \ NH
HO-N N
H
145 (O)
N
' N S N
HO-NN N
H N
NO2
146 (O)
N
N S N
HO H N
Y-C
N I \
CN
147 co)
N
N S ~N
N' \
HO_ N
H N I \
CI
CA 02795952 2012-10-09
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148 EN)
0~~ / Oy--(' ~ N S L N
HO-NH N
N
149 EN)
N S ~N
0Y-C
HO H N
N I \
S02
150 (0)
N
0 Y-C \ N g
N
HN\ N
OH N
NH2
151 EN)
0O N ff(' \>- N S N
HO-H /
N
N I \
NH
152 (0)
N
~- Off(' \ N S N
HO N N
H N
Nel
56
CA 02795952 2012-10-09
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153 (O)
0 N N
-
N S N
HO-N N
H
N
NH
154 EN)
O)-
N S N
Y-C
HO-N N
N I \
Nl ~ '
155 (O)
N
N S N
HO-N N L~\
H N
Nc
156 (O)
N
N S N
Y-C
HO H N L~\
N \ NH
NH2
157 (O)
N
N S N
HO-N N
H N I \ ~\
N
H
57
CA 02795952 2012-10-09
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158
OH g
N
HN/ / \ \ N
N NH
\ N/ \
0 - N
159 N
HOH g N N NH
N / ~-CN \ \
~NN/
O U
160
HN
OH g
HN/ / \ \N NH
N
) N- N/
0 N
161 HO
N N
-O~ C ~N/ S \
HO NH N \ I NH
N/
100
162
H N / HO N OH
OH
~-N /
S
N N
0 N NH
N
58
CA 02795952 2012-10-09
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163
0 fu
N
>___)___N(<SJ.L
H
N\ N NH
OH N
164
HN \
OH S N N
HN/LN' NH
O \
165 / ~
~~ HN \
N N
-O~ ~-NS
HO NH N NH
166
HN \
O ~N S N N
HO-NH N NH
N/
/
167
H N\
~\yy----((~////~~
-O -N/ S N N
HO NH N NH
N
59
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168 1
HN N
H S N N
HN N NH
N N/
0 N \
169 / N
I
HN
S N _N
N
O C NrN N
_ "
HO H N
170 N
HN
S N _N
O N N
~ N C
jar N
HO-N N
H
171
O (0)
HO, N N / 1 \
H NO S NZ
, N _N
,S N
O N
172 (0)
HO,N 1 \ N
H NI/0 S ~N _N
~S ~N
O N
CA 02795952 2012-10-09
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173 (0)
N
O ~N _N
N
O C Nr N N
_ "
HO H N
174 EN)
Os l ~N _N
O N ,- 4
N \ N N N
HO-N N
175 OH
HN
O
O
N
()
r N N
N S N _N
\ I NH
N , I
176
END
OS I
O N N
`\ \ N \ NH
N
HO- H N /
177
CO
)
N
S ",N
-O`\ CN NH
HO -N
N 61
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178 0
END
O
S N
O / N N NH
r N
HO-N -N
H
179 (0)
N
S H
N N
O / \N N/
HO-N
N
180 O
CD
N
S O H
N N
O
~N N~
Y-C
HO_H N
181
(o)
N
S H
O N N
~~N Ni O
HO_N N
H
182 O
C~
N
S ~N
Y_C , N Ni \ N
O
_
HO N N
H O
62
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183 o11
CNJ
S I ~N
O ~N \ Ni N O
HO_N N
H
184
CO
)
N
S N
\ N \ N~ i
_ O
Y-C
HO N N
H N
H
O
185 CNI
S N
c /
_O \~N Ni N
HO -N N
Y-C
H N
H
186 (0)
N
N S N
HO H N N N
I / ~O
N
H
187 0)
CN
N S N
Y-C
HO -N
N
N
N
H
63
CA 02795952 2012-10-09
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188 (O)
N
N
O(' > N S N
HO-N N /\ i N
N N I \
N O
H
189 (O)
O N
-\>-N S N
HO N N H
H N a N
N O
H
190 (O)
N
~~ff(' \>- N S N
HO-0N N /\ N i O
N O
H
191 (O)
N
-O -
Y (,SLN
C
HO -N
N i O
N I
/ N
H
192 (O)
N
N S N
Y-C \
HO H N
N
N
H
64
CA 02795952 2012-10-09
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193 EN)
OO ~~ N / (' \> N S N
HO- H ~N
N
N
H
194 EN)
ON
/ O- N S N
HO H ~~ C\>-
N
N
O
H
195 (0)
N
N S N
HO- N H N , N
N
N
H
196 (0)
N
N S N
HO- N H N
N I N
N
H
197 0
EN)
Y-~/- \>- N S ~N
HO H N
N
N
H
CA 02795952 2012-10-09
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198 EN)
OY-C
N S N
N
HO -N
N NO2
N
H
199 (0)
N
6 N
O
N
N \ I l N
N
HO-NH N N
200 (0)
N
0 -N N S N
HO-N N L~\ I
N
F
201 0
EN)
0 -N ~-c \>- - N S N
HO- N N
N
202 0
EN)
N S N
HO-N N i
N I \
/ CF3
66
CA 02795952 2012-10-09
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203 ON N, NH
O N
HO-HN S
H N
N
204 ~~ N
, NH
N
O N
HO-N S / \
H H
O
205 CN'
O N/ S ~N
HO-H -N
N
NH2
206 (0)
O / N
N S ~N
HO -H
N
NH2
207 HO 0
H
N-
//N EN)
N pNN
I~Zllt
NH2
67
CA 02795952 2012-10-09
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208 EN)
OR
S N
O / \ N
)- N )--' \ N \
HO-N N
NH2
209 HO 0
N H (0)
N
H
N S N
N
NH2
210 HO 0
N
H
(0)
N
H
N S N
N
NH2
211 (0)
O H N
N S N
HO_ H N N
N
NH2
212 (0)
N
0 N
N S \ N
HO H N
N
NH2
68
CA 02795952 2012-10-09
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213 (0)
O N
~ -N S N
HO H N
N
NH2
214 0
EN)
/ \ -N S N
HO H N N
N
NH2
215 0
EN)
O N S \ N
HO H N
N
NH2
216 (0)
0
HN N
HO S
N N
S N \ NH
O O
217 co)
N
O
II s
s N
N O H N
/
~ I ~N
HO
0 NH2
69
CA 02795952 2012-10-09
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218 HO
NH
O N
COD
S N
O/ O N ON
N NH2
219 COD
N
S
O w \ I N/
O O
HO-NH 0
220
0 co)
HN N
HO S N
N \
// p N I \~
O
N
221 (0)
0
HN N
HO S
N
# \O N N
O
O
222 co)
N
S N
O is N/
O
O
HO-NH
CA 02795952 2012-10-09
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223 CO/
N
S N
O
~- / / / \ SHN \ I N/ N
H
HO-NH N
224 O
co)
HN N
HO S
N
~ N N
O
NH
225 co)
N
S
H N
N
HOB N
H N
N
O
N NH2
226 COD
N
O S
N
HOB
H H \ N/ N
N:~ NHZ
227 (0)
N
O S
HOB
H H \ N/ I ~N
N~N
H
71
CA 02795952 2012-10-09
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228 co)
N
O S
HOB
H H N au
N
H
229 (0)
N
HOB
H H N/ IN
230 co)
N
O S
HOB
H H
N
O
231 (0)
N
HOB
H H 1 OH
N
F
232 co)
S N
HN
N , ,N
H N NH2
HO_N
0
72
CA 02795952 2012-10-09
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233 CO/
s
N
HOB N \
H N I ~N
o
234 COD
OH
HN
N
H
N NH2
235 off co)
N
O N
\
N
~-N Lcl
HN N OH N/ N
/ NH2
236 (0)
N
-O ~-N S
HO NH N
N/ N
O
237 CO/
N
S N
O
~_CN
N N N
HN N
OH N
H
73
CA 02795952 2012-10-09
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238 co)
-- 11
o N
\\ N
HO-NH N~N
N NH
239 CO)
O N -N I
e,N N
HO-NH N
N NH
240 CO/
O ~-NH S
N
HN\ N
OH N/ I ~N
H
241 CO"
N
o~ ~N/ 5
HO-NH N
N~ N
NH
242 (0)
NJ
0 N
\NH <JN
HO-NH N 0/ ~\
0 N N
N
H
74
CA 02795952 2012-10-09
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243 HO 0
//o
HN f 1
N N
\--~ S \
N
N \
N
H
244 HO 0 O1
N CN
HN \ I /
N N
NH
245 OH co)
N
0 C~N 9
N
HN N \
OH N/ I \ N
H
246 (0)
N
~_C
/~ I
HO NH N L(cll
N
N
247 (0)
N
S \ N
N N N
HN\OH -N / il,
O
CA 02795952 2012-10-09
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248 (O)
O N -N/ S \
N
HO-NH N \
N I N
/
O
249 (0)
N
OC\-N S
N
HO-NH N
N I ~N
/ O
250 CO/
N
9 N
O`/\\JU-\/~ NH N N
HN `
OH 0
251 CO)
N
O N -N/ 3
N
HO-NH N
N/ \N
0
o
252 END
O N
S
% C N 1LN
HO-NH N Oi
O N I ~N
O
76
CA 02795952 2012-10-09
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253 HO` 0
N (0)
NN
N~
S ~ N
HN x
N I N
"' O
254
/H co)
HN \ N
-N S Nk, N
O N
N/ I ~N
/ OH
255 (O)
OH IL\
HN \
-N 5
N
O N
N/ I ~N
0
256 co)
OH
~N
HNC; Lo,
N
O N
N- N
OI
OH
257
OH (0)
HN \ N
~-N S
N
O N
N N
O- V N~
77
CA 02795952 2012-10-09
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258 co)
N
S N
N N
HN -N
\ \ I / H
OH p~ V N~
259 co)
N
S N
Y_C ~ ~- 'o
N N --- N
H N \ N 'NHZ
OH p ^v
260
OH (0)
HN \
~-N S
N
O N
N/ N
261 ( )
s N
NrN ,
N N
HORN -11 N_ O
0
262 COD
N
9 N
N N N
HN -N
O H N O
78
CA 02795952 2012-10-09
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263 co)
N
S N
O / NH N N
HNN
OH N O
264 co)
N
O ~ \~ N
N 3
HO-NH N
N/ I '
O
265 OH CO/
N
ONN
I N
HO-NH N
N/ I ~N
O
266 CO/
p>
~ / S N
HN \ N \
OH N/ I \N
N O
267
C
)
N
S N
O / \ Nom'
N
HO-NH -N
NH2
79
CA 02795952 2012-10-09
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268 (O)
N
S N
O \~N \ N /
HO-NH N
NH2
269 COD
~ N
N
O' L- \NH S
HO_NH N
NH2
270 CO/
N
O~ \~N S
N
HO-NH N
N/
/ NH2
271 COD
N
0 NH
// I N
HN\ N \\ /
OH 0 0 N
/ NH2
272 0 HN
N
HO COD
N
N- S \
N
N \
~ N/ aNH2
CA 02795952 2012-10-09
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273 HO 0
O
N
N CNJ
HN \ I /
N / N
N I ~
NH2
274
OH
N
O~-~-C N
N
HN N
OH N
aNH2
275 co)
N
S
N
O
\\-N _ N
\ N \ /
OH NH2
CI
276 (0)
N
/
-\\r N
O, C _ NL<cl~ \
HN N OH N/
NH2
277 OH co)
N
0, ~-N N
N
-C N
F
D,
OH N
NH2
81
CA 02795952 2012-10-09
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278 (0)
N
N
N s ~l
HN N
~-C
OH N/ O_
NHz
279 (0)
N
S
O N
/ )--N OH
N/
HO-NH -N
NH2
280
co)
N
O -N ~
I S ~N
\
N
HN N
OH N
NH2
281 (0)
O N N
N/ S
~N 0
HN\ \
OH N
NH2
282 co)
N
S N
O / / \ I F
N/
HO-NH N NHz
F
82
CA 02795952 2012-10-09
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283 (0)
N
O / ~ N S
N F
H N N \
OH N
NH2
284 (0)
N
O Y-C/- N/ S
N Cl
HN\ N
OH N
NH2
285 co)
N
OY-C~-NS
HN N
OH N OH
NH2
286 co)
N
S N
O -N N/
HN N
I /
NH
OH
287 co)
N
pN
/ S I N
HN \
OH N/
H
83
CA 02795952 2012-10-09
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288 co)
N
S N
O \ O
-N\ N
N\II/
HN
N
\ O
OH
289 co)
O N N
N S
I N
HN\
O` ^
OH N/ I v `OH
290 OH (0)
H N/ N
N/ S
N
O N \
DI / S
HO
291 (O)
N
O CN -N S
HO-NH N \ I \ N II
N S
I I ,
OH
292 co)
N
O, N N/ S
HN \ \
OH
OH N/
F
84
CA 02795952 2012-10-09
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293 COD
N
O ~~~ S ~N
\\--(~ // OH
/y
N
HN N O
OH N
0-
294 (0)
N
O ~N 9
H N \ N
OH N/ OH
295 (0)
N
O >-c N~N S
N OH
H N \ N
OH N/
F
296 CO/
N
ON~N/
N OH
HN\ N
OH N
297
C
)
N
O,
N OH
HN\
OH N
CA 02795952 2012-10-09
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298 C /
N
N HO"N
0 -NLc
HY-0
OH N
F. 299 co)
N
S HOB
_N N/
HO NH N
300 co)
N
S LNO
/ N
HO-NH -N\ N
301 (0)
N
O Y-C/- N S
N O
HN N
OH N
F
302 C /
N
N
O ~_ N \ N/
HO-NH N
OH
86
CA 02795952 2012-10-09
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303 co)
N
N 0
~-C / N S
HN N
OH N/ OH
U
304
/H co)
HN \ N
~-N/ S
N
O N
N/
O O
305 COD
N
O -N S
HO-NH N I N II
S
N I II~NHZ
306 (0)
N
N/ S
N
HN\ \
OH N
OH
307 ()
N
O
N
NH \ N/
NH,
N
N
N
HO
HN
87
CA 02795952 2012-10-09
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308 (0)
N
O g
N
NH N ~H
N
N~
N
HO
HN
309
%H co)
NHN \
NCX 0 s
HN-SI N
O N
pl \ I N/
/ NHZ
310
OH CO/
HN N
~~ S N
p / ~N~/\//\/HN-SI -11
N -- N
N
H
311 co)
N
i H S N
HN \
N N Y~ \
I ~N
O N g<
312 CO/
N
/OH S N
HN \
N N" Y/ \
O N
OH
88
CA 02795952 2012-10-09
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313 OH 0
~-N -O N N
S
N
HO NH N
N/ N
N
314 co)
N
HO-NH N / O
O /N S N HN~
~~~~~ N H
N
315
C
)
N
OY S
N HN
-C~-N S
HN N
NH
OH N
316 (0)
S
OY N N
N HN
~-N S
HN N
NH
OH N
317 OH co)
HN ~-c N O
/>- S N S~
O N / NH
N
89
CA 02795952 2012-10-09
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318 co)
N
S
N
0 / /-N
N \ N N F
HO-NH -N F
N F
H
319 co)
s
N
N\ N/ I ~ O
/J /OH
NJ O
O4tz, S/ H
/ -l-O
320 co)
I "
N N
NJ / O
~5~ HNII
O OH
321 co)
N
N\-\ N/ O
OH
NJ O
044" 5 H
/-10
322 CO/
O N
HOB
N N~ \ I \ N
N I
N/
H
CA 02795952 2012-10-09
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323 0
0
HOB N CN
H
N
k~l
N\
`NH N/ I N
/ NH
324 N (0)
N
HORN / \ 1 S ~N
N \ I N/
I / NHz
325 O N
co)
HO-
N / \ I N
S N
N N/
H
/ NH2
326 CN/
s
I N
NH \ / I \
Nom(
N~ / NHz
O
HO/NH
327 CNl
\
I "
N N-
N- / NHp
O
HO/NH
91
CA 02795952 2012-10-09
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o
328 C /
HORN / aN 5 \N
H
N N \ I N/ \N
H
N
H
329 ( )
N S N
HNOH ~ \ I \ N N/
I I / N
H
330 CO/
S \N
HN N"-- N
O
HN
HO O
331 (O)
N
HN
N"--
N
H
HN
1 4
HO O
In one embodiment, the P13 kinase and inhibitor and the HDAC inhibitor, or the
bifunctional compound is administered as a pharmaceutical composition
comprising the
active compound and a pharmaceutically acceptable carrier or excipient.
Suitable
pharmaceutical compositions comprise any solid or liquid physical form of the
active
compound. For example, the compounds can be in a crystalline form, in
amorphous form,
and have any particle size. The particles may be micronized, or may be
agglomerated,
particulate granules, powders, oils, oily suspensions or any other form of
solid or liquid
physical form. Suitable pharmaceutical compositions typically comprise a
therapeutically
effective amount of an active compound, and a pharmaceutically acceptable
carrier.
92
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
Compounds of use in the methods of the invention may be administered by any
suitable means, including, without limitation, parenteral, intravenous,
intramuscular,
subcutaneous, implantation, oral, sublingual, buccal, nasal, pulmonary,
transdermal,
topical, vaginal, rectal, and transmucosal administrations or the like.
Topical
administration can also involve the use of transdermal administration such as
transdermal
patches or iontophoresis devices. Pharmaceutical preparations include a solid,
semisolid or
liquid preparation (tablet, pellet, troche, capsule, suppository, cream,
ointment, aerosol,
powder, liquid, emulsion, suspension, syrup, injection etc.) containing a
compound of the
invention as an active ingredient, which is suitable for selected mode of
administration. In
one embodiment, the pharmaceutical compositions are administered orally, and
are thus
formulated in a form suitable for oral administration, i.e., as a solid or a
liquid preparation.
Suitable solid oral formulations include tablets, capsules, pills, granules,
pellets, sachets
and effervescent, powders, and the like. Suitable liquid oral formulations
include solutions,
suspensions, dispersions, emulsions, oils and the like. In one embodiment of
the present
invention, the composition is formulated in a capsule. In accordance with this
embodiment,
the compositions of the present invention comprise in addition to the active
compound and
the inert carrier or diluent, a hard gelatin capsule.
Any inert excipient that is commonly used as a carrier or diluent may be used
in the
formulations of the present invention, such as for example, a gum, a starch, a
sugar, a
cellulosic material, an acrylate, or mixtures thereof. A preferred diluent is
microcrystalline
cellulose. The compositions may further comprise a disintegrating agent (e.g.,
croscarmellose sodium) and a lubricant (e.g., magnesium stearate), and may
additionally
comprise one or more additives selected from a binder, a buffer, a protease
inhibitor, a
surfactant, a solubilizing agent, a plasticizer, an emulsifier, a stabilizing
agent, a viscosity
increasing agent, a sweetener, a film forming agent, or any combination
thereof.
Furthermore, the compositions of the present invention may be in the form of
controlled
release or immediate release formulations.
For liquid formulations, pharmaceutically acceptable carriers may be aqueous
or
non-aqueous solutions, suspensions, emulsions or oils. Examples of non-aqueous
solvents
are propylene glycol, polyethylene glycol, and injectable organic esters such
as ethyl oleate.
Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or
suspensions,
including saline and buffered media. Examples of oils are those of petroleum,
animal,
vegetable, or synthetic origin, for example, peanut oil, soybean oil, mineral
oil, olive oil,
sunflower oil, and fish-liver oil. Solutions or suspensions can also include
the following
93
CA 02795952 2012-10-09
WO 2011/130628 PCT/US2011/032683
components: a sterile diluent such as water for injection, saline solution,
fixed oils,
polyethylene glycols, glycerine, propylene glycol or other synthetic solvents;
antibacterial
agents such as benzyl alcohol or methyl parabens; antioxidants such as
ascorbic acid or
sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid
(EDTA); buffers
such as acetates, citrates or phosphates, and agents for the adjustment of
tonicity such as
sodium chloride or dextrose. The pH can be adjusted with acids or bases, such
as
hydrochloric acid or sodium hydroxide.
In addition, the compositions may further comprise binders (e.g., acacia,
cornstarch,
gelatin, carbomer, ethyl cellulose, guar gum, hydroxypropyl cellulose,
hydroxypropyl
methyl cellulose, povidone), disintegrating agents (e.g., cornstarch, potato
starch, alginic
acid, silicon dioxide, croscarmellose sodium, crospovidone, guar gum, sodium
starch
glycolate, Primogel), buffers (e.g., tris-HCI., acetate, phosphate) of various
pH and ionic
strength, additives such as albumin or gelatin to prevent absorption to
surfaces, detergents
(e.g., Tween 20, Tween 80, Pluronic F68, bile acid salts), protease
inhibitors, surfactants
(e.g., sodium lauryl sulfate), permeation enhancers, solubilizing agents
(e.g., glycerol,
polyethylene glycerol), a glidant (e.g., colloidal silicon dioxide), anti-
oxidants (e.g.,
ascorbic acid, sodium metabisulfite, butylated hydroxyanisole), stabilizers
(e.g.,
hydroxypropyl cellulose, hydroxypropylmethyl cellulose), viscosity increasing
agents (e.g.,
carbomer, colloidal silicon dioxide, ethyl cellulose, guar gum), sweeteners
(e.g., sucrose,
aspartame, citric acid), flavoring agents (e.g., peppermint, methyl
salicylate, or orange
flavoring), preservatives (e.g., Thimerosal, benzyl alcohol, parabens),
lubricants (e.g.,
stearic acid, magnesium stearate, polyethylene glycol, sodium lauryl sulfate),
flow-aids
(e.g., colloidal silicon dioxide), plasticizers (e.g., diethyl phthalate,
triethyl citrate),
emulsifiers (e.g., carbomer, hydroxypropyl cellulose, sodium lauryl sulfate),
polymer
coatings (e.g., poloxamers or poloxamines), coating and film forming agents
(e.g., ethyl
cellulose, acrylates, polymethacrylates) and/or adjuvants.
In one embodiment, the active compounds are prepared with carriers that will
protect the compound against rapid elimination from the body, such as a
controlled release
formulation, including implants and microencapsulated delivery systems.
Biodegradable,
biocompatible polymers can be used, such as ethylene vinyl acetate,
polyanhydrides,
polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for
preparation
of such formulations will be apparent to those skilled in the art. The
materials can also be
obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc.
Liposomal
suspensions (including liposomes targeted to infected cells with monoclonal
antibodies to
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viral antigens) can also be used as pharmaceutically acceptable carriers.
These can be
prepared according to methods known to those skilled in the art, for example,
as described
in U.S. Pat No. 4,522,811.
It is especially advantageous to formulate oral compositions in unit dosage
form for
ease of administration and uniformity of dosage. Dosage unit form as used
herein refers to
physically discrete units suited as unitary dosages for the subject to be
treated; each unit
containing a predetermined quantity of active compound calculated to produce
the desired
therapeutic effect in association with the required pharmaceutical carrier.
The specification
for the dosage unit forms of the invention are dictated by and directly
dependent on the
unique characteristics of the active compound and the particular therapeutic
effect to be
achieved, and the limitations inherent in the art of compounding such an
active compound
for the treatment of individuals.
The pharmaceutical compositions can be included in a container, pack, or
dispenser
together with instructions for administration.
The preparation of pharmaceutical compositions that contain an active compound
is
well understood in the art, for example, by mixing, granulating, or tablet-
forming
processes. The active therapeutic ingredient is often mixed with excipients
that are
pharmaceutically acceptable and compatible with the active ingredient. For
oral
administration, the active agents are mixed with additives customary for this
purpose, such
as vehicles, stabilizers, or inert diluents, and converted by customary
methods into suitable
forms for administration, such as tablets, coated tablets, hard or soft
gelatin capsules,
aqueous, alcoholic or oily solutions and the like as detailed above.
The amount of the compound administered to the patient is preferably less than
an
amount that would cause toxicity in the patient. In certain embodiments, the
amount of the
compound that is administered to the patient is less than the amount that
causes a
concentration of the compound in the patient's plasma to equal or exceed the
toxic level of
the compound. Preferably, the concentration of the compound in the patient's
plasma is
maintained at about 10 nM. In one embodiment, the concentration of the
compound in the
patient's plasma is maintained at about 25 nM. In one embodiment, the
concentration of the
compound in the patient's plasma is maintained at about 50 nM. In one
embodiment, the
concentration of the compound in the patient's plasma is maintained at about
100 nM. In
one embodiment, the concentration of the compound in the patient's plasma is
maintained at
about 500 nM. In one embodiment, the concentration of the compound in the
patient's
plasma is maintained at about 1000 nM. In one embodiment, the concentration of
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compound in the patient's plasma is maintained at about 2500 nM. In one
embodiment, the
concentration of the compound in the patient's plasma is maintained at about
5000 nM. The
optimal amount of the compound that should be administered to the patient in
the practice
of the present invention will depend on the particular compound used and the
type of
cancer being treated.
DEFINITIONS
Listed below are definitions of various terms used to describe this invention.
These
definitions apply to the terms as they are used throughout this specification
and claims,
unless otherwise limited in specific instances, either individually or as part
of a larger
group.
An "aliphatic group" or "aliphatic" is non-aromatic moiety that may be
saturated
(e.g. single bond) or contain one or more units of unsaturation, e.g., double
and/or triple
bonds. An aliphatic group may be straight chained, branched or cyclic, contain
carbon,
hydrogen or, optionally, one or more heteroatoms and may be substituted or
unsubstituted.
An aliphatic group, when used as a linker, preferably contains between about 1
and about
24 atoms, more preferably between about 4 to about 24 atoms, more preferably
between
about 4-12 atoms, more typically between about 4 and about 8 atoms. An
aliphatic group,
when used as a substituent, preferably contains between about 1 and about 24
atoms, more
preferably between about 1 to about 10 atoms, more preferably between about 1-
8 atoms,
more typically between about 1 and about 6 atoms. In addition to aliphatic
hydrocarbon
groups, aliphatic groups include, for example, polyalkoxyalkyls, such as
polyalkylene
glycols, polyamines, and polyimines, for example. Such aliphatic groups may be
further
substituted. It is understood that aliphatic groups may include alkyl,
substituted alkyl,
alkenyl, substituted alkenyl, alkynyl, substituted alkynyl groups described
herein.
The term "substituted carbonyl" includes compounds and moieties which contain
a
carbon connected with a double bond to an oxygen atom, and tautomeric forms
thereof.
Examples of moieties that contain a substituted carbonyl include aldehydes,
ketones,
carboxylic acids, amides, esters, anhydrides, etc. The term "carbonyl moiety"
refers to
groups such as "alkylcarbonyl" groups wherein an alkyl group is covalently
bound to a
carbonyl group, "alkenylcarbonyl" groups wherein an alkenyl group is
covalently bound to
a carbonyl group, "alkynylcarbonyl" groups wherein an alkynyl group is
covalently bound
to a carbonyl group, "arylcarbonyl" groups wherein an aryl group is covalently
attached to
the carbonyl group. Furthermore, the term also refers to groups wherein one or
more
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heteroatoms are covalently bonded to the carbonyl moiety. For example, the
term includes
moieties such as, for example, aminocarbonyl moieties, (wherein a nitrogen
atom is bound
to the carbon of the carbonyl group, e.g., an amide).
The term "acyl" refers to hydrogen, alkyl, partially saturated or fully
saturated
cycloalkyl, partially saturated or fully saturated heterocycle, aryl, and
heteroaryl substituted
carbonyl groups. For example, acyl includes groups such as (Ci-C6)alkanoyl
(e.g., formyl,
acetyl, propionyl, butyryl, valeryl, caproyl, t-butylacetyl, etc.), (C3-
C6)cycloalkylcarbonyl
(e.g., cyclopropylcarbonyl, cyclobutylcarbonyl, cyclobetylcarbonyl,
cyclohexylcarbonyl,
etc.), heterocyclic carbonyl (e.g., pyrrolidinylcarbonyl, pyrrolid-2-one-5-
carbonyl,
piperidinylcarbonyl, piperazinylcarbonyl, tetrahydrofuranylcarbonyl, etc.),
aroyl (e.g.,
benzoyl) and heteroaroyl (e.g., thiophenyl-2-carbonyl, thiophenyl-3-carbonyl,
furanyl-2-
carbonyl, furanyl-3-carbonyl, 1H-pyrroyl-2-carbonyl, 1H-pyrroyl-3-carbonyl,
benzo[b]thiophenyl-2-carbonyl, etc.). In addition, the alkyl, cycloalkyl,
heterocycle, aryl
and heteroaryl portion of the acyl group may be any one of the groups
described in the
respective definitions. When indicated as being "optionally substituted", the
acyl group may
be unsubstituted or optionally substituted with one or more substituents
(typically, one to
three substituents) independently selected from the group of substituents
listed below in the
definition for "substituted" or the alkyl, cycloalkyl, heterocycle, aryl and
heteroaryl portion
of the acyl group may be substituted as described above in the preferred and
more preferred
list of substituents, respectively.
The term "alkyl" embraces linear or branched radicals having one to about
twenty
carbon atoms or, preferably, one to about twelve carbon atoms. More preferred
alkyl
radicals are "lower alkyl" radicals having one to about ten carbon atoms. Most
preferred are
lower alkyl radicals having one to about eight carbon atoms. Examples of such
radicals
include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-
butyl, pentyl,
iso-amyl, hexyl and the like.
The term "alkenyl" embraces linear or branched radicals having at least one
carbon-
carbon double bond of two to about twenty carbon atoms or, preferably, two to
about
twelve carbon atoms. More preferred alkenyl radicals are "lower alkenyl"
radicals having
two to about ten carbon atoms and more preferably about two to about eight
carbon atoms.
Examples of alkenyl radicals include ethenyl, allyl, propenyl, butenyl and 4-
methylbutenyl.
The terms "alkenyl", and "lower alkenyl", embrace radicals having "cis" and
"trans"
orientations, or alternatively, "E" and "Z" orientations.
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The term "alkynyl" embraces linear or branched radicals having at least one
carbon-
carbon triple bond of two to about twenty carbon atoms or, preferably, two to
about twelve
carbon atoms. More preferred alkynyl radicals are "lower alkynyl" radicals
having two to
about ten carbon atoms and more preferably about two to about eight carbon
atoms.
Examples of alkynyl radicals include propargyl, 1-propynyl, 2-propynyl, 1-
butyne, 2-
butynyl and 1-pentynyl.
The term "cycloalkyl" embraces saturated carbocyclic radicals having three to
about
twelve carbon atoms. The term "cycloalkyl" embraces saturated carbocyclic
radicals having
three to about twelve carbon atoms. More preferred cycloalkyl radicals are
"lower
cycloalkyl" radicals having three to about eight carbon atoms. Examples of
such radicals
include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.
The term "cycloalkenyl" embraces partially unsaturated carbocyclic radicals
having
three to twelve carbon atoms. Cycloalkenyl radicals that are partially
unsaturated
carbocyclic radicals that contain two double bonds (that may or may not be
conjugated) can
be called "cycloalkyldienyl". More preferred cycloalkenyl radicals are "lower
cycloalkenyl"
radicals having four to about eight carbon atoms. Examples of such radicals
include
cyclobutenyl, cyclopentenyl and cyclohexenyl.
The term "alkoxy" embraces linear or branched oxy-containing radicals each
having
alkyl portions of one to about twenty carbon atoms or, preferably, one to
about twelve
carbon atoms. More preferred alkoxy radicals are "lower alkoxy" radicals
having one to
about ten carbon atoms and more preferably having one to about eight carbon
atoms.
Examples of such radicals include methoxy, ethoxy, propoxy, butoxy and tert-
butoxy.
The term "alkoxyalkyl" embraces alkyl radicals having one or more alkoxy
radicals
attached to the alkyl radical, that is, to form monoalkoxyalkyl and
dialkoxyalkyl radicals.
The term "aryl", alone or in combination, means a carbocyclic aromatic system
containing one, two or three rings wherein such rings may be attached together
in a pendent
manner or may be fused. The term "aryl" embraces aromatic radicals such as
phenyl,
naphthyl, tetrahydronaphthyl, indane and biphenyl.
The terms "heterocyclyl", "heterocycle" "heterocyclic" or "heterocyclo"
embrace
saturated, partially unsaturated and unsaturated heteroatom-containing ring-
shaped radicals,
which can also be called "heterocyclyl", "heterocycloalkenyl" and "heteroaryl"
correspondingly, where the heteroatoms may be selected from nitrogen, sulfur
and oxygen.
Examples of saturated heterocyclyl radicals include saturated 3 to 6-membered
heteromonocyclic group containing 1 to 4 nitrogen atoms (e.g. pyrrolidinyl,
imidazolidinyl,
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piperidino, piperazinyl, etc.); saturated 3 to 6-membered heteromonocyclic
group
containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms (e.g. morpholinyl,
etc.); saturated
3 to 6-membered heteromonocyclic group containing 1 to 2 sulfur atoms and 1 to
3
nitrogen atoms (e.g., thiazolidinyl, etc.). Examples of partially unsaturated
heterocyclyl
radicals include dihydrothiophene, dihydropyran, dihydrofuran and
dihydrothiazole.
Heterocyclyl radicals may include a pentavalent nitrogen, such as in
tetrazolium and
pyridinium radicals. The term "heterocycle" also embraces radicals where
heterocyclyl
radicals are fused with aryl or cycloalkyl radicals. Examples of such fused
bicyclic radicals
include benzofuran, benzothiophene, and the like.
The term "heteroaryl" embraces unsaturated heterocyclyl radicals. Examples of
heteroaryl radicals include unsaturated 3 to 6 membered heteromonocyclic group
containing 1 to 4 nitrogen atoms, for example, pyrrolyl, pyrrolinyl,
imidazolyl, pyrazolyl,
pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazolyl (e.g., 4H-1,2,4-
triazolyl, 1H-1,2,3-
triazolyl, 2H-1,2,3-triazolyl, etc.) tetrazolyl (e.g. 1H-tetrazolyl, 2H-
tetrazolyl, etc.), etc.;
unsaturated condensed heterocyclyl group containing 1 to 5 nitrogen atoms, for
example,
indolyl, isoindolyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl,
indazolyl,
benzotriazolyl, tetrazolopyridazinyl (e.g., tetrazolo[1,5-b]pyridazinyl,
etc.), etc.;
unsaturated 3 to 6-membered heteromonocyclic group containing an oxygen atom,
for
example, pyranyl, furyl, etc.; unsaturated 3 to 6-membered heteromonocyclic
group
containing a sulfur atom, for example, thienyl, etc.; unsaturated 3- to 6-
membered
heteromonocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen
atoms, for
example, oxazolyl, isoxazolyl, oxadiazolyl (e.g., 1,2,4-oxadiazolyl, 1,3,4-
oxadiazolyl,
1,2,5-oxadiazolyl, etc.) etc.; unsaturated condensed heterocyclyl group
containing 1 to 2
oxygen atoms and 1 to 3 nitrogen atoms (e.g. benzoxazolyl, benzoxadiazolyl,
etc.);
unsaturated 3 to 6-membered heteromonocyclic group containing 1 to 2 sulfur
atoms and 1
to 3 nitrogen atoms, for example, thiazolyl, thiadiazolyl (e.g., 1,2,4-
thiadiazolyl, 1,3,4-
thiadiazolyl, 1,2,5-thiadiazolyl, etc.) etc.; unsaturated condensed
heterocyclyl group
containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms (e.g.,
benzothiazolyl,
benzothiadiazolyl, etc.) and the like.
The term "heterocycloalkyl" embraces heterocyclo-substituted alkyl radicals.
More
preferred heterocycloalkyl radicals are "lower heterocycloalkyl" radicals
having one to six
carbon atoms in the heterocyclo radicals.
The term "alkylthio" embraces radicals containing a linear or branched alkyl
radical,
of one to about ten carbon atoms attached to a divalent sulfur atom. Preferred
alkylthio
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radicals have alkyl radicals of one to about twenty carbon atoms or,
preferably, one to
about twelve carbon atoms. More preferred alkylthio radicals have alkyl
radicals are
"lower alkylthio" radicals having one to about ten carbon atoms. Most
preferred are
alkylthio radicals having lower alkyl radicals of one to about eight carbon
atoms. Examples
of such lower alkylthio radicals are methylthio, ethylthio, propylthio,
butylthio and
hexylthio.
The terms "aralkyl" or "arylalkyl" embrace aryl-substituted alkyl radicals
such as
benzyl, diphenylmethyl, triphenylmethyl, phenylethyl, and diphenylethyl.
The term "aryloxy" embraces aryl radicals attached through an oxygen atom to
other radicals.
The terms "aralkoxy" or "arylalkoxy" embrace aralkyl radicals attached through
an
oxygen atom to other radicals.
The term "aminoalkyl" embraces alkyl radicals substituted with amino radicals.
Preferred aminoalkyl radicals have alkyl radicals having about one to about
twenty carbon
atoms or, preferably, one to about twelve carbon atoms. More preferred
aminoalkyl
radicals are "lower aminoalkyl" that have alkyl radicals having one to about
ten carbon
atoms. Most preferred are aminoalkyl radicals having lower alkyl radicals
having one to
eight carbon atoms. Examples of such radicals include aminomethyl, aminoethyl,
and the
like.
The term "alkylamino" denotes amino groups which are substituted with one or
two
alkyl radicals. Preferred alkylamino radicals have alkyl radicals having about
one to about
twenty carbon atoms or, preferably, one to about twelve carbon atoms. More
preferred
alkylamino radicals are "lower alkylamino" that have alkyl radicals having one
to about ten
carbon atoms. Most preferred are alkylamino radicals having lower alkyl
radicals having
one to about eight carbon atoms. Suitable lower alkylamino may be
monosubstituted N-
alkylamino or disubstituted N,N-alkylamino, such as N-methylamino, N-
ethylamino, N,N-
dimethylamino, N,N-diethylamino or the like.
The term "linker" means an organic moiety that connects two parts of a
compound.
Linkers typically comprise a direct bond or an atom such as oxygen or sulfur,
a unit such as
NR8, C(O), C(O)NH, SO, SO2, SO2NH or a chain of atoms, such as substituted or
unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or
unsubstituted
alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl,
heteroarylalkenyl,
heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl,
heterocyclylalkynyl, aryl,
heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl,
alkylarylalkenyl,
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alkylarylalkynyl, alkenylarylalkyl, alkenylarylalkenyl, alkenylarylalkynyl,
alkynylarylalkyl,
alkynylarylalkenyl, alkynylarylalkynyl, alkylheteroarylalkyl,
alkylheteroarylalkenyl,
alkylheteroarylalkynyl, alkenylheteroarylalkyl, alkenylheteroarylalkenyl,
alkenylheteroarylalkynyl, alkynylheteroarylalkyl, alkynylheteroarylalkenyl,
alkynylheteroarylalkynyl, alkylheterocyclylalkyl, alkylheterocyclylalkenyl,
alkylhererocyclylalkynyl, alkenylheterocyclylalkyl,
alkenylheterocyclylalkenyl,
alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl,
alkynylheterocyclylalkenyl,
alkynylheterocyclylalkynyl, alkylaryl, alkenylaryl, alkynylaryl,
alkylheteroaryl,
alkenylheteroaryl, alkynylhereroaryl, which one or more methylenes can be
interrupted or
terminated by 0, S, S(O), SO2, N(R8), C(O), substituted or unsubstituted aryl,
substituted
or unsubstituted heteroaryl, substituted or unsubstituted heterocyclic; where
Rs is hydrogen,
acyl, aliphatic or substituted aliphatic. In one embodiment, the linker B is
between 1-24
atoms, preferably 4-24 atoms, preferably 4-18 atoms, more preferably 4-12
atoms, and most
preferably about 4-10 atoms. In some embodiments, the linker is a
C(O)NH(alkyl) chain or
an alkoxy chain. It is to be understood that an asymmetric linker, such as
alkylaryl, can
connect two structurally distinct moieties in either of its two possible
orientations.
The term "substituted" refers to the replacement of one or more hydrogen
radicals in
a given structure with the radical of a specified substituent including, but
not limited to:
halo, alkyl, alkenyl, alkynyl, aryl, heterocyclyl, thiol, alkylthio, arylthio,
alkylthioalkyl,
arylthioalkyl, alkylsulfonyl, alkylsulfonylalkyl, arylsulfonylalkyl, alkoxy,
aryloxy,
aralkoxy, aminocarbonyl, alkylaminocarbonyl, arylaminocarbonyl,
alkoxycarbonyl,
aryloxycarbonyl, haloalkyl, amino, trifluoromethyl, cyan, nitro, alkylamino,
arylamino,
alkylaminoalkyl, arylaminoalkyl, aminoalkylamino, hydroxy, alkoxyalkyl,
carboxyalkyl,
alkoxycarbonylalkyl, aminocarbonylalkyl, acyl, aralkoxycarbonyl, carboxylic
acid, sulfonic
acid, sulfonyl, phosphonic acid, aryl, heteroaryl, heterocyclic, and
aliphatic. It is
understood that the substituent may be further substituted.
For simplicity, chemical moieties are defined and referred to throughout can
be
univalent chemical moieties (e.g., alkyl, aryl, etc.) or multivalent moieties
under the
appropriate structural circumstances clear to those skilled in the art. For
example, an "alkyl"
moiety can be referred to a monovalent radical (e.g. CH3-CH2-), or in other
instances, a
bivalent linking moiety can be "alkyl," in which case those skilled in the art
will understand
the alkyl to be a divalent radical (e.g., -CH2-CH2-), which is equivalent to
the term
"alkylene." Similarly, in circumstances in which divalent moieties are
required and are
stated as being "alkoxy", "alkylamino", "aryloxy", "alkylthio", "aryl",
"heteroaryl",
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"heterocyclic", "alkyl" "alkenyl", "alkynyl", "aliphatic", or "cycloalkyl",
those skilled in
the art will understand that the terms alkoxy", "alkylamino", "aryloxy",
"alkylthio", "aryl",
"heteroaryl", "heterocyclic", "alkyl", "alkenyl", "alkynyl", "aliphatic", or
"cycloalkyl"
refer to the corresponding divalent moiety.
The terms "halogen" or "halo" as used herein, refers to an atom selected from
fluorine, chlorine, bromine and iodine.
As used herein, the term "aberrant proliferation" refers to abnormal cell
growth.
The phrase "adjunctive therapy" encompasses treatment of a subject with agents
that
reduce or avoid side effects associated with the combination therapy of the
present
invention, including, but not limited to, those agents, for example, that
reduce the toxic
effect of anticancer drugs, e.g., bone resorption inhibitors, cardioprotective
agents; prevent
or reduce the incidence of nausea and vomiting associated with chemotherapy,
radiotherapy
or operation; or reduce the incidence of infection associated with the
administration of
myelosuppressive anticancer drugs.
The term "apoptosis" as used herein refers to programmed cell death as
signaled by
the nuclei in normally functioning human and animal cells when age or state of
cell health
and condition dictates. An "apoptosis inducing agent" triggers the process of
programmed
cell death.
The term "cancer" as used herein denotes a class of diseases or disorders
characterized by uncontrolled division of cells and the ability of these cells
to invade other
tissues, either by direct growth into adjacent tissue through invasion or by
implantation into
distant sites by metastasis.
The term "compound" is defined herein to include pharmaceutically acceptable
salts, solvates, hydrates, polymorphs, enantiomers, diastereoisomers,
racemates and the like
of the compounds having a formula as set forth herein.
The term "device" refers to any appliance, usually mechanical or electrical,
designed to perform a particular function.
As used herein, the term "dysplasia" refers to abnormal cell growth, and
typically
refers to the earliest form of pre-cancerous lesion recognizable in a biopsy
by a pathologist.
As used herein, the term "a therapeutically effective amount" of a compound or
combination of compounds with respect to the subject method of treatment,
refers to an
amount of the subject compound or the amounts of two or more compounds in
combination, which, when delivered as part of desired dose regimen, brings
about, e.g. a
change in the rate of cell proliferation and/or state of differentiation
and/or rate of survival
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of a cell to clinically acceptable standards. This amount may further relieve
to some extent
one or more of the symptoms of a neoplasia disorder, including, but is not
limited to: 1)
reduction in the number of cancer cells; 2) reduction in tumor size; 3)
inhibition (i.e.,
slowing to some extent, preferably stopping) of cancer cell infiltration into
peripheral
organs; 4) inhibition (i.e., slowing to some extent, preferably stopping) of
tumor metastasis;
5) inhibition, to some extent, of tumor growth; 6) relieving or reducing to
some extent one
or more of the symptoms associated with the disorder; and/or 7) relieving or
reducing the
side effects associated with the administration of anticancer agents.
The term "hyperplasia," as used herein, refers to excessive cell division or
growth.
The phrase an "immunotherapeutic agent" refers to agents used to transfer the
immunity of an immune donor, e.g., another person or an animal, to a host by
inoculation.
The term embraces the use of serum or gamma globulin containing performed
antibodies
produced by another individual or an animal; nonspecific systemic stimulation;
adjuvants;
active specific immunotherapy; and adoptive immunotherapy. Adoptive
immunotherapy
refers to the treatment of a disease by therapy or agents that include host
inoculation of
sensitized lymphocytes, transfer factor, immune RNA, or antibodies in serum or
gamma
globulin.
The term "inhibition," in the context of neoplasia, tumor growth or tumor cell
growth, may be assessed by delayed appearance of primary or secondary tumors,
slowed
development of primary or secondary tumors, decreased occurrence of primary or
secondary tumors, slowed or decreased severity of secondary effects of
disease, arrested
tumor growth and regression of tumors, among others. In the extreme, complete
inhibition,
is referred to herein as prevention or chemoprevention.
The term "metastasis," as used herein, refers to the migration of cancer cells
from
the original tumor site through the blood and lymph vessels to produce cancers
in other
tissues. Metastasis also is the term used for a secondary cancer growing at a
distant site.
The term "neoplasm," as used herein, refers to an abnormal mass of tissue that
results from excessive cell division. Neoplasms may be benign (not cancerous),
or
malignant (cancerous) and may also be called a tumor. The term "neoplasia" is
the
pathological process that results in tumor formation.
As used herein, the term "pre-cancerous" refers to a condition that is not
malignant,
but is likely to become malignant if left untreated.
The term "proliferation" refers to cells undergoing mitosis.
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The phrase "P13 kinase related disease or disorder" refers to a disease or
disorder
characterized by inappropriate phosphoinositide-3-kinase activity or over-
activity of the
phosphoinositide-3-kinase. Inappropriate activity refers to either; (i) P13
kinase expression
in cells which normally do not express P13 kinase; (ii) increased P13 kinase
expression
leading to unwanted cell proliferation, differentiation and/or growth; or,
(iii) decreased P13
kinase expression leading to unwanted reductions in cell proliferation,
differentiation
and/or growth. Over-activity of P13 kinase refers to either amplification of
the gene
encoding a particular P13 kinase or production of a level of P13 kinase
activity which can
correlate with a cell proliferation, differentiation and/or growth disorder
(that is, as the level
of the P13 kinase increases, the severity of one or more of the symptoms of
the cellular
disorder increases).
The phrase a "radio therapeutic agent" refers to the use of electromagnetic or
particulate radiation in the treatment of neoplasia.
The term "recurrence" as used herein refers to the return of cancer after a
period of
remission. This may be due to incomplete removal of cells from the initial
cancer and may
occur locally (the same site of initial cancer), regionally (in vicinity of
initial cancer,
possibly in the lymph nodes or tissue), and/or distally as a result of
metastasis.
The term "treatment" refers to any process, action, application, therapy, or
the like,
wherein a mammal, including a human being, is subject to medical aid with the
object of
improving the mammal's condition, directly or indirectly.
The term "vaccine" includes agents that induce the patient's immune system to
mount an immune response against the tumor by attacking cells that express
tumor
associated antigens (Teas).
As used herein, the term "pharmaceutically acceptable salt" refers to those
salts
which are, within the scope of sound medical judgment, suitable for use in
contact with the
tissues of humans and lower animals without undue toxicity, irritation,
allergic response
and the like, and are commensurate with a reasonable benefit/risk ratio.
Pharmaceutically
acceptable salts are well known in the art. For example, S. M. Berge, et at.
describes
pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66:
1-19 (1977).
The salts can be prepared in situ during the final isolation and purification
of the
compounds of the invention, or separately by reacting the free base function
with a suitable
organic acid or inorganic acid. Examples of pharmaceutically acceptable
nontoxic acid
addition salts include, but are not limited to, salts of an amino group formed
with inorganic
acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid and
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WO 2011/130628 PCT/US2011/032683
perchloric acid or with organic acids such as acetic acid, maleic acid,
tartaric acid, citric
acid, succinic acid lactobionic acid or malonic acid or by using other methods
used in the
art such as ion exchange. Other pharmaceutically acceptable salts include, but
are not
limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate,
benzoate, bisulfate,
borate, butyrate, camphorate, camphorsulfonate, citrate,
cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate,
glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide,
2-hydroxy-
ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate,
maleate, malonate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,
oxalate, palmitate,
pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,
pivalate, propionate,
stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate,
undecanoate, valerate
salts, and the like. Representative alkali or alkaline earth metal salts
include sodium,
lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically
acceptable
salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and
amine
cations formed using counterions such as halide, hydroxide, carboxylate,
sulfate,
phosphate, nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and aryl
sulfonate.
As used herein, the term "pharmaceutically acceptable ester" refers to esters
which
hydrolyze in vivo and include those that break down readily in the human body
to leave the
parent compound or a salt thereof. Suitable ester groups include, for example,
those
derived from pharmaceutically acceptable aliphatic carboxylic acids,
particularly alkanoic,
alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl or alkenyl
moiety
advantageously has not more than 6 carbon atoms. Examples of particular esters
include,
but are not limited to, formates, acetates, propionates, butyrates, acrylates
and
ethylsuccinates.
The term "pharmaceutically acceptable prodrugs" as used herein refers to those
prodrugs of the compounds of the present invention which are, within the scope
of sound
medical judgment, suitable for use in contact with the tissues of humans and
lower animals
with undue toxicity, irritation, allergic response, and the like, commensurate
with a
reasonable benefit/risk ratio, and effective for their intended use, as well
as the zwitterionic
forms, where possible, of the compounds of the present invention. "Prodrug",
as used
herein means a compound which is convertible in vivo by metabolic means (e.g.
by
hydrolysis) to a compound of the invention. Various forms of prodrugs are
known in the
art, for example, as discussed in Bundgaard, (ed.), Design of Prodrugs,
Elsevier (1985);
Widder, et al. (ed.), Methods in Enzymology, vol. 4, Academic Press (1985);
Krogsgaard-
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Larsen, et al., (ed). "Design and Application of Prodrugs, Textbook of Drug
Design and
Development, Chapter 5, 113-191 (1991); Bundgaard, et at., Journal of Drug
Deliver
Reviews, 8:1-38(1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285 et
seq. (1988);
Higuchi and Stella (eds.) Prodrugs as Novel Drug Delivery Systems, American
Chemical
Society (1975); and Bernard Testa & Joachim Mayer, "Hydrolysis In Drug And
Prodrug
Metabolism: Chemistry, Biochemistry And Enzymology," John Wiley and Sons, Ltd.
(2002).
As used herein, "pharmaceutically acceptable carrier" is intended to include
any and
all solvents, dispersion media, coatings, antibacterial and antifungal agents,
isotonic and
absorption delaying agents, and the like, compatible with pharmaceutical
administration,
such as sterile pyrogen-free water. Suitable carriers are described in the
most recent edition
of Remington's Pharmaceutical Sciences, a standard reference text in the
field, which is
incorporated herein by reference. Preferred examples of such carriers or
diluents include,
but are not limited to, water, saline, finger's solutions, dextrose solution,
and 5% human
serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also
be used.
The use of such media and agents for pharmaceutically active substances is
well known in
the art. Except insofar as any conventional media or agent is incompatible
with the active
compound, use thereof in the compositions is contemplated. Supplementary
active
compounds can also be incorporated into the compositions.
As used herein, the term "pre-cancerous" refers to a condition that is not
malignant,
but is likely to become malignant if left untreated.
The term "subject" as used herein refers to an animal. Preferably the animal
is a
mammal. More preferably the mammal is a human. A subject also refers to, for
example,
dogs, cats, horses, cows, pigs, guinea pigs, fish, birds and the like.
The compounds of this invention may be modified by appending appropriate
functionalities to enhance selective biological properties. Such modifications
are known in
the art and may include those which increase biological penetration into a
given biological
system (e.g., blood, lymphatic system, central nervous system), increase oral
availability,
increase solubility to allow administration by injection, alter metabolism and
alter rate of
excretion.
The synthesized compounds can be separated from a reaction mixture and further
purified by a method such as column chromatography, high pressure liquid
chromatography, or recrystallization. As can be appreciated by the skilled
artisan, further
methods of synthesizing the compounds of the formulae herein will be evident
to those of
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ordinary skill in the art. Additionally, the various synthetic steps may be
performed in an
alternate sequence or order to give the desired compounds. Synthetic chemistry
transformations and protecting group methodologies (protection and
deprotection) useful in
synthesizing the compounds described herein are known in the art and include,
for
example, those such as described in R. Larock, Comprehensive Organic
Transformations,
VCH Publishers (1989); T.W. Greene and P.G.M. Wuts, Protective Groups in
Organic
Synthesis, 2d. Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser,
Fieser and
Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L.
Paquette, ed.,
Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995),
and
subsequent editions thereof.
Certain of the compounds described herein contain one or more asymmetric
centers
and thus give rise to enantiomers, diastereomers, and other stereoisomeric
forms that may
be defined, in terms of absolute stereochemistry, as (R)- or (S)-, or as (D)-
or (L)- for amino
acids. The present invention is meant to include all such possible isomers, as
well as their
racemic and optically pure forms. Optical isomers may be prepared from their
respective
optically active precursors by the procedures described above, or by resolving
the racemic
mixtures. The resolution can be carried out in the presence of a resolving
agent, by
chromatography or by repeated crystallization or by some combination of these
techniques
which are known to those skilled in the art. Further details regarding
resolutions can be
found in Jacques, et al., Enantiomers, Racemates, and Resolutions (John Wiley
& Sons,
1981). When the compounds described herein contain olefinic double bonds,
other
unsaturation, or other centers of geometric asymmetry, and unless specified
otherwise, it is
intended that the compounds include both E and Z geometric isomers and/or cis-
and trans-
isomers. Likewise, all tautomeric forms are also intended to be included. The
configuration of any carbon-carbon double bond appearing herein is selected
for
convenience only and is not intended to designate a particular configuration
unless the text
so states; thus a carbon-carbon double bond or carbon-heteroatom double bond
depicted
arbitrarily herein as trans may be cis, trans, or a mixture of the two in any
proportion.
Pharmaceutical Compositions
The pharmaceutical compositions of the present invention comprise a
therapeutically effective amount of a compound of the present invention
formulated
together with one or more pharmaceutically acceptable carriers or excipients.
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As used herein, the term "pharmaceutically acceptable carrier or excipient"
means a
non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating
material or
formulation auxiliary of any type. Some examples of materials which can serve
as
pharmaceutically acceptable carriers are sugars such as lactose, glucose and
sucrose;
cyclodextrins such as alpha- (a), beta- (0) and gamma- (y) cyclodextrins;
starches such as
corn starch and potato starch; cellulose and its derivatives such as sodium
carboxymethyl
cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt;
gelatin; talc;
excipients such as cocoa butter and suppository waxes; oils such as peanut
oil, cottonseed
oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols
such as propylene
glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents
such as
magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water;
isotonic
saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as
well as other
non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium
stearate, as
well as coloring agents, releasing agents, coating agents, sweetening,
flavoring and
perfuming agents, preservatives and antioxidants can also be present in the
composition,
according to the judgment of the formulator.
The pharmaceutical compositions of this invention may be administered orally,
parenterally, by inhalation spray, topically, rectally, nasally, buccally,
vaginally or via an
implanted reservoir, preferably by oral administration or administration by
injection. The
pharmaceutical compositions of this invention may contain any conventional non-
toxic
pharmaceutically-acceptable carriers, adjuvants or vehicles. In some cases,
the pH of the
formulation may be adjusted with pharmaceutically acceptable acids, bases or
buffers to
enhance the stability of the formulated compound or its delivery form. The
term parenteral
as used herein includes subcutaneous, intracutaneous, intravenous,
intramuscular,
intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal,
intralesional and
intracranial injection or infusion techniques.
Liquid dosage forms for oral administration include pharmaceutically
acceptable
emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In
addition to the
active compounds, the liquid dosage forms may contain inert diluents commonly
used in
the art such as, for example, water or other solvents, solubilizing agents and
emulsifiers
such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate,
benzyl alcohol,
benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide,
oils (in
particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame
oils), glycerol,
tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of
sorbitan, and
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mixtures thereof. Besides inert diluents, the oral compositions can also
include adjuvants
such as wetting agents, emulsifying and suspending agents, sweetening,
flavoring, and
perfuming agents.
Injectable preparations, for example, sterile injectable aqueous or oleaginous
suspensions, may be formulated according to the known art using suitable
dispersing or
wetting agents and suspending agents. The sterile injectable preparation may
also be a
sterile injectable solution, suspension or emulsion in a nontoxic parenterally
acceptable
diluent or solvent, for example, as a solution in 1,3-butanediol. Among the
acceptable
vehicles and solvents that may be employed are water, Ringer's solution,
U.S.P. and
isotonic sodium chloride solution. In addition, sterile, fixed oils are
conventionally
employed as a solvent or suspending medium. For this purpose any bland fixed
oil can be
employed including synthetic mono- or diglycerides. In addition, fatty acids
such as oleic
acid are used in the preparation of injectables.
The injectable formulations can be sterilized, for example, by filtration
through a
bacterial-retaining filter, or by incorporating sterilizing agents in the form
of sterile solid
compositions which can be dissolved or dispersed in sterile water or other
sterile injectable
medium prior to use.
In order to prolong the effect of a drug, it is often desirable to slow the
absorption of
the drug from subcutaneous or intramuscular injection. This may be
accomplished by the
use of a liquid suspension of crystalline or amorphous material with poor
water solubility.
The rate of absorption of the drug then depends upon its rate of dissolution,
which, in turn,
may depend upon crystal size and crystalline form. Alternatively, delayed
absorption of a
parenterally administered drug form is accomplished by dissolving or
suspending the drug
in an oil vehicle. Injectable depot forms are made by forming microencapsule
matrices of
the drug in biodegradable polymers such as polylactide-polyglycolide.
Depending upon the
ratio of drug to polymer and the nature of the particular polymer employed,
the rate of drug
release can be controlled. Examples of other biodegradable polymers include
poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also
prepared by
entrapping the drug in liposomes or microemulsions that are compatible with
body tissues.
Compositions for rectal or vaginal administration are preferably suppositories
which
can be prepared by mixing the compounds of this invention with suitable non-
irritating
excipients or carriers such as cocoa butter, polyethylene glycol or a
suppository wax which
are solid at ambient temperature but liquid at body temperature and therefore
melt in the
rectum or vaginal cavity and release the active compound.
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Solid dosage forms for oral administration include capsules, tablets, pills,
powders,
and granules. In such solid dosage forms, the active compound is mixed with at
least one
inert, pharmaceutically acceptable excipient or carrier such as sodium citrate
or dicalcium
phosphate and/or: a) fillers or extenders such as starches, lactose, sucrose,
glucose,
mannitol, and silicic acid, b) binders such as, for example,
carboxymethylcellulose,
alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants
such as
glycerol, d) disintegrating agents such as agar-agar, calcium carbonate,
potato or tapioca
starch, alginic acid, certain silicates, and sodium carbonate, e) solution
retarding agents
such as paraffin, f) absorption accelerators such as quaternary ammonium
compounds, g)
wetting agents such as, for example, cetyl alcohol and glycerol monostearate,
h) absorbents
such as kaolin and bentonite clay, and i) lubricants such as talc, calcium
stearate,
magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and
mixtures
thereof. In the case of capsules, tablets and pills, the dosage form may also
comprise
buffering agents.
Solid compositions of a similar type may also be employed as fillers in soft
and
hard-filled gelatin capsules using such excipients as lactose or milk sugar as
well as high
molecular weight polyethylene glycols and the like.
The solid dosage forms of tablets, dragees, capsules, pills, and granules can
be
prepared with coatings and shells such as enteric coatings and other coatings
well known in
the pharmaceutical formulating art. They may optionally contain opacifying
agents and can
also be of a composition that they release the active ingredient(s) only, or
preferentially, in
a certain part of the intestinal tract, optionally, in a delayed manner.
Examples of
embedding compositions that can be used include polymeric substances and
waxes.
Dosage forms for topical or transdermal administration of a compound of this
invention include ointments, pastes, creams, lotions, gels, powders,
solutions, sprays,
inhalants or patches. The active component is admixed under sterile conditions
with a
pharmaceutically acceptable carrier and any needed preservatives or buffers as
may be
required. Ophthalmic formulation, ear drops, eye ointments, powders and
solutions are also
contemplated as being within the scope of this invention.
The ointments, pastes, creams and gels may contain, in addition to an active
compound of this invention, excipients such as animal and vegetable fats,
oils, waxes,
paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols,
silicones,
bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.
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Powders and sprays can contain, in addition to the compounds of this
invention,
excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium
silicates and
polyamide powder, or mixtures of these substances. Sprays can additionally
contain
customary propellants such as chlorofluorohydrocarbons.
Transdermal patches have the added advantage of providing controlled delivery
of a
compound to the body. Such dosage forms can be made by dissolving or
dispensing the
compound in the proper medium. Absorption enhancers can also be used to
increase the
flux of the compound across the skin. The rate can be controlled by either
providing a rate
controlling membrane or by dispersing the compound in a polymer matrix or gel.
For pulmonary delivery, a therapeutic composition of the invention is
formulated
and administered to the patient in solid or liquid particulate form by direct
administration
e.g., inhalation into the respiratory system. Solid or liquid particulate
forms of the active
compound prepared for practicing the present invention include particles of
respirable size:
that is, particles of a size sufficiently small to pass through the mouth and
larynx upon
inhalation and into the bronchi and alveoli of the lungs. Delivery of
aerosolized
therapeutics, particularly aerosolized antibiotics, is known in the art (see,
for example U.S.
Pat. No. 5,767,068 to VanDevanter et at., U.S. Pat. No. 5,508,269 to Smith et
at., and WO
98/43650 by Montgomery, all of which are incorporated herein by reference). A
discussion
of pulmonary delivery of antibiotics is also found in U.S. Pat. No. 6,014,969,
incorporated
herein by reference.
Preferably, a therapeutically effective amount of a compound or combination of
compounds is an amount which confers a therapeutic effect on the treated
subject, at a
reasonable benefit/risk ratio applicable to any medical treatment. The
therapeutic effect
may be objective (i.e., measurable by some test or marker) or subjective
(i.e., subject gives
an indication of or feels an effect). An effective amount of the compound
described above
may range from about 0.1 mg/Kg to about 500 mg/Kg, preferably from about 1 to
about 50
mg/Kg. Effective doses will also vary depending on route of administration, as
well as the
possibility of co-usage with other agents. It will be understood, however,
that the total
daily usage of the compounds and compositions of the present invention will be
decided by
the attending physician within the scope of sound medical judgment. The
specific
therapeutically effective dose level for any particular patient will depend
upon a variety of
factors including the disorder being treated and the severity of the disorder;
the activity of
the specific compound employed; the specific composition employed; the age,
body
weight, general health, sex and diet of the patient; the time of
administration, route of
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administration, and rate of excretion of the specific compound employed; the
duration of
the treatment; drugs used in combination or contemporaneously with the
specific
compound employed; and like factors well known in the medical arts.
The total daily dose of the compounds of this invention administered to a
human or
other animal in single or in divided doses can be in amounts, for example,
from 0.01 to 50
mg/kg body weight or more usually from 0.1 to 25 mg/kg body weight. Single
dose
compositions may contain such amounts or submultiples thereof to make up the
daily dose.
In general, treatment regimens according to the present invention comprise
administration
to a patient in need of such treatment from about 10 mg to about 1000 mg of
the
compound(s) of this invention per day in single or multiple doses.
The compounds of the formulae described herein can, for example, be
administered
by injection, intravenously, intraarterially, subdermally, intraperitoneally,
intramuscularly,
or subcutaneously; or orally, buccally, nasally, transmucosally, topically, in
an ophthalmic
preparation, or by inhalation, with a dosage ranging from about 0.1 to about
500 mg/kg of
body weight, alternatively dosages between 1 mg and 1000 mg/dose, every 4 to
120 hours,
or according to the requirements of the particular drug. The methods herein
contemplate
administration of an effective amount of compound or compound composition to
achieve
the desired or stated effect. Typically, the pharmaceutical compositions of
this invention
will be administered from about 1 to about 6 times per day or alternatively,
as a continuous
infusion. Such administration can be used as a chronic or acute therapy. The
amount of
active ingredient that may be combined with pharmaceutically excipients or
carriers to
produce a single dosage form will vary depending upon the host treated and the
particular
mode of administration. A typical preparation will contain from about 5% to
about 95%
active compound (w/w). Alternatively, such preparations may contain from about
20% to
about 80% active compound.
Lower or higher doses than those recited above may be required. Specific
dosage
and treatment regimens for any particular patient will depend upon a variety
of factors,
including the activity of the specific compound employed, the age, body
weight, general
health status, sex, diet, time of administration, rate of excretion, drug
combination, the
severity and course of the disease, condition or symptoms, the patient's
disposition to the
disease, condition or symptoms, and the judgment of the treating physician.
Upon improvement of a patient's condition, a maintenance dose of a compound,
composition or combination of this invention may be administered, if
necessary.
Subsequently, the dosage or frequency of administration, or both, may be
reduced, as a
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function of the symptoms, to a level at which the improved condition is
retained when the
symptoms have been alleviated to the desired level. Patients may, however,
require
intermittent treatment on a long-term basis upon any recurrence of disease
symptoms.
Synthetic Methods
The compounds and processes of the present invention will be better understood
in
connection with the following synthetic schemes that illustrate the methods by
which the
compounds of the invention may be prepared, which are intended as an
illustration only and
not limiting of the scope of the invention.
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Scheme 1
HN McSOZCI ^ CF3COOH McSOzN~
1 Me
SOZN I CF3COOH
NBoc ~IINBoc ~IINH
0101 0102 0103 \ \ /
Br 1 V,
Ac20 Br o o _
oe"ao O~ O
Br NH2 iso-arnyl nitrite I \ N 6N HCI 30 x, \H B'
R O R I/ N Pd -12(dPP02 \ \ N
R 0105 0106 R H
0104 0107
C1 CO)
S CO Me O
2 Hz S NH PO S N morph N LDA/DMF N 30 NH 1900C I reflux Z I S N 77% 0 S N
2 H O N CI \
0108 0109 0110 N CI N CI
0111 0112
4O 0
\ i 0 O iS C O
0 N I 1 0 ON Br +~n
0103 N 01 N S N _N
N S N NH
N
N CI
0114
0113
4O 0 \ ~O (0)
N
C O < N
`N N N S O OH
\ I N O N N % N
N 4in \`/\ NH
N 4+n
NH2OH
0115
O \ 4O O
\SN O r 1 0 0~ N S C) O OH
CN N yam/ O H
S N I NN/L J N N%
N
N
0116
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Scheme 2
l H 0 0
COJ Q N I 0~ 4N HG N C) Br O
N NBoe N Dioxane N L J
N
O& S ~N S ~N ON
\`i\ J N G
N Cl N Cl
0112 0201 0202
0>
DO
S N S NJ /\0 0 ~N I 0107 /\0 0 N
NH2OH
N N
N~ n N
Cl N
NH
0203 0204
R
~O>
S NJ
O
N I N
HORN N
H " 4 + n N
!N
NH
R
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Scheme 3
Urea/HCI ^ IT COOEt COOEt
EtO'~'CO Et Na/EtOH OMe EtOH T Br2/HAc 3POCI3 N
~IrI J
z HCOOEt EtO COzEt O H O H Cl N
0301 0302 0303 0304 0305
H N ^l
N
ONH H ONYN
DMF
0
0306
0 0
O 0-_N N
CO`
N 0306 N-~ N rp) 0107 N~N~ `0)
/
O\ S I N N
NCI N S N N S DI: N N
\ x NH
0112 NCI 0308 I /
0307
HO 0
N
H
~N
NHZOH N~ O
`N S ~N ~N
NH
N \
O
O
CN1
H
C
NR yM NR S N 0107
p\ S I ~N + ~,puM Y
\ NCI IOI O y i
NCI
0112
0309
O /p1
CN/ H X R N
X R N yM
~M \>-N LS NHZOH HOB \)_N S
-
N -N y N N
p Y \ I NH O NH
N N
0310
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WO 2011/130628 PCT/US2011/032683
Sdien-e 4
nl OOCEt I GOC~
0l IT
N N
Ad\tv z
0305 0401
0 (0) N0401 0 \ N 0107 ~N (D i N, N
S ~ N T~ ~ -N ~ ~J~ N
N N R1 S O N N R1 S N
N 0 N
R
0112 0402 0403
0
0
N
N / N
Gi
N N
OH I N-~N T S N
N
R
0 0 OJ O
CN) NH~O
R3 L J~ S ~N 0107 S N -N,
~ N \ I \ -I
0 N N O 0 N N \
N a R3 ,R3
0112 `0 0404 `0 0405
R
COJ
N-6CH N N
3 0 N N \ H
9f f ,R3 1 14
HO-N
H R
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Scheme 5
(O) (O) (O) (O)
N N
NaBH4 NBS, PPh3 R NHz R1-N H
S
O\ S N HO S N Br S LN I LN
N CI N CI N CI N CI
0112 0501 0502 0503
(O) (O)
0305 S 0107 NHZOH S 0 0
\ N \ I N \ I iN NNH
N CI ~ N
N R1 0504 ~O N R1 0505
R
(O)
0 N \ NH
N NNN~
HO-N N~ R1
H
R
Scheme 6
0 0
'B-B 0
Br O O BOO
R- AcOK,dioxane, 80 C R y,
0601 0602
CO) 0602 CO)
NS 'N NaHCO3, (Ph3P)2PdCI2, toluene, N
O N \ EtOH, H2O, M.W., 120 C O / N N X
N N L~
Cl Y
N R
O N R /-O -N~ R1
1
0504 0603
C~)
NH2OH/MeOH S N
0 NX
N N
HO-NH N R1 Y R
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Scheme 7
/
i Rt Rt
Cl
CH3CN HN THF Boc,N
S N PhNH2, Et3N S N (Boc)20, DMAP S L N LDA, THF, DMF
NCI 50 C, overnight r.t., overnight -30 C, 30 min
N Cl N Cl
0110 0701 0702
Boc.N \ I R1 Boc, \ R, Boc, R1
NaBH4 N N
p S McOH/THF Hp S N MsCI, Et3N, THF Ms-O S N
r.t., 30 min r.t., 1h
N Cl N Cl N Cl
0703 0704 0705
11/ ~ i Rj
Ri
HN T \ EtO2C-(N
`)-CI HN
McNH2 in MeOH N S N
-NH N N
r.t., 1 h Et3N, CH3CN EtOzC N NCI
N Cl r.t., overnight N \
0706 0707
R
0107 HN
(Ph3P)3PdCI2, NaHCO3 S N N NH2OH in MeOH
Toluent/EtOH/Hz0 N \
Et02C ANN NH r.t., 20 min
MW, 120 C, lh {N N
0708
R
Rj
HN
OH S N _N
N
HN 7-N I \ I i NH
O N
N
R
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Scheme 8
Boc Boc
N,Boc N N
Cl H~ C) C)
'N N s 1) LDA, THF, -70 C N NBH4, MeOH
\ I ~N
NCI MeOH, NEt3, r.t. 2) DMF, -65 C 0\ \ N
N Cl NCI
0110 0801 0802
Boc Boc -S02 111009 Boc
N~
1
CN) PPh3, (N) N CF3COOH -502 I
N N N
CH2CI2, r.t.
HO S N Br N S ~N
N K2CO3, CH3CN, DMF,
N Cl ~N Cl N Cl
0803 0804 0805
0
H OEt
_ N n 0
HCI / THF S0 CN) Br " 1 n OEt -SN2 (N) 111013
S N
N Et3N, DMF, 50 C ~-N S N (Ph3P)2PdCI2, NaHCO30NCI toluene, EtOH, H2O, MW
0806 N Cl
0807
0
0
OEtNOH
-SO2 CN NH2OH N H
N -SO2 ~N)
\J N MeOH CN D
N S N S N
NH _N
N I \ I N NH
/
0808
Scheme 9
CO) CN 0CN
ON R1NH2, NaBH4 S \ n-BuLi/DMF S R -NH S
O\ S I I N
N CI N CI N CI
0111 0112 0503
120
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Scheme 10
0 0 Cl COQ
S NH Br2 S NH p S N H
\-I
H'ZO HOAc I N~O N CI MeOH
Br Br
0109 1001 1002
COD ( )
CND N N
xLN fuming HNO3 Sn, con. S N S N
con. H2SO4 02N \ I N Cl MeOH H2N NCI
N Cl Br Br
1003 1004 1005
o
ci C-,,,o R R-O
s
02
CN 06
0 1006 pN \
S I N
H NCI
1007
O 0
R-O CNJ NH2OH HO-NH CD
Oi \\ ~0 Oi \ '0 N
\~ pS~ S N X 0N S
N N X
H N~ H NY~J
R R
1008
121
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Scheme 11
O o co)
Boc2O CNN
CN
S \N S O Boc S ~N
H BocHN \ ON \ I NCI
N Cl N Cl
~O n
1005 1101 1102
Co) co)
0602 N TFA N
Boc S S N 0 O/"` N \ l- ' HN X
n N Y R O n N ~Y R
1103 1104
NH Co)
2OH N
S ~
0 N
HN \ I -X
HO-N n N
H Y R
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Scheme 12
COJ 0 0 ~O (0) 0305 O N CON)
/ N 0
/~ 0 /N HN \ I 0 N-N \
N Cl N Cl N Cl , N Cl
0111 1201 1202 1203
O 0
0602 O N / CNJ NH2OH O N
Y-C >\ 10 CNJ
\-N S N ~ \ N S N
O N fl X HO-NH `N~ N~X
N N NCJ
Y R Y R
1204
O O O" O
acetone N N TMSCN~0 ~N conc. HCI ~N~
S / N THE HO S / N H2SO4~NH S N Me0 HH H2O HZN S N
N Cl N Cl N Cl N Cl
1205 1206 1207
0111
O
0305 O N (0) NNaHDMS O N CNJ
NH S MeC S N
EtO N \ I Et0 N \
N Cl CI
1208 1209
I 0602 0602
COQ N
O, ~ N N O_, N /
~NH S )-l /~(/~~-N S - N
N~ X
O N X O N \ I N
I~j `~R
1210 R 1211
0 O
CNJ
0 ~N (N" O / N
\>-NH S 'N \>-N S N
HO-NH N X HO-NH N X
N `Y~j Ni\` y~R
R
123
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Scheme 13
Nco) N
co) Cpl
S02C12 NH3 p 0305
S I ~N
S N
DW 11
O S /01
CI_S \ HZN-SO
N CI O N Cl N Cl
0111 1301 1302
co) co)
N N
p 0602 p
N H u S N N H S N 10 EtOOC~N N- 0 \ EtOOC~N NSO N~"
X
N Cl Y R
1303 1304
co)
N
NH2OH p N O S /I N
\N-S \ I~X
H2N-O N 0 N J
Y R
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Scheme 14
O CO
cN) (O) O S N
mCPBA 0401 H
S
OHC \ I N HOOC S I \ N N\ N
Cl
N Cl
N Cl EtOOC ~ N
0112 1401 1402
O
0602 0 0 N NH2OH
S
N N
~I / ~rN
N ~^ X
EtOOC" v N
1403 Y R
0
O N
S
N NH N
HI ~~~/// N
HORN II N
O Y
R
~ O
0
N OSO S N
O S \ N 0401 H X/ 0602
11 T"* CI-S NYN N
\
0
1301 N Cl I IN Cl
EtOOC 1404
0) 0 N ~rN 0S0 SNH OH ^^ OSO S
\ NHN 2 N\ Nr T `H/ N
Y N X HrI Y ~~~/// N~~X
DOOC IN 1 HO"N II N Ni
1405 CYR O Y
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Scheme 15
0 O O
Br I \ Br Br B\
O O
/ OH
l l n~ PdCl2(dppf)2, KOAc
0 dioxane
1501 1502
0'/
o s. N
(0)
0N S N
n ~
I/ \
1503
1504 N O'O,
0
O~ N CO)
NH2OH ON N
S N
H
N -~\
/ O n~ N-OH
0
Scheme 16
OH
Br
xnBuLi/B(O-iPr)3 B- OH
R` R`
Y y
0601 1601
C / O \ N CI (0)
N
R,-NH S L N - O N S L N 0602
N Cl - >-N N Cl or 1601
O N
0503 R,
1602
CO)
CND
S I N ^X NHZOH/MeOH S N
- y--(i \ N
N N II I \j O N \ I I I r^ XI
0 N Rt Y. R \N. N ::~J
HO-NH N R1 R
1603
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EXAMPLES
The compounds and processes of the present invention will be better understood
in
connection with the following examples, which are intended as an illustration
only and not
limiting of the scope of the invention. Various changes and modifications to
the disclosed
embodiments will be apparent to those skilled in the art and such changes and
modifications including, without limitation, those relating to the chemical
structures,
substituents, derivatives, formulations and/or methods of the invention may be
made
without departing from the spirit of the invention and the scope of the
appended claims.
EXAMPLE 1: Preparation of N-hydroxy-5-(4-(6-((4-(methylsulfonyl) piperazin-l-
yl)methyl)-4-morpholinothieno [3,2-d] pyrimidin-2-yl)-1H-indazol-l-
yl)pentanamide
(Compound 3)
Step la: 1-(Methylthioperoxy)piperazine trifluoroacetic acid salt (Compound
0103)
A mixture of compound 0101 (10.0 g, 54 mmol), methanesulfonyl chloride (6.5 g,
57 mmol) and triethylamine in CH2Cl2 (50 mL) was stirred at reflux overnight.
The
reaction was cooled to room temperature and filtered. The filtrate was
concentrated to give
compound 0102 which was used to the next step without further purification.
A mixture of compound 0102 and trifluoroacetic acid (15 mL) in CH2Cl2 (100 mL)
was
stirred at room temperature for 3 h. The reaction was filtered and the
filtrate was
concentrated to give the title compound 0103 (9.7 g, 66%) as a white solid.
LCMS: 165
[M+1]+; 'H NMR (400 MHz, CDC13): 6 2.99 (s, 3H), 3.21 (m, 4H), 3.33 (m, 4H),
8.95 (br
s, 2H).
Step 1b: 4-Bromo-]H-indazole (Compound 0106-3)
To a solution of 3-bromo-2-methyl aniline (0104) (0.50 g, 2.69 mmol) in
chloroform (5 mL) was added potassium acetate (0.28 g, 2.82 mmol). The mixture
was
cooled with ice-water bath and then acetic anhydride (0.50 mL, 5.37 mmol) was
added to it.
Ice-water bath was then removed and the resulting mixture was stirred at room
temperature
for 10 minutes after which time a white gelatinous solid formed. 18-Crown-6
(0.14 g, 0.54
mmol) was then added followed by isoamyl nitrite (0.80 mL, 5.90 mmol). The
mixture was
then heated under reflux for 18 hours. The reaction mixture was allowed to
cool, and was
partitioned between chloroform (3 x 10 mL) and saturated aqueous sodium
hydrogen
carbonate (10 mL). The combined organic extracts were washed with brine (10
mL), dried
over Na2SO4, filtered and evaporated to give a crude product which was
purified by column
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chromatography (ethyl acetate in petroleum ether, 10% v/v) to give 1-(4-
bromoindrazol-l-
yl)-ethanone (0105) as an orange solid (0.31 g, 49%), and 4-bromo-]H-indazole
(0106-3)
as a pale orange solid (0.21 g, 40%). Compound 0105: LCMS: 239 [M+1]+;'H NMR
(400
Hz, CDC13) 6 2.80 (s, 3H), 7.41 (t, J= 6.8 Hz, 1H), 7.50 (d, J= 6.0 Hz, 1H),
8.15 (s, 1H),
8.40 (d, J= 6.8 Hz, 1H). Compound 0106: LCMS: 197 [M+1]+;'H NMR (400 Hz,
CDC13)
6 7.25 (t, J= 6.0 Hz, I H), 7.34 (d, J= 6.4 Hz, I H), 7.46 (d, J= 6.8 Hz, I
H), 8.12 (s, I H).
To a solution of compound 0105 (0.30g, 1.29 mmol) in methanol (5.0 mL) was
added 6 N aqueous HC1(3.0 mL). The mixture was stirred at room temperature for
7 h.
Methanol was evaporated and the mixture partitioned between EtOAc (2 x 50 mL)
and
water (5.0 mL). The combined organic layers were washed with brine (5.0 mL),
dried over
Na2SO4, filtered and evaporated to give 4-bromo-]H -indazole (0106-3) (0.24 g,
94%).
Step 1c: 4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-]H-indazole (Compound
0107-3)
To a stirred solution of compound 0106 (500 mg, 2.54 mmol) and
bis(pinacolato)diboron (968 mg, 3.81 mmol) in DMSO (20 mL) was added potassium
acetate (747 mg, 7.61 mmol) and PdC12(dppf)2 (3 mol%, 62 mg, 0.076 mmol). The
mixture
was degassed with argon and heated at 80 C for 40 hours. The reaction mixture
was
allowed to cool and partitioned between water (50 mL) and ether (3 x 50 mL).
The
combined organic layers were separated, washed with brine (50 mL), dried over
MgSO4,
filtered and evaporated to give crude material which was purified by column
chromatography (ethyl acetate in petroleum ether, 20% v/v) to give compound
0107-3 as an
off-white solid (370 mg, 60%): LCMS: 245 [M+1]+; 'H NMR (400 Hz, CDC13) 6 1.41
(s,
12H), 7.40 (dd, J= 6.8 Hz, 8.4 Hz, 1 H), 7.62 (d, J = 8.8 Hz, 1 H), 7.90 (d, J
= 6.8 Hz, 1 H),
8.50 (s, 1H).
Step 1d: Thieno[3,2-d]pyrimidine-2,4(IH,3H)-dione (Compound 0109)
A mixture of methyl 3-amino-2-thiophenecarboxylate (0108) (13.48 g, 85.85
mmol)
and urea (29.75 g, 0.43 mol) was heated at 190 C for 2 h. The hot reaction
mixture was
poured into sodium hydroxide solution and insoluble material was removed by
filtration.
The mixture was then acidified by 2 N HC1 solution. The resulting solid was
collected by
filtration, dried to give title compound 0109 (9.62 g, 67%) as a white solid:
LCMS: 169
[M+l ]+; 'H NMR (400 MHz, DMSO-d6): 6 6.92 (d, J = 4.0 Hz, I H), 8.04 (d, J =
4.0 Hz,
1 H), 11.19 (d, J = 14.0 Hz , 1 H), 11.60 (s, 1 H).
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Step le: 2,4-Dichlorothieno[3,2-d]pyrimidine (Compound 0110)
A mixture of compound 0109 (9.49 g, 56.49 mmol) and phosphorous oxychloride
(150 mL) was heated at reflux for 10 h. The solvent was then removed and the
residue was
poured onto ice/water with vigorous stirring to give title compound 0110 (8.62
g, 74%) as a
white solid: LCMS: 205 [M+1]; 'H NMR (400 MHz, CDC13): 5 7.48 (d, J= 5.6 Hz,
1H),
8.05(d,J=5.6Hz,1H).
Step 1f: 4-(2-Chlorothieno[3,2-d]pyrimidin-4-yl)morpholine (Compound 0111)
A mixture of compound 0110 (8.68 g, 42.34 mmol) and morpholine (8.11 mL,
93.15 mmol) in methanol (150 mL) was stirred at room temperature for 1 h. The
reaction
mixture was then filtered, washed with water (50 mL x 3) and methanol (50 mL x
1) to give
the title compound 0111 (11.04 g, 100%) as a white solid: LCMS: 256 [M+1]+; 1H
NMR
(400 MHz, DMSO-d6): 6 3.76 (t, J= 4.8 Hz, 4H), 3.91 (t, J= 4.8 Hz, 4H),
7.41(d, J= 5.6
Hz , 1 H), 8.31 (d, J = 5.6 Hz , 1 H).
Step 1g: 2-Chloro-4-morpholinothieno[3,2-d]pyrimidine-6-carbaldehyde (Compound
0112)
To a suspension of compound 0111 (1.75 g, 6.85 mmol) in dry tetrahydrofuran
(40
mL) at -78 C was added a 2.0 M solution of LDA in THF/hexane(20.55 mL, 41.1
mmol).
After stirring for 1 h, dry N,N-dimethylformamide (3.2 mL, 41.1 mmol) was
added. The
reaction mixture was stirred for 1 h at -78 C and then warmed slowly to room
temperature.
After a further stir for 10 h at room temperature, the reaction mixture was
poured onto
NH4C1 saturated solution, extracted with ethyl acetate (100 mL x 3), dried
over Na2SO4 and
filtered. The filtrate was concentrated to leave a residue which was washed
with ethyl
acetate (10 mL x 2) to give the title compound 0112 (0.66 g, 35%) as a yellow
solid:
LCMS: 284 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 653.76 (t, J= 4.8 Hz, 4H), 4.10
(t, J=
4.8 Hz, 4H), 8.29(s, 1H), 10.21 (s, 1H).
Step 1h: 4-(2-Cloro-6-((4-(methylthioperoxy)piperazin-1-yl)methyl)thieno[3,2-
d]
pyrimidin-4-yl)morpholine (Compound 0113)
A mixture of compound 0112 (1.10 g, 3.89 mmol), 0103 (2.20 g, 7.78 mmol),
triethylamine (471 mg, 4.7 mmol) and titanium tetraisopropanolate (1.30 g,
4.67 mmol) in
chloroform (30 ml) was stirred at reflux overnight. The solvent was then
removed, and 1,2-
dichloroethane (40 mL) and sodium cyanborohydride (368 mg, 5.84 mmol) were
added.
The reaction mixture was then stirred at room temperature for 12 h. The
reaction was
concentrated and the resulting solid was recrystallized with ethanol to give
the title
compound 0113 (800 mg, 48%) as a yellow solid: LCMS: 432 [M+1]+; 1H NMR (400
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MHz, DMSO-d6): 6 2.57 (t, J= 4.4 Hz, 4H), 2.89(s, 3H), 3.13 (t, J= 4.4 Hz,
4H), 3.74 (t, J
= 5.2 Hz , 4H), 3.88 (t, J= 5.2 Hz, 4H), 3.91 (s, 2H), 7.31 (s, 1H).
Step 1i: 4-(2-Chloro-6-((4-(methylthioperoxy)piperazin-1-yl)methyl)thieno[3,2-
d]
pyrimidin-4-yl)morpholine (Compound 0114)
A mixture of compound 0113 (800 mg, 1.86 mmol), 0107-3 (500 mg, 2.04 mmol),
sodium hydrogen carbonate (470 mg, 5.58 mmol) and bis(triphenylphosphine)
palladium(0)
chloride (80 mg, 0.093 mmol) in toluene (20 mL), ethanol (12 mL) and water
(5.6 mL) was
flushed with nitrogen and heated under microwave irradiation at 120 C for 1
h. The
reaction mixture was partitioned between dichloromethane and water. The
organic layer
was separated and washed with brine, dried over magnesium sulfate, filtered
and
evaporated. The resulting residue was purified by column chromatography
(silica gel,
dichloromethane, 2%, v/v, to give title compound 0114 (350 mg, 37%) as a white
solid. mp
148-149 C. LCMS: 514 [M+1]+; 'H NMR (400 MHz, CDC13): 6 2.70 (t, J= 4.4 Hz,
4H),
2.81(s, 3H), 3.13 (t, J = 4.4 Hz , 4H), 3.92 (m, 6H), 4.09 (t, J = 5.6 Hz,
4H), 7.41 (s, 1 H),
7.50 (m, I H), 7.59 (d, J= 8.4 Hz, I H), 8.28 (d, J= 6.8 Hz, I H), 9.00 (s, I
H), 10.32 (br s,
I H).
Step 1j: Ethyl 5-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholinothieno[3,2-d]pyrimidin-2-yl)-2H-indazol-2-yl)pentanoate (Compound
0116-3)
and ethyl 5-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholinothieno[3,2-
d]pyrimidin-2-yl)-1H-indazol-l-yl)pentanoate (Compound 0115-3)
A mixture of compound 0114 (370 mg, 0.72 mmol), ethyl 5-bromopentanoate (181
mg, 0.87 mmol) and potassium carbonate (199 mg, 1.44 mmol) in acetonitrile (50
mL) was
refluxed for 58 hours. Solvent was removed and the residue was partitioned
between
dichloromethane and water. The organic layer was separated and washed with
brine, dried
over magnesium sulfate, filtered and evaporated to give a crude product which
was purified
by prep-HPLC to give the title compound 0115-3 (80 mg, 17%) and 0116-3 (60 mg,
13%).
Compound 0115-3: a white solid; LCMS: 642 [M+1]+; 'H NMR (400 MHz, CDC13): 6
1.15 (t, J= 7.2 Hz, 3H), 1.61 (m, 2H), 1.94 (m, 2H), 2.26 (t, J= 7.2 Hz, 2H),
2.62 (t, J=
4.4 Hz, 4H), 2.74 (s, 3H), 3.23 (t, J= 4.4 Hz, 4H), 3.84 (m, 6H), 4.01 (m,
6H), 4.38 (t, J=
6.8 Hz, 2H), 7.33 (s, 1H), 7.42 (m, 2H), 8.17 (m, 1H), 8.81 (s, 1H). 13C NMR
(100 MHz,
CDC13): 6 172.2, 161.6, 159.5, 157.1, 147.8, 139.3, 133.7, 131.2, 125.0,
123.0, 121.4,
120.8, 112.1, 109.7, 65.8 (2C), 59.3, 56.3, 51.4, 47.5 (2C), 45.6 (2C), 44.8
(2C), 33.5, 32.8,
28.6, 21.2, 13.2.
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Compound 0116-3: a white solid; LCMS: 642 [M+1]+; 1H NMR (400 MHz, CDC13): 6
1.15
(t, J = 7.2 Hz, 3H), 1.60 (m, 2H), 2.06 (m, 2H), 2.29 (t, J = 7.2 Hz, 2H),
2.63 (s, 4H), 2.74
(s, 3H), 3.24 (s, 4H), 3.84 (m, 6H), 4.04 (m, 6H), 4.43 (t, J= 6.8 Hz, 2H),
7.33 (m, 2 H),
7.74 (d, J= 8.4 Hz, 1H), 8.19 (d, J= 6.8 Hz, 1H), 8.82 (s, 1H).13C NMR (100
MHz,
CDC13): (S 172.1, 161.5, 159.5, 156.9, 148.8, 147.4, 130.0, 124.6, 123.1,
122.2, 119.0,
118.8, 112.0, 65.8 (2C), 59.4, 56.3, 52.4, 51.4 (2C), 45.5 (2C), 44.8 (2C),
33.5, 32.7, 28.6,
21.1, 13.1.
Step 1k: N-Hydroxy-5-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholinothien-[3,2-d]pyrimidin-2-yl)-1H-indazol-1-yl)pentanamide (Compound
3)
To a stirred solution of hydroxylamine hydrochloride (4.67 g, 67 mmol) in
methanol
(24 mL) at 0 C was added a solution of potassium hydroxide (5.61 g, 100 mmol)
in
methanol (14 mL). After addition, the mixture was stirred for 30 minutes at 0
C. The
resulting precipitate was filtered off and the filtrate was prepared as free
hydroxylamine
solution.
The above freshly prepared hydroxylamine solution (4.00 mL) was placed in 10
mL
flask. Compound 0115-3 (80 mg, 0.12 mmol) was added to this solution and
stirred at 0-10
C for 15 minutes. The reaction process was monitored by TLC. After the
reaction was
completed, the reaction was filtered. The collected solid was washed with
water and
methanol, dried to give compound 3 (45 mg, 58%) as a white solid: mp 139-143
C. LCMS:
629 [M+1]+; 1H NMR (400 MHz, DMSO-d6) (51.47 (m, 2H), 1.83 (m, 2H), 1.98 (t,
J=
7.4 Hz, 2H), 2.62 (s, 4H), 2.91 (s, 3H), 3.17 (s, 4H), 3.84 (s, 4H), 3.86 (s,
2H), 4.01 (m,
4H), 4.47 (t, J = 6.6 Hz, 2H), 7.52 (m, 2H), 7.82 (d, J = 8.4 Hz, 1 H), 8.24
(d, J = 6.8 Hz,
1H), 8.67 (s, 1H), 8.86 (s, 1H), 10.36 (s, 1H).
EXAMPLE 2: Preparation of N-hydroxy-3-(4-(6-((4-(methylsulfonyl)piperazin -1-
yl)methyl)-4-morpholinothieno [3,2-d] pyrimidin-2-yl)-1H-indazol-1-
yl)propanamide
(Compound 4)
Step 2a: Ethyl 6-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholinothieno [3,2-d]pyrimidin-2-yl)-2H-indazol-2-yl)hexanoate (Compound
0116-4)
and ethyl 3-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholinothieno[3,2-
d]pyrimidin-2-yl)-1H-indazol-l-yl)propanoate (Compound 0115-4)
A mixture of compound 0114 (160 mg, 0.31 mmol) , ethyl 6-bromohexanoate (83
mg, 0.37 mmol) and potassium carbonate (85 mg, 0.62 mmol) in acetonitrile (50
mL) was
refluxed overnight. Solvent was removed and the residue was partitioned
between
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dichloromethane and water. The organic layer was separated and washed with
brine, dried
over magnesium sulfate, filtered and evaporated in vacuo. The resulting
residue was
purified by prep-HPLC to give the title compound 0116-4 (40 mg, 20%) and 0115-
4 (70
mg, 34%).
Compound 0116-4: an oil; LCMS: 657 [M+1]+; 1H NMR (400 MHz, CDC13): 6 1.14 (t,
J=
7.2 Hz, 3H), 1.34 (m, 2H), 1.60 (m, 2H), 2.02 (m, 2H), 2.22 (t, J= 7.2 Hz,
2H), 2.62 (s,
4H), 2.73 (s, 3H), 3.23 (m, 4H), 3.82 (s, 2H), 3.84 (m, 4H), 4.00 (m, 6H),
4.40 (t, J= 7.2
Hz, 2H), 7.33 (m, 2 H), 7.74 (d, J= 8.4 Hz, 1H), 8.19 (d, J= 6.8 Hz, 1H), 8.82
(s, 1H).13C
NMR (100 MHz, CDC13): 6 172.4, 161.7, 159.6, 156.7, 148.6, 147.5, 130.1,
124.6, 123.1,
122.1, 119.2, 118.8, 112.0, 65.8, 59.4, 56.4, 52.6, 51.2, 45.3, 44.8, 33.3,
32.9, 29.3, 25.1,
23.4, 13.4.
Compound 0115-4: an oil; LCMS: 657 [M+1]+; 1H NMR (400 MHz, CDC13): 61.14 (t,
J=
7.2 Hz, 3H), 1.27 (m, 2H), 1.59 (m, 2H), 1.90 (m, 2H), 2.19 (t, J= 7.2 Hz,
2H), 2.61 (m,
4H), 2.72 (s, 3H), 3.22 (m, 4H), 3.83 (m, 6H), 4.00 (m, 6H), 4.35 (t, J= 7.2
Hz, 2H), 7.31
(s, 1H), 7.41 (m, 2H), 8.16 (m, 1H), 8.81 (s, 1H).13C NMR (100 MHz, CDC13): 6
172.6,
161.7, 159.8, 157.1, 147.7, 139.2, 133.5, 131.2, 124.9, 123.2, 121.4, 120.7,
112.1, 109.7,
65.8 (2C), 59.2, 56.3, 51.4, 47.7, 45.6, 44.8, 33.5, 33.0, 28.6, 23.5, 19.8,
13.2.
Step 2b: N-hydroxy-3-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholinothieno[3,2-d]pyrimidin-2-yl)-1H-indazol-1-yl)propanamide (Compound
4)
The title compound 4 was prepared as a yellow solid (45 mg, 22%) from 0115-4
(210 mg, 0.32 mmol) and freshly prepared hydroxylamine methanol solution (5.0
mL)
using a procedure similar to that described for compound 3 (Example 1): mp 186-
187 C.
LCMS: 643 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 1.22 (m, 2H), 1.52 (m, 2H), 1.84-
1.93 (m, 4H), 2.61 (m, 4H), 2.91 (s, 3H), 3.17 (m, 4H), 3.82-3.85 (m, 4H),
3.95 (s, 2H),
3.99- 4.01 (m, 4H), 4.44 (t, J= 6.8 Hz, 2H), 7.51 (m, 2 H), 7.80 (d, J= 8.8
Hz, 1H), 8.23
(d, J= 8.8 Hz, I H), 8.63 (s, I H), 8.85 (s, I H), 10.30 (s, I H).
EXAMPLE 3: Preparation of N-hydroxy-7-(4-(6-((4-(methylsulfonyl)piperazin- 1-
yl)methyl)-4-morpholinothieno [3,2-d] pyrimidin-2-yl)-1H-indazol-1-
yl)heptanamide
(Compound 5)
Step 3a: Ethyl 7-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)- 4-
morpholinothieno[3,2-d]pyrimidin-2-yl)-2H-indazol-2-yl)heptanoate (Compound
0116-5)
and ethyl 7-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholinothieno[3,2-
d]pyrimidin-2-yl)-1H-indazol-l-yl)heptanoate (Compound 0115-5)
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The title compound 0115-5 (110 mg, 27%) and 0116-5 (60 mg, 16%) were prepared
from 0114 (280 mg, 0.55 mmol), ethyl 7-bromoheptanoate (133 mg, 0.65 mmol) and
potassium carbonate (152 mg, 1.10 mmol) in acetonitrile (25 mL) using a
procedure similar
to that described for compound 0115-3 and compound 0 116-3 (Example 1):
Compound 0115-3: a white solid; LCMS: 670 [M+1]+; 'H NMR (400 MHz, CDC13): 6
1.15 (t, J= 7.2 Hz, 3H), 1.29 (m, 4H), 1.46 (m, 2H), 1.84 (m, 2H), 2.23 (t, J=
7.2 Hz, 2H),
2.61 (s, 4H), 2.91 (s, 3H), 3.16 (s, 4H), 3.83 (m, 4H), 3.95 (s, 2H), 4.00 (m,
6H), 4.45 (t, J=
6.6 Hz, 2H), 7.51 (m, 2H), 7.79 (d, J = 8.8 Hz, 1 H), 8.23 (d, J = 6.8 Hz 1
H), 8.86 (s, 1 H).
Compound 0116-3: a white solid; LCMS: 670 [M+1]+; 'H NMR (400 MHz, CDC13): 6
1.14 (t, J= 7.2 Hz, 3H), 1.31 (m, 4H), 1.53 (m, 2H), 1.97 (m, 2H), 2.25 (t, J=
7.6 Hz, 2H),
2.61 (s, 4H), 2.91 (s, 3H), 3.17 (s, 4H), 3.83 (m, 4H), 3.95 (s, 2H), 4.00 (m,
6H), 4.50 (t, J=
7.0 Hz, 2H), 7.3 7 (t, J = 7.8 Hz, 1 H), 7.52 (s, 1 H), 7.46 (d, J = 8.8 Hz, 1
H), 8.17 (d, J = 6.8
Hz I H), 9.02 (s, 1 H).
Step 3b: N-hydroxy-7-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholinothieno[3,2-d]pyrimidin-2-yl)-1H-indazol-1-yl)hentanamide (Compound
5)
The title compound 5 was prepared as a white solid (70 mg, 71%) from 0115-5
(100 mg,
0.15 mmol) and freshly prepared hydroxylamine methanol solution (4.0 mL) using
a
procedure similar to that described for compound 3 (Example 1): mp 127-130 C.
LCMS:
657 [M+1]; 'H NMR (400 MHz, DMSO-d6) 5 1.26 (s, 4H), 1.44 (m, 2H), 1.84 (m,
2H),
1.91 (t, J= 7.2 Hz, 2H), 2.61 (s, 4H), 2.91 (s, 3H), 3.16 (s, 4H), 3.83 (m,
4H), 3.95 (s, 2H),
4.00 (m, 4H), 4.45 (t, J = 6.8 Hz, 2H), 7.5 (m, 2 H), 7.80 (d, J = 8.4 Hz, 1
H), 8.23 (d, J =
7.6 Hz, I H), 8.65 (s, I H), 8.86 (s, I H), 10.32 (s, I H).
EXAMPLE 4: Preparation of N-hydroxy-5-(4-(6-((4-(methylsulfonyl)piperazin- 1-
yl)methyl)-4-morpholinothieno[3,2-d]pyrimidin-2-yl)-2H-indazol-2-
yl)pentanamide
(Compound 7)
The title compound 7 was prepared as a white solid (35 mg, 47%) from 0 116-3
(60
mg, 0.12 mmol) and freshly prepared hydroxylamine methanol solution (4.0 mL)
using a
procedure similar to that described for compound 3 (Example 1): mp 146-169 C.
LCMS:
629 [M+l]+; 'H NMR (400 MHz, DMSO-d6) 6 1.57 (m, 2H), 2.00 (m, 2H), 2.07 (t,
J= 7.2
Hz, 2H), 2.67 (s, 4H), 2.97 (s, 3H), 3.23 (s, 4H), 3.89 (s, 4H), 4.01 (s, 2H),
4.04 (m, 4H),
4.57 (t, J = 7.0 Hz, 2H), 7.43 (t, J = 7.8 Hz, 1 H), 7.5 8 (s, 1 H), 7.81 (d,
J = 8.4 Hz, 1 H),
8.28 (d, J= 6.8 Hz, 1H), 9.08 (s, 1H).
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EXAMPLE 5: Preparation of N-hydroxy-6-(4-(6-((4-(methylsulfonyl)piperazin- 1-
yl)methyl)-4-morpholinothieno [3,2-d] pyrimidin-2-yl)-2H-indazol-2-
yl)hexanamide
(Compound 8)
The title compound 8 was prepared as a yellow solid (15 mg, 11%) from 0116-4
(140 mg, 0.21 mmol) and freshly prepared hydroxylamine methanol solution (5.0
mL)
using a procedure similar to that described for compound 3 (Example 1): mp 124-
125 C.
LCMS: 643 [M+1]+;'H NMR (400 MHz, DMSO-d6) 6 1.23 (m, 2H), 1.55 (m, 2H), 1.92-
1.97 (m, 4H), 2.61 (m, 4H), 2.91 (s, 3H), 3.17 (m, 4H), 3.82-3.85 (m, 4H),
3.95 (s, 2H),
3.99- 4.01 (m, 4H), 4.50 (t, J= 6.8 Hz, 2H), 7.37 (m, I H), 7.53 (s, 2 H),
7.74 (d, J= 8.4 Hz,
I H), 8.21 (d, J= 6.8 Hz, I H), 8.64 (s, I H), 9.02 (s, I H), 10.31 (s, I H).
EXAMPLE 6: Preparation of N-hydroxy-7-(4-(6-((4-(methylsulfonyl)piperazin- 1-
yl)methyl)-4-morpholinothieno [3,2-d] pyrimidin-2-yl)-2H-indazol-2-
yl)heptanamide
(Compound 9)
The title compound 9 was prepared as a white solid (45 mg, 76%) from 0 116-5
(60
mg, 0.09 mmol) and freshly prepared hydroxylamine methanol solution (4.0 mL)
using a
procedure similar to that described for compound 3 (Example 1): mp 123-126 C.
LCMS:
657 [M+1 ]+; 'H NMR (400 MHz, DMSO-d6) 6 1.29 (s, 4H), 1.48 (m, 2H), 1.93 (m,
4H),
2.61 (s, 4H), 2.91 (s, 3H), 3.16 (s, 4H), 3.83 (m, 4H), 3.95 (s, 2H), 3.98 (m,
4H), 4.50 (t, J=
7.0 Hz, 2H), 7.3 7 (t, J = 7.8 Hz, 1 H), 7.52 (s, 1 H), 7.75 (d, J = 8.8 Hz, 1
H), 8.22 (d, J = 6.8
Hz, I H), 8.65 (s, I H), 9.03 (s, I H), 10.32 (s, I H).
Example 7: Preparation of 5-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)piperazin-1-yl)-N-hydroxypentanamide (Compound 11)
Step 7a: Tert-butyl 4-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)
piperazine-l-carboxylate (Compound 0201)
To a mixture of 0112 (4.0 g, 14.10 mmol) and tent-butyl piperazine-l-
carboxylate
(3.94 g, 21.15 mmol) in chloroform (50 mL) was added tetraisopyl titanate
(4.81 g, 16.92
mmol). The mixture was stirred at room temperature overnight. The solvent was
removed
under reduced pressure. The residue was dissolved in C1CH2CH2C1(60 mL) and
NaBH3CN
(1.33 g, 21.15 mmol) was added to the mixture. The mixture was stirred at room
temperature for 4 hours and was diluted with NaHCO3 solution. The mixture was
then
extracted with ethyl acetate. The organic phase was separated, dried and
concentrated to
afford the product 0201 (5.2 g, 81%): LCMS: 454 [M+1]+; 1H NMR (400 Hz, CDC13)
6
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1.46 (s, 9H), 2.49 (s, 4H), 3.47 (t, J = 4.4 Hz, 4H), 3.80 (s, 2H), 3.84 (t, J
= 5.2 Hz, 4H),
3.99 (t, J= 4.8 Hz, 4H), 7.17 (s, 1H).
Step 7b: 4-(2-Chloro-6-(piperazin-1-ylmethyl)thieno[3,2-d]pyrimidin-4-yl)
morpholine
(Compound 0202)
To a mixture of 0201 (5.2 g, 11.45 mmol) in dioxane was added 4 N HC1/dioxane
(30 mL) under N2. The reaction mixture was stirred at room temperature for 5
h. The
mixture was poured into water (30 mL), adjusted pH 7 with saturated NaHCO3
solution,
extracted with ethyl acetate, dried and concentrated to afford the product
0202 (3.0 g,
74%): LCMS: 354 [M+1]+; 1H NMR (400 Hz, CDC13) 6 2.52 (s, 4H), 2.93 (t, J= 4.8
Hz,
4H), 3.78 (s, 2H), 3.84 (t, J = 4.8 Hz, 4H), 3.99 (t, J = 4.4 Hz, 4H), 7.16
(s, 1 H).
Step 7c: Ethyl 5-(4-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)
piperazin-l-yl)pentanoate (Compound 0203-11)
To a mixture of 0202 (0.3 g, 0.85 mmol) in DMF (3 mL) was added Cs2CO3 (0.61
g, 1.87 mmol) and ethyl 5-bromopentanoate (0.2 g, 0.93 mmol). The reaction
mixture was
stirred at room temperature overnight and then poured into water (10 mL). The
mixture was
extracted with ethyl acetate. The organic phase was separated and washed with
water (10
mL x 5) and brine, dried over Na2SO4, filtered and concentrated to afford the
product 0203-
11 (0.36 g, 80%) as a gray solid: LCMS: 482 [M+1]+; 1H NMR (400 Hz, CDC13) 6
1.25 (t,
J= 7.2 Hz, 3H), 1.52-1.69 (m, 6H), 2.30-2.37 (s, 2H), 2.55 (m, 6H), 3.79 (s,
2H), 3.84 (t, J
= 5.2 Hz, 4H), 3.98 (t, J= 4.4 Hz, 4H), 4.13 (q, J= 6.8 Hz, 2H), 7.16 (s, 1H).
Step 7d: Ethyl 5-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d] pyrimidin-
6-
yl)methyl)piperazin-l-yl)pentanoate (Compound 0204-11)
A mixture of 0203-11 (250 mg, 0.52 mmol), 0107-3 (140 mg, 0.57 mmol), NaHCO3
(131 mg, 1.56 mmol) and Pd(dppf)2C12 (18 mg, 0.026 mmol) in toluene (4.8 mL),
ethanol
(2.5 mL) and water (1.3 mL) was flushed with N2 and heated under microwave
irradiation
at 130 C for 2 h. To the mixture was added water (10 mL) and extracted with
ethyl acetate.
The organic phase was separated and washed with brine, dried over Na2SO4,
filtered and
concentrated to give the crude product which was purified by silica gel column
(methanol
in dichloromethane 5% v/v) to afford the title product 0204-11 as a white
solid (62 mg,
21%): LCMS: 565 [M+2]+; 1H NMR (400 Hz, CDC13) 6 1.23 (m, 3H), 1.27 (m, 2H),
1.55
(m, 2H), 1.65 (m, 2H), 2.32 (t, J= 7.6 Hz, 2H), 2.40 (m, 2H), 2.54 (m, 2H),
2.64 (m, 4H),
3.86 (s, 2H), 3.92 (t, J= 4.8 Hz, 4H), 4.09 (t, J= 5.2 Hz, 4H), 4.13 (q, J=
7.2 Hz, 2H), 7.38
(s, 1 H), 7.5 0 (t, J = 7.2 Hz, 1 H), 7.5 9 (d, J = 8.0 Hz, 1 H), 8.28 (dd, J
= 7.2 Hz, 0.8 Hz, 1 H),
9.01 (d, J= 1.2 Hz, 1H).
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Step 7e: 5-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d] pyrimidin-6-
yl)methyl)piperazin-l-yl)-N-hydroxypentanamide (Compound 11)
The title compound 11 was prepared as a white solid (21 mg, 17%) from 0204-11
(129 mg, 0.23 mmol) and freshly prepared hydroxylamine methanol solution (1.0
mL, 1.77
mol/L) using a procedure similar to that described for compound 3 (Example 1):
m.p. 125-127 C, LCMS: 552 [M+2]+; 1H NMR (400 Hz, DMSO-d6) 6 1.23 (s, 2H),
1.39
(m, 2H), 1.48 (m, 2H), 1.94 (t, J = 7.6 Hz, 2H), 2.09 (s, 2H), 2.26 (t, J =
6.8 Hzõ 2H), 2.50
(m, 4H), 2.64 (m, 4H), 3.86 (m, 6H), 4.00 (m, 4H), 7.47 (t, J = 6.8 Hz, 1 H),
7.67 (d, J = 8.4
Hz, 1H), 8.22 (d, J= 6.8 Hz, 1H), 8.88 (s, 1H), 10.27 (s, 1H), 13.20 (s, 1H).
Example 8: Preparation of 6-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno [3,2-
d]pyrimidin-6-yl)methyl)piperazin-1-yl)-N-hydroxyhexanamide (Compound 12)
Step 8a: Ethyl 6-(4-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
methyl)piperazin-1-yl)hexanoate (Compound 0203-12)
The title compound 0203-12 was prepared as a gray solid (0.57 g, 82%) from
0202
(0.5 g, 1.41 mmol), Cs2CO3 (0.92 g, 2.82 mmol) and ethyl 6-bromohexanoate
(0.35 g, 1.55
mmol) using a procedure similar to that described for compound 0203-11
(Example 7):
LCMS: 496 [M+1]+; 1H NMR (400 Hz, CDC13) 6 1.25 (m, 4H), 1.34 (m, 2H), 1.64
(m, 2H),
2.30 (t, J= 7.6 Hz, 2H), 2.38 (m, 2H), 2.60 (m, 8H), 3.80 (s, 2H), 3.84 (t, J=
4.4 Hz, 4H),
3.40 (t, J= 4.8 Hz, 4H), 4.11 (q, J= 7.2 Hz, 2H), 7.16 (s, 1H).
Step 8b: Ethyl6-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin -6-
yl)methyl)piperazin-1-yl)hexanoate (Compound 0204-12)
The title compound 0204-12 was prepared as a white solid (56 mg, 16%) from
0203-12 (295 mg, 0.61 mmol), 0107-3 (164 mg, 0.67 mmol), NaHCO3 (150 mg, 1.79
mmol) and Pd(dppf)2C12 (23 mg, 0.031 mmol) in toluene (5.6 mL), ethanol (3 mL)
and
water (1.5 mL) using a procedure similar to that described for compound 0204-
11
(Example 7): LCMS: 579 [M+2]+; 1H NMR (400 Hz, CDC13) 6 1.25 (t, J= 7.6 Hz,
3H),
1.41 (m, 2H), 1.67 (m, 2H), 1.81 (m, 2H), 2.32 (t, J= 7.2 Hz, 2H), 3.02 (m,
8H), 3.61 (m,
2H), 3.96 (m, 5H), 4.13 (q, J= 14.4 Hz, 2H), 4.19 (m, 2H), 7.39 (t, J= 7.2 Hz,
1H), 7.55 (s,
1H), 7.60 (d, J= 8.8 Hz, 1H), 7.97 (d, J= 6.8 Hz, 1H), 8.50 (s, 1H).
Step 8c: 6-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin- 6-
yl)methyl)piperazin-1-yl)-N-hydroxyhexanamide (Compound 12)
The title compound 12 was prepared as a white solid (15 mg, 13%) from 0204-12
(120 mg, 0.21 mmol) and freshly prepared hydroxylamine methanol solution (1.0
mL, 1.77
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mol/L) using a procedure similar to that described for compound 3 (Example 1):
m.p. 123-
124 C, LCMS: 565 [M+1]+; 'H NMR (400 Hz, DMSO-d6) 6 1.23 (m, 2H), 1.26 (m,
2H),
1.36 (m, 2H), 1.93 (t, J= 7.2 Hz, 2H), 2.25 (t, J= 6.8 Hz, 2H), 2.39 (m, 4H),
3.30 (m, 4H),
3.85 (m, 6H), 4.00 (t, J = 5.2 Hz, 4H), 7.47 (d, J = 15.2 Hz, 2H), 7.66 (d, J
= 8.4 Hz, 1 H),
8.22 (d, J= 6.8Hz, 1H), 8.65 (s, 1H), 8.88 (s, 1H).
Example 9: Preparation of 7-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)piperazin-1-yl)-N-hydroxyheptanamide (Compound 13)
Step 9a: Ethyl 7-(4-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)
piperazin-1-yl)heptanoate (Compound 0203-13)
The title compound 0203-13 was prepared as a gray solid (0.55 g, 76%) from
0202
(0.5 g, 1.41 mmol), Cs2CO3 (0.92 g, 2.82 mmol) and ethyl 6-bromohexanoate
(0.35 g, 1.55
mmol) using a procedure similar to that described for compound 0203-11
(Example 7): 512
[M+2]+; 'H NMR (400 Hz, DMSO-d6) 6 1.17 (t, J= 6.8 Hz, 3H), 1.28 (m, 4H), 1.52
(m,
2H), 1.61 (m, 2H), 2.28 (t, J= 7.2 Hz, 2H), 2.49 (m, 4H), 3.05 (m, 6H), 3.75
(t, J= 4.4 Hz,
4H), 3.88 (t, J= 4.8 Hz, 2H), 3.97 (m, 2H), 4.04 (q, J= 14 Hz, 2H), 7.34 (s,
1H).
Step 9b: Ethyl7-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d] pyrimidin-6-
yl)methyl)piperazin-1-yl)heptanoate (Compound 0204-13)
The title compound 0204-13 was prepared as a white solid (56 mg, 16%) from
0203-13 (300 mg, 0.60 mmol), 0107-3 (162 mg, 0.67 mmol), NaHCO3 (151 mg, 1.80
mmol) and Pd(dppf)2C12 (21 mg, 0.03 mmol) in toluene (5.5 mL), EtOH (3 mL) and
water
(1.5 mL) using a procedure similar to that described for compound 0204-11
(Example 7):
LCMS: 592 [M+1]+; 'H NMR (400 Hz, CDC13) 6 1.25 (m, 3H), 1.33 (m, 4H), 1.51
(m, 2H),
1.63 (m, 2H), 2.29 (t, J= 7.6 Hz, 2H), 2.37 (m, 2H), 2.52 (m, 4H), 2.64 (mõ
4H), 3.86 (s,
2H), 3.92 (t, J= 4.4 Hz, 4H), 4.09 (t, J= 6.0 Hz, 4H), 4.12 (q, J= 14.4 Hz,
2H), 7.38 (s,
1 H), 7.5 0 (t, J = 8.4 Hz, 1 H), 7.5 9 (d, J = 8.4 Hz, 1 H), 8.28 (dd, J =
7.6 Hz, 0.8 Hz, 1 H),
9.02 (d, J = 0.8 Hz, 1 H).
Step 9c: 7-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)piperazin-1-yl)-N-hydroxyheptanamide (Compound 13)
The title compound 13 was prepared as a white solid (25 mg, 37%) from 0204-13
(68 mg, 0.11 mmol) and freshly prepared hydroxylamine methanol solution (0.5
mL, 1.77
mol/L) using a procedure similar to that described for compound 3 (Example 1):
m.p. 119-
122 C, LCMS: 580 [M+2]+; 'H NMR (400 Hz, DMSO-d6) 6 1.27 (m, 4H), 1.50 (m,
2H),
1.62 (m, 2H), 1.94 (t, J = 6.8 Hz, 2H), 2.47 (m, 4H), 3.06 (m, 6H), 3.85 (t, J
= 4.0 Hz, 4H),
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4.02 (m, 6H), 4.00 (t, J = 5.2 Hz, 4H), 7.49 (t, J = 8.4 Hz, 1 H), 7.5 6 (s, 1
H), 7.69 (d, J = 8.0
Hz, 1H), 8.21 (d, J= 7.2 Hz, 1H), 8.87 (s, 1H), 9.39 (s, 1H), 10.35 (s, 1H).
EXAMPLE 10: Preparation of 2-(4-((2-(1H-indazol-4-yl)-4- morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)piperazin-1-yl)-N-hydroxypyrimidine-5-carboxamide
(Compound 14)
Step 10a: (Z)-Ethyl-2-(ethoxymethyl)-3-methoxyacrylate (Compound 0302)
Sodium (13.8 g) was added to a mixture of benzene (200 mL) and ethanol (27 g)
at
room temperature. To the above mixture was added a mixture of ethyl formate
(45.0 g, 0.61
mol) and ethyl 3-ethoxypropionate (44.0 g, 0.30 mol) slowly at 0 C. The
resulting reaction
mixture was stirred for 2 hours and then dimethyl sulfate (76.0 g, 0.61 mol)
was added and
stirred at 50 C for 3 h. The mixture was filtered, and the filtrate was
washed with water.
The organic layer was separated and was added triethylammonium chloride (40.0
g, 0.29
mol) and sodium hydroxide (7.00 g, 0.175 mol). The resulting mixture was
stirred for 4 h.
and then filtered. The filtrate was washed with water, dried over Na2SO4,
filtered and
evaporated to give a residue which was distilled under vacuum to provide
compound 0302
(18.8 g, 33%) which was used directly to the next step without further
purification: 1H
NMR (400 MHz, CDC13): 6 1.26 (m, 6H), 3.48 (m, 3H), 3.63 (m, 3H), 4.20 (m,
2H).
Step 10b: Ethyl 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate (Compound
0303)
A mixture of compound 0302 (21.4 g, 0.11 mol), urea (5.70 g, 0.095 mol), and
concentrated hydrochloric acid (36%-38%, 5 mL) in ethanol (300 mL) was heated
at reflux
overnight. After evaporation, the residue was recrystallized from ethanol to
give compound
0303 (7.80 g, 65%) as a colorless prisms: LCMS: 171 [M+1]+, 1H NMR (400 MHz,
CDC13): 6 1.27 (t, J = 7.2 Hz , 3H), 4.19 (m, 4H), 5.28 (s, 1 H), 7.21 (d, J =
5.6 Hz, 1 H),
7.40 (s, 1H).
Step 10c: Ethyl 2-oxo-1,2-dihydropyrimidine-5-carboxylate (Compound 0304)
A solution of compound 0303 (2.50 g, 14.7 mmol) and bromine (2.40 g, 15 mmol)
in acetic acid (55 mL) was heated at reflux for 1.5 h. Removal of the solvent
afforded crude
compound 0304 (3.60 g, 99%) which was used directly to the next step without
further
purification:. LCMS: 169 [M+1]+, 1H NMR (400 MHz, CDC13): 6 1.27 (t, J= 7.2 Hz
, 3H),
4.28 (q, J= 7.2 Hz , 2H), 8.85 (s, 2H), 12.19 (ds, 2H).
Step 10d: Ethyl 2-chloropyrimidine-5-carboxylate (Compound 0305)
A mixture of compound 0304 (3.60 g, 21 mmol), phosphorus oxychloride ( 25 mL),
and N,N-dimethylaniline (2.5 mL) was heated at reflux for 1.5 h. After removal
of the
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solvent, ice water (10 mL) was added to the residue. The mixture was added to
2 N NaOH
(90 ml), and extracted with EtOAc. The organic layer was evaporated and
purified by
column chromatography (ethyl acetate in petroleum ether, 5% v/v) to give
compound 0305
(1.20 g, 30%): LCMS: 187 [M+1]+, 'H NMR (300 MHz, CDC13): 6 1.42 (t, J= 7.5 Hz
,
3H), 4.48 (q, J= 7.5 Hz, 2H), 9.15 (s, 2H); 'H NMR (400 MHz, DMSO-d6): 6 1.33
(t, J=
6.8 Hz, 3H); 4.37 (q, J= 6.8 Hz, 2H), 9.18 (s, 2H).
Step 10e: Ethyl 2-(piperazin-1-yl)pyrimidine-5-carboxylate (Compound 0306)
A mixture of compound 0305 (1.10 g, 5.9 mmol) and piperazine (1.02 g, 11.8
mmol) in DMF (50 mL) was stirred at room temperature for 1.5 h. The mixture
was diluted
with water and extracted with ethyl acetate. The organic layer was washed with
water and
dried, concentrate to give compound 0306 (1.20 g, 86%): LCMS: 237 [M+1]+, 'H
NMR
(400 MHz, DMSO-d6): 6 1.29 (t, J= 7.2 Hz, 3H), 2.73 (t, J= 5.2 Hz, 4H), 3.77
(t, J= 5.2
Hz, 4H), 4.25 (q, J= 7.2 Hz, 2H), 8.76 (s, 2H).
Step 10f: Ethyl 2-(4-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
methyl)piperazin-1-yl)pyrimidine-5-carboxylate (Compound 0307)
To a mixture of compound 0112 (500 mg, 1.77 mmol) and compound 0306 (376
mg, 1.59 mmol) in chloroform (40 mL) was added tetraisopyl titanate (754 mg,
2.66 mmol).
The mixture was stirred at reflux overnight. Solvent was evaporated and then
1,2-
dichloroethane (50 mL) and sodium cyanborohydride (168 mg, 2.66 mmol) were
added.
The resulting mixture was stirred at room temperature for 12 h. The mixture
was poured
into saturated NaHCO3 and extract with ethyl acetate ( 2 x 50 mL). The organic
layer was
separated and evaporated to afford a mixture which was purified by silica gel
column
chromatography (ethyl acetate in petroleum ether 10% v/v) to geve compound
0307 (270
mg, 34%) as a yellow solid: LCMS: 504 [M+1]+; 'H NMR (400 MHz, CDC13) 6 1.35
(t, J
6.8 Hz, 3H), 2.62 (s, 4H), 3.85 (m, 6H), 4.00 (m, 8H), 4.33 (q, J= 6.8 Hz,
2H), 7.26 (s,
I H), 8.84 (s, 2H).
Step 10g: Ethyl 2-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d] pyrimidin-
6-
yl)methyl)piperazin-1-yl)pyrimidine-5-carboxylate (Compound 0308)
The title compound 0308 was prepared as a yellow solid (60 mg, 23%) from 0307
(220 mg, 0.44 mmol), 0107-3 (161 mg, 0.66 mmol), NaHCO3 (111 mg, 1.32 mmol)
and
Pd(dppf)2C12 (19 mg, 0.022 mmol) in toluene (4.7 mL), ethanol (2.8 mL) and
water (1.2
mL) using a procedure similar to that described for compound 0204-11 (Example
7):
LCMS: 586 [M+1]+; 'H NMR (400 MHz, CDC13): 6 1.35 (t, J= 6.8 Hz, 3H), 2.66 (s,
4H),
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3.93 (m, 6H), 4.01 (m, 4H), 4.11 (m, 4H), 4.33 (q, J = 6.8 Hz, 2H), 7.41 (s, 1
H), 7.51 (m,
I H), 7.69 (d, J= 8.4 Hz, I H), 8.28 (d, J= 6.8 Hz, I H), 8.84 (s, 2H), 9.01
(s, I H).
Step 10h: 2-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d] pyrimidin-6-
yl)methyl)piperazin-1-yl)-N-hydroxypyrimidine-5-carboxamide (Compound 14)
The title compound 14 was prepared as a yellow solid (30 mg, 52%) from 0308
(60
mg, 0.10 mmol) and freshly prepared hydroxylamine methanol solution (3.0 mL,
1.77
mol/L) using a procedure similar to that described for compound 3 (Example 1):
mp 209-
221 C. LCMS: 573 [M+1]; 'H NMR (400 MHz, DMSO-d6) (52.59 (s, 4H), 3.85 (m,
8H), 3.95 (s, 2H), 4.01 (m, 4H), 7.47 (m, 1 H), 7.52 (m, 1 H), 7.67 (d, J =
8.4 Hz, 1 H), 8.23
(d, J= 6.8 Hz, I H), 8.68 (s, 2H), 8.89 (s, I H), 9.00 (s, I H), 11.07 (s, I
H), 13.19(s, 1 H).
EXAMPLE 11: Preparation of 2-(4-(((2-(1H-indazol-4-yl)-4-morpholinothieno [3,2-
d] pyrimidin-6-yl)methylamino)methyl)piperidin-1-yl)-N-hydroxypyrimidine-5-
carboxamide (Compound 15)
Step 11 a: Ethyl 2-(4-(aminomethyl)piperidin-l-yl)pyrimidine-5-carboxylate
(Compound 0401)
A mixture of 0405 (1.10 g, 5.9 mmol), piperidin-4-ylmethanamine (1.35 g, 11.8
mmol) in 2-(dimethylamino)acetamide (50 mL) was stirred at room temperature
for 1.5 h.
After removal of the solvent, the residue was purified by column
chromatography on silica
gel (CH3OH in CH2C12 6% v/v) to give desired product 0401 (1.27 g, 81%): LCMS:
265
[M+1]+, 'H NMR (400 MHz, CDC13): 6 1.16 (m, 2H), 1.22(m, 5H), 1.36 (m, 1H),
1.64 (m,
IH), 1.85 (d, J= 12 Hz, 2H), 2.62 (d, J= 6.42 Hz, 2H), 2.94 (ds, J= 12.8 Hz,
J= 2.4 Hz
,2H), 4.91 (d, J= 11.2 Hz, 2H), 7.26(s, 1H) , 8.82 (s, 2H).
Step 11b: Ethyl 2-(4-(((2-chloro-4-morpholinothieno[3,2-d]pyrimidin- 6-
yl)methylamino)me thyl)piperidin-1-yl)pyrimidine-5-carboxylate (Compound 0402-
15)
To a mixture of compound 0112 (589 mg, 2.08 mmol) and compound 0401 (500
mg, 1.89 mmol) in chloroform (50 mL) was added tetraisopyl titanate (644 mg,
2.26 mmol).
The mixture was stirred at reflux overnight. The solvent was removed and 1,2-
dichloroethane (30 mL) and sodium cyanborohydride (179 mg, 2.84 mmol) were
then
added. The mixture was stirred at room temperature for 12 h. The mixture was
poured into
saturated NaHCO3 and extract with ethyl acetate ( 2 x 50 mL. The organic layer
was
separated evaporated. The residue was purified by column on silica gel (ethyl
acetate in
petroleum ether 10% v/v) to give compound 0402 (630 mg, 57%). LCMS: 533
[M+1]+. 'H
NMR (400 MHz, DMSO-d6) 6 1.05-1.14 (m, 2H), 1.29 (t, J= 7.2 Hz, 3H), 1.78-1.85
(m,
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3H), 2.46 (d, J= 6.0 Hz, 2H), 2.68 (brs, 1H), 2.98 (t, J= 11 Hz, 2H), 3.75 (t,
J= 4.8 Hz,
4H), 3.88 (t, J = 4.8 Hz, 4H), 4.02 (s, 2H), 4.26 (t, J = 7.2 Hz, 2H), 4.74
(d, J = 13 Hz, 2H),
7.23 (s, 1H), 8.74 (s, 2H).
Step 11c: Ethyl2-(4-(((2-(1H-indazol-4-yl)-4-morpholinothieno [3,2-d]pyrimidin-
6-yl)me
thylamino)methyl)piperidin-1-yl)pyrimidine-5-carboxylate (Compound 0403-15)
The title compound 0403 was prepared as a white solid (120 mg, 17%) from 0402
(630 mg, 1.18 mmol), 0107-3 (580 mg, 2.37 mmol), NaHCO3 (297 mg, 3.54 mmol)
and
Pd(dppf)2C12 (25 mg, 0.036 mmol) in toluene (11 mL), ethanol (6.6 mL) and
water (3.1
mL) using a procedure similar to that described for compound 0204-11 (Example
7):
LCMS: 614 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 6 1.09-1.14 (m, 2H), 1.28 (t, J=
7.2
Hz, 3H), 1.82-1.87 (m, 3H), 3.00 (t, J= 12 Hz, 2H), 3.83 (t, J= 4.6 Hz, 4H),
4.01 (t, J= 4.6
Hz, 4H), 4.07 (s, 2H), 4.26 (t, J= 7.2 Hz, 2H), 4.75 (d, J= 13 Hz, 2H), 7.47
(t, J= 7.6 Hz,
2H), 7.66 (d, J = 8.0 Hz, 1 H), 8.22 (d, J = 7.2 Hz, 1 H), 8.75 (s, 2H), 8.89
(s, 1 H), 13.18 (s,
I H).
Step 11d: 2-(4-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)methyl)piperidin-1-yl)-N-hydroxypyrimidine-5-carboxamide
(Compound
15)
The title compound 15 was prepared as a white solid (30 mg, 63%) from 0403 (50
mg, 0.08 mmol) and freshly prepared hydroxylamine methanol solution (3.0 mL,
1.77
mol/L) using a procedure similar to that described for compound 3 (Example 1):
mp 170-
172 C. LCMS: 601 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 1.05-1.13 (m, 2H), 1.23
(m,
1H), 1.82-1.85 (m, 3H), 2.95 (t, J= 12 Hz, 2H), 3.84 (s, 4H), 4.01 (s, 4H),
4.07 (s, 2H),
4.71 (d, J = 13 Hz, 2H), 7.47 (t, J = 7.6 Hz, 2H), 7.66 (d, J = 8.0 Hz, 1 H),
8.22 (d, J = 7.2
Hz, I H), 8.65 (s, 2H), 8.88 (s, I H), 8.98 (s, I H), 11.03 (s, I H), 13.18
(s, I H).
EXAMPLE 12: Preparation of 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno [3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 54)
Step 12a: (2-Chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methanol (Compound
0501)
To a mixture of compound 0112 (500 mg, 1.77 mmol) in methanol (10 mL) was
added sodium borohydride (200 mg, 5.3 mmol). The mixture was stirred at room
temperature for 1 hour. The reaction was quenched with water and extracted
with ethyl
acetate, dried over Na2SO4 and concentrated to obtain crude compound 0501 (500
mg,
99%) as a yellow solid: LCMS: 286 [M+1]+; 1H NMR (400 MHz, DMSO-d6). 5 3.74
(t, J
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= 4.4 Hz, 4H), 3.88 (t, J= 4.4 Hz, 4H), 4.80 (d, J= 5.6 Hz, 2H), 5.93 (t, J=
5.6 Hz, 1H),
7.21 (s, 1H).
Step 12b: 4-(6-(Bromomethyl)-2-chlorothieno[3,2-d]pyrimidin-4-yl)morpholine
(Compound 0502)
To a solution of compound 0501 (1.6 g, 5.6 mmol) in dichloromethane (100 mL)
was added N-bromosuccinimide (1.2 g, 6.7 mmol) and triphenylphosphine (1.75 g,
6.7
mmol). The mixture was stirred at 25 C for 3 hours. Solvent was removed the
residue was
purified by column chromatography (ethyl acetate in petroleum ether 20% v/v)
to give title
compound 0502 (1.16 mg, 60%) as a yellow solid: LCMS: 348 [M+1]+; 'H NMR (400
MHz, DMSO-d6). 6 3.74 (t, J= 4.8 Hz, 4H), 3.88 (t, J= 4.4 Hz, 4H), 4.79 (s,
2H), 7.21
(s, 1H).
Step 12c: (2-Chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)-N-
methylmethanamine
(Compound 0503-54)
A mixture of compound 0502 (190 mg) and methanamine alcohol solution (50 mL)
was stirred at reflux temperature for 1 hour. The solvent was removed at
reduce pressure
and the residue was purified by column chromatography (methanol in
dichloromethane,
12% v/v) to give title compound 0503-54 (190 mg, 54%) as a yellow solid: LCMS:
299
[M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 2.06 (s, 3H), 2.93 (s, 1H), 3.45 (t, J=
4.4 Hz,
4H), 3.57 (t, J = 4.4 Hz, 4H), 3.73 (s, 2H), 7.02 (s, 1 H).
Step 12d: Ethyl 2-(((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)
(methyl)amino)pyrimidine-5-carboxylate (Compound 0504-54)
A mixture of compound 0503-54 (215 mg, 0.72 mmol), compound 0305 (336 mg,
1.8 mmol) and N, N-diisopropylethylamine(20 mL) in acetonitrile (30 mL) was
stirred at
room temperature overnight. The solvent was removed at reduce pressure and the
resulting
precipitation was washed with ethyl acetate and dried to provide the title
compound 0504-
54 (210 mg, 65%) as a yellow solid: LCMS: 531 [M+1]+; 1H NMR (400 MHz, DMSO-
d6):
6 1.4 (t, J= 6.8 Hz, 3H), 3.35 (s, 3H), 3.81 (t, J= 4 Hz, 4H), 3.93 (t, J= 4
Hz, 4H), 4.38
(q, J = 7.2 Hz, 2H), 5.31 (s, 2H), 7.05 (s, 1 H), 8.97 (s, 2H).
Step 12e: Ethyl 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)
methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0505)
A mixture of compound 0504 (210 mg, 0.47 mmol), 0107-3 (171 mg, 0.7 mmol),
sodium hydrogen carbonate (118 mg, 1.4 mmol) and bis(triphenylphosphine)
palladium(a)
chloride (16 mg, 0.02 mmol) in toluene (5 mL), ethanol (3 mL) and water (1.3
mL) was
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flushed with nitrogen and heated under microwave irradiation at 120 C for 1
h. The
reaction mixture was added water and extracted with ethyl acetate. The ethyl
acetate layer
was collected and washed with brine, dried over magnesium sulfate, filtered
and evaporated
to give a residue which was washed with dichloromethane to obtain the title
compound
0505-54 (130 mg, 52%) as a white solid: LCMS: 449 [M+1]+; 1H NMR (400 MHz,
DMSO-d6): 6 1.37 (t, J= 6.8 Hz, 3H), 3.36 (s, 3H), 3.86 (t, J= 4.8 Hz, 4H),
4.02 (t, J= 4.8
Hz, 4H), 4.35 (q, J = 6.8 Hz, 2H), 5.33 (s, 2H), 7.53 (m, 1 H), 7.66 (s, 1 H),
7.73 (d, J = 8
Hz, 1 H), 8.28 (d, J = 7.2 Hz, 1 H), 8.95 (dd, JI = 8 Hz, J2 = 7.6 Hz, I H),
13.28 (s, I H).
Step 12f: 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin- 6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 54)
The title compound 54 was prepared as a white solid (17 mg, 15%) from 0505-54
(120 mg, 0.22 mmol) and freshly prepared hydroxylamine methanol solution (4.0
mL, 1.77
mol/L) using a procedure similar to that described for compound 3 (Example 1):
m.p. 197-200 C. LCMS: 518 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 6 3.26 (s, 3H),
3.86 (t, J= 4.0 Hz, 4H), 3.96 (t, J= 4.0 Hz, 4H), 5.23 (s, 2H), 7.47 (t, J=
7.6 Hz, 1H), 7.58
(s, 1 H), 7.67 (d, J = 8 Hz, 1 H), 8.22 (d, J = 7.2 Hz, 1 H), 8.76 (s, 2H),
8.87 (s, 1 H), 9.09 (s,
1 H), 11.16 (s, 1 H), 13.22 (s, 1 H).
Example 13: Preparation of 2-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methylamino)-N-hydroxypyrimidine-5-carboxamide (Compound 53)
Step 13a: (2-Chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methanamine
(Compound
0503-53)
To a solution of compound 0502 (1.5g, 4.3 mmol) in methanol (20 mL) was added
NH3.H20 (2OmL). The mixture was stirred overnight at 65 C. The solvent was
removed at
reduce pressure and the resulting residue was purified by column
chromatography (ethyl
acetate in petroleum ether, 50% v/v) to give title compound 0503-53 (270 mg,
22%) as a
yellow solid: LCMS: 285 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 6 3.75 (t, J= 4.8
Hz,
4H), 3.89 (t, J = 4.4 Hz, 4H), 4.06 (s, 2H), 7.22 (s,1 H).
Step 13b: Ethyl 2-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)
pyrimidine-5-carboxylate (Compound 0504-53)
A mixture of compound 0503-53 (270 mg, 0.95 mmol), compound 0305 (353 mg,
1.9 mmol) and N, N-Diisopropylethylamine(2 mL) in acetonitrile (10 mL) was
stirred at
room temperature overnight. The solvent was removed at reduce pressure and the
precipitation was washed with dichloromethane, dried to provide the title
compound 0504-
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53 (160 mg, 39%) as a white solid: LCMS: 435 [M+1]+; 1H NMR (400 MHz, DMSO-
d6): 6
1.29 (t, J= 7.2 Hz, 3H), 3.72 (t, J= 5.2 Hz, 4H), 3.84 (t, J= 4.8 Hz, 4H),
4.27 (dd, J 1=
14.0 Hz, J2= 6.8 Hz,2H), 4.88 (d, J= 6 Hz, 2H), 7.30 (s, 1H), 8.79 (s, 2H),
8.85 (t, J= 6
Hz, 1H).
Step 13c: Ethyl 2-((2-(IH-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)
methylamino)pyrimidine-5-carboxylate (Compound 0505-53)
A mixture of compound 0504-53 (160 mg, 0.37 mmol), 0107-3 (135 mg, 0.55
mmol), sodium hydrogen carbonate (93 mg, 1.11 mmol) and
bis(triphenylphosphine)
palladium(a) chloride (13 mg, 0.02 mmol) in toluene (5 mL), ethanol (3 mL) and
water
(1.3 mL) was flushed with nitrogen and heated under microwave irradiation at
120 C for 1
h. The reaction mixture was added water and extracted with ethyl acetate. The
organic
layer was collected and washed with brine, dried over magnesium sulfate,
filtered and
evaporated to give a residue which was washed with dichloromethane to give
title
compound 0505-53 (80 mg, 42%) as a white solid: LCMS: 517 [M+1]+; 1H NMR (400
MHz, DMSO-d6): 6 1.29 (t, J= 6.8 Hz, 3H), 3.81 (t, J= 4.8 Hz, 4H), 3.97 (t, J=
4.0 Hz,
4H), 4.27 (dd, J,= 14.0 Hz, J2= 6.8 Hz,2H), 4.93 (d, J= 6.0 Hz, 2H), 7.45-7.50
(m,2H),
7.66 (d, J = 8.0 Hz, 1 H), 8.22 (d, J = 7.6 Hz, 1 H), 8.81 (d, J = 6.4 Hz,
2H), 8.8 8 (s, 2H),
13.2 (s, 1H).
Step 13d: 2-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
methylamino)-N-hydroxypyrimidine-5-carboxamide (Compound 53)
The title compound 53 was prepared as a light yellow solid (19 mg, 24%) from
0505-53 (80 mg, 0.15 mmol) and freshly prepared hydroxylamine methanol
solution (5.0
mL, 1.77 mol/L) using a procedure similar to that described for compound 3
(Example 1):
m.p. 234-237 C. LCMS: 504 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 5 2.09 (s, 2H),
3.81 (t, J= 4.8 Hz, 4H), 3.97 (t, J= 4.4 Hz, 4H), 4.90 (d, J= 6.0 Hz, 2H),
7.47 (t, J= 9.2
Hz, 2H), 7.66 (d, J= 8.4 Hz, I H), 8.22 (d, J= 6.8 Hz, I H), 8.56 (t, J= 6.0
Hz, I H), 8.68 (s,
2H), 8.88 (s, I H), 9.04 (s, I H), 11.09 (s, I H), 13.21 (s, I H).
Example 14: Preparation of 2-(4-((((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)methyl)piperidin-1-yl)-N-
hydroxypyrimidine-
5-carboxamide (Compound 16)
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Step 14a: Ethyl 2-(4-((((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)methyl)piperidin-1-yl)pyrimidine-5-carboxylate
(Compound
0402-16)
A compound of 0402-15 (510 mg, 0.96 mmol) and paraformaldehyde (58 mg, 1.92
mmol) was solved in methanol (20 mL), then NaBH3CN (121 mg, 1.92 mmol) was
added,
the mixture was stirred at room temperature overnight. Methanol was removed
and the
residue was added ethyl acetate and water. The organic layer was washed by
water and
brine water, dried with anhydrous Na2SO4. Filtered, concentrated and the
residue was
purified by column (ethyl acetate in petroleum ether 40% v/v) to get compound
0402-16
(265 mg, 51%) as a yellow solid. LCMS: 546 [M+1]+. 'H NMR (400 MHz, DMSO-d6):
6
1.04 (m, 2H), 1.28 (t, J= 7.2, 3H), 1.84-1.88 (m, 3H), 2.24 (s, 3H), 2.28 (d,
J= 6.8 Hz,
2H), 3.01 (t, J= 11.6 Hz, 2H), 3.75 (m, 4H), 3.83 (m, 2H), 3.88 (m, 4H), 4.25
(q, J= 7.2
Hz, 2H), 4.74 (d, J = 13.2 Hz, 2H), 7.27 (s, 1 H), 8.75 (s, 2H).
Step 14b: Ethyl 2-(4-((((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)methyl)piperidin-1-yl)pyrimidine-5-carboxylate
(Compound
0403-16)
A mixture of compound 0402-16 (246 mg, 0.45 mmol), 0107-3 (221 mg, 0.90
mmol), sodium hydrogen carbonate (12.6 mg, 1.5 mmol) and
bis(triphenylphosphine)
palladium(II) chloride (19 mg, 0.023 mmol) in toluene (5 mL), ethanol (2.9 mL)
and water
(1.3 mL) was flushed with nitrogen and heated under microwave irradiation at
120 C for 2
h. The reaction mixture was partitioned between dichloromethane and water,
organic layer
was washed with brine, dried over magnesium sulfate, filtered and evaporated
in vacuo.
The resulting residue was purified using column chromatography eluting
methanol in
dichloromethane (2%, v/v), to give title compound 0403-16 (200 mg, 71%) as a
yellow
solid. LCMS: 628 [M+1]+; 'H NMR (400 MHz, CDC13): 6 1.18 (m, 2H), 1.35 (t, J=
7.2
Hz, 3H), 1.94-1.97 (m, 3H), 2.34- 2.36 (m, 5H), 2.97 (t, J= 12.8 Hz, 2H), 3.84
(m, 2H),
3.94 (m, 4H), 4.10 (m, 4H), 4.32 (q, J= 7.2 Hz, 2H), 4.90 (d, J= 13.2 Hz, 2H),
7.36 (s,
I H), 7.49 (t, J= 8.0 Hz, I H), 7.57 (d, J= 8.0 Hz, I H), 8.27 (d, J= 7.2 Hz,
I H), 8.82 (s,
2H), 9.01 (s, I H), 10.42 (s, I H).
Step 14c: 2-(4-((((2-(1 H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)methyl)piperidin-1-yl)-N-hydroxypyrimidine-5-
carboxamide
(Compound 16)
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To a stirred solution of hydroxylamine hydrochloride (4.67 g, 67 mmol) in
methanol
(24 mL) at 0 C was added a solution of potassium hydroxide (5.61 g, 100 mmol)
in
methanol (14 mL). After addition, the mixture was stirred for 30 minutes at 0
C, and was
allowed to stand at low temperature. The resulting precipitate was isolated,
and the solution
was prepared to give free Hydroxylamine.
The above freshly prepared hydroxylamine solution (10.00 mL) was placed in 100
mL flask. Compound 111-47-2 (200 mg, 0.32 mmol) was added to this solution and
degassed at 0 C for 15 minutes. The reaction process was monitored by TLC.
The mixture
was neutralized with dry ice, filtered and washed with water, methanol and DCM
to give
the title compound 111-47 (130 mg, 66%) as a yellow solid: mp 174-175 C.
LCMS: 616
[M+1]+; 'H NMR (400 MHz, DMSO-d6) 6 1.03 (m, 2H), 1.85-1.94 (m, 3H), 2.27-
2.31
(m, 5H), 2.97 (t, J= 12.0 Hz, 2H), 3.84 (m, 6H), 4.00 (m, 4H), 4.71 (d, J=
12.8 Hz, 2H),
7.45-7.49 (m, 2H), 7.66 (d, J = 8.4 Hz, 1 H), 8.22 (d, J = 7.2 Hz, 1 H), 8.65
(s, 2H), 8.89 (s,
I H), 9.00 (s, I H), 11.06 (s, I H), 13.22 (s, I H).
Example 15: Preparation of 4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methylamino)-N-hydroxybutanamide (Compound 41)
Step 15a: Ethyl4-(tert-butoxycarbonyl((2-chloro-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)amino)butanoate (Compound 0404-41)
15a-1: Ethyl 4-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)butanoate
Ethyl 4-aminobutanoate hydrochloride (1.97 g, 11.77 mmol) was solved in
chloroform (30 mL) and the pH of mixture was adjusted to 8-9 with
triethylamine, then
compound 0112 (1.66 g, 5.88 mmol) and tetraisopyl titanate (2.01 g, 7.06 mmol)
were
added and the mixture was stirred at reflux overnight. Removed the solvent,
then 1,2-
dichloroethane (50 mL) and sodium cyanborohydride (1.48 g, 23.53 mmol) were
added and
then stirred at room temperature for 12 h. The mixture was poured into
saturated NaHCO3
solution and extract with ethyl acetate ( 3 x 50 mL) and purified by column
(ethyl acetate
in petroleum ether 50% v/v) to get 4-((2-chloro-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methylamino)butanoate (1.38 g, 59%) as a yellow solid. LCMS: 399 [M+1]+; 'H
NMR
(400 MHz, CDC13) 6 1.15 (t, J= 7.2 Hz, 3H), 1.68 (m, 2H), 2.35 (t, J= 7.2 Hz,
2H), 2.55 (t,
J = 7.2 Hz, 2H), 3.74 (t, J = 4.8 Hz, 4H), 3.89 (t, J = 4.8 Hz, 4H), 4.00 (s,
2H), 4.03 (q, J =
7.2 Hz, 2H), 7.25 (s, 1H).
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15a-2: Ethyl 4-(tert-butoxycarbonyl((2-chloro-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)amino)butanoate (0404-41)
To a solution of 4-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)butanoate (400 mg, 1.0 mmol) in THE (10 mL) was added (Boc)20
(218
mg, 1.0 mmol).Then the mixture was stirred at room temperature overnight. The
reaction
mixture was extracted with ethyl acetate and washed with water, brine and
dried over
sodium sulfate. The crude product was purified using column chromatography
(ethyl
acetate in petroleum ether 75% v/v), to give title compound 0404-41 (330 mg,
66%) as a
colorless liquid. LCMS: 499 [M+1]+; 'H NMR (400 MHz, DMSO-d6) 6 1.16 (t, J=
7.2 Hz,
3H), 1.42 (s, 9H), 1.74 (m, 2H), 2.26 (t, J = 7.2 Hz, 2H), 3.23 (t, J = 7.2
Hz, 2H), 3.74 (t, J
= 4.4 Hz, 4H), 3.87 (t, J= 4.4 Hz, 4H), 4.03 (q, J= 7.2 Hz, 2H), 4.64 (s, 2H),
7.30 (s, 1H).
Step 15b. Ethyl 4-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(tert-butoxycarbonyl)amino)butanoate (Compound 0405-41)
A mixture of compound 0404-41 (386 mg, 0.78 mmol), 0107-3 (378 mg, 1.55
mmol), sodium hydrogen carbonate (196 mg, 2.33 mmol) and
bis(triphenylphosphine)palladium(a) chloride (27 mg, 0.05 mmol) in toluene (8
mL),
ethanol (5 mL) and water (2 mL) was flushed with nitrogen and heated under
microwave
irradiation at 120 C for 1 h. The reaction mixture was partitioned between
dichloromethane and water, organic layer was washed with brine, dried over
sodium
sulfate, filtered and evaporated in vacuum. The resulting residue was purified
using column
chromatography eluting methanol in dichloromethane (2-5%, v/v), to give title
compound
0405-41 (396 mg, 79%) as a white solid. LCMS: 581 [M+1]+; 1H NMR (400 MHz,
DMSO-d6): 651.16 (t, J= 7.2 Hz, 3H), 1.44 (s, 9H), 1.78 (m, 2H), 2.28 (t, J=
7.2 Hz, 2H),
3.28 (t, J = 6.8 Hz, 2H), 3.83 (m, 4H), 4.03 (m, 6H), 4.70 (s, 2H), 7.47 (t, J
= 8.0 Hz, 1 H),
7.51 (s, I H), 7.67 (d, J= 8.0 Hz, I H), 8.22 (d, J= 8.0 Hz, I H), 8.88 (s, I
H), 13.22 (s, I H).
Step 15c: 4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)-N-hydroxybutanamide (Compound 41)
To a stirred solution of hydroxylamine hydrochloride (4.67 g, 67 mmol) in
methanol
(24 mL) at 0 C was added a solution of potassium hydroxide (5.64 g, 100 mmol)
in
methanol (14 mL). After addition, the mixture was stirred for 30 minutes at 0
C, and was
allowed to stand at low temperature. The resulting precipitate was isolated,
and the solution
was prepared to give free Hydroxylamine.
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The freshly prepared hydroxylamine solution (6.00 mL) was placed in 50 mL
flask.
Compound 0405-41 (300 mg, 0.51 mmol) was added to this solution and degassed
at room
temperature for 30 minutes. The reaction process was monitored by TLC. The
mixture was
neutralized with dry ice, filtered, and the precipitation was washed with
methanol and water
to give tert-butyl (2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl(4-
(hydroxyamino)-4-oxobutyl)carbamate (267 mg, 91%) as a white solid. LCMS: 568
[M+1]; 'H NMR (400 MHz, DMSO-d6): (51.45 (s, 9H), 1.74 (m, 2H), 1.96 (t, J=
7.2 Hz,
2H), 3.23 (m, 2H), 3.83 (m, 4H), 4.00 (m, 4H), 7.47 (t, J= 8.0 Hz, 1H), 7.50
(s, 1H), 7.67
(d, J = 8.0 Hz, 1 H), 8.22 (d, J = 8.0 Hz, 1 H), 8.70 (s, 1 H), 8.88 (s, 1 H),
10.39 (s, 1 H), 13.22
(s, 1H).
The above prepared compound was then added to a freshly prepared isopropanol
hydrogen chloride solution (7.00 mL). The reaction mixture was stirred at room
temperature for 2 h. The mixture was evaporated and dissolved in water. Then
the mixture
was neutralized with ammonia at 0 C, filtered and the precipitation was
washed with
methanol and water to give the crude product which was purified with pre-HPLC.
Compound 41 was got (50 mg, 24%) as an orange solid: m.p. 149-152 C. LCMS:
468
[M+1]; 'H NMR (400 MHz, DMSO-d6): 651.55 (m, 2H), 1.87 (t, J= 7.2 Hz, 2H),
2.45 (t,
J= 7.2 Hz, 2H), 3.68 (m, 4H), 3.86 (m, 6H), 3.96 (s, 2H), 7.32 (m, 2H), 7.51
(d, J= 8.0 Hz,
I H), 7.98 (s, I H), 8.07 (d, J= 8.0 Hz, I H), 8.56 (s, I H), 8.73 (s, I H),
13.06 (s, I H).
Example 16: Preparation of 5-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methylamino)-N-hydroxypentanamide (Compound 42)
Step 16a: Methyl 5-(tert-butoxycarbonyl((2-chloro-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)amino)pentanoate (Compound 0404-42)
The title compound 0404-42 (0.75 g, 33.3%) was prepared as a white solid
without
further purification from 0112 (1.2 g, 4.24 mmol), 5-aminopentanoate
hydrochloride (1.416
g, 8.48 mmol) using a procedure similar to that described for compound 0404-41
(Example
15): LCMS: 399 [M+l]+; 1H NMR (400 MHz, DMSO-d6) 6 1.39-1.47 (m, 2H), 1.52-
1.60
(m, 2H), 2.30 (t, J= 7.2 Hz, 2H), 2.53 (t, J= 6.8 Hz, 2H), 3.58 (s, 3H), 3.74
(t, J= 4.8 Hz,
4H), 3.88 (t, J= 4.8 Hz, 4H), 3.99 (s, 2H), 7.23 (s, 1H). Compound 0404-42:
LCMS: 499
[M+1]+; 1H NMR (400 MHz, DMSO-d6) 61.23 (s, 9H),1.47-1.49 (m, 4H), 2.30 (t, J=
6.8
Hz, 2H), 3.20 (s, 2H), 3.56 (s, 3H), 3.74 (t, J= 4.8 Hz, 4H), 3.88 (t, J= 4.8
Hz, 4H), 4.64
(s, 2H), 7.31 (s, 1H).
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Step 16b: Methyl 5-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(tert-butoxycarbonyl)amino)pentanoate(Compound 0405-42)
The title compound 0405-42 (260 mg, 55.8%) was prepared as a yellow solid from
0404-42 (400 mg, 0.803 mmol), 0107-3 (216 mg, 0.884 mmol), sodium hydrogen
carbonate
(202.4 mg, 2.41 mmol) and bis(triphenylphosphine)palladium(a) chloride (30 mg,
0.0402
mmol) in toluene (8 mL), ethanol (5 mL) and water (2 mL) using a procedure
similar to that
described for compound 0404-42 (Example 15): LCMS: 581 [M+1]+; 1H NMR (400
MHz,
DMSO-d6): 6 1.44 (s, 9H), 1.50-1.52 (m, 4H), 3.25 (s, 2H), 3.56 (s, 3H), 3.83
(d, J= 5.2
Hz, 4H), 4.00-4.10 (m, 6H), 4.69 (s, 2H), 7.47-7.51 (m, 2H), 7.67 (d, J= 8.4
Hz , 1H), 8.23
(d, J = 6.8 Hz, 1 H), 8.88 (s, 1 H), 13.20 (s, 1 H).
Step 16c: 5-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)-N-hydroxypentanamide(Compound 42)
The title compound 42 was prepared as a white solid (23 mg, 13.9%) from 0405-
42
(260 mg, 0.45 mmol) and freshly prepared hydroxylamine methanol solution (8.0
mL)
followed by deprotection using a procedure similar to that described for
compound 41
(Example 15): m.p 145-147 C. LCMS: 482 [M+1]+; 1H NMR (400 MHz, DMSO-d6):
61.54-1.63 (m, 4H), 1.99 (t, J=7.2 Hz, 2H), 3.02 (s, 2H), 3.87 (m, 4H), 4.05
(m, 4H), 4.58
(t, J =4.4 Hz, 2H), 7.49 (t, J =8.4 Hz, 1 H), 7.70 (d, J =8 Hz, 1 H), 7.78 (s,
1 H), 8.24 (d, J
=10.4 Hz, I H), 8.87 (s, I H), 9.11 (s, 2H),10.42 (s, I H), 13.28 (s, 1 H).
Example 17: Preparation of 6-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methylamino)-N-hydroxyhexanamide (Compound 43)
Step 17a: Ethyl 6-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)hexanoate (Compound 0404-43)
The title compound 0404-43 (343 mg, 38%) was prepared as a yellow solid from
0112 (0.6 g, 2.12 mmol) and ethyl 6-aminohexanoate hydrochloride (0.83 g, 4.24
mmol)
using a procedure similar to that described for compound 0404-41 (Example 15):
LCMS:
427 [M+1]+; 1H NMR (400 MHz, CDC13) 651.17 (t, J= 6.4 Hz, 3H), 1.24-1.33 (m,
2H),
1.40-1.45 (m, 2H), 1.48-1.55 (m, 2H), 2.09 (s, 1H), 2.27 (t, J= 7.2 Hz, 2H),
2.53 (t, J= 6.8
Hz 2H), 3.74 (t, J= 5.2 Hz, 4H), 3.88 (t, J= 4.8 Hz, 4H), 3.99(s, 2H), 4.03
(q, J= 6.4 Hz,
2H), 7.23 (s, I H).
Step 17b: Ethyl6-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)hexanoate (Compound 0405-43)
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A mixture of compound 0404-43 (343 mg, 0.80 mmol), 0107-3 (294 mg, 1.2
mmol), sodium hydrogen carbonate (294 mg, 2.4 mmol) and
bis(triphenylphosphine)palladium(a) chloride (29 mg, 0.05 mmol) in toluene (8
mL),
ethanol (5 mL) and water (2 mL) was flushed with nitrogen and heated under
microwave
irradiation at 120 C for 1 h. The reaction mixture was partitioned between
dichloromethane
and water, organic layer was washed with brine, dried over magnesium sulfate,
filtered and
evaporated in vacuum. The resulting residue was purified using column
chromatography
eluting methanol in dichloromethane (2-5%, v/v), to give title compound 0405-
43 (120 mg,
29%) as a yellow solid. LCMS: 509 [M+1]+; 1H NMR (400 MHz, CDC13): 6 1.17(t,
J= 7.2
Hz, 3H), 1.28-1.35(m, 2H), 1.42-1.57 (m, 4H), 2.28 (t, J= 7.2 Hz, 2H), 2.57
(t, J= 6.8 Hz,
2H), 3.83 (t, , J = 4.0 Hz, 4H), 3.97-4.06 (m, 6H), 7.45 (s, 1 H), 7.47 (d, J
= 7.6 Hz , 1 H),
7.66 (d, J = 8.0 Hz , 1 H), 8.22 (d, J = 7.2 Hz, 1 H), 8.8 8 (s, 1 H), 13.21
(s, 1 H).
Step 17c: 6-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)-N-hydroxyhexanamide (43)
The title compound 43 was prepared (17 mg, 15%) as a yellow solid from 0405-43
(120 mg, 0.24 mmol) and freshly prepared hydroxylamine methanol solution (4.0
mL)
using a procedure similar to that described for compound 3 (Example 1): m.p.
128-130 C.
LCMS: 496 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 5 1.23-1.32 (m, 2H), 1.42-1.53
(m,
4H), 1.95 (t, J = 6.8 Hz, 2H), 2.57(t, J = 7.2 Hz, 2H), 3.84 (t, J = 4.0 Hz,
4H), 4.01 (t, J =
4.0 Hz, 4H), 4.06 (s, 2H), 7.45 (s, 1H), 7.48 (d, J= 7.6 Hz, 1H), 7.66 (d, J=
8 Hz, 1H),
8.22 (d, J= 7.2 Hz, I H), 8.66 (s, I H), 8.88 (s, I H), 10.34 (s, I H), 13.21
(s, 1 H).
Example 18: Preparation of 7-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methylamino)-N-hydroxyheptanamide (Compound 44)
Step 18a: Ethyl 7-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)heptanoate (Compound 0404-44)
The title compound 0404-44 (700 mg, 45%) was prepared as a yellow solid from
0112 (1 g, 3.5 mmol) and 7-aminoheptanoate hydrochloride (1.5 g, 7.0 mmol)
using a
procedure similar to that described for compound 0404-41 (Example 15): LCMS:
442
[M+1]+; 1H NMR (400 MHz, CDC13) 651.16 (t, J= 7.2 Hz, 3H), 1.21-1.27 (m, 4H),
1.45 (m,
2H), 1.50 (m, 2H), 2.21-2.28 (m, 2H), 2.52 (t, J= 7.2 Hz, 2H), 2.66 (s, 1H),
3.74 (t, J= 5.2
Hz, 4H), 3.88 (t, J= 4.8 Hz, 4H), 4.00-4.06 (m, 4H), 7.23 (s, 1H).
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Step 18b: Ethyl7-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)heptanoate (Compound 0405-44)
The title compound 0405-44 (260 mg, 63%) was prepared as a yellow solid from
0404-44 (350 mg, 0.79 mmol) and 0107-3 (290 mg, 1.19 mmol) using a procedure
similar
to that described for compound 0405-43 (Example 17): LCMS: 523 [M+1]+; 'H NMR
(400
MHz, CDC13): 6 1.16(t, J= 6 Hz, 3H), 1.26-1.31(m, 4H), 1.44 (m, 2H), 1.52 (m,
2H), 2.21-
2.28 (m, 2H), 2.56 (t, , J= 7.2 Hz, 2H), 3.83 (t, , J= 4.8 Hz, 4H), 4.01 (t,
J= 4.8 Hz, 4H),
4.04 (s, 2H), 7.46 (m, 2H), 7.66 (d, J = 8.4Hz , 1 H), 8.22 (d, J = 7.2Hz, 1
H), 8.8 8 (s, 1 H),
13.20 (s, 1H).
Step 18c: 7-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)-N-hydroxyheptanamide (Compound 44)
The title compound 44 was prepared (37 mg, 24%) as a yellow solid from 0405-44
(160 mg, 0.31 mmol) and freshly prepared hydroxylamine methanol solution (3.0
mL)
using a procedure similar to that described for compound 3 (Example 1): m.p.
188-190 C.
LCMS: 510 [M+1]+;'H NMR (400 MHz, DMSO-d6): 6 1.19-1.33 (m, 4H), 1.41-1.52 (m,
4H), 1.93 (t, J = 7.2 Hz, 2H), 2.56(t, J = 6.8 Hz, 2H), 3.83 (t, J = 4.4 Hz,
4H), 4.01 (t, J =
4.4 Hz, 4H), 4.05 (s, 2H), 7.44 (s, 1H), 7.48 (d, J= 7.6 Hz, 1H), 7.66 (d, J=
8 Hz, 1H),
8.22 (d, J= 6.8 Hz, 2H), 8.68 (s, 1H), 8.88 (s, 1H), 10.34 (s, 1H), 13.21 (s,
1 H).
Example 19: Preparation of 4-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxybutanamide (Compound 101)
Step 19a: Ethyl 4-(((2-(1 H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)butanoate (Compound 0404-101)
To a solution of ethyl 4-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)butanoate
(100 mg, 0.25 mmol) in methanol (5 mL) was added poly-formaldehyde (15 mg,
0.50
mmol). After stirred for 30 min at room temperature, NaBH3CN (32 mg, 0.50
mmol) was
added slowly, and the mixture was stirred for another 30 min. The reaction was
terminated
by adding water (5 mL) at 0 C and stirred. The resulting precipitate was
filtered and
washed with water to give 0404-101 (85 mg, 83%) as a yellow solid. LCMS: 413
[M+1]+;
iH NMR (400 MHz, DMSO-d6) 6 1.14 (t, J= 7.2 Hz, 3H), 1.72 (m, 2H), 2.22 (s,
3H), 2.32
(t, J = 7.2 Hz, 2H), 2.41 (t, J = 7.2 Hz, 2H), 3.75 (t, J = 4.4 Hz, 4H), 3.82
(s, 2H), 3.88 (t, J
= 4.4 Hz, 4H), 4.02 (q, J= 7.2 Hz, 2H), 7.27 (s, 1H).
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Step 19b: Ethyl 4-(((2-(1 H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)butanoate (Compound 111-48-3)
The title compound 0405-101 (200 mg, 56%) was prepared as a yellow solid from
0404-101 (300 mg, 0.73 mmol) and 0107-3 (356 mg, 1.46 mmol) using a procedure
similar
to that described for compound 0405-43 (Example 17): LCMS: 495 [M+1]+; 'H NMR
(400
MHz, CDC13): 61.15 (t, J= 7.2 Hz, 3H), 1.75 (m, 2H), 2.25 (s, 3H), 2.35 (t, J=
7.2 Hz,
2H), 2.44 (t, J= 7.2 Hz, 2H), 3.83 (m, 6H), 4.02 (m, 6H), 7.47 (m, 2H), 7.67
(d, J= 8.0 Hz,
I H), 8.22 (d, J= 8.0 Hz, I H), 8.89 (s, I H), 13.21 (s, I H).
Step 19c: 4-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxybutanamide (Compound 101)
The title compound 101 was prepared (160 mg, 88%) as a yellow solid from 0405-
101 (187 mg, 0.38 mmol) and freshly prepared hydroxylamine methanol solution
(6.0 mL)
using a procedure similar to that described for compound 3 (Example 1): m.p.
115-118 C.
LCMS: 482 [M+1]; 'H NMR (400 MHz, DMSO-d6): 5 1.72 (m, 2H), 2.02 (t, J= 6.0
Hz,
2H), 2.25 (s, 3H), 2.42 (t, J= 6.0 Hz, 2H), 3.85 (m, 6H), 4.01 (m, 4H), 7.47
(m, 2H), 7.67
(d, J = 7.6 Hz, 1 H), 8.22 (d, J = 6.8 Hz, 1 H), 8.89 (s, 1 H), 13.26 (s, 1
H).
Example 20: Preparation of 5-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypentanamide (Compound 102)
Step 20a: Methyl 5-(((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pentanoate (Compound 0404-102)
The title compound 0404-102 (0.62 g, 86%) was prepared as a white solid from
Methyl 5-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methylamino)
pentanoate
(700 mg, 1.76 mmol), paraformaldehyde (106 mg, 3.52 mmol) in methanol (30 mL)
and
NaBH3CN (221 mg, 3.52 mmol) using a procedure similar to that described for
compound
0404-101 (Example 19): LCMS: 413 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 1.45-1.59
(m, 4H), 2.21 (s, 3H), 2.32 (t, J= 6.8 Hz, 2H), 2.40 (t, J= 7.2 Hz, 2H), 3.58
(s, 3H), 3.75 (t,
J = 5.2 Hz, 4H), 3.81 (s, 2H), 3.88 (t, J = 4.8 Hz, 4H), 7.26 (s, 1 H).
Step 20b: Methyl 5-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pentanoate (Compound 0405-102)
The title compound 0405-102 (305 mg, 61.7%) was prepared as a yellow solid
from
0404-102 (350 mg, 0.85 mmol) and 0107-3 (311 mg, 1.27 mmol) using a procedure
similar
to that described for compound 0405-43 (Example 17): LCMS: 495 [M+1]+; 1H NMR
(400
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MHz, DMSO-d6): 6 1.48-1.57 (m, 4H), 2.24 (s, 3H), 2.34 (t, J= 6.8 Hz, 2H),
2.44 (t, J=
6.8 Hz, 2H), 3.58 (s, 3H),3.84 (d, J= 8.4 Hz, 6H), 3.97-4.04 (m, 4H), 7.49 (m,
2H), 7.67 (d,
J = 8.4 Hz , 1 H), 8.22 (d, J = 7.2 Hz, 1 H), 8.88 (s, 1 H), 13.20 (s, 1 H).
Step 20c: 5-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypentanamide (Compound 102)
The title compound 102 was prepared (60 mg, 25%) as a yellow solid from 0405-
102 (240 mg, 0.48 mmol) and freshly prepared hydroxylamine methanol solution
(8.0 mL)
using a procedure similar to that described for compound 3 (Example 1): m.p.
120-122 C.
LCMS: 496 [M+1]; 'H NMR (400 MHz, DMSO-d6) (51.48-1.54 (m, 4H), 1.98 (t, J=
6.8
Hz, 2H), 2.24 (s, 3H), 2.44 (t, J= 7.2 Hz, 2H), 3.85 (d, J= 6.8 Hz, 4H), 4.01
(s, 4H), 7.46
(s, 2H), 7.66 (d, J= 7.2 Hz, I H), 8.22 (d, J=7.6 Hz, I H), 8.88 (s, I H),
10.31 (s, I H), 13.18
(s, 1 H).
Example 21: Preparation of 6-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxyhexanamide (Compound 103)
Step 21a: Ethyl 6-(((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)hexanoate (Compound 0404-103)
The title compound 0404-103 (0.62 g, 86%) was prepared as a white solid from
0404-43 (0.67 g, 1.57 mmol), paraformaldehyde (94 mg, 3.14 mmol) and NaBH3CN
(197
mg, 3.14 mmol) using a procedure similar to that described for compound 0404-
101
(Example 19): LCMS: 441 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 6 1.16 (t, J= 7.2
Hz,
3H), 1.24-1.32 (m, 2H), 1.43-1.55 (m, 4H), 2.21 (s, 3H), 2.27 (t, J= 7.2 Hz,
2H), 2.39 (t, J
= 7.6 Hz, 2H), 3.75 (t, J= 4.8 Hz, 4H), 3.81 (s, 2H), 3.88 (t, J= 4.4 Hz, 4H),
4.03 (q, J
7.2 Hz, 2H), 7.26 (s, 1H).
Step 21b: Ethyl6-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)hexanoate (Compound 0405-103)
The title compound 0405-103 (190 mg, 54%) was prepared as a yellow solid from
0404-103 (300 mg, 0.68 mmol) and 0107-3 (199 mg, 0.82 mmol) using a procedure
similar
to that described for compound 0405-43 (Example 17): LCMS: 523 [M+1]+; 'H NMR
(400
MHz, DMSO-d6): 6 1.16(t, J= 7.2 Hz, 3H), 1.28-1.35(m, 2H), 1.47-1.57 (m, 4H),
2.27 (m,
5H), 2.41 (t, J= 6.4 Hz, 2H), 3.84 (m, 6H), 4.04 (m, 6H), 7.47 (m, 2H), 7.67
(d, J= 8 Hz,
1 H), 8.23 (d, J = 6.8 Hz , 1 H), 8.89 (s, 1 H), 13.21(s, 1 H).
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Step 21c: 6-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxyhexanamide (Compound 103)
The title compound 103 was prepared (75 mg, 41%) as a white solid from 0405-
103
(190 mg, 0.22 mmol) and freshly prepared hydroxylamine methanol solution (4.0
mL)
using a procedure similar to that described for compound 3 (Example 1): m.p.
115-118 C.
LCMS: 510 [M+1]; 'H NMR (400 MHz, DMSO-d6): 6 1.23-1.27 (m, 2H), 1.45-1.55 (m,
4H), 1.95 (t, J= 7.2 Hz, 2H), 2.24 (s, 3H), 2.42(t, J= 7.2 Hz, 2H), 3.84 (m,
6H), 4.00 (m,
4H), 7.47 (m, 2H), 7.66 (d, J = 8.4 Hz, 1 H), 8.22 (d, J = 7.6 Hz, 1 H), 8.68
(s, 1 H), 8.89 (s,
1H), 10.35 (s, 1H), 13.21 (s, 1 H).
Example 22: Preparation of 7-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxyheptanamide (Compound 104)
Step 22a: Ethyl 7-(((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)heptanoate (Compound 0404-104)
The title compound 0404-104 (670 mg, 95%) was prepared as a yellow solid from
0404-44 (0.67 g, 1.52 mmol), paraformaldehyde (91 mg, 3.04 mmol) and NaBH3CN
(191
mg, 3.04 mmol) using a procedure similar to that described for compound 0404-
101
(Example 19): LCMS: 455 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 61.16 (t, J = 7.2
Hz,
3H), 1.23-1.32 (m, 4H), 1.42-1.54 (m, 4H), 2.21 (s, 3H), 2.25 (t, J = 7.6 Hz,
2H), 2.39 (t, J
= 7.2 Hz, 2H), 3.74 (t, J = 4 Hz, 4H), 3.80 (s, 2H), 3.88 (t, J = 4.8 Hz, 4H),
4.03 (q, J = 7.2
Hz, 2H), 7.25 (s, 1H).
Step 22b: Ethyl7-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)heptanoate (Compound 0405-104)
The title compound 0405-104 (260 mg, 67%) was prepared as a yellow solid from
0404-104 (330 mg, 0.73 mmol) and 0107-3 (176 mg, 0.73 mmol) using a procedure
similar
to that described for compound 0405-43 (Example 17): LCMS: 537 [M+1]+; 'H NMR
(400
MHz, DMSO-d6): 6 1.14 (t, J= 7.2 Hz, 3H), 1.28-1.41 (m, 4H), 1.49-1.55 (m,
4H), 2.25
(m, 5H), 2.41 (t, J= 6.8 Hz, 2H), 3.83 (m, 6H), 4.01 (m, 6H), 7.46 (m, 2H),
7.65 (d, J= 7.6
Hz , 1 H), 8.21 (d, J = 6.8 Hz , 1 H), 8.87 (s, 1 H), 13.17 (s, 1 H).
Step 22c: 7-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxyheptanamide (Compound 104)
The title compound 104 was prepared (65 mg, 55%) as a white solid from 0405-
104
(120 mg, 0.22 mmol) and freshly prepared hydroxylamine methanol solution (4.0
mL)
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using a procedure similar to that described for compound 3 (Example 1): m.p.
131-133 C.
LCMS: 524 [M+1]; 'H NMR (400 MHz, DMSO-d6): (51.21-1.27 (m, 4H), 1.34-1.43 (m,
4H), 1.94 (t, J= 7.6 Hz, 2H), 2.24 (s, 3H), 2.43(t, J= 7.6 Hz, 2H), 3.84 (m,
6H), 4.00 (m,
4H), 7.47 (m, 2H), 7.67 (d, J = 8 Hz, 1 H), 8.22 (d, J = 7.2 Hz, 1 H), 8.68
(s, 1 H), 8.89 (s,
1H), 10.34 (s, 1H), 13.21 (s, 1 H).
Example 23: Preparation of 2-(((2-(6-fluoro-lH-indazol-4-yl)-4-
morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 69)
Step 23a: 3-Bromo-5-fluoro-2-methylbenzenamine (Compound 0104-69)
To a solution of 4-fluoro-2-nitrotoluene (10.0 g, 64.4 mmol) in
trifluoroacetic acid
(40 mL) was added con. sulfuric acid (12.5 mL) followed by NBS (17.2 g, 96.6
mmol) and
the reaction mixture was stirred at room temperature for 16 h. Then the
reaction mixture
was poured into ice and water and stirred for 15 min. Extracted with ethyl
acetate and the
organic layer was washed with brine, dried, concentrated to get compound 1-
bromo-5-
fluoro-2-methyl-3-nitrobenzene (15.0 g, 100%) as a yellow oil. 1H NMR (400
MHz,
DMSO-d6) 6 2.41 (s, 3H), 7.96 (dd, J= 8.0, 2.4 Hz, 1H), 8.02 (dd, J= 8.0, 2.4
Hz, 1H). A
mixture of above prepared compound (15.0 g, 64.4 mmol), Fe (18.0 g, 0.32 mol),
con. HC1
(2 mL) in MeOH (150 mL) and water (30 mL) was stirred at reflux for 4 h. Then
the
mixture was adjusted to pH 12 with aqueous NaOH solution and filtered. The
solvent was
removed and diluted with water. Extracted with ethyl acetate, dried,
concentrated. The
residue was purified by column chromatograph (ethyl acetate in petroleum
ether, 7%) to get
compound 0104-69 (5.8 g, 44%) as yellow oil. LCMS: 204 [M+1]+, 1H NMR (400
MHz,
DMSO-d6) 6 2.12 (s, 3H), 5.57 (s, 2H), 6.45 (dd, J= 11.2, 2.4 Hz, 1H), 6.60
(dd, J= 8.0,
2.4 Hz, 1H).
Step 23b: 4-Bromo-6-fluoro-lH-indazole (Compound 0106-69)
The title compound 0 106-69 (3.7 g, 61 %) was prepared as a yellow solid from
0104-69 (5.8 g, 28.4 mmol), potassium acetate (2.93 g, 29.8 mmol), Ac20 (5.8
g, 56.8
mmol) and iso-amyl nitrite (7.32 g, 62.5 mmol) followed by hydrolysis by
aqueous
hydrochloric acid (6N, 35 mL) using a procedure similar to that described for
compound
0106-3 (Example 1): LCMS: 215 [M+1]+, 1H NMR (400 MHz, DMSO-d6) 6 7.37-7.44
(m,
2H), 8.07 (s, I H), 13.54 (s, I H).
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Step 23c: 6-Fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole
(Compound 107-69)
The title compound 0107-69 (700 mg, 57%) was prepared as a yellow solid from
0106-69 (1.0 g, 4.65 mmol), bis(pinacolato)diboron (1.77 g, 6.98 mmol),
PdC12(dppf)2
(380 mg, 0.47 mmol) and dried potassium acetate (1.37 g, 14.0 mmol) in dioxane
(40 mL)
using a procedure similar to that described for compound 0 107-3 (Example 1):
LCMS: 263
[M+1 ]+.
Step 23d: Ethyl 2-(((2-(6-fluoro-1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0505-69)
A mixture of compound 0504-54 (200 mg, 0.45 mmol), 0107-69 (135 mg, 0.5
mmol), sodium hydrogen carbonate (115 mg, 1.3 mmol) and
bis(triphenylphosphine)palladium(II) chloride (15 mg, 0.02 mmol) in toluene (8
mL),
ethanol (5mL) and water (2 mL) was flushed with nitrogen and heated under
microwave
irradiation at 120 C for 5 h. The reaction mixture was partitioned between
ethyl acetate
and water, organic layer was washed with brine, dried over sodium sulfate,
filtered,
concentrated and washed with ethyl acetate to obtain the title compound 0505-
69 (100 mg,
41%) as a yellow solid. LCMS: 549 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 5 1.30
(t, J=
7.2 Hz, 3H), 3.29 (s, 3H), 3.79 (t, J= 4.4 Hz, 4H), 3.96 (t, J= 4.4 Hz, 4H),
4.29 (q, J= 7.2
Hz, 2H), 5.27 (s, 2H), 7.46 (d, J = 7.2 Hz, 1 H), 7.62 (s, 1 H), 7.98 (dd, J =
10.4 Hz, 2.4 Hz,
1H), 8.94 (m, 3H), 13.27 (s, 1H).
Step 23e: 2-(((2-(6-Fluoro-1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-
6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 69)
The title compound 69 was prepared (24 mg, 25%) as a yellow solid from 0505-69
(100 mg, 0.18 mmol) and freshly prepared hydroxylamine methanol solution (10.0
mL)
using a procedure similar to that described for compound 3 (Example 1): m.p.
215-217 C.
LCMS: 536 [M+1]+; 'H NMR (400 MHz, DMSO-d6) 5 3.26 (s, 3H), 3.79 (m, 4H), 3.96
(m, 4H), 5.24 (s, 2H), 7.46 (d, J= 7.6 Hz, 1H), 7.59 (s, 1H), 7.98 (dd, J=
10.8, 2.0 Hz, 1H),
8.76 (s, 2H), 8.89 (s, 1 H), 9.08 (s, 1 H), 11.14 (s, 1 H), 13.28 (s, 1 H).
Example 24: Preparation of 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(neopentyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 83)
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Step 24a: N-((2-Chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)-2,2-
dimethylpropan-l-amine (Compound 0503-83)
The solution of compound 0503-53 (600 mg, 2.1 mmol), pivalaldehyde (912 mg,
10.6 mmol) and Ti(OEt)4 (958 mg, 4.2 mmol) in CHC13/MeOH (8 mL/4 mL) was
stirred at
35 C for 20 hr. Then NaBH3CN (530 mg, 8.4 mmol) was added and stirred for 3
hr at 45
C. This mixture was diluted with H20, extracted with CH2C12, dried by Na2SO4,
and
concentrated to obtain 0503-83 (631 mg, 85%) as a yellow solid. LCMS: 355
[M+1]+. 'H-
NMR (400 MHz, DMSO-d6) 6 0.95 (s, 9H), 2.29 (s, 2H), 3.74 (m, 4H), 3.88 (m,
4H), 4.02
(s, 2H), 7.23 (s, 1H).
Step 24b: Ethyl 2-(((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(neopentyl) amino)pyrimidine-5-carboxylate (Compound 0504-83)
A mixture of compound 0503-83 (400 mg, 1.13 mmol), 0305 (846 mg, 4.52 mmol)
and DIPEA (1.5 g, 11.3 mmol) in MeCN (8 mL) was stirred at 70 C for 24 hr,
concentrated, purified by column chromatograph (ethyl acetate in petroleum
ether, 10%
v/v) to provide compound 0504-83 (530 mg, 93%) as a yellow solid. LCMS: 505
[M+1]+,
iH NMR (400 MHz, DMSO-d6) 6 0.97 (s, 9H), 1.29 (m, 3H), 3.67 (s, 2H), 3.72 (m,
4H),
3.87 (m, 4H), 4.27 (m, 2H), 5.19 (s, 2H), 7.35 (s, 1H), 8.84 (s, 2H).
Step 24c: Ethyl 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)
(neopentyl)amino)pyrimidine-5-carboxylate (Compound 0505-83)
A mixture of compound 0504-83 (300 mg, 0.6 mmol), 0107-3 (176 mg, 0.72
mmol), NaHCO3 (152 mg, 1.8 mmol) and bis(triphenylphosphine)palladium(II)
chloride
(22 mg, 0.03 mmol) in toluene (4 mL), ethanol (2 mL) and water (1 mL) was
flushed with
nitrogen and heated under microwave irradiation at 120 C for 1 h. The
reaction mixture
was partitioned between dichloromethane and water, organic layer was washed
with brine,
dried over Na2SO4, filtered and evaporated in vacuum. The resulting residue
was purified
using column chromatography (methanol in dichloromethane, 2-5% v/v) to give
title
compound 0505-83 (300 mg, 85%) as a white solid. LCMS: 587 [M+1]+; 1H NMR (400
MHz, DMSO-d6) 6 0.99 (s, 9H), 1.29 (t, J= 7.2 Hz, 3H), 3.70 (s, 2H), 3.79 (m,
4H), 3.95
(m, 4H), 4.27 (q, J= 7.6 Hz, 2H), 5.24 (s, 2H), 7.46 (t, J= 7.6 Hz, 1H), 7.55
(s, 1H), 7.67
(d, J = 8.4 Hz, 1 H), 8.21 (d, J = 7.2 Hz, 1 H), 8.85 (s, 2H), 8.89 (s, 1 H),
13.21 (s, 1 H).
Step 24d: 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(neopentyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 83)
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The title compound 83 was prepared (191 mg, 65%) as a yellow solid from 0505-
83
(300 mg, 0.51 mmol) and freshly prepared hydroxylamine methanol solution (20.0
mL)
using a procedure similar to that described for compound 3 (Example 1): m.p.
240-242 C.
LCMS: 574 [M+1]; 'H NMR (400 MHz, DMSO-d6) 5 0.99 (s, 9H), 3.67 (s, 2H), 3.80
(m, 4H), 3.95 (m, 4H), 5.22 (s, 2H), 7.46 (t, J= 8.0 Hz, I H), 7.54 (s, I H),
7.67 (d, J= 8.0
Hz, 1 H), 8.21 (d, J = 7.2 Hz, 1 H), 8.73 (s, 2H), 8.89 (s, 1 H), 9.07 (s, 1
H), 11.12 (s, 1 H),
13.20 (s, 1H).
Example 25: Preparation of 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(propyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 84)
Step 25a: N-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)propan-l-
amine
(Compound 0503-84)
Compound 0502 (500 mg, 1.43 mmol) was dissolved in methanol (30 mL) and then
propan-l-amine (5 mL) was added. The mixture was stirred at 65 C and the
reaction
process was monitored by TLC. Then the solvent was removed at reduce pressure
and the
precipitation was partitioned between dichloromethane and water, organic layer
was
washed with brine and dried over sodium sulfate anhydrous, filtered,
evaporated in vacuum
and then purified by column chromatography (methanol in dichloromethane, 1.7%
v/v) to
give title compound 0503-84 (412 mg, 88%) as a light yellow solid. LCMS: 327
[M+1]+;
iH NMR (400 MHz, DMSO-d6) 6 0.86 (t, J= 7.4 Hz, 3H), 1.39-1.48 (m, 2H), 2.49
(t, J
2.0 Hz, 2H), 2.82 (s, 1H), 3.73 (t, J= 4.8 Hz, 4H), 3.87 (t, J= 4.8 Hz, 4H),
4.00 (s, 2H),
7.23 (s, 1H).
Step 25b: Ethyl 2-(((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(propyl)amino)pyrimidine-5-carboxylate (Compound 0504-84)
The title compound 0504-84 was prepared (477 mg, 79%) as a yellow solid from
0503-84 (412 mg, 1.26 mmol) and 0305 (353 mg, 1.89 mmol) in acetonitrile (30
ML), and
N, N-Diisopropylethylamine(3 mL) using a procedure similar to that described
for
compound 0504-83 (Example 24): LCMS: 477 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6
0.86 (t, J= 7.6 Hz, 3H), 1.30 (t, J= 7.2 Hz, 3H), 1.57-1.66 (m, 2H), 3.65 (t,
J= 7.6 Hz,
2H), 3.71 (t, J = 4.8 Hz, 4H), 3.83 (t, J = 4.6 Hz, 4H),4.28 (q, J = 7.2 Hz,
2H), 5.17 (s, 2H),
7.42 (s, 1H), 8.86 (s, 2H).
Step 25c: Ethyl 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(propyl)amino)pyrimidine-5-carboxylate (Compound 0505-84)
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The title compound 0505-84 was prepared (240 mg, 82%) as a white solid from
0504-84 (250 mg, 0.52 mmol), 0107-3 (154 mg, 0.63 mmol), sodium hydrogen
carbonate
(132 mg, 1.57 mmol) and bis(triphenylphosphine)palladium(II) chloride (18.5
mg, 0.026
mmol) in toluene (8 mL), ethanol (5 mL) and water (2 mL) using a procedure
similar to that
described for compound 0505-83 (Example 24): LCMS: 559 [M+1]+; 1H NMR (400
MHz,
DMSO-d6) 6 0.88 (t, J= 7.2 Hz, 3H), 1.30 (t, J= 7.0 Hz, 3H), 1.61-1.71 (m,
2H), 3.70 (t, J
= 7.4 Hz, 2H), 3.80 (t, J= 4.6 Hz, 4H), 3.96 (t, J = 4.6 Hz, 4H), 4.29 (q, J =
7.2 Hz, 2H),
5.23 (s, 2H), 7.47 (t, J = 7.8 Hz, 1 H), 7.61 (s, 1 H), 7.67 (d, J = 8.4 Hz, 1
H), 8.22 (d, J = 6.8
Hz, 1H), 8.88 (s, 3H), 13.20 (s, 1H).
Step 25d: 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(propyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 84)
The title compound 84 was prepared (189 mg, 81%) as a white solid from 0505-84
(240 mg, 0.43 mmol) and freshly prepared hydroxylamine methanol solution (16.0
mL)
using a procedure similar to that described for compound 3 (Example 1): m.p.
224-226 C.
LCMS: 546 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 0.88 (t, J= 7.4 Hz, 3H), 1.60-
1.69
(m, 2H), 3.67 (t, J = 7.6 Hz, 2H), 3.80 (t, J = 4.4 Hz, 4H), 3.96 (t, J = 4.8
Hz, 4H), 5.20 (s,
2H), 7.47 (t, J = 7.8 Hz, 1 H), 7.5 9 (s, 1 H), 7.67 (d, J = 8.0 Hz, 1 H),
8.22 (d, J = 6.8 Hz,
1 H), 8.75 (s, 2H), 8.88 (s, 1 H), 9.07 (s, 1 H), 11.12 (s, 1 H), 13.20 (s, 1
H).
Example 26: Preparation of 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(butyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 85)
Step 26a: N-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)butan-l-
amine
(Compound 0503-85)
The title compound 0503-85 was prepared (430 mg, 88%) as a light yellow solid
from 0502 (500mg, 1.43 mmol) and butan-l-amine (5mL) in methanol (30 mL) using
a
procedure similar to that described for compound 0503-84 (Example 25): LCMS:
341
[M+1]+; 1H NMR (400 MHz, DMSO-d6): 6 0.93 (t, J= 7.4 Hz, 3H), 1.33-1.42 (m,
2H),
1.45-1.52 (m, 2H), 2.61 (t, J= 7.0 Hz, 2H), 2.90 (s, 1H), 3.81-3.82 (m, 4H),
3.94-3.95 (d,
4H), 4.07 (s, 2H), 7.30 (s, 1H).
Step 26b: Ethyl 2-(butyl((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0504-84)
The title compound 0504-85 was prepared (519 mg, 84%) as a light yellow solid
from 0503-85 (430 mg, 1.26 mmol) and (353 mg, 1.89 mmol) in acetonitrile (30
mL) and
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N, N-Diisopropylethylamine(3 mL) using a procedure similar to that described
for
compound 0504-83 (Example 24): LCMS: 491 [M+1]+;'H NMR (400 MHz, DMSO-d6): 5
0.88 (t, J= 7.4 Hz, 3H), 1.26-1.34 (m, 5H), 1.55-1.62 (m, 2H), 3.67-3.72 (m,
6H), 3.83 (t, J
= 4.8 Hz, 4H), 4.28 (q, J= 7.1 Hz, 2H), 5.17 (s, 2H), 7.41 (s, 1H), 8.86 (s,
2H).
Step 26c: Ethyl2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(butyl)amino)pyrimidine-5-carboxylate (Compound 0505-85)
The title compound 0505-85 was prepared (255 mg, 87%) as a light yellow solid
from 0504-85 (250 mg, 0.51 mmol), 0107-3 (149 mg, 0.61 mmol), sodium hydrogen
carbonate (128 mg, 1.53 mmol) and bis(triphenylphosphine)palladium(a) chloride
(17.8
mg, 0.025 mmol) in toluene (8 mL), ethanol (5 mL) and water (2 mL)using a
procedure
similar to that described for compound 0505-83 (Example 24): LCMS: 573 [M+1]+;
1H
NMR (400 MHz, DMSO-d6): 6 1.04 (t, J= 7.4 Hz, 3H), 1.43-1.48 (m, 5H), 1.73-
1.81 (m,
2H), 3.88 (t, J = 7.6 Hz, 2H), 3.94 (t, J= 4.6 Hz, 4H), 4.10 (t, J = 4.6 Hz,
4H), 4.43 (q, J =
7.1 Hz, 2H), 5.37 (s, 2H), 7.61 (t, J= 7.8 Hz, I H), 7.75 (s, I H), 7.81 (d,
J= 8.0 Hz, I H),
8.36 (d, J= 6.8 Hz, 1H), 9.02 (s, 3H), 13.34 (s, 1H).
Step 26d: 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(butyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 85)
The title compound 85 was prepared (131 mg, 53%) as an off-white solid from
0505-85 (255 mg, 0.45 mmol) and freshly prepared hydroxylamine methanol
solution (16.0
mL) using a procedure similar to that described for compound 3 (Example 1):
m.p. 234-236
C. LCMS: 560 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 6 0.96 (t, J= 7.4 Hz, 3H),
1.32-
1.42 (m, 2H), 1.64-1.72 (m, 2H), 3.77 (t, J= 7.2 Hz, 2H), 3.86 (t, J= 4.2 Hz,
4H), 4.02 (t, J
= 4.4 Hz, 4H), 5.26 (s, 2H), 7.53 (t, J= 7.8 Hz, 1H), 7.66 (s, 1H), 7.73 (d,
J= 8.4 Hz, 1H),
8.28 (d, J = 7.2 Hz, 1 H), 8.81 (s, 2H), 8.94 (s, 1 H), 9.14 (s, 1 H), 11.18
(s, 1 H), 13.27 (s,
1H).
Example 27: Preparation of 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(2-hydroxyethyl)amino)-N-hydroxypyrimidine-5-
carboxamide (Compound 86)
Step 27a: 2-((2-Chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)ethanol
(Compound 0503-86)
The title compound 0503-86 was prepared (230 mg, 41%) as a light yellow solid
from 0502 (600 mg, 1.72 mmol) and 2-aminoethanol (6 mL) in methanol (60 mL)
using a
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procedure similar to that described for compound 0503-84 (Example 25): LCMS:
329
[M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 2.62 (t, J= 5.8 Hz, 2H), 2.74 (s, 1H),
3.47 (dd,
J 1= 11.2 Hz, J2= 6.0 Hz,2H), 3.73 (t, J= 4.8 Hz, 4H), 3.88 (t, J= 5.0 Hz,
4H), 4.04
(s,2H), 4.53 (t, J = 5.2 Hz, 1 H), 7.24 (s,1 H).
Step 27b: Ethyl 2-(((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(2-
hydroxyethyl)amino)pyrimidine-5-carboxylate (Compound 0504-86)
The title compound 0504-86 was prepared (170 mg, 51%) as a white solid from
0503-86 (230 mg, 0.7 mmol) and 0305 (157 mg, 0.84 mmol).in acetonitrile (20
mL) and N,
N-Diisopropylethylamine (4 mL) using a procedure similar to that described for
compound
0504-83 (Example 24): LCMS: 479 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 6 1.29 (t,
J=
7.2 Hz, 3H), 3.62 (dd, J 1= 11.2 Hz, J 2= 6.0 Hz,2H), 3.70 (t, J= 4.6 Hz, 4H),
3.76 (t, J=
6.0 Hz, 2H), 3.82 (t, J= 4.6 Hz, 4H), 4.27 (dd, J 1= 13.6 Hz, J2= 6.8 Hz,2H),
4.86 (t, J=
5.2 Hz, 1H), 5.23 (s, 2H), 7.39 (s, 1H), 8.85 (s, 2H).
Step 27c: Ethyl 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(2-hydroxyethyl)amino)pyrimidine-5-carboxylate (0505-86)
The title compound 0505-86 was prepared (120 mg, 60%) as a white solid from
0504-86 (170 mg, 0.35 mmol), 0107-3 (104 mg, 0.43 mmol), sodium hydrogen
carbonate
(89 mg, 1.06 mmol) and bis(triphenylphosphine)palladium(a) chloride (13 mg,
0.02 mmol)
in toluene (4 mL), ethanol (2.5 mL) and water (1 mL) using a procedure similar
to that
described for compound 0505-83 (Example 24): LCMS: 561 [M+1]+; 1H NMR (400
MHz,
DMSO-d6): 6 1.29 (t, J= 6.8 Hz, 3H), 3.66 (dd, J 1= 10.8 Hz, Jz= 5.6 Hz,2H),
3.78-3.83
(m, 6H), 3.95 (t, J= 4.6 Hz, 4H), 4.28 (dd, J i = 14.4 Hz, J2= 7.2 Hz,2H),
4.88 (t, J= 5.4
Hz, 1 H), 5.29 (s, 2H), 7.46 (t, J = 7.8 Hz, 1 H), 7.5 9 (s, 1 H), 7.66 (d, J
= 8.4 Hz, 1 H), 8.21
(d, J= 7.2 Hz, 1H), 8.87 (s, 3H), 13.2 (s, 1H).
Step 27d: 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(2-
hydroxyethyl)amino)-N-hydroxypyrimidine-5-carboxamide (86)
The title compound 86 was prepared (42 mg, 36%) as an off-white solid from
0505-
86 (120 mg, 0.21 mmol) and freshly prepared hydroxylamine methanol solution
(8.0 mL)
using a procedure similar to that described for compound 3 (Example 1): m.p.
190-194 C.
LCMS: 548 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 6 3.64 (dd, J 1= 10.8 Hz, J2= 5.6
Hz
,2H), 3.79 (dd, J 1= 8.4 Hz, J 2= 4.4 Hz ,6H), 3.95 (t, J = 4.4 Hz, 4H), 4.85
(t, J = 5.2 Hz,
1 H), 5.25 (s, 2H), 7.46 (t, J = 7.8 Hz, 1 H), 7.5 7 (s, 1 H), 7.66 (d, J =
8.4 Hz, 1 H), 8.21 (d, J
= 6.8 Hz, 1 H), 8.74 (s, 2H), 8.87 (s, 1 H), 9.07 (s, 1 H), 11.13 (s, 1 H),
13.20 (s, 1 H).
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Example 28: 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(2-methoxyethyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound
90)
Step 28a: N-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)-2-
methoxyethanamine (Compound 0503-90)
The title compound 0503-90 was prepared (410 mg, 80%) as oil from 0502 (520
mg, 1.5 mmol) and 2-methoxyethanamine (1.1 g. 10.0 mmol) in methanol (20 mL)
using a
procedure similar to that described for compound 0503-84 (Example 25): LCMS:
343
[M+1]+; 'H NMR (400 MHz, CDC13) 6 2.858 (t, J= 7.2 Hz, 3H), 3.37 (s, 3H), 3.53
(t, J=
5.2 Hz, 2H), 3.53 (t, J= 5.2 Hz, 2H), 3.83 (t, J= 5.2 Hz, 4H), 3.99 (t, J= 4.8
Hz, 4H), 4.12
(s, 2H), 7.16 (s, 1H).
Step 28b: Ethyl 2-(((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(2-
methoxyethyl)amino)pyrimidine-5-carboxylate (Compound 0504-90)
The title compound 0504-90 was prepared (400 mg, 81%) as a yellow solid from
0503-90 (342 mg, 1.0 mmol) and 0305 (205 mg, 1.1 mmol) in acetonitrile (20 mL)
and N,
N-Diisopropylethylamine (400mg, 3.3 mmol) using a procedure similar to that
described
for compound 0504-83 (Example 24): LCMS: 493 [M+1]+; 'H NMR (400 MHz, DMSO-
d6) 6 1.29 (t, J= 6.8 Hz, 3H), 3.22 (s, 3H), 3.56 (t, J= 5.2 Hz, 2H), 3.70
(brs, 4H), 3.82
(brs, 4H), 3.88 (t, J= 5.2 Hz, 2H), 4.27 (q, J= 6.8 Hz, 2H), 5.19 (s, 2H),
7.39 (s, 1H), 8.86
(s, 1H).
Step 28c: Ethyl 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(2-methoxyethyl)amino)pyrimidine-5-carboxylate (Compound 0505-90)
The title compound 0505-90 was prepared (260 mg, 90%) as a white solid from
0504-90 (246 mg, 0.5 mmol), 0107-3 (146 mg, 0.6 mmol), sodium hydrogen
carbonate
(126 mg, 1.5 mmol) and bis(triphenylphosphine)palladium(a) chloride (18 mg,
0.025
mmol) in toluene (8.0 mL), ethanol (5 mL) and water (3 mL) using a procedure
similar to
that described for compound 0505-83 (Example 24): LCMS: 575 [M+1]+ 'H NMR (400
MHz, DMSO6) 6 1.38 (t, J= 7.2 Hz, 3H), 3.44 (s, 3H), 3.69 (t, J= 5.6 Hz, 2H),
3.87 (m,
4H), 4.02 (m, 6H), 4.27 (q, J= 7.2 Hz, 2H), 5.34 (s, 2H), 7.55 (t, J= 7.6 Hz,
1H), 7.67 (s,
I H), 7.75 (d, J= 8.0 Hz, I H), 8.30 (d, J= 6.8 Hz, I H), 8.97 (s, 2H), 13.29
(s, I H).
Step 28d: 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(2-
methoxyethyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 90)
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The title compound 90 was prepared (180 mg, 71%) as a white solid from 0505-90
(260 mg, 0.45 mmol) and freshly prepared hydroxylamine methanol solution (15.0
mL)
using a procedure similar to that described for compound 3 (Example 1): m.p.
219-222 C.
LCMS: 482 [M+l]+. 'H-NMR (400 MHz. DMSO-d6) 6 3.26 (s, 3H), 3.59 (t, J= 5.6
Hz,
2H), 3.79 (m, 4H), 3.90 (t, J = 5.6 Hz, 2H), 3.95 (m, 4H), 5.22 (s, 2H), 7.46
(t, J = 7.2 Hz,
1 H), 7.5 6 (s, 1 H), 7.65 (d, J = 8.4 Hz, 1 H), 8.21 (d, J = 7.2 Hz, 1 H),
8.74 (s, 2H), 8.87 (s,
1 H), 9.07 (brs, 1 H), 11.12 (s, 1 H), 13.19 (s, 1 H).
Example 29: Preparation of 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(isobutyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 93)
Step 29a: N-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)-2-
methylpropan-
1-amine (Compound 0503-93)
The title compound 0503-93 was prepared (0.6 g, 88%) as a yellow solid from
0502
(694 g, 2.0 mmol), 2-methylpropan-l-amine (1.5 g, 20 mmol) and DIPEA (2.6 g,
20 mmol)
in MeOH (5 mL) using a procedure similar to that described for compound 0503-
84
(Example 25): LCMS: 341 [M+1]+. 'H-NMR (400 MHz, DMSO-d6) 6 0.87 (d, J= 6.8
Hz,
6H), 1.69 (m, I H), 2.35 (d, J= 6.8 Hz, 2H), 2.60 (s, I H), 3.74 (m, 4H), 3.88
(m, 4H), 4.00
(s, 2H), 7.23 (s, 1H).
Step 29b: Ethyl 2-(((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(isobutyl)
amino)pyrimidine-5-carboxylate (Compound 0504-93)
The title compound 0504-93 was prepared (500 mg, 57%) as a yellow solid from
0503-93 (613 mg, 1.8 mmol) and 0305 (675 mg, 3.6 mmol) in acetonitrile (8 mL)
and N,
N-Diisopropylethylamine (1.2 g, 9 mmol) using a procedure similar to that
described for
compound 0504-83 (Example 24): LCMS: 491 [M+1]+, 1H NMR (400 MHz, DMSO-d6) 6
0.87 (d, J= 6.8 Hz, 6H), 1.29 (t, J= 7.2 Hz, 3H), 2.17 (m, 1H), 3.58 (d, J=
7.6 Hz, 2H),
3.71 (m, 4H), 3.83 (m, 4H), 4.28 (q, J= 7.2 Hz, 2H), 5.17 (s, 2H), 7.41 (s,
1H), 8.85 (s,
2H).
Step 29c: Ethyl 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)
(isobutyl)amino)pyrimidine-5-carboxylate (Compound 0505-93)
The title compound 0505-93 was prepared (257 mg, 91%) as a white solid from
0504-93 (245 mg, 0.5 mmol), 0107-3 (147 mg, 0.6 mmol), sodium hydrogen
carbonate
(126 mg, 1.5 mmol) and bis(triphenylphosphine)palladium(a) chloride (18 mg,
0.025
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mmol) in toluene (4.0 mL), ethanol (2 mL) and water (1 mL) using a procedure
similar to
that described for compound 0505-83 (Example 24): LCMS: 573 [M+1]+;'H NMR (400
MHz, DMSO-d6): 6 0.89 (d, J= 6.8 Hz, 6H), 1.29 (m, 3H), 2.20 (m, 1H), 3.60 (d,
J= 7.6
Hz, 2H), 3.79 (m, 4H), 3.95 (m, 4H), 4.27 (m, 2H), 5.21 (s, 2H), 7.47 (t, J=
8.0 Hz, 1H),
7.60 (s, I H), 7.68 (d, J= 8.4 Hz, I H), 8.22 (d, J= 8.0 Hz, I H), 8.86 (s,
2H), 8.90 (s, I H),
13.22(s, 1H).
Step 29d: 2-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(isobutyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 93)
The title compound 93 was prepared (90mg, 26 %) as a white solid from 0505-93
(357 mg, 0.6 mmol) and freshly prepared hydroxylamine methanol solution (20.0
mL)
using a procedure similar to that described for compound 3 (Example 1): m.p.
196-198 C.
LCMS: 560 [M+1]; 'H NMR (400 MHz, DMSO-d6) (50.89 (d, J= 6.4 Hz, 6H), 2.20 (m,
1H), 3.59 (d, J= 7.6 Hz, 2H), 3.80 (m, 4H), 3.96 (m, 4H), 5.20 (s, 2H), 7.47
(t, J= 8.0 Hz,
I H), 7.59 (s, I H), 7.68 (d, J= 8.0 Hz, I H), 8.22 (d, J= 7.2 Hz, I H), 8.75
(s, 2H), 8.90 (s,
1 H), 9.12 (s, 1 H), 11.14 (s, 1 H), 13.23 (s, 1 H).
Example 30: Preparation of 6-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methylamino)-N-hydroxynicotinamide (Compound 76)
Step 30a: Isopropyl 6-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)nicotinate (Compound 0309-76)
To a suspension of 0112 (3.4 g, 12 mmol) and ethyl 6-aminonicotinate (912 mg,
6
mmol) in toluene (50 mL) was added tetraisopropyl titanate (2 g, 7.2 mmol) and
the mixture
was stirred at 120 C overnight. NaBH(OAc)3 (1.9 g, 9 mmol) was added to the
reaction
mixture, then the mixture was cooled to room temperature and stirred for
additional 4
hours, extracted with dichloromethane (10 mL X 2).The combined organic layer
was
washed with saturated NaHCO3 (aq., 20 mL), brine (20 mL X 2), dried and
concentrated.
The residue was purified by column chromatography on silica gel (ethyl acetate
in
petroleum ether, 20% v/v) to obtain 0309-76 (1.5 g, 28%) as a white solid.
LCMS: 448
[M+1]+; 'H-NMR (400 MHz. DMSO-d6) 6 1.29 (d, J= 6 Hz, 6H) 3.73 (m, 4H), 3.85
(m,
4H), 4.89 (d, J = 5.6 Hz, 2H), 5.09 (m, 1 H), 6.64 (d, J = 8.8 Hz, 1 H), 6.81
(s, 1 H), 7.31 (s,
1 H), 7.86 (d, J = 2 Hz, 1 H), 8.5 8 (s, 1 H).
Step 30b: Methyl 6-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)nicotinate (Compound 0310-76)
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The isopropyl 6-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)- nicotinate was prepared (200 mg, 82%) as a yellow solid from
0309-76
(200 mg, 0.462 mmol), 0107-3 (124 mg, 0.51 mmol), sodium hydrogen carbonate
(120 mg,
1.4 mmol) and bis(triphenylphosphine)palladium(II) chloride (16 mg, 0.0231
mmol) in
toluene (8 mL), ethanol (5 mL) and water (2 mL) using a procedure similar to
that
described for compound 0505-83 (Example 24): LCMS: 530 [M+1]+'H-NMR (400 MHz.
DMSO-d6) 6 1.28 (d, J= 6 Hz, 6H) 3.81 (m, 4H), 3.98 (m, 4H), 4.94 (d, J= 5.6
Hz, 2H),
5.08 (m, 1 H), 6.67 (d, J = 8.8 Hz, 1 H), 7.47 (t, J = 8.4 Hz, 1 H), 7.51 (s,
1 H), 7.677 (d, J =
8.4 Hz, 1 H), 7.90 (d, J = 2.4 Hz, 1 H), 8.21 (d, J = 7.6 Hz, 2H), 8.60 (s, 1
H), 8.87 (s, 1 H),
13.20 (s, 1H).
To a mixture of above compound (200 mg, 0.378 mmol) in MeOH (8 mL) was
added dropwise conc. H2SO4 (2 ml). The mixture was refluxed overnight.
Evaporated to
give the crude methyl ester, 0310-76 (140 mg, 75%) which was used in next step
directly
without further purification. LCMS: 502 [M+1]-'-'H-NMR (400 MHz. DMSO-d6) 6
3.77 (s,
3H), 3.81 (m, 4H), 3.98 (m, 4H), 4.94 (d, J = 5.6 Hz, 2H), 6.67 (d, J = 8.8
Hz, 1 H), 7.47 (t,
J = 8.4 Hz, 1 H), 7.51 (s, 1 H), 7.677 (d, J = 8.4 Hz, 1 H), 7.91 (d, J = 2.4
Hz, 1 H), 8.21 (d, J
= 7.6 Hz, 2H), 8.62 (s, I H), 8.87 (s, I H), 13.21 (s, I H).
Step 30c: 6-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)-N-hydroxynicotinamide (Compound 76)
The title compound 76 was prepared (23 mg, 16%) as a brown solid from 0310-76-
2
(140 mg, 0.28 mmol) and freshly prepared hydroxylamine methanol solution (10.0
mL)
using a procedure similar to that described for compound 3 (Example 1). m.p
218-220 C.
LCMS: 503 [M+1]+.'H-NMR (400 MHz. DMSO-d6): 6 3.81 (m, 4H), 3.98 (m, 4H), 4.90
(d, J = 4.8 Hz, 2H), 6.62 (d, J = 6.8 Hz, 1 H), 7.46 (t, J = 8 Hz, 1 H), 7.5 0
(s, 1 H), 7.62 (d, J
= 8 Hz, 1 H), 7.78 (d, J = 4 Hz, 1 H), 7.89 (t, J = 4.8 Hz, 1 H), 8.21 (d, J =
6.4 Hz, 1 H), 8.43
(s, I H), 8.85 (s, I H), 10.95 (s, I H), 13.19 (s, I H).
Example 31: Preparation of 4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methylamino)-N-hydroxybenzamide (Compound 78)
Step 31a: Ethyl 4-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)benzoate (Compound 0309-78)
The title compound, 0309-78 was prepared (580 mg, 95%) as an orange solid from
ethyl 4-aminobenzoate (256 mg, 1.55 mmol), 0112 (400 mg, 1.41 mmol) and
tetraisopyl
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titanate (480 mg, 1.69 mmol) using a procedure similar to that described for
compound
0309-76 (Example 30). LCMS: 433 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 1.26 (t,
J=
7.2 Hz, 3H), 3.71 (t, J= 4.8 Hz, 4H), 3.83 (t, J= 4.6 Hz, 4H), 4.20 (q, J= 5.3
Hz, 2H), 4.71
(d, J = 6.0 Hz, 2H), 6.68 (d, J = 9.2 Hz, 2H), 7.35 (t, J = 6.4 Hz, 1 H), 7.36
(s, 1 H), 7.69 (d,
J = 8.4 Hz, 2H).
Step 31b: Ethyl4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)benzoate (Compound 0310-78)
The title compound, 0310-78 was prepared (85 mg, 33%) as a white solid from
0309-78 (216 mg, 0.5 mmol), 0107-3 (256 mg, 0.53 mmol), sodium hydrogen
carbonate
(126 mg, 1.5 mmol) and bis(triphenylphosphine)palladium(II) chloride (18 mg,
0025
mmol) in toluene (4.0 mL), ethanol (2.5 mL) and water (1.5 mL) using a
procedure similar
to that described for compound 0310-76 (Example 30). LCMS: 515 [M+1]+ 1H NMR
(400
MHz, DMSO-d6) 6 1.25 (t, J= 7.2 Hz, 3H), 3.80 (t, J= 4.6 Hz, 4H), 3.97 (t, J=
4.6 Hz,
4H), 4.20 (q, J = 5.3 Hz, 2H), 4.75 (d, J = 6.0 Hz, 2H), 6.73 (d, J = 8.8 Hz,
2H), 7.39 (t, J =
6.0 Hz, 1H), 7.47 (t, J= 8.0 Hz, 1H), 7.56 (s, 1H), 7.67 (d, J= 8.4 Hz, 1H),
7.71 (d, J= 8.8
Hz, 2H), 8.22 (d, J= 7.2 Hz, 1H), 8.88 (s, 1H).
Step 31c: 4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)-N-hydroxybenzamide (Compound 78)
The title compound 78 was prepared (41 mg, 28%) as a yellow solid from 0310-78
(150 mg, 0.29 mmol) and freshly prepared hydroxylamine methanol solution (10.0
mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
180-183 C.
LCMS: 502 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 3.80 (br s, 4H), 3.96 (t, 4H),
4.72
(d, J = 5.6 Hz, 2H), 6.68 (d, J = 8.8 Hz, 2H), 7.09 (t, J = 6.0 Hz, 1 H), 7.47
(t, J = 8.0 Hz,
1H), 7.54 (d, J= 8.4 Hz, 2H), 7.57 (s, 1H), 7.67 (d, J= 8.0 Hz, 1H), 8.22 (d,
J= 7.2 Hz,
I H), 8.72 (s, I H), 8.87 (s, I H), 10.81 (s, I H), 13.22 (s, I H).
Example 32: Preparation of (E)-3-(4-((2-(1H-indazol-4-yl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methylamino)phenyl)-N-hydroxyacrylamide (Compound 80)
Step 32a: (E)-ethyl 3-(4-((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)phenyl)acrylate (Compound 0309-80)
The title compound, 0309-80 was prepared (968 mg, 71%) as a light yellow solid
from (E)-ethyl 3-(4-aminophenyl)acrylate (623 mg, 3.26 mmol), 0112 (840 mg,
2.96
mmol), and tetraisopyl titanate (1 g, 3.55 mmol) using a procedure similar to
that described
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for compound 0309-76 (Example 30). LCMS: 459 [M+1]+;'H NMR (400 MHz, DMSO-d6)
6 1.22 (t, J= 7.2 Hz, 3H), 3.71 (t, J= 4.6 Hz, 4H), 3.83 (t, J= 4.6 Hz, 4H),
4.13 (q, J= 7.2
Hz, 2H), 4.69 (d, J = 6.0 Hz, 2H), 6.27 (d, J = 16.0 Hz, 1 H), 6.65 (d, J =
8.8 Hz, 2H), 7.17
(t, J = 6.0 Hz,1 H), 7.36 (s, 1 H), 7.44 (d, J = 8.8 Hz, 2H), 7.48 (d, J =
16.0 Hz, 1 H).
Step 32b: (E)-ethyl 3-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methylamino)phenyl)acrylae (Compound 0310-80)
The title compound, 0310-80 was prepared (490 mg, 69%) as a light yellow solid
from 0309-80 (600 mg, 1.31 mmol), 0107-3 (383 mg, 1.57 mmol), sodium hydrogen
carbonate (329 mg, 3.92 mmol) and bis(triphenylphosphine)palladium(11)
chloride (46 mg,
0.065 mmol) in toluene (16.0 mL), ethanol (10 mL) and water (4 mL) using a
procedure
similar to that described for compound 0310-76 (Example 30). LCMS: 541 [M+1]+
'H
NMR (400 MHz, DMSO-d6) 6 1.22 (t, J= 7.2 Hz, 3H), 3.82 (t, J= 4.0 Hz, 4H),
3.97 (t, J=
4.4 Hz, 4H), 4.12 (q, J = 7.2 Hz, 2H), 4.74 (d, J = 5.2 Hz, 2H), 6.27 (d, J =
15.6 Hz, 1 H),
6.70 (d, J= 8.8 Hz, 2H), 7.21 (t, J= 5.8 Hz, 1H), 7.45-7.50 (m, 4H), 7.57 (s,
1H), 7.67 (d, J
= 8.0 Hz, I H), 8.22 (d, J= 6.8 Hz, I H), 8.87 (s, I H), 13.21 (s, I H).
Step 32c: (E)-3-(4-((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methylamino)phenyl)-N-hydroxyacrylamide (Compound 80)
The title compound 80 was prepared (71 mg, 15%) as a light yellow solid from
0310-80 (490 mg, 0.91 mmol) and freshly prepared hydroxylamine methanol
solution (20.0
mL) using a procedure similar to that described for compound 3 (Example 1).
m.p. >300
C. LCMS: 528 [M+1]+; 1H NMR (400 MHz, DMSO-d6) :6S 3.87 (t, J= 4.6 Hz, 4H),
4.03 (t,
J= 4.6 Hz, 4H), 4.78 (d, J= 5.6 Hz, 2H), 6.21 (d, J= 15.6 Hz, 1H), 6.76 (d, J=
8.8 Hz,
2H), 7.11 (t, J= 5.8 Hz, 1H), 7.34-7.39 (m, 3H), 7.53 (t, J= 7.8 Hz, 1H),
7.63(s, 1H), 7.73
(d, J = 8.0 Hz, 1 H), 8.29 (d, J = 7.6 Hz, 1 H), 8.93 (d, J = 4.8 Hz, 2H),
10.60 (s, 1 H), 13.27
(s, 1H).
Example 33: Preparation of (E)-3-(4-(((2-(1H-indazol-4-yl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)phenyl)-N-hydroxyacrylamide (Compound
81)
Step 33a: (E)-ethyl 3-(4-(((2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)
(methyl)amino)phenyl)acrylate (Compound 0309-81)
The solution of compound 0309-80 (1.0 g, 2.2 mmol), CH3I (6.2 g, 44 mmol), and
Cs2CO3 (1.44 g, 4.4 mmol) in dry CH3CN / DMF solution (5 mL /5 mL) was stirred
at
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room temperature for 3 days. CH3I and CH3CN was removed in vacuo and the
residue was
diluted with H20, extracted with ethyl acetate, dried over Na2SO4, and the
crude product
was purified by column chromatography on silica gel (ethyl acetate in
petroleum ether,
20% v/v) to obtain 0309-81 (0.3 g, 30%) as a yellow solid. LCMS: 473 [M+1]+.
'H-NMR
(400 MHz, DMSO-d6) 6 1.23 (t, J= 7.2 Hz, 3H), 3.12 (s, 3H), 3.70 (m, 4H), 3.82
(m, 4H),
4.14 (q, J= 7.2 Hz, 2H), 4.97 (s, 2H), 6.33 (d, J= 16.4 Hz, 1H), 6.82 (d, J=
8.8 Hz, 2H),
7.31 (s, 1H), 7.53 (m, 3H).
Step 33b: (E)-ethyl 3-(4-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)
methyl)(methyl)amino)phenyl)acrylate (Compound 0310-81)
The title compound, 0310-81 was prepared (200 mg, 71 %) as a white solid from
0309-81 (240 mg, 0.5 mmol), 0107-3 (135 mg, 0.55 mmol), NaHCO3 (126 mg, 1.5
mmol),
and bis(triphenylphosphine)palladium( II) chloride (18 mg, 0.025 mmol) in
toluene (4 mL),
ethanol (2 mL) and water (1 mL) using a procedure similar to that described
for compound
0310-76 (Example 30). LCMS: 555 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 1.23 (m,
3H), 3.17 (s, 3H), 3.80 (m, 4H), 3.95 (m, 4H), 4.15 (m, 2H), 5.01 (s, 2H),
6.33 (d, J= 15.6
Hz, 1 H), 6.86 (d, J = 8.8 Hz, 2H), 7.46 (m, 1 H), 7.54 (m, 4H), 7.66 (d, J =
8.0 Hz , 1 H),
8.21 (d, J= 7.2 Hz , 1H), 8.87 (s, 1H), 13.21(s, 1H).
Step 33c: (E)-3-(4-(((2-(1H-indazol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)phenyl)-N-hydroxyacrylamide (Compound 81)
The title compound 81 was prepared (24 mg, 10 %) as a light yellow solid from
0310-81 (250 mg, 0.45 mmol) and freshly prepared hydroxylamine methanol
solution (20.0
mL) using a procedure similar to that described for compound 3 (Example 1).
m.p. 188-190
C. LCMS: 542 [M+1]+; 'H NMR (400 MHz, DMSO-d6) (53.14 (s, 3H), 3.80 (m, 4H),
3.95 (m, 4H), 4.98 (s, 2H), 6.21 (d, J= 15.2 Hz, 1H), 6.86 (d, J= 8.8 Hz, 2H),
7.32 (d, J=
15.2 Hz, 1 H), 7.40 (d, J = 8.4 Hz, 2H), 7.46 (t, J = 7.6 Hz, 1 H), 7.5 0 (s,
1 H), 7.66 (d, J =
8.0 Hz, I H), 8.21 (d, J= 7.2 Hz, I H), 8.34 (s, I H), 8.87 (s, I H), 10.58
(s, I H), 13.23 (s,
I H).
Example 34: Preparation of (2-(((2-(3-acetamidophenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 107)
Step 34a: N-(3-bromophenyl)acetamide (Compound 0601-107)
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To the solution of 3-bromoaniline (6.3 g, 63.7mmol) in CH2C12 (50 mL) was
added
acetyl chloride (3.75 g, 47.7 mmol) and TEA (7.4 g, 73.4 mmol) at 0 C,
stirred for 2 hours.
The mixture was washed with water, brine, dried over Na2SO4, filtered, and
concentrated
under reduced pressure to give the title compound 0601-107 (7.8 g, 99.3%) as a
brown
solid. LCMS: 215 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 2.05 (s, 3H), 7.22 (m,
2H),
7.46 (d, J = 7.6 Hz, 1 H), 7.95 (s, 1 H), 10.11 (s, 1 H).
Step 34b: N-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetamide
(Compound
0602-107)
To a solution of compound 0601-107 (2.5 g, 11.6 mmol) and
bis(pinacolato)diboron
(4.4 g, 17.5 mmol) in dioxane (100 mL) was added potassium acetate (3.4 g, 35
mmol) and
PdC12(dppf)2 (0.95 g, 1.1 mmol). The mixture was degassed with nitrogen and
heated at 85
C for overnight. The reaction mixture was concentrated under reduced pressure
to afford
the crude product, which purified by column chromatography (ethyl acetate in
petroleum
ether, 15% v/v) to give the compound 0602-107 (1.55 g, 51%) as a pink solid.
LCMS: 262
[M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.29 (s, 12H), 2.03 (s, 3H), 7.30 (s, 1H),
7.31 (d,
J= 2.0 Hz I H), 7.73 (d, J= 2.0 Hz, I H), 7.89 (d, J= 1.6 Hz, I H), 9.93 (s, I
H).
Step 34c: Ethyl 2-(((2-(3-acetamidophenyl)-4-morpholinothieno[3,2-d]pyrimidin-
6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-107)
The title compound, 0603-107 was prepared (160 mg, 99%) as a gray solid from
0504-54 (130 mg, 0.30 mmol), 0602-107 (84 mg, 0.7 mmol), sodium hydrogen
carbonate
(74 mg, 0.88 mmol) and bis(triphenylphosphine)palladium(II) chloride (12 mg,
0.014
mmol) in toluene (2.5 mL), ethanol (1.6 mL) and water (0.7 mL) using a
procedure similar
to that described for compound 0310-76 (Example 30). LCMS: 548 [M+1]+; 'H NMR
(400
MHz, DMSO-d6): 6 1.31 (t, J= 6.8 Hz, 3H), 2.07 (s, 3H), 3.27 (s, 1H), 3.77 (t,
J= 5.2 Hz,
4H), 3.94 (t, J= 5.2 Hz, 4H), 4.29 (q, J= 7.2 Hz, 2H), 5.24 (s, 2H), 7.39 (t,
J= 8.4 Hz,
1 H), 7.49 (s, 1 H), 7.82 (d, J = 8.0 Hz, 1 H), 8.05 (d, J= 8.8 Hz, 1 H), 8.52
(s, 1 H), 8.89 (s,
2H), 10.08 (s, 1H).
Step 34d: 2-(((2-(3-acetamidophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 107)
The title compound 107 was prepared (64 mg, 50%) as a white solid from 0603-
107
(130 mg, 0.23 mmol) and freshly prepared hydroxylamine methanol solution (4.0
mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
183-185 C.
LCMS: 535 [M+1]; 'H NMR (400 MHz, DMSO-d6): 6 2.07 (s, 3H), 3.24 (s, 3H), 3.77
(t,
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J = 4.0 Hz, 4H), 3.94 (t, J = 4.0 Hz, 4H), 5.21 (s, 2H), 7.39 (t, J = 8 Hz, 1
H), 7.46 (s, 1 H),
7.82 (d, J = 7.6 Hz, 1 H), 8.05 (d, J = 8 Hz, 1 H), 8.51 (s, 1 H), 8.75 (s,
2H), 9.07 (s, 1 H),
10.08 (s, 1H), 11.13 (s, 1H).
Example 35: Preparation of 2-(((2-(3-(dimethylamino)phenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)-N-
hydroxypyrimidine-5-carboxamide (Compound 108)
Step 35a: Ethyl 6-(2-chloro-4-morpholinothieno [3, 2-d] pyrimidin-6-ylamino)
hexanoate
(Compound 0602-108)
The title compound, 0602-108 was prepared (600 mg, 80%) as oil from 3-bromo-
N,N-dimethylaniline (600 mg, 3.0 mmol), bis(pinacolato)diboron (1.14 g, 4.5
mmol),
potassium acetate (882 g, 9.0 mmol), and PdC12(dppf)2 (245 mg, 0.3 mmol) using
a
procedure similar to that described for compound 0602-107 (Example 34). LCMS:
248
[M+1 ]+. 'H NMR (400 MHz, DMSO-d6) 6 1.34 (s, 12H), 2.97 (s, 6H), 7.19 (m,
2H), 7.26
(m 2H).
Step 35b: Ethyl 2-(((2-(3-(dimethylamino)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-108)
The title compound 0603-108 was prepared (245 mg, 91%) as a white solid from
0504-54 (224 mg, 0.5 mmol), 0602-108 (490 mg, 2.0 mmol), NaHCO3 (126 mg, 1.5
mmol)
and bis(triphenylphosphine)palladium( II) chloride (18 mg, 0.025 mmol) in
toluene (4 mL),
ethanol (2 mL) and water (1 mL) using a procedure similar to that described
for compound
0603-107 (Example 30). LCMS: 534 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 6 1.30 (t,
J
= 6.8 Hz, 3H), 2.97 (s, 3H), 3.27 (s, 2H), 3.76 (m, 4H), 3.92 (m, 4H), 4.28
(q, J= 6.8 Hz,
2H), 5.23 (s, 2H), 6.85 (m, I H), 7.28 (t, J= 8.0 Hz, I H), 7.49 (s, I H),
7.71 (d, J= 7.2 Hz,
I H), 7.79 (br s, I H), 8.79 (s, I H).
Step 35c: 2-(((2-(3-(dimethylamino)phenyl)-4-morpholinothieno[3,2-d]pyrimidin-
6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 108)
The title compound 108 was prepared (35 mg, 15 %) as a yellow solid from 0603-
108 (130 mg, 0.23 mmol) and freshly prepared hydroxylamine methanol solution
(20 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
172-175 C.
LCMS: 521 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 2.96 (s, 3H), 3.23 (s, 2H), 3.75
(m,
4H), 3.91 (m, 4H), 5.19 (s, 2H), 6.84 (m, 1 H), 7.27 (t, J = 8.0 Hz, 1 H),
7.46 (s, 1 H), 7.70
(d, J = 7.2 Hz, 1 H), 7.75 (br s, 1 H), 8.74 (s, 1 H), 9.11 (br s, 1 H), 11.16
(br s, 1 H).
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Example 36: Preparation of N-hydroxy-2-(methyl((4-morpholino-2-(pyridin-3-
yl)thieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound
109)
Step 36a: Ethyl 2-(methyl((4-morpholino-2-(pyridin-3-yl)thieno[3,2-d]pyrimidin-
6-
yl)methyl)amino)pyrimidine-5-carboxylate (compound 0603-109)
The title compound, 0603-109 was prepared (140 mg, 94%) as a yellow solid from
0504-54 (135 mg, 0.30 mmol), 3-pyridylboronic acid (41 mg, 0.60 mmol), NaHCO3
(76
mg, 0.90 mmol) and Pd(dppf)2C12 (11 mg, 0.015 mmol) in toluene (2.5 mL),
ethanol (1.6
mL) and water (0.7 mL) using a procedure similar to that described for
compound 0603-
107 (Example 30). LCMS: 492 [M+1]+. 'HNMR (400 MHz, DMSO-d6) 6 1.30 (t, J= 7.2
Hz, 3H), 3.28 (s, 3H), 3.76 (m, 4H), 3.95 (m, 4H), 5.25 (s, 2H), 7.53 (m, 2H),
8.66 (m, 2H),
8.88 (s, 2H), 9.51 (s, 1H).
Step 36b: N-hydroxy-2-(methyl((4-morpholino-2-(pyridin-3-yl)thieno[3,2-
d]pyrimidin-6-
yl)methyl)amino)pyrimidine-5-carboxamide (Compound 109)
The title compound 109 was prepared (30 mg, 44%) as a yellow solid from 0603-
109 (70 mg, 0.14 mmol) and freshly prepared hydroxylamine methanol solution
(10 mL)
using a procedure similar to that described for compound 3 (Example 1). mp:
160-164 C.
LCMS: 479 [M+1]. 'H NMR (400 MHz, DMSO-d6) 5 3.24 (s, 3H), 3.77 (s, 4H), 3.94
(s,
4H), 5.21 (s, 2H), 7.52 (m, 2H), 8.67 (m, 2H), 8.76 (s, 2H), 9.09 (s, 1H),
9.52 (s, 1H), 11.15
(s, H).
Example 37: Preparation of 2-(((2-(6-aminopyridin-3-yl)-4-morpholinothieno
[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 110)
Step 37a: 5-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine
(Compound
0602-110)
The title compound, 0602-110 was prepared (500 mg, 23%) as an oil from 2-amino-
5-bromopyridine (1.73 g, 10 mmol), bis(pinacolato)diboron (3.81 g, 15 mmol),
potassium
acetate (3 g, 30 mmol), and PdC12(dppf)2 (408 mg, 5 mmol) using a procedure
similar to
that described for compound 0602-107 (Example 34). LCMS: 221 [M+1]+. 1H NMR
(400
MHz, DMSO-d6) 6 1.25 (s, 12H), 6.30 (s, 2H), 6.39 (d, J= 8.0 Hz, 1H), 7.54 (d,
J= 10.0
Hz, I H), 8.16 (s, I H).
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Step 37b: Ethyl 2-(((2-(6-aminopyridin-3-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5 -carboxylate (Compound 0603-110)
The title compound, 0603-110 was prepared (200 mg, 59%) as a white solid from
0602-110 (300 mg, 0.67 mmol), 0504-54 (176 mg, 0.8 mmol), NaHCO3 (172 mg,
2mmol)
and bis(triphenylphosphine)palladium( II) chloride (23 mg, 0.0335 mmol) in
toluene (8
mL), ethanol (5 mL) and water (2 mL) using a procedure similar to that
described for
compound 0603-107 (Example 30). LCMS: 507 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6
1.29 (t, J= 6.8 Hz, 3H), 3.29 (s, 3H), 3.81 (m, 4H), 3.95 (m, 4H), 4.28 (m,
2H), 5.24 (s,
2H), 6.42 (s, 2H), 6.56 (d, J = 8.8 Hz, 1 H), 7.44 (s, 1 H), 8.327 (d, J = 8.8
Hz,1 H), 8.81 (s,
2H), 8.99 (s, I H), 9.13 (s, I H), 11.69 (s, I H).
Step 37c: 2-(((2-(6-Aminopyridin-3-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 110)
The title compound 110 was prepared (25 mg, 13 %) as a yellow solid from 0603-
110 (200 mg, 0.4 mmol) and freshly prepared hydroxylamine methanol solution
(20 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
175-181 C.
LCMS: 494 [M+1]+; 'H NMR (400 MHz, DMSO-d6) 6 3.29 (s, 3H), 3.81 (m, 4H), 3.95
(m,
4H), 5.24 (s, 2H), 6.42 (s, 2H), 6.56 (d, J = 8.8 Hz, 1 H), 7.44 (s, 1 H),
8.327 (d, J = 8.8
Hz,1 H), 8.81 (s, 2H), 8.99 (s, I H), 9.13 (s, I H), 11.195 (s, I H).
Example 38: Preparation of 2-(((2-(2-aminopyrimidin-5-yl)-4-
morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 115)
Step 38a: 5-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine
(Compound
0602-115)
The title compound, 0602-115 was prepared (120 mg, 11%) as an oil from 2-amino-
5-bromopyrimidine (865 mg, 5.0 mmol) and bis(pinacolato)diboron (2.54 g, 10
mmol),
potassium acetate (1.47 g, 15 mmol), and PdC12(dppf)2 (204 mg, 0.25 mmol)
using a
procedure similar to that described for compound 0602-107 (Example 34). LCMS:
222
[M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.26 (s, 12H), 7.04 (s, 2H), 8.37 (s, 2H).
Step 38b: Ethyl 2-(((2-(2-aminopyrimidin-5-yl)-4-morpholinothieno[3,2-
d]pyrimidin- 6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-115)
The title compound, 0603-115 was prepared (110 mg, 51%) as a white solid from
0602-115 (120 mg, 0.54 mmol), 0504-54 (200 mg, 0.45 mmol), NaHCO3 (114 mg,
1.35
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mmol), and bis(triphenylphosphine)palladium(II) chloride (16 mg, 0.0225 mmol)
in
toluene (8 mL), ethanol (5 mL) and water (2 mL) using a procedure similar to
that
described for compound 0603-107 (Example 30). LCMS: 508 [M+1]+; 1H NMR (400
MHz,
DMSO-d6) 6 1.39 (t, J= 6.8 Hz, 3H), 3.32 (s, 3H), 3.89 (m, 4H), 4.02 (m, 4H),
4.36 (q, J=
6.8 Hz, 2H), 5.20 (s, 2H), 5.43 (s, 2H), 7.44 (s, 1H), 8.94 (s, 2H), 9.31 (s,
2H).
Step 38c: 2-(((2-(2-Aminopyrimidin-5-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 115)
The title compound 115 was prepared (25 mg, 23 %) as a yellow solid from 0603-
115 (110 mg, 0.2 mmol) and freshly prepared hydroxylamine methanol solution
(20 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
175-181 C.
LCMS: 495 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 3.21 (s, 3H), 3.75 (m, 4H), 3.91
(m,
4H), 5.19 (s, 2H), 7.13 (s, 2H), 7.41 (s, 1 H), 8.74 (s, 2H), 9.02 (br s, 1
H), 9.10 (s, 2H),
11.13 (br s, 1H).
Example 39: Preparation of N-hydroxy-2-(methyl((2-(2-(methylamino)pyrimidin-5-
yl)-4-morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)amino)pyrimidine-5-
carboxamide (Compound 116)
Step 39a: 5-Bromo-N-methylpyrimidin-2-amine (compound 0601-116) and 5-bromo-
N,N-
dimethylpyrimidin-2-amine (compound 0601-117)
A mixture of 5-bromopyrimidin-2-amine (3.48 g, 20 mmol) and DMF (20 mL) was
cooled to 0 C. To the mixture NaH (60%, 1.44 g, 36 mmol) was added. After 15
minutes,
iodomethane (5 mL, 80 mmol) was added and stirred at 0 C for 0.5 h and the
mixture was
warmed to room temperature for additional 4 hours. Water (30 mL) was added and
extracted with ethyl acetate (3 x 30 mL). The combined organic layers was
washed with
brine, dried over Na2SO4, concentrated and purified by column chromatograph on
silica
gel (ethyl acetate in petroleum ether, 10% v/v) to give two compounds:
compound 0601-
116 (0.76 g, 20%) as a white solid, LCMS: 190 [M+2]+; 'HNMR (400 MHz, DMSO-d6)
6
2.75 (d, J= 4.8 Hz, 3H), 7.35 (d, J= 4.0 Hz, 1H), 8.34 (s, 2H); compound 0601-
117 (1.96
g, 49%) as a yellow solid, LCMS: 202 [M+1]+; 'HNMR (400 MHz, DMSO-d6) 6 3.12
(s,
6H), 8.43 (s, 2H).
Step 39b: N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-
amine
(Compound 0602-116)
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The title compound, 0602-116 was prepared (350 mg, 50%) as yellow solid from 5-
bromo-N-methylpyrimidin-2-amine (0.56 g, 3 mmol), bis(pinacolato)diboron (1.14
g, 4.5
mmol), potassium acetate (0.88 g, 9 mmol), and Pd(dppf)2C12 (490 mg, 0.6 mmol)
using a
procedure similar to that described for compound 0602-107 (Example 34). LCMS:
236
[M+1]+; 'HNMR (400 MHz, DMSO-d6) 6 1.27 (s, 12H), 2.82 (d, J= 4.8 Hz, 3H),
7.47 (m,
I H), 8.38 (m, I H), 8.45 (m, I H).
Step 39c: Ethyl 2-(methyl((2-(2-(methylamino)pyrimidin-5-yl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-116)
The title compound, 0603-116 was prepared (100 mg, 64%) as a yellow solid from
0504-54 (135 mg, 0.30 mmol), 0602-116 (106 mg, 0.45 mmol), NaHCO3 (76 mg, 0.90
mmol) and Pd(dppf)2C12 (11 mg, 0.015 mmol) in toluene (2.5 mL), ethanol (1.6
mL) and
water (0.7 mL) using a procedure similar to that described for compound 0603-
107
(Example 30). LCMS: 522 [M+1]+; 'HNMR (400 MHz, CDC13) 6 1.39 (t, J= 7.2 Hz,
3H),
3.09 (d, J= 5.2 Hz, 3H), 3.31 (s, 3H), 3.85 (t, J= 4.8 Hz, 4H), 3.98 (t, J=
4.8 Hz, 4H),
4.36 (q, J= 7.2 Hz, 2H), 5.19 (s, 2H), 5.47 (d, J= 4.8 Hz, 1H), 7.34 (s, 1H),
8.93 (s, 2H),
9.26 (s, 2H).
Step 39d: N-hydroxy-2-(methyl((2-(2-(methylamino)pyrimidin-5-yl)-4-
morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound 116)
The title compound 116 was prepared (50 mg, 54%) as a yellow solid from 0603-
116 (96 mg, 0.13 mmol) and freshly prepared hydroxylamine methanol solution (5
mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.:
183-187 C.
LCMS: 509 [M+1]+; 'HNMR (400 MHz, DMSO-d6) 6 2.88 (d, J= 4.4 Hz, 3H), 3.23 (s,
3H), 3.74 (s, 4H), 3.90 (s, 4H), 5.19 (s, 2H), 7.40 (s, I H), 7.55 (d, J= 4.4
Hz, I H), 8.75 (s,
2H), 9.07 (s, 1 H), 9.13 (m, 2H), 11.13 (s, 1 H).
Example 40: Preparation of 2-(((2-(2-(dimethylamino)pyrimidin-5-yl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)-N-
hydroxypyrimidine-5-carboxamide (Compound 117)
Step 40a: N,N-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
yl)pyrimidin-2-amine
(Compound 0602-117)
The title compound, 0602-117 was prepared (194 mg, 26%) as yellow solid from 5-
bromo-N,N-dimethylpyrimidin-2-amine (0.61 g, 3 mmol), bis(pinacolato)diboron
(1.14 g,
4.5 mmol), potassium acetate (0.88 g, 9 mmol), and Pd(dppf)2C12 (490 mg, 0.6
mmol) using
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a procedure similar to that described for compound 0602-107 (Example 34).
LCMS: 168
[M-81]+; 'HNMR (400 MHz, DMSO-d6) 6 1.27 (s, 12H), 3.14 (s, 6H), 8.47 (s, 2H).
Step 40b: Ethyl 2-(((2-(2-(dimethylamino)pyrimidin-5-yl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (0603-117)
The title compound, 0603-117 was prepared (100 mg, 64%) as a yellow solid from
0504-54 (135 mg, 0.30 mmol), 0602-117 (112 mg, 0.45 mmol), NaHCO3 (76 mg, 0.90
mmol) and Pd(dppf)2C12 (11 mg, 0.015 mmol) in toluene (2.5 mL), ethanol (1.6
mL) and
water (0.7 mL) using a procedure similar to that described for compound 0603-
107
(Example 30). LCMS: 536 [M+1]+; 'HNMR (400 MHz, CDC13) 6 1.38 (t, J= 7.2 Hz,
3H),
2.37 (s, 6H), 3.30 (s, 3H), 3.83 (t, J= 4.8 Hz, 4H), 3.97 (t, J= 4.8 Hz, 4H),
4.36 (q, J= 7.2
Hz, 2H), 5.18 (s, 2H), 7.35 (s, 1H), 8.93 (s, 2H), 9.30 (s, 2H).
Step 40c: 2-(((2-(2-(Dimethylamino)pyrimidin-5-yl)-4-morpholinothieno[3,2-
d]pyrimidin-
6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 117)
The title compound 117 was prepared (60 mg, 66%) as a yellow solid from 0603-
117 (93 mg, 0.17 mmol) and freshly prepared hydroxylamine methanol solution (5
mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.:
200-206 C.
LCMS: 523 [M+1]+; 'HNMR (400 MHz, DMSO-d6) 6 3.18 (d, J= 8.8 Hz, 6H), 3.23 (s,
3H), 3.74 (d, J = 4.8 Hz, 4H), 3.90 (d, J = 4.4 Hz, 4H), 5.17 (s, 2H), 7.39
(s, 1 H), 8.75 (s,
2H), 9.19 (s, 2H).
Example 41: Preparation of N-hydroxy-2-(methyl((4-morpholino-2-(pyrimidin-5-
yl)thieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound
119)
Step 41a: Ethyl 2-(methyl((4-morpholino-2-(pyrimidin-5-yl)thieno[3,2-
d]pyrimidin-6-
yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-119)
The title compound, 0603-119 was prepared (160 mg, 46%) as a white solid from
0504-54 (314 mg, 0.7 mmol), pyrimidin-2-ylboronic acid (175 mg, 1.4 mmol),
NaHCO3
(176 mg, 2.1 mmol) and bis(triphenylphosphine)palladium(II) chloride (24 mg,
0.03
mmol) in toluene (8 mL), ethanol (5 mL) and water (3 mL) using a procedure
similar to that
described for compound 0603-107 (Example 30). LCMS: 493 [M+1]+; 'H NMR (400
MHz,
DMSO-d6): 6 1.35 (t, J= 7.2 Hz, 3H), 3.31 (d, J= 8.4 Hz, 3H), 3.76 (m, 4H),
3.92 (m, 4H),
4.33 (q, J= 6.8 Hz, 2H), 5.28 (d, J= 12.0 Hz, 2H), 7.47 (s, 0.5H), 7.59 (s,
0.5H), 8.53 (s,
0.5H), 8.92 (d, J= 6.0 Hz, 2H), 9.34 (s, 0.5H), 9.67 (s, 1H).
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Step 41b: N-hydroxy-2-(methyl((4-morpholino-2-(pyrimidin-5-yl)thieno[3,2-
d]pyrimidin-
6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 119)
The title compound 119 was prepared (60 mg, 40%) as a white solid from 0603-
119
(150 mg, 0.3 mmol) and freshly prepared hydroxylamine methanol solution (5 mL)
using a
procedure similar to that described for compound 3 (Example 1). m.p. 159-160
C. LCMS:
480 [M+1 ]+; 'H NMR (400 MHz, DMSO-d6) 6 3.40 (s, 3H), 3.92 (m, 4H), 4.13 (m,
4H),
5.19 (s, 2H), 6.84 (m, 1 H), 7.27 (t, J = 8.0 Hz, 1 H), 7.46 (s, 1 H), 7.70
(d, J = 7.2 Hz, 1 H),
7.75 (br s, 1 H), 8.74 (s, 1 H), 9.11 (br s, 1 H), 11.16 (br s, 1 H).
Example 42: Preparation of N-hydroxy-2-(methyl((2-(2-methylpyrimidin-5-yl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound 120)
Step 42a: 5-Bromo-2-methylpyrimidine (Compound 0601-120)
Sodium (356 mg, 15.5 mmol) was carefully added to ethanol (5.9 mL) to prepare
sodium ethoxide solution in ethanol. The above freshly prepared sodium
ethoxide in
ethanol solution (3.5 mL) was added to a stirred suspension of acetamidine
hydrochloride
(0.91 g, 9.69 mmol). The mixture was warmed to 50 C, then the heating bath
was removed
and a solution of mucobromic acid (1 g, 3.87 mmol) in ethanol was added
dropwise at a
rate which maintained a constant temperature, followed by a further sodium
ethoxide in
ethanol solution (2 mL). After cooling, the mixture was filtered and
evaporated to a residue
which was shaken vigorously with hydrochloric acid (2 M x 2.4 mL). The brown
precipitate was filtered and washed with cold water, then freeze-dried to give
5-bromo-2-
methylpyrimidine-4-carboxylic acid (350 mg, 42%) as a brown solid. LCMS: 218
[M+1]+,
iH NMR (400 MHz, DMSO-d6): 6 2.62 (s, 3H), 9.03 (s, 1H).
A mixture of compound 5-bromo-2-methylpyrimidine-4-carboxylic acid (350 mg,
1.6 mmol) in xylene (5 mL) was refluxed for 2 h. After cooling, the mixture
was applied
directly to a silica column, which was eluted with petroleum ether, then ethyl
acetate in
petroleum ether (5% v/v) to give compound 0601-120 (170 mg, 61%) as a white
solid.
LCMS: 173 [M+1]+, 1H NMR (400 MHz, DMSO-d6): 6 2.59 (s, 3H), 8.87 (s, 2H).
Step 42b: 2-Methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine
(Compound
0602-120)
The title compound, 0602-120 was prepared (100 mg, 52%) as a yellow oil from
0601-120 (150 mg, 0.87 mmol), bis(pinacolato)diboron (331 mg, 1.3 mmol),
PdC12(dppf)2
(21 mg, 0.026 mmol) and dried potassium acetate (256 mg, 2.62 mmol)using a
procedure
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similar to that described for compound 0602-107 (Example 34). LCMS: 221
[M+1]+;'H
NMR (400 MHz, DMSO-d6): 6 1.32 (s, 12H), 2.64 (s, 3H), 8.81 (s, 2H).
Step 42c: Ethyl 2-(methyl((2-(2-methylpyrimidin-5-yl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-120)
The title compound, 0603-120 was prepared (210 mg, 74%) as a off-white solid
from 0504-54 (250 mg, 0.56 mmol), 0602-120 (880 mg, 4 mmol), sodium hydrogen
carbonate (168 mg, 2 mmol), and bis(triphenylphosphine)palladium(11) chloride
(23 mg,
0.03 mmol) in toluene (8 mL), ethanol (5mL) and water (2 mL) using a procedure
similar
to that described for compound 0603-107 (Example 30). LCMS: 507 [M+1]+; 'H NMR
(400 MHz, CDC13): 6 1.31 (t, J= 7.2 Hz, 3H), 2.74 (s, 3H), 3.25 (s, 3H), 3.79
(t, J= 4.4 Hz,
4H), 3.94 (t, J= 4.4 Hz, 4H), 4.29 (q, J= 7.2 Hz, 2H), 5.14 (s, 2H), 7.32 (s,
1H), 8.86 (s,
2H), 9.48 (s, 2H).
Step 42d: N-hydroxy-2-(methyl((2-(2-methylpyrimidin-5-yl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 120)
The title compound 120 was prepared (150 mg, 73%) as a white solid from 0603-
120 (210 mg, 0.41 mmol) and freshly prepared hydroxylamine methanol solution
(5 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
184-186 C.
LCMS: 494 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 2.69 (s, 3H), 3.24 (s, 3H),
3.76 (t, J
= 4.4 Hz, 4H), 3.94 (t, J= 4.4 Hz, 4H), 5.21 (s, 2H), 7.49 (s, 1H), 8.76 (s,
2H), 9.06 (s,
1H), 9.9.48 (s, 2H), 11.14 (s, 1H).
Example 43: Preparation of 2-(((2-(2-ethylpyrimidin-5-yl)-4-
morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 121)
Step 43a: 5-Bromo-2-ethylpyrimidine (0601-121)
After dissolving propionitrile (38 g, 0.69 mol) in anhydrous ethanol (100 mL),
HCI
gas was bubbled in at 0 C for 4 h. The mixture was stirred overnight at room
temperature
and the excess HCI gas and ethanol were removed in vacuum. Ether (100 mL) was
added in
and the solid substance were filtered and washed with ether (100 mL). The
solid was dried
and then dissolved in ethanol (100 mL) and NH3 gas was bubbled in at 0 C for
an hour, the
solution was filtered and the filtrate was concentrated to half of the
original volume, the
solid was filtered off. The solid substance thus obtained was filtered off
again and the
filtrate was concentrated to give propionimidamide hydrochloride (34 g, 45%)
as a white
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solid. GCMS: 71 [M-1]+, 'H NMR (400 MHz, DMSO-d6): 6 1.17 (t, J= 7.6 Hz, 3H),
2.40
(q, J= 7.6 Hz, 2H), 8.79 (s, 2H), 9.09 (s, 2H).
Sodium (356 mg, 15.5 mmol) was carefully added to ethanol (5.9 mL) to prepare
sodium ethoxide solution in ethanol. The above freshly prepared ethanol
solution (3.5 mL)
was added to a stirred suspension of propionimidamide hydrochloride (1.05 g,
9.69 mmol).
The mixture was warmed to 55 C, then the heating bath was removed and a
solution of
mucobromic acid (1 g, 3.87 mmol) in ethanol was added dropwise at a rate which
maintained a constant temperature, followed by a further sodium ethoxide
solution (2 mL).
After cooling, the mixture was filtered and evaporated to a residue which was
shaken
vigorously with hydrochloric acid (2 M x 2.4 mL). The brown precipitate was
filtered off
and washed with cold water, then freeze-dried to give 5-Bromo-2-
ethylpyrimidine-4-
carboxylic acid (330 mg, 37%) as a yellow solid. LCMS: 231 [M+1]+, 'H NMR (400
MHz,
DMSO-d6): 6 1.25 (t, J= 7.6 Hz, 3H), 2.88 (q, J= 7.6 Hz, 2H), 9.05 (s, 1H).
A mixture of 5-Bromo-2-ethylpyrimidine-4-carboxylic acid (5.6 g, 24.3 mmol) in
xylene (50 mL) was refluxed for 2h. After cooling, the mixture was applied
directly to a
silica column, which was eluted with petroleum ether, then ethyl acetate in
petroleum ether
(5%) to give compound 0601-121 (1.7 g, 38%) as a yellow liquid. 'H NMR (400
MHz,
DMSO-d6): 1.26 (t, J= 7.6 Hz, 3H), 2.87 (q, J= 7.6 Hz, 2H), 8.90 (s, 2H).
Step 43b: 2-Ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine
(Compound
0602-121)
The title compound, 0602-121 was prepared (crude 3.7 g) as a yellow oil from
0601-121 (1.7 g, 9.1 mmol), bis(pinacolato)diboron (3.5 g, 13.6 mmol),
PdC12(dppf)2 (222
mg, 0.27 mmol) and potassium acetate (2.7 g, 27 mmol) using a procedure
similar to that
described for compound 0602-107 (Example 34). LCMS: 235 [M+1]+; 1H NMR (400
MHz,
DMSO-d6): 6 1.27 (t, J= 7.6 Hz, 3H), 1.32 (s, 12H), 2.91 (q, J= 7.6 Hz, 2H),
8.84 (s, 2H).
Step 43c: Ethyl 2-(((2-(2-ethylpyrimidin-5-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-121)
The title compound, 0603-121 was prepared (120 mg, 41%) as a yellow solid from
0504-54 (250 mg, 0.56 mmol), 0602-121 (3.7 g, crude), sodium hydrogen
carbonate (168
mg, 2 mmol) and bis(triphenylphosphine)palladium(II) chloride (23 mg, 0.03
mmol) in
toluene (8 mL), ethanol (5mL) and water (2 mL) using a procedure similar to
that described
for compound 0603-107 (Example 30). LCMS: 521 [M+1]+; 1H NMR (400 MHz, CDC13):
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6 1.29-1.36 (m, 6H), 3.00 (q, J= 8 Hz, 2H), 3.25 (s, 3H), 3.79 (t, J= 4.4 Hz,
4H), 3.94 (t, J
= 4.4 Hz, 4H), 4.29 (q, J= 7.2 Hz, 2H), 5.14 (s, 2H), 7.32 (s, 1H), 8.86 (s,
2H), 9.51(s, 2H).
Step 43d: N-Hydroxy-2-(methyl((2-(2-methylpyrimidin-5-yl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 121)
The title compound 121 was prepared (66 mg, 56%) as a white solid from 0603-
121
(120 mg, 0.23 mmol) and freshly prepared hydroxylamine methanol solution (8
mL) using
a procedure similar to that described for compound 3 (Example 1). m.p. 153-156
C.
LCMS: 508 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 1.32 (t, J= 7.2 Hz, 3H), 2.97
(q, J
= 7.2 Hz, 2H), 3.24 (s, 3H), 3.76 (t, J = 4.4 Hz, 4H), 3.95 (t, J = 4.4 Hz,
4H), 5.21 (s, 2H),
7.49 (s, I H), 8.75 (s, 2H), 9.06 (s, I H), 9.52 (s, 2H), 11.13 (s, I H).
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Example 44: Preparation of 2-(((2-(2-amino-4-methylpyrimidin-5-yl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)-N-
hydroxypyrimidine-5-carboxamide (Compound 125)
Step 44a: 5-Bromo-4-methylpyrimidin-2-amine (Compound 0601-125)
A mixture of 2-amino-4-methylpyrimidine (4.0 g, 36.7 mmol), NBS (7.18 g, 40.3
mmol) in chloroform (100 mL) was stirred for 2 h at room temperature, then the
solvent
was removed in vacuum. Water (100 mL) was added and stirred for 30 min at room
temperature, filtered. The solid was washed with water and dried to get
compound 0601-
125 (6.3 g, 91%) as a white solid. LCMS: 188 [M+1]+, 'H NMR (400 MHz, DMSO-d6)
6
2.32 (s, 3H), 6.79 (s, 2H), 8.21 (s, 1H).
Step 44b: 4-Methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-
amine
(Compound 0602-125)
The title compound, 0602-125 was prepared (430 mg, 69%) as a grey solid from
0601-125 (500 mg, 2.66 mmol), bis(pinacolato)diboron (1.01 g, 4.0 mmol),
PdC12(dppf)2
(217.2 mg, 0.27 mmol), potassium acetate (783 mg, 7.98 mmol) using a procedure
similar
to that described for compound 0602-107 (Example 34). LCMS: 236 [M+1]+, 'H NMR
(400 MHz, DMSO-d6) 6 1.27 (s, 12H), 2.37 (s, 3H), 6.89 (s, 2H), 8.30 (s, 1H).
Step 44c: Ethyl2-(((2-(2-amino-4-methylpyrimidin-5-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-
125)
The title compound, 0603-125 was prepared (160 mg, 55%) as a white solid from
0504-54 (376 mg, 0.84 mmol), 0602-125 (130 mg, 0.56 mmol), CsF (256 mg, 1.68
mmol)
and bis(triphenylphosphine)palladium(II) chloride (59 mg, 0.084 mmol) in 1,4-
dioxane (5
mL) and water (1 mL) using a procedure similar to that described for compound
0603-107
(Example 30). LCMS: 522 [M+1]+; 'H NMR (400 MHz, CDC13): 6 1.38 (d, J= 7.2 Hz,
3H), 2.73 (s, 3H), 3.31 (s, 3H), 3.84 (m, 4H), 3.96 (m, 4H), 4.37 (q, J= 7.2
Hz, 2H), 5.20
(s, 2H), 5.23 (s, 2H), 7.35 (s, 1H), 8.89 (s, 1H), 8.93 (s, 2H).
Step 44d: 2-(((2-(2-Amino-4-methylpyrimidin-5-yl)-4-morpholinothieno[3,2-
d]pyrimidin-
6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 125)
The title compound 125 was prepared (92 mg, 62 %) as a white solid from 0603-
125 (150 mg, 0.29 mmol) and freshly prepared hydroxylamine methanol solution
(20 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
195-198 C.
LCMS: 509 [M+1]; 'H NMR (400 MHz, DMSO-d6) 5 2.62 (s, 3H), 3.23 (s, 3H), 3.74
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(m, 4H), 3.86 (m, 4H), 5.20 (s, 2H), 6.86 (s, 2H), 7.42 (s, 1H), 8.77 (s, 2H),
8.81 (s, 1H),
9.10 (s, I H), 11.21 (s, I H).
Example 45: Preparation of N-hydroxy-2-(((2-(3-methoxyphenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide (Compound 130)
Step 45a: 2-(3-Methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
(Compound
0602-130)
The title compound, 0602-130 was prepared (800 mg, 68%) as an oil from 1-bromo-
methoxybenzene (930 mg, 5.0 mmol), bis(pinacolato)diboron (2.54 g, 10 mmol),
potassium
acetate (1.47 g, 15 mmol), and PdC12(dppf)2 (204 mg, 0.25 mmol) using a
procedure similar
to that described for compound 0602-107 (Example 34). LCMS: 235 [M+1]+. 'H NMR
(400 MHz, DMSO-d6) 6 1.34 (s, 12H), 3.83 (s, 3H), 7.0 (d, J= 6.4 Hz, 2H), 7.29
(m, 2H),
7.41 (d, J= 5.6 Hz, 1H).
Step 45b: Ethyl 2-(((2-(3-methoxyphenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-130)
The title compound, 0603-130 was prepared (120 mg, 51%) as a white solid from
0602-130 (126 mg, 0.54 mmol), 0504-54 (200 mg, 0.45 mmol), NaHCO3 (114 mg,
1.35
mmol), and bis(triphenylphosphine)palladium(II) chloride (16 mg, 0.0225 mmol)
in
toluene (8 mL), ethanol (5 mL), and water (2 mL) using a procedure similar to
that
described for compound 0603-107 (Example 30). LCMS: 521 [M+1]+; 1H NMR (400
MHz,
DMSO-d6): 6 1.30 (t, J= 6.8 Hz, 3H), 3.27 (s, 3H), 3.76 (m, 4H), 3.83 (s, 3H),
3.92 (m,
4H), 3.94 (s, 2H), 4.29(q, J = 6.8 Hz, 2H), 5.23 (s, 2H), 7.06 (d, J = 7.2 Hz
1 H), 7.39 (t, J =
10.4 Hz, I H), 7.49 (s, I H), 7.92 (s, I H), 7.99 (d, J= 8 Hz,1 H), 8.82 (s, I
H).
Step 45c: N-hydroxy-2-(((2-(3-methoxyphenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 130)
The title compound 130 was prepared (15 mg, 15 %) as a yellow solid from 0603-
130 (110 mg, 0.2 mmol) and freshly prepared hydroxylamine methanol solution
(20 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
179-181 C.
LCMS: 508 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 3.23 (s, 3H), 3.77 (m, 4H), 3.83
(s,
3H), 3.93 (m, 4H), 5.21 (s, 2H), 7.07 (d, J= 9.2 Hz, 1H), 7.39 (t, J= 10.4 Hz,
1H), 7.42 (s,
1 H), 7.91 (s, 1 H), 8.0 (d, J = 16.4 Hz, 1 H), 8.75 (s, 2H), 9.02 (br s, 1
H), 11.14 (br s, 1 H).
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Example 46: Preparation of N-hydroxy-2-(((2-(3-hydroxyphenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide (Compound 132)
Step 46a: 3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (Compound 0602-
132)
The title compound, 0602-132 was prepared (600 mg, 68%) as an oil from 3-
bromophenol (700 mg, 4.0 mmol), bis(pinacolato)diboron (1.5 g, 6 mmol),
potassium
acetate (1.2 g, 12 mmol), and PdC12(dppf)2 (163 mg, 0.2 mmol) using a
procedure similar
to that described for compound 0602-107 (Example 34). LCMS: 221 [M+1]+. 1H NMR
(400 MHz, CDC13) 6 1.34 (s, 12H), 5.37 (s, 1H), 6.96 (d, J= 4.0 Hz, 1H), 7.26
(m, 2H),
7.36 (d, J= 7.2 Hz, 1H).
Step 46b: Ethyl 2-(((2-(3-hydroxyphenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-132)
The title compound, 0603-132 was prepared (160 mg, 47%) as a white solid from
0602-132 (300 mg, 0.67 mmol), 0504-54 (176 mg, 0.8 mmol), NaHCO3 (172 mg,
2mmol),
and bis(triphenylphosphine)palladium( II) chloride (23 mg, 0.0335 mmol) in
toluene (8
mL), ethanol (5 mL) and water (2 mL) using a procedure similar to that
described for
compound 0603-107 (Example 30). LCMS: 507 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6
1.30 (t, J= 6.8 Hz, 3H), 3.27 (s, 3H), 3.76 (m, 4H), 3.92 (m, 4H), 4.29 (q, J=
6.8 Hz, 2H),
5.24 (s, 2H), 6.84 (d, J= 16.8 Hz, 2H), 7.24 (t, J= 8 Hz, 1H), 7.48 (s, 1H),
7.83 (m, 2H),
8.88 (s, 2H), 9.49 (s, 1H).
Step 46c: N-hydroxy-2-(((2-(3-hydroxyphenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 132)
The title compound 132 was prepared (53 mg, 34 %) as a yellow solid from 0603-
132 (160 mg, 0.32 mmol) and freshly prepared hydroxylamine methanol solution
(20 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
175-181 C.
LCMS: 494 [M+1]+; 'H NMR (400 MHz, DMSO-d6) 6 3.23 (s, 3H), 3.76 (m, 4H), 3.92
(m,
4H), 5.19 (s, 2H), 6.85 (d, J = 10.4 Hz, 1 H), 7.24 (t, J = 6.8 Hz, 1 H), 7.44
(s, 1 H), 7.82 (m,
2H), 8.74 (s, 2H), 9.51 (br s, 1H).
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Example 47: Preparation of 2-(((2-(3-aminophenyl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 134)
Step 47a: Ethyl-2-(((2-(3-aminophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (compound 0603-134)
To the solution of 0603-107 (170 mg, 0.31 mmol) in THE (10 mL) was added
aqueous HC1 solution (6M, 10 mL) at 50 C and the mixture was stirred for 2
hours at this
temperature. The reaction mixture was neutralized with saturated aqueous
NaHCO3 and
extracted with ethyl acetate. The separated organic layer was washed with
water, brine,
dried over Na2SO4, filtered, and concentrated under reduced pressure to give
the title
compound 0603-134 (130 mg, 83%) as a white solid. LCMS: 506 [M+1]+. 'H NMR
(400
MHz, DMSO-d6) 6 1.30 (t, J= 7.6 Hz, 3H), 3.27 (s, 3H), 3.40(s, 2H), 3.76 (t,
J= 5.2 Hz,
4H), 3.93 (t, J = 4.8 Hz, 4H), 7.89 (q, J = 7.6 Hz, 2H), 5.24(s, 1 H), 6.79
(d, J = 7.6 Hz, 1 H),
7.19 (t, J= 8.0 Hz, I H), 7.47 (s, I H), 7.69 (d, J= 8.0 Hz, I H), 7.77 (s, I
H), 8.88 (s, I H).
Step 47b: 2-(((2-(3-Aminophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 134)
The title compound 134 was prepared (35 mg, 28%) as a white solid from 0603-
134
(130 mg, 0.25 mmol) and freshly prepared hydroxylamine methanol solution (4
mL) using
a procedure similar to that described for compound 3 (Example 1). m.p. 179-182
C.
LCMS: 493 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 63.24 (s, 3H), 3.77 (t, J= 4.0
Hz,
4H), 3.96 (t, J = 4.0 Hz, 4H), 5.22 (s, 2H), 6.98 (d, J = 7.6 Hz, 1 H), 7.32
(t, J = 7.6 Hz, 1 H),
7.47 (s, 1 H), 7.86 (d, J = 7.6 Hz, 1 H), 7.91 (s, 1 H), 8.76 (s, 2H), 9.07
(s, 1 H), 11.15 (s, 1 H).
Example 48: Preparation of 2-(((2-(4-(aminomethyl)phenyl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 135)
Step 48a: N-(4-bromobenzyl)acetamide (Compound 0601-135)
To the solution of 4-bromobenzylamine hydrochloride (1.2 g, 5.4 mmol) and Et3N
(5.5 g, 54 mmol) in dichloromethane (10 mL) was added CH30001(555 mg, 7.02
mmol) at
0 C and stirred for 2 hr at 30 C. Then the mixture was concentrated and the
residue was
dissolved in CH2C12 (30 mL), washed with water, dried over Na2SO4 and
concentrated to
obtain 0601-135 (1.3 g, 100%) as a yellow solid. LCMS: 228 [M+1]+, 1H NMR (400
MHz,
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DMSO-d6) 6 2.00 (s, 3H), 4.33 (d, J= 6.4 Hz, 2H), 6.26 (s, 1H), 7.13 (d, J=
8.4 Hz, 2H),
7.43 (d, J= 8.4 Hz, 2H).
Step 48b: N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)acetamide
(Compound 0602-135)
The title compound, 0602-135 was prepared (825 mg, 60%) as a yellow solid from
0601-135 (1.2 g, 5 mmol), bis(pinacolato)diboron (1.9 g, 7.5 mmol), potassium
acetate
(1.47 g, 15 mmol), and Pd(dppf)2C12 (410 mg, 0.5 mmol) using a procedure
similar to that
described for compound 0602-107 (Example 34). LCMS: 276 [M+1]+, 'H NMR (400
MHz,
DMSO-d6) 6 1.27 (s, 12H), 1.87 (s, 3H), 4.26 (d, J= 6.0 Hz, 2H), 7.25 (d, J=
8.0 Hz, 2H),
7.62 (d, J= 8.0 Hz, 2H), 8.36 (t, J= 5.6 Hz, 1H).
Step 48c: Methyl 2-(((2-(4-(aminomethyl)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl) (methyl)amino)pyrimidine-5-carboxylate (Compound 0603-135)
A mixture of compound 0602-135 (200 mg, 0.73 mmol), 0504-54 (261 mg, 0.58
mmol), NaHCO3 (184 mg, 2.2 mmol) and bis(triphenylphosphine)palladium(II)
chloride
(52 mg, 0.073 mmol) in toluene (4 mL), ethanol (2 mL) and water (0.5 mL) was
flushed
with nitrogen and heated under microwave irradiation at 130 C for 2 h. The
reaction
mixture was partitioned between dichloromethane and water, organic layer was
washed
with brine, dried over Na2SO4, filtered and evaporated in vacuum. The
resulting residue
was purified by column chromatography (methanol in dichloromethane, 2-5% v/v)
to give
ethyl 2-(((2-(4-(acetamidomethyl)phenyl)-4- morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carbo- xylate (300 mg, 92%) as a white
solid.
LCMS: 562 [M+1]+;'H NMR (400 MHz, DMSO-d6): 61.36 (m, 3H), 1.96 (s, 3H), 3.33
(s,
3H), 3.83 (m, 4H), 3.94 (m, 4H), 4.36 (m, 4H), 5.29 (s, 2H), 7.41 (d, J= 7.6
Hz, 2H), 7.54
(s, 1H), 8.39 (d, J= 8.0 Hz, 2H), 8.47 (m, 1H), 8.94 (s, 2H).
The above ethyl ester (250 mg, 0.45 mmol) was dissolved in THE (8 mL), then
aqueous HCI solution (6M, 12 mL) was added and stirred for 12 hr at 85 C.
Then the
mixture was adjusted pH 4 with NaOH at 0 C, filtered and washed with CH2C12 to
get 2-
(((2-(4-(aminomethyl)phenyl)-4-morpholinothieno [3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylic acid (200 mg, 91%) as a white
solid.
The solid was directly used in next step without further purification. LC-MS:
492 [M+1]+.
The above acid (230 mg, 0.47 mmol) was dissolved in MeOH (l OmL). SOC12 (5
mL) was added to above solution at 0 C and stirred for 1.5 hr at reflux. Then
the mixture
was concentrated, added water, adjusted to pH8 with saturated aqueous NaHCO3
solution,
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extracted with CH2C12 and evaporated in vacuum to get compound 0603-135 (210
mg,
88%) as a yellow solid. LC-MS: 506 [M+1]+.'H NMR (400 MHz, DMSO-d6) 6 3.28 (s,
3H), 3.83 (m, 9H), 3.97 (m, 4H), 5.20 (m, 2H), 7.38 (m, 3H), 8.34 (m, 2H),
8.86 (m, 2H).
Step 48d:
2-(((2-(4-(aminomethyl)phenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 135)
The title compound 135 was prepared (60 mg, 30 %) as a light yellow solid from
0603-135 (200 mg, 0.40 mmol) and freshly prepared hydroxylamine methanol
solution (20
mL) using a procedure similar to that described for compound 3 (Example 1).
m.p. 184-186
C. LCMS: 507 [M+1]; 'H NMR (400 MHz, DMSO-d6) (53.23 (s, 3H), 3.76 (m, 4H),
3.93 (m, 6H), 5.20 (s, 2H), 7.45 (s, 1 H), 7.49 (d, J = 8.4 Hz, 2H), 8.36 (d,
J = 8.0 Hz, 2H),
8.75 (s, 2H).
Example 49: Preparation of 2-(((2-(3-(aminomethyl)phenyl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 137)
Step 49a: N-(3-bromobenzyl)acetamide (Compound 0601-137)
To the solution of 3-bromobenzylamine hydrochloride (1.2 g, 5.4 mmol) and Et3N
(5.5 g, 54 mmol) in CH2C12 (10 mL) was added CH30001(555mg, 7.02 mmol) at 0 C
and
stirred for 2 hr at 30 C. Then the mixture was concentrated and the residue
was dissolved
in CH2C12, washed with water, dried over Na2SO4, and concentrated to obtain
0601-137
(1.2 g, 98%) as a yellow solid. LCMS: 228 [M+1]+. 'H NMR (400 MHz, DMSO-d6)
61.88
(s, 3H), 4.25 (d, J= 6.0 Hz, 2H), 7.28 (m, 2H), 7.43 (m, 2H), 8.39 (s, 1H).
Step 49b: N-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)acetamide
(Compound 0602-137)
The title compound, 0602-137 was prepared (1.0 g, 72%) as a yellow solid from
0601-137 (1.2 g, 5 mmol), bis(pinacolato)diboron (1.9 g, 7.5 mmol), potassium
acetate
(1.47 g, 15 mmol), and Pd(dppf)2C12 (410 mg, 0.5 mmol) using a procedure
similar to that
described for compound 0602-107 (Example 34). LCMS: 276 [M+1]+, 'H NMR (400
MHz,
DMSO-d6) 6 1.28 (s, 12H), 1.85 (s, 3H), 4.23 (d, J= 6.0 Hz, 2H), 7.33 (m, 2H),
7.54 (d, J
7.2 Hz, 1H), 7.57 (s, 1H), 8.35 (t, J= 5.6 Hz, 1H).
Step 49c: Methyl 2-(((2-(3-(aminomethyl)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl) methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-137)
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A mixture of 0602-137 (400 mg, 1.46 mmol), 0504-54 (522 mg, 1.16 mmol),
NaHCO3 (368 mg, 4.4 mmol), and bis(triphenylphosphine)palladium(II) chloride
(104 mg,
0.146 mmol) in toluene (4 mL), ethanol (2 mL) and water (0.5 mL) was flushed
with
nitrogen and heated under microwave irradiation at 130 C for 2 h. The
reaction mixture
was partitioned between dichloromethane and water, organic layer was washed
with brine,
dried over Na2SO4, filtered and evaporated in vacuum. The resulting residue
was purified
by column chromatography (methanol in dichloromethane, 2-5% v/v) to give ethyl
2-(((2-
(3-(acetamidomethyl)phenyl)- 4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carbo-xylate (580 mg, 89%) as a white
solid.
LCMS: 562 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 6 1.28 (m, 3H), 1.88 (s, 3H),
3.27 (s,
3H), 3.76 (m, 4H), 3.92 (m, 4H), 4.27 (m, 4H), 5.23 (s, 2H), 7.38 (m, 2H),
7.49 (m, 1H),
8.30 (m, 2H), 8.43 (m, 1H), 8.87 (s, 2H).
The above ethyl ester (300 mg, 0.53 mmol) was dissolved in THE (8 mL), then
aqueous HCI solution (6M, 12 mL) was added and stirred for 12 hr at 85 C.
Then the
above mixture was adjusted pH 4 with NaOH at 0 C, filtered and washed by
CH2C12 to get
2-(((2-(3-(Aminomethyl)phenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
methyl)(methyl)amino)pyrimidine-5-carboxylic acid (245 mg, 93%) as a white
solid. LC-
MS: 492 [M+1]+.
The acid (300 mg, 0.61 mmol) was dissolved in MeOH (l OmL). SOCI2 (5 mL) was
added to above solution at 0 C and stirred for 1.5 hr at reflux. Then the
mixture was
concentrated, added water, adjusted PH 8 with NaHCO3, extracted with CH2C12
and
evaporated in vacuum to get compound 0603-137 (260 mg, 84%) as a yellow solid.
LC-
MS: 506 [M+1]+. 1H NMR (400 MHz, DMSO-d6) 6 3.27 (s, 3H), 3.76 (m, 4H), 3.82
(s,
3H), 3.95 (m, 6H), 5.24 (s, 2H), 7.45 (m, 3H), 8.31 (m, 1H), 8.41 (s, 1H),
8.88 (m, 2H).
Step 49d: 2-(((2-(3-(aminomethyl)phenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 137)
The title compound 137 was prepared (46 mg, 18%) as a light yellow solid from
0603-137 (250 mg, 0.5 mmol) and freshly prepared hydroxylamine methanol
solution (20
mL) using a procedure similar to that described for compound 3 (Example 1).
m.p. 173-176
C. LCMS: 507 [M+1]+; 'H NMR (400 MHz, DMSO-d6) 6 3.23 (s, 3H), 3.76 (m, 4H),
3.94 (m, 4H), 3.99 (s, 2H), 5.20 (s, 2H), 7.45 (s, 1H), 7.50 (m, 2H), 8.32 (m,
1H), 8.38 (s,
2H), 8.43 (s, I H), 8.75 (s, 2H).
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Example 50: preparation of N-hydroxy-2-(((2-(3-(hydroxymethyl)phenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide (Compound 138)
Step 50a: (3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanol
(Compound
0602-138)
The title compound, 0602-138 was prepared (300 mg, 43%) as a yellow oil from m-
bromobenzyl alcohol (0.56 g, 3 mmol), bis(pinacolato)diboron (1.14 g, 4.5
mmol),
potassium acetate (1.32 g, 9 mmol), and Pd(dppf)2C12 (490 mg, 0.6 mmol) using
a
procedure similar to that described for compound 0602-107 (Example 34). LCMS:
252
[M+18]+; 'HNMR (400 MHz, CDC13) 6 1.35 (s, 12H), 1.66 (s, 1H), 4.70 (s, 2H),
7.38 (t, J
= 7.2 Hz, 1 H), 7.49 (d, J = 7.6 Hz, 1 H), 7.74 (d, J = 7.2 Hz, 1 H), 7.80 (s,
1 H).
Step 50b: Ethyl 2-(((2-(3-(hydroxymethyl)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (0603-138)
The title compound, 0603-138 was prepared (140 mg, 90%) as a white solid from
0504-54 (135 mg, 0.30 mmol), 0602-138 (105 mg, 0.45 mmol), NaHCO3 (76 mg, 0.90
mmol), and Pd(dppf)2C12 (11 mg, 0.015 mmol) in toluene (2.5 mL), ethanol (1.6
mL), and
water (0.7 mL) using a procedure similar to that described for compound 0603-
107
(Example 30). LCMS: 521 [M+1]+; 'HNMR (400 MHz, DMSO-d6) 6 1.30 (t, J= 7.2 Hz,
3H), 3.27 (s, 3H), 3.77 (m, 4H), 3.93 (q, J= 7.2 Hz, 2H), 4.59 (d, J= 6.0 Hz,
2H), 5.24 (s,
2H), 5.29 (t, J= 5.6 Hz, 1H), 7.43 (m, 2H), 7.50 (s, 1H), 8.26 (m, 1H), 8.37
(s, 1H), 8.88 (s,
2H).
Step 50c: N-hydroxy-2-(((2-(3-(hydroxymethyl)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 138)
The title compound 138 was prepared (30 mg, 44%) as a light yellow solid from
0603-138 (70 mg, 0.13 mmol) and freshly prepared hydroxylamine methanol
solution (10
mL) using a procedure similar to that described for compound 3 (Example 1). mp
160-164
C. LCMS: 508 [M+1]+; 'HNMR (400 MHz, DMSO-d6) 6 3.24 (s, 3H), 3.77 (s, 4H),
3.94
(s, 2H), 4.59 (d, J= 5.6 Hz, 2H), 5.21 (s, 2H), 5.28 (t, J= 5.6 Hz, 1H), 7.44
(m, 3H), 8.27
(m, I H), 8.37 (s, I H), 8.75 (s, 2H), 9.07 (s, I H).
Example 51: Preparation of N-hydroxy-2-(((2-(3-(methoxymethyl)phenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide (Compound 139)
Step 51a: 1-Bromo-3-(methoxymethyl)benzene (Compound 0601-139)
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To a solution of m-bromobenzyl alcohol (1.0 g, 5.3 mmol) in THE (10 mL) was
added NaH (0.26 g, 10.6 mmol) at 0 C, stirred for 10 minutes, followed by
addition of
iodomethane (1.1 g, 7.9 mmol). The resulting reaction mixture was stirred for
1 hour. To
the mixture ethyl acetate (30 mL) was added, washed with water, brine, dried
over Na2SO4,
filtered, and concentrated under reduced pressure to give the title compound
0601-139 (1.0
g, 93%) as an oil. 1H NMR (400 MHz, DMSO-d6) 6 3.30 (s, 3H), 4.41 (s, 2H),
7.29 (t, J
= 8.0 Hz, 1H), 7.40 (t, J= 7.6 Hz, 1H), 7.47 (d, J= 8.0 Hz, 1H), 7.64 (d, J=
8.0 Hz, 1H).
Step 51b: 2-(3-(methoxymethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
(Compound 0602-139)
The title compound, 0602-139 was prepared (1.2 g, 97%) as an oil from 0601-139
(1.1 g, 5.4 mmol), bis(pinacolato)diboron (2.1 g, 8.1 mmol), potassium acetate
(1.6 g, 16.3
mmol), and PdC12(dppf)2 (45 mg, 0.05 mmol) using a procedure similar to that
described
for compound 0602-107 (Example 34). 1H NMR (400 MHz, DMSO-d6) 6 1.28 (s, 12H),
3.28 (s, 3H), 4.40 (s, 2H), 7.35 (t, J = 7.2 Hz, I H), 7.42 (d, J = 7.2 Hz, I
H), 7.58 (d, J = 7.2
Hz, I H), 7.63 (s, I H).
Step 51c: Ethyl-2-(((2-(3-(methoxymethyl)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-139)
The title compound, 0603-139 was prepared (180 mg, 71%) as a white solid from
0504-54 (210 mg, 0.46 mmol), 0602-139 (174 mg, 0.7 mmol), sodium hydrogen
carbonate
(118 mg, 1.4 mmol), and bis(triphenylphosphine)palladium(II) chloride (16 mg,
0.02
mmol) in toluene (8 mL), ethanol (2 mL), and water (1 mL) using a procedure
similar to
that described for compound 0603-107 (Example 30). LCMS: 535 [M+1]+; 1H NMR
(400
MHz, DMSO-d6): 6 1.30 (t, J= 7.2 Hz, 3H), 3.27 (s, 3H), 3.33 (s, 3H), 3.76 (m,
4H), 3.93
(m, 4H), 4.28 (q, J= 7.2 Hz, 2H), 4.50 (s, 2H), 5.23 (s, 2H), 7.45 (q, J= 7.2
Hz, 1H), 7.49
(s, 1H), 8.31 (d, J= 7.6 Hz, 1H), 8.34 (s, 1H), 8.87 (m, 2H).
Step 51d: N-hydroxy-2-(((2-(3-(methoxymethyl)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 139)
The title compound 139 was prepared (56 mg, 47%) as an orange solid from 0603-
139 (120 mg, 0.22 mmol) and freshly prepared hydroxylamine methanol solution
(8 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
178-181 C.
LCMS: 522 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 5 3.24 (s, 3H), 3.33(s, 3H), 3.77
(m,
4H), 3.93 (m, 4H), 4.51 (s, 2H), 5.21 (s, 2H), 7.43 (q, J= 7.6 Hz, 2H), 7.47
(s, 1H), 8.31 (d,
J= 7.2 Hz, I H), 8.34 (s, I H), 8.75 (s, 2H), 9.05 (s, I H), 11.12 (s, I H).
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Example 52: Preparation of N-hydroxy-2-(((2-(4-(hydroxymethyl)phenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide (Compound 140)
Step 52a: (4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanol
(compound
0602-140)
The title compound, 0602-140 was prepared (670 mg, 94%) as an oil from (4-
bromophenyl)methanol (0.56 g, 3 mmol), bis(pinacolato)diboron (1.14 g, 4.5
mmol),
potassium acetate (1.32 g, 9 mmol), and PdC12(dppf)2 (490 mg, 0.6 mmol) using
a
procedure similar to that described for compound 0602-107 (Example 34). LCMS:
217 [M-
OH]+; 'HNMR (400 MHz, DMSO-d6) 6 1.28 (s, 12H), 4.51(d, , J=5.6 Hz 2H), 5.23
(t,
J=6.0 Hz, 1H), 7.32 (d, J=8.0 Hz, 2H), 7.62 (d, J=7.6 Hz, 2H).
Step 52b: Ethyl 2-(((2-(4-(hydroxymethyl)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (compound 0603-140)
The title compound, 0603-140 was prepared (120 mg, 77%) as a white solid from
0504-54 (135 mg, 0.30 mmol), 0602-140 (105 mg, 0.45 mmol), NaHCO3 (76 mg, 0.90
mmol), and (PPh3)PdC12 (11 mg, 0.015 mmol) in toluene (2.5 mL), ethanol (1.6
mL), and
water (0.7 mL) using a procedure similar to that described for compound 0603-
107
(Example 30). LCMS: 521 [M+1]+; 'HNMR (400 MHz, DMSO-d6) 6 1.28 (t, J=7.2 Hz,
3H), 3.25 (s, 3H), 3.74 (t, J=4.4 Hz, 4H), 3.90 (t, J=7.2 Hz, 4H), 4.26 (q,
J=7.2 Hz, 2H),
4.56 (d, J=6.4 Hz, 2H), 5.21 (s, 2H), 5.26 (t, J=5.6 Hz, 1H), 7.41 (d, J=8.0
Hz, 2H), 7.46 (s,
1H), 8.34 (d, J=8.8 Hz, 2H), 8.85 (s, 2H).
Step 52c: N-Hydroxy-2-(((2-(4-(hydroxymethyl)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (compound 140)
The title compound 140 was prepared (61 mg, 63%) as a yellow solid from 0603-
140 (100 mg, 0.19 mmol) and freshly prepared hydroxylamine methanol solution
(10 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
218-223 C.
LCMS: 508 [M+1]+; 'HNMR (400 MHz, DMSO-d6) 6 3.23 (s, 3H), 3.76 (d, J= 4.4 Hz,
4H), 3.91 (d, J= 4.0 Hz, 4H), 4.56 (d, J= 4.8 Hz, 2H), 5.19 (s, 2H), 5.27 (t,
J= 5.6 Hz,
1H), 7.42 (t, J= 8.8 Hz, 3H), 8.34 (d, J= 8.0 Hz, 2H), 8.74 (s, 2H), 9.08 (s,
1H), 11.13 (s,
I H).
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Example 53: Preparation of N-hydroxy-2-(((2-(2-(hydroxymethyl)phenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide (Compound 141)
Step 53a: 2-Bromobenzyl acetate (Compound 0601-141)
To a solution of o-bromobenzyl alcohol (2.0 g, 10.7 mmol) in CH2C12 (20 mL)
was
added acetyl chloride (1.1 g, 13.9 mmol) and TEA (2.16 g, 21.4 mmol) at 0 C,
then stirred
for 2 hours. The reaction mixture was washed with water, brine, dried over
Na2SO4,
filtered, and concentrated under reduced pressure to give the title compound
0601-141 (2.4
g, 97%) as a yellow solid. 1H NMR (400 MHz, DMSO-d6) 5 2.09 (s, 3H), 5.10 (s,
2H),
7.29 (t, J= 8.0 Hz, I H), 7.40 (t, J= 7.6 Hz, I H), 7.47 (d, J= 8.0 Hz, I H),
7.64 (d, J= 8.0
Hz, 1H).
Step 53b: 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl acetate
(Compound 0602-
141)
The title compound, 0602-141 was prepared (2.3 g, 80%) as an oil from 0601-141
(2.4 g, 10.5 mmol), bis(pinacolato)diboron (4.1 g, 16.3 mmol), potassium
acetate (3.2 g,
32.7 mmol), and PdC12(dppf)2 (89 mg, 0.11 mmol) using a procedure similar to
that
described for compound 0602-107 (Example 34). LCMS: 277 [M+1]+. 1H NMR (400
MHz,
DMSO-d6) 6 1.28 (s, 12H), 2.03 (s, 3H), 5.23 (s, 2H), 7.34 (t, J= 7.2 Hz, 1H),
7.37 (d, J=
7.2 Hz, 1H), 7.89 (t, J= 7.6 Hz, 1H), 7.70 (d, J= 7.2 Hz, 1H).
Step 53c: Ethyl 2-(((2-(2-(hydroxymethyl)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-141)
The title compound, 0603-141 was prepared (80 mg, 17%) as a white solid from
0504-54 (420 mg, 0.92 mmol), 0602-141 (386 mg, 1.4 mmol), sodium hydrogen
carbonate
(236 mg, 2.8 mmol), and bis(triphenylphosphine)palladium(II) chloride (32 mg,
0.04
mmol) in toluene (8 mL), ethanol (4 mL), and water (2 mL) using a procedure
similar to
that described for compound 0603-107 (Example 30). LCMS: 521 [M+1]+; 1H NMR
(400
MHz, DMSO-d6): 6 1.29 (t, J= 7.2 Hz, 3H), 3.26 (s, 3H), 3.74 (m, 4H), 3.88 (m,
4H), 4.28
(q, J = 6.8 Hz, 2H), 4.75 (d, J = 6.0 Hz, 2H), 5.24 (s, 2H), 5.44 (t, J = 6.0
Hz, 1 H), 7.3 5 (t, J
= 7.2 Hz, 1 H), 7.44 (t, J = 7.6 Hz, 1 H), 7.45 (s, 1 H), 7.62 (m, 1 H), 7.92
(d, J = 6.8 Hz, 1 H),
8.87 (s, 1H).
Step 53d: N-hydroxy-2-(((2-(2-(hydroxymethyl)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 141)
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The title compound 141 was prepared (58 mg, 37%) as a yellow solid from 0603-
141 (160 mg, 0.3 mmol) and freshly prepared hydroxylamine methanol solution (4
mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
173-176 C.
LCMS: 508 [M+1]; 'H NMR (400 MHz, DMSO-d6): 5 3.23 (s, 3H), 3.73 (m, 4H), 3.88
(m, 4H), 4.75 (d, J = 6.0 Hz, 2H), 5.20 (s, 2H), 5.45 (t, J = 6.0 Hz, 1 H),
7.35 (t, J = 6.8 Hz,
1 H), 7.42 (d, J = 7.6 Hz, 1 H), 7.46 (s, 1 H), 7.61 (d, J = 7.6 Hz, 1 H),
7.92 (d, J = 7.6 Hz,
1 H), 8.74 (s, 2H), 9.05 (s, 1 H), 11.12 (s, 1 H).
Example 54: Preparation of 2-(((2-(3-carbamoylphenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 142)
Step 54a: 3-Bromobenzamide (Compound 0601-142)
To a solution of m-bromobenzonitrile (2 g, 10 mol) in DMSO (6 mL) was added
30% H202 (5 g, 13 mmol) and K2CO3 at 0 C, and stirred at room temperature for
30 min.
The mixture was poured into water and filtered, the solid was washed with
water, dried to
got the compound 0601-142 (1.8 g, 82%) as a white solid. LCMS: 200 [M+1]+; 1H
NMR
(400 MHz, DMSO-d6) 67.41 (t, J= 8.0 Hz, 1H), 7.50 (s, 1H), 7.70(dd, J,2 = 8.0,
0.8 Hz,
1H), 7.85 (d, J= 8.0 Hz, 1H), 8.03 (t, J= 1.6 Hz, 1H), 8.06 (s, 1H).
Step 54b: 3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (Compound
0602-
142)
The title compound, 0602-142 was prepared (450 mg, 73%) as a solid from 0601-
142 (500 mg, 2.5 mmol), bis(pinacolato)diboron (952 mg, 3.75 mmol), potassium
acetate
(735 mg, 7.5 mmol), and PdC12(dppf)2 (61 mg, 0.075 mmol) using a procedure
similar to
that described for compound 0602-107 (Example 34). LCMS: 248 [M+1]+; 1H NMR
(400
MHz, DMSO-d6) 61.31 (s, 12H), 7.34 (s, 1H), 7.46 (t, J= 7.2 Hz, 1H), 7.79 (d,
J= 7.2
Hz, I H), 7.98 (m, I H), 8.04 (s, I H), 8.19 (s, I H).
Step 54c: Ethyl 2-(((2-(3-carbamoylphenyl)-4-morpholinothieno[3,2-d]pyrimidin-
6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-142)
The title compound, 0603-142 was prepared (180 mg, 50%) as a yellow solid from
0504-54 (305 mg, 0.68 mmol), 0602-142 (200 mg, 0.81 mmol), Pd(PPh3)2C12 (21
mg, 0.03
mmol), and NaHCO3 (171 mg, 2.04 mmol) in toluene (5 mL), ethanol (3 ml), and
water
(1.3 ml) using a procedure similar to that described for compound 0603-107
(Example 30).
LCMS: 534 [M+1]+. 1H NMR (400 MHz, CDC13) 6 1.38 (t, J = 7.6 Hz, 3H), 3.32 (s,
3H),
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3.87 (t, J = 4.6 Hz, 4H), 4.03 (t, J = 4.8 Hz, 4H), 4.37 (q, J = 7.6 Hz, 2H),
5.20 (s, 2H),
5.77 (br, 1H), 6.37 (br, 1H), 7.39(s, 1H), 7.55 (t, J= 7.8 Hz, 1H), 7.96 (d, J
= 7.6 Hz, 4H),
8.59 (d, J = 7.6 Hz, I H), 8.84 (s, I H), 8.93 (s, 2H).
Step 54d: 2-(((2-(3-carbamoylphenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 142).
The title compound 142 was prepared (65 mg, 44%) as a yellow solid from 0603-
142 (150 mg, 0.28 mmol) and freshly prepared hydroxylamine methanol solution
(8 mL)
using a procedure similar to that described for compound 3 (Example 1). LCMS:
521
[M+1]+; 'H NMR (400 MHz, DMSO-d6) 6 3.25 (s, 3H), 2.77-3.79 (m, 4H), 3.95-3.97
(m,
4H), 5.22 (s, 2H), 7.44 (s, 1 H), 7.50 (s, 1 H), 7.56 (t, J = 7.6 Hz, 1 H),
7.97 (d, J = 8.0 Hz,
I H), 8.13 (s, I H), 8.52 (d, J= 7.6 Hz, I H), 8.76 (s, 2H), 8.96 (s, I H),
9.03 (br, I H), 10.93
(br, 1 H).
EXAMPLE 55: Preparation of N-hydroxy-2-(methyl((2-(3-(methylcarbamoyl)phenyl)-
4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound 144)
Step 55a: 3-Bromo-N-methylbenzamide (Compound 0601-144)
A suspension of McNH2.HCl (1.85 g, 27 mmol) and Et3N (4.6 g, 45 mmol) in
anhydrous CH2C12 (50 mL) was cooled to 0 C and treated with m-bromobenzoyl
chloride (2
g, 9 mmol). The mixture was warmed to room temperature and stirred for 4h. To
the
reaction mixture was added ethyl acetate, washed with water, brine, and dried
over Na2SO4,
concentrated to give compound 0601-144 (1.9 g, 97%) as a white solid. LCMS:
214
[M+1]; 'H NMR (400 MHz, DMSO-d6) 62.78 (d, J= 4.4 Hz, 3H), 7.43 (t, J= 7.6 Hz,
1H), 7.72 (d, J= 8.8 Hz, 1H), 7.83 (d, J= 7.6 Hz, 1H), 8.00 (s, 1H), 8.57 (s,
1H).
Step 55b: N-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide
(Compound
0602-144)
The title compound, 0602-144 was prepared (480 mg, 82%) as a white solid from
0601-144 (500 mg, 2.3 mmol), bis(pinacolato)diboron (890 mg, 3.5 mmol),
potassium
acetate (687 mg, 7 mmol), and PdC12(dppf)2 (57.2 mg, 0.07 mmol) using a
procedure
similar to that described for compound 0602-107 (Example 34). LCMS: 262
[M+1]+; 1H
NMR (400 MHz, DMSO-d6) 61.21 (s, 12H), 2.77 (d, J= 4.8 Hz, 3H), 7.47 (t, J=
7.6 Hz,
1 H), 7.79 (d, J = 7.6 Hz, 1 H), 7.94 (d, J = 8.0 Hz, 1 H), 8.14 (s, 1 H),
8.51 (m, 1 H).
Step 55c: Ethyl 2-(methyl((2-(3-(methylcarbamoyl)phenyl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-144)
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The title compound, 0603-144 was prepared (250 mg, 68%) as a yellow solid from
0504-54 (300 mg, 0.67 mmol), compound 0602-144 (349 mg, 1.34 mmol), NaHCO3
(168
mg, 2.0 mmol), (Ph3P)2PdC12 (23 mg, 0.03 mmol) in toluene (5 mL), ethanol (3
mL) and
water (1.3 mL) using a procedure similar to that described for compound 0603-
107
(Example 30). LCMS: 548 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.30 (t, J= 6.8Hz,
3H), 2.82 (d, J= 4.8 Hz, 3H), 3.28 (s, 3H), 3.77 (m, 4H), 3.95 (m, 4H), 4.28
(q, J= 6.8 Hz,
2H), 5.24 (s, 2H), 7.52 (s, 1 H), 7.56 (t, J = 8.0 Hz, 1 H), 7.91 (d, J = 8.0
Hz, 1 H), 8.51 (d, J
= 8.0 Hz, I H), 8.56 (m, I H), 8.82 (s, I H), 8.88 (s,1 H).
Step 55d: N-hydroxy-2-(methyl((2-(3-(methylcarbamoyl)phenyl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 144)
The title compound 144 was prepared (116 mg, 48%) as a yellow solid from 0603-
144 (250 mg, 0.45 mmol) and freshly prepared hydroxylamine methanol solution
(8 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.:
215-217 C.
LCMS: 535 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 2.81 (d, J= 4.4 Hz, 3H), 3.25
(s,
3H), 3.78 (m, 4H), 3.95 (m, 4H), 5.22 (s, 2H), 7.50 (s, 1H), 7.56 (t, J= 7.6
Hz, 1H), 7.91
(d, J = 7.6 Hz, 1 H), 8.51 (d, J = 7.6 Hz, 1 H), 8.56 (m, 1 H), 8.76 (s, 2H),
8.82 (s,1 H).
Example 56: Preparation of 2-(((2-(4-aminophenyl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 150)
Step 56a: N-(4-bromophenyl)acetamide (Compound 0601-150)
To the solution of 4-bromoaniline (6.3 g, 63.7mmol) in CH2C12 (50 mL) was
added
acetyl chloride (3.75 g, 47.7 mmol) and TEA (7.4 g, 73.4 mmol) at 0 C,
stirred for 2 hours.
The reaction mixture was washed with water, brine dried over Na2SO4, filtered,
and
concentrated under reduced pressure to give the title compound 0601-150 (3.6
g, 46%) as a
brown solid. LCMS: 214 [M+1]; 'H NMR (400 MHz, DMSO-d6). (52.05 (s, 3H), 7.46
(d, J = 8.8 Hz, 2H), 7.57 (d, J = 8.8 Hz, 2H), 10.12 (s, 1 H).
Step 56b: N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetamide
(Compound
0602-150)
The title compound, 0602-150 was prepared (2.3 g, 94%) as a white solid from
0601-150 (2.0 g, 9.3 mmol), bis(pinacolato)diboron (4.4 g, 17.5 mmol),
potassium acetate
(3.5 g, 14 mmol), and PdC12(dppf)2 (76 mg, 0.088 mmol) using a procedure
similar to that
described for compound 0602-107 (Example 34). LCMS: 262 [M+1]+. 'H NMR (400
MHz,
DMSO-d6) 6 1.27 (d, J= 6.8 Hz, 12H), 2.04 (s, 3H), 7.58 (s, 4H), 10.03 (s,
1H).
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Step 56c: Ethyl 2-(((2-(4-aminophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-150)
A mixture of compound 0504-54 (210 mg, 0.46 mmol), 0602-150 (159 mg, 0.60
mmol), sodium hydrogen carbonate (118 mg, 1.4 mmol), and
bis(triphenylphosphine)palladium(II) chloride (17 mg, 0.02 mmol) in toluene (4
mL),
ethanol (2 mL) and water (1 mL) was flushed with nitrogen and heated under
microwave
irradiation at 120 C for 2 h. The reaction mixture was partitioned between
ethyl acetate
and water, organic layer was washed with brine, dried over magnesium sulfate,
filtered and
evaporated in vacuum. The residue was washed with dichloromethane to obtain
ethyl 2-
(((2-(4-acetamidophenyl)-4-morpholinothieno- [3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (136 mg, 53%) as a white
solid.
LCMS: 548 [M+1]+,'H NMR (400 MHz, DMSO-d6): 5 1.29 (t, J= 7.2 Hz, 3H), 2.06
(s,
6H), 3.26 (s, 3H), 3.75 (m, 4H), 3.91 (m, 4H), 4.28 (q, J= 7.2 Hz, 2H), 5.22
(s, 2H), 7.45
(s, 1 H),7.67 (d, J = 8.8 Hz, 1 H), 8.31 (d, J = 8.8 Hz, 1 H), 8.87 (s, 1 H),
10.10 (s, 1 H).
To the solution of above ethyl ester (280 mg, 0.51 mmol) in THE (10 mL) was
added aqueous HCI solution (6M, 15 mL) at 40 C, stirred for 2 hours, the
reaction mixture
was neutralized with NaHCO3 and extracted with CH2C12, the organic layer was
washed
with water, brine dried over Na2SO4, filtered, and concentrated, purified by
column
chromatography (methanol in dichloromethane, 2% v/v), to give title compound
0603-150
(180 mg, 48%) as a white solid. LCMS: 506 [M+l]+. 'H NMR (400 MHz, DMSO-d6) 6
1.29 (t, J= 7.6 Hz, 3H), 3.24 (s, 3H), 3.73(m, 4H), 3.86 (m, 4H), 4.27 (q, J=
6.8 Hz, 2H),
5.20 (s, 2H), 6.59 (d, J= 8.8 Hz, 2H), 7.36 (s, 1H), 8.07 (d, J= 8.0 Hz, 2H),
8.86(s, 1H).
Step 56d: 2-(((2-(4-aminophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 150)
The title compound 150 was prepared (43 mg, 26%) as a yellow solid from 0603-
150 (170 mg, 0.3 mmol) and freshly prepared hydroxylamine methanol solution (4
mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
183-186 C.
LCMS: 493 [M+1]; 'H NMR (400 MHz, DMSO-d6): 5 3.22 (s, 3H), 3.74 (m, 4H), 3.87
(m, 4H), 4.27 (q, J= 6.8 Hz, 2H), 5.20 (s, 2H), 5.50 (s, 2H), 6.59 (d, J= 8.8
Hz, 2H), 7.36
(s, 1H), 8.07 (d, J= 8.0 Hz, 2H), 8.86 (s, 2H).
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Example 57: Preparation of 2-(((2-(4-acetamidophenyl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 157)
Step 57a: 2-(((2-(4-Acetamidophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 157)
The title compound 157 was prepared (104 mg, 61 %) as a gray solid from ethyl
2-
(((2-(4-acetamidophenyl)-4-morpholinothieno [3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (170 mg, 0.3 mmol) and
freshly
prepared hydroxylamine methanol solution (10 mL) using a procedure similar to
that
described for compound 3 (Example 1). m.p. 228-230 C. LCMS: 535 [M+1]+; 1H
NMR
(400 MHz, DMSO-d6) 6 2.07 (s, 3H), 3.24 (s, 3H), 3.77 (m, 4H), 3.91 (m, 4H),
5.19 (s,
2H), 7.44 (s, 1H), 7.68 (d, J= 8.8 Hz, 2H), 8.31 (d, J= 8.8 Hz, 2H), 8.75 (s,
2H), 9.06 (s,
1H),10.12(s,1H),11.13(s,1H).
Example 58: Preparation of 2-(((2-(1H-indazol-4-yl)-4-(phenylamino)thieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 164)
Step 58a: 2-Chloro-N-phenylthieno[3,2-d]pyrimidin-4-amine (Compound 0701-164)
To a suspension of compound 0110 (4.00 g, 19.608 mmol) in CH3CN (100 mL) was
added Et3N (4.00 g, 39.216 mmol) and aniline (2.00 g, 21.581 mmol) at room
temperature.
The mixture was stirred at 50 C overnight, evaporated. The residue was
purified by column
chromatography on silica gel (ethyl acetate in petroleum ether, 20%-30% v/v)
to obtained
compound 0701-164 (2.00 g, 39%) as a pale yellow solid. LCMS: 262 [M+1]+; 1H
NMR
(400 MHz, DMSO-d6): 6 7.20 (t, J= 7.6 Hz, 1H), 7.42 (m, 3H), 7.69 (d, J= 7.6
Hz, 1H),
8.27 (d, J= 5.2 Hz, 1H), 10.16 (s, 1H).
Step 58b: Tert-butyl 2-chlorothieno[3,2-d]pyrimidin-4-yl(phenyl)carbamate
(Compound
0702-164)
To a solution of compound 0701-164 (5.60 g, 21.456 mmol) and (Boc)20 (5.60 g,
25.747 mmol) in THE (100 mL) was added DMAP (130 mg, 1.073 mmol) at room
temperature and stirred for 3 h. Solvent was removed and the residue was
purified by
column chromatography (ethyl acetate in petroleum ether, 15%-30% v/v) to
obtained
compound 0702-164 (6.20 g, 80%) as a white solid. LCMS: 362 [M+1]+; 1H NMR
(400
MHz, DMSO-d6): 6 1.44 (s, 9H), 7.31 (d, J= 7.6 Hz, 2H), 7.39 (m, 1H), 7.43 (m,
2H), 7.56
(d, J = 5.6 Hz, 1 H), 8.48 (d, J = 5.2 Hz, 1 H).
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Step 58c: Tert-butyl 2-chloro-6-formylthieno[3,2-d]pyrimidin-4-
yl(phenyl)carbamate
(Compound 0703-164)
Compound 0702-164 (2.90 g, 8.033 mmol) was suspended in THE (80 mL) and
cooled to -50 C. To the mixture LDA solution (2M, 12 mL, 24.099 mmol) was
added
dropwise while temperature was kept below -30 C and stirred for 1 h followed
by the
addition of DMF (2 mL) at -50 C. The mixture was stirred for additional 30
min. A
saturated aqueous NH4C1(20 mL) was added dropwise at -50 - -60 C. Ethyl
acetate (300
mL) was added to the mixture, washed with saturated aqueous NH4C1(2 x 100 mL),
water
(2 x 100 mL), brine (200 mL), dried over anhydrous Na2SO4, concentrated and
the residue
was purified by column chromatography on silica gel (ethyl in petroleum ether,
20%-50%
v/v) to obtain compound 0703-164 (900 mg, 29%) as a yellowish solid. LCMS: 390
[M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 1.46 (s, 9H), 7.30 (d, J= 6.8 Hz, 1H),
7.36 (m,
2H), 7.43 (m, 2H), 8.43 (s, I H), 10.21 (s, I H).
Step 58d: Tert-butyl 2-chloro-6-(hydroxymethyl)thieno[3,2-d]pyrimidin-4-
yl(phenyl)carbamate (Compound 0704-164)
To a solution of 0703-164 (900 mg, 2.314 mmol) in MeOH/THF (10/5 mL) was
added NaBH4 (176 mg, 4.627 mmol) slowly at room temperature and stirred for 30
min.
After solvent removed, the residue was washed with water and filtered to
obtain 0704-164
(800 g, 88%) as a white solid. LCMS: 392 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6
1.44
(s, 9H), 4.80 (d, J= 5.6 Hz, 2H), 6.02 (t, J = 5.6 Hz, 1H), 7.28 (d, J= 4.8
Hz, 2H), 7.37 (m,
2H), 7.45 (m, 2H).
Step 58e: (4-(Tent-butoxycarbonyl(phenyl)amino)-2-chlorothieno[3,2-d]pyrimidin-
6-
yl)methyl methanesulfonate (Compound 0705-164)
To a solution of 0704-164 (500 mg, 1.279 mmol) and Et3N (2 mL) in THE (20 mL)
was added MsC1(175 mg, 1.534 mmol) at ice bath temperature. The mixture with
white
solid was stirred at room temperature for 30 min. After solvent removed, the
residue was
washed with water and filtered to obtain 0705-164 (600 mg, 83%) as a white
solid. LCMS:
470 [M+1 ]+.
Step 58f: 2-Chloro-6-((methylamino)methyl)-N-phenylthieno[3,2-d]pyrimidin-4-
amine
(Compound 0706-164)
Compound 0705-164 (600 mg, 1.279 mmol) was dissolved in a solution of 32%
MeNH2 in MeOH (5 mL). This solution was stirred at room temperature for 1 h.
The
solvent was removed under reduced pressure and the residue was washed with
water,
filtered to obtained 0706-164 (300 mg, 77%) as a white solid. LCMS: 305
[M+1]+.
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Step 58g: Ethyl 2-(((2-chloro-4-(phenylamino)thieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0707-164)
To a solution of compound 0706-164 (260mg, 0.855 mmol) and compound 0305
(159 mg, 0.855 mmol) in CH3CN (5 mL) was added Et3N (0.5 mL) at room
temperature
and stirred overnight. After concentrated, the residue was washed with water,
filtered to
obtained compound 0707-164 (200 mg, 52%) as a white solid. LCMS: 455 [M+1]+;
'H
NMR (400 MHz, DMSO-d6): 6 1.31 (t, J= 4.2 Hz, 3H), 3.25 (s, 3H), 4.29 (m, 2H),
5.23 (s,
2H), 7.15 (t, J = 6.0 Hz. 1 H), 7.3 7 (t, J = 6.4 Hz, 2H), 7.42 (s, 1 H), 7.64
(d, J = 6.4 Hz, 2H),
8.88 (s, 2H), 9.98 (s, 1H).
Step 58h: Ethyl 2-(((2-(1H-indazol-4-yl)-4-(phenylamino)thieno[3,2-d]pyrimidin-
6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0708-164)
The title compound, 0708-164 was prepared (210 mg, 89%) as a white solid from
0706-164 (200 mg, 0.440 mmol), compound 0107-3 (118 mg, 0.484 mmol), NaHCO3
(110
mg, 1.320 mmol), and bis(triphenylphosphine)- palladium(II) chloride (15 mg,
0.022
mmol) in toluene (5.6 mL), ethanol (3.5 mL), and water (1.4 mL) using a
procedure similar
to that described for compound 0603-107 (Example 30). LCMS: 537 [M+1]+; 'H NMR
(400 MHz, DMSO-d6): 6 1.33 (t, J = 7.2 Hz, 3H), 3.31 (s, 3H), 4.31 (m, 2H),
5.31 (s, 2H),
7.16 (t, J = 7.2 Hz, 1 H), 7.41 (t, J = 4.4 Hz, 2H), 7.47 (t, J = 8.0 Hz, 1
H), 7.62 (s, 1 H), 7.65
(d, J = 8.4 Hz, 1 H), 7.79 (d, J = 7.6 Hz, 2H), 8.21 (d, J = 7.2 Hz, 1 H),
8.63 (s, 1 H), 8.92 (s,
2H), 9.66 (s, I H), 13.16 (s, I H).
Step 58i: 2-(((2-(1H-indazol-4-yl)-4-(phenylamino)thieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 164)
The title compound 164 was prepared (100 mg, 49%) as a pale white solid from
0708-164 (210 mg, 0.392 mmol) and freshly prepared hydroxylamine methanol
solution (8
mL) using a procedure similar to that described for compound 3 (Example 1).
m.p.: 232-
235 C; LCMS: 524 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 3.27 (s, 3H), 5.27 (s,
2H),
7.16 (t, J= 7.2 Hz, 1H), 7.41 (t, J= 4.4 Hz, 2H), 7.47 (t, J= 8.0 Hz, 1H),
7.59 (s, 1H), 7.65
(d, J = 8.4 Hz, 1 H), 7.78 (d, J = 7.6 Hz, 2H), 8.21 (d, J = 7.2 Hz, 1 H),
8.78 (s, 2H), 8.81 (s,
1 H), 9.07 (s, 1 H), 9.64 (s, 1 H), 11.15 (s, 1 H), 13.14 (s, 1 H).
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Example 59: Preparation of 2-(((2-(1H-indazol-4-yl)-4-(pyridin-2-
ylamino)thieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 168)
Step 59a: 2-Chloro-N-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-amine (Compound
0701-
168)
To a suspension of compound 0110 (5.00 g, 24.5 10 mmol) in THE (200 mL) was
added t-BuOK (4.19 g, 36.765 mmol) and pyridin-2-amine (2.30 g, 24.5 10 mmol)
at room
temperature. The mixture was stirred at 50 C overnight, then the mixture was
purified by
column chromatography on silica gel eluting with ethyl acetate in petroether
(20% to 40%,
v/v) to obtained compound 0701-168 (3.00 g, 47%) as a pale yellow solid. LCMS:
263
[M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 7.18 (m, 1H), 7.43 (d, J = 5.2 Hz, 1H),
7.87 (m,
2H), 8.3 3 (d, J = 5.6 Hz, 1 H), 8.40 (d, J = 4.8 Hz, 1 H), 10.94 (s, 1 H).
Step 59b: Tert-butyl 2-chlorothieno[3,2-d]pyrimidin-4-yl(pyridin-2-
yl)carbamate
(Compound 0702-168)
The title compound 0702-168 was prepared (2.80 g, 100%) as a white solid from
0701-168 (2.40 g, 9.160 mmol), (Boc)20 (3.00 g, 13.740 mmol), and DMAP (56 mg,
0.458
mmol) using a procedure similar to that described for compound 0702-164
(Example 58).
LCMS: 363 [M+1]+;'H NMR (400 MHz, DMSO-d6): 5 1.44 (s, 9H), 7.36 (m, 1H), 7.64
(m, 2H), 7.97 (t, J = 7.6 Hz, 1 H), 8.34 (m, 1 H), 8.54 (d, J = 5.6 Hz, 1 H).
Step 59c: Tert-butyl 2-chloro-6-formylthieno[3,2-d]pyrimidin-4-yl(pyridin-2-
yl)carbamate
(Compound 0703-168)
The title compound 0703-168 was prepared (1.90 g, 48%) as a light yellow solid
from 0702-168 (3.69 g, 10.199 mmol), LDA solution (2M, 15.3 mL, 30.597 mmol),
and
DMF (4 mL) using a procedure similar to that described for compound 0703-164
(Example
58). LCMS: 391 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 1.46 (s, 9H), 7.41 (m,
1H),
77.66 (d, J = 8.0 Hz, 1 H), 8.00 (m, 1 H), 8.40 (m, 1 H), 8.49 (s, 1 H), 10.12
(s, 1 H).
Step 59d: Tert-butyl 2-chloro-6-(hydroxymethyl)thieno[3,2-d]pyrimidin-4-
yl(pyridin-2-
yl)carbamate (Compound 0704-168)
The title compound 0704-168 was prepared (1.9 g, 100%) as a white solid from
0703-168 (1.9 g, 4.870 mmol) and NaBH4 (277 mg, 7.31 mmol) using a procedure
similar
to that described for compound 0704-164 (Example 58). LCMS: 393 [M+1]+; 'H NMR
(400 MHz, DMSO-d6): 6 1.44 (s, 9H), 4.84 (d, J= 5.4 Hz, 2H), 6.05 (t, J= 6.0
Hz, 1H),
7.34 (m, I H), 7.43 (s, I H), 7.65 (d, J= 8.0 Hz, I H), 7.96 (t, J= 7.6 Hz, I
H), 8.33 (m, I H).
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Step 59e: (4-(Tent-butoxycarbonyl(pyridin-2-yl)amino)-2-chlorothieno[3,2-
d]pyrimidin-6-
yl)methyl methanesulfonate (Compound 0705-168)
The title compound 0705-168 was prepared (2.10 g, 92%) as a white solid from
0704-168 (1.90 g, 4.847 mmol), Et3N (2 mL), and MsC1(1.10 g, 9.694 mmol) using
a
procedure similar to that described for compound 0705-164 (Example 58). LCMS:
471
[M+1]+.'H NMR (400 MHz, DMSO-d6): 6 1.44 (s, 9H), 3.31 (s, 3H), 5.68 (s, 2H),
7.37 (m,
I H), 7.65 (d, J= 8.0 Hz, I H), 7.74 (s, I H), 7.99 (t, J= 7.6 Hz, I H), 8.36
(m, I H).
Step 59f: 2-Chloro-6-((methylamino)methyl)-N-(pyridin-2-yl)thieno[3,2-
d]pyrimidin-4-
amine (Compound 0706-168)
The title compound 0706-168 was prepared (1.80 g, 100%) as a white solid from
0705-168 (2.00 g, 4.255 mmol) and 32% MeNH2 in MeOH (5 mL) using a procedure
similar to that described for compound 0706-164 (Example 58). LCMS: 306
[M+1]+.'H
NMR (400 MHz, DMSO-d6): 6 2.32 (s, 3H), 3.97 (s, 2H), 7.15 (m, 1H), 7.26 (s,
1H), 7.83
(m, 2H), 8.38 (m, I H), 9.71 (s, I H).
Step 59g: Ethyl 2-(((2-chloro-4-(pyridin-2-ylamino)thieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0707-168)
The title compound 0707-168 was prepared (410 mg, 69%) as a white solid from
0706-168 (400 mg, 1.311 mmol) and compound 0305 (244 mg, 1.311 mmol) using a
procedure similar to that described for compound 0707-164 (Example 58). LCMS:
456
[M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 1.31 (t, J= 7.2 Hz, 3H), 3.25 (s, 3H),
4.29 (m,
2H), 5.21 (s, 2H), 7.13 (m, I H), 7.40 (s, I H), 7.84 (m, 2H), 8.30 (m, I H),
8.88 (s, 2H),
10.84 (s, 1H).
Step 59h: Ethyl 2-(((2-(1H-indazol-4-yl)-4-(pyridin-2-ylamino)thieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0708-168)
The title compound, 0708-168 was prepared (200 mg, 85%) as a white solid from
0707-168 (200 mg, 0.440 mmol), compound 0107-3 (118 mg, 0.484 mmol), NaHCO3
(110
mg, 1.320 mmol), and bis(triphenylphosphine)-palladium(II) chloride (15 mg,
0.022
mmol) in toluene (8 mL), ethanol (5 mL), and water (2 mL) using a procedure
similar to
that described for compound 0603-107 (Example 30). LCMS: 538 [M+1]+; 'H NMR
(400
MHz, DMSO-d6): 6 1.33 (t, J= 7.2 Hz, 3H), 3.29 (s, 3H), 4.30 (m, 2H), 5.26 (s,
2H), 7.12
(m, I H), 7.7.57 (s, I H), 7.84 (m, I H), 8.23 (d, J = 7.2 Hz, I H), 8.33 (m,
I H), 8.89 (s, 2H),
10.39 (s, 1H), 13.19 (s, 1H).
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Step 59i: 2-(((2-(1H-indazol-4-yl)-4-(pyridin-2-ylamino)thieno[3,2-d]pyrimidin-
6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 168)
The title compound 168 was prepared (80 mg, 82%) as a pale white solid from
0708-168 (100 mg, 0.186 mmol) and freshly prepared hydroxylamine methanol
solution (8
mL) using a procedure similar to that described for compound 3 (Example 1).
m.p.: 200-
203 C; LCMS: 525 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 3.34 (s, 3H), 5.30 (s,
2H),
7.19 (t, J= 6.0 Hz, I H), 7.55 (t, J= 8.0 Hz, I H), 7.62 (s, I H), 7.47 (d, J=
8.0 Hz, I H), 7.91
(t, J = 8.8 Hz, 1 H), 8.00 (d, J = 8.4 Hz, 1 H), 8.30 (d, J = 7.6 Hz, 1 H),
8.41 (s, 1 H), 8.84 (s,
2H), 9.04 (s, I H), 9.14 (s, I H), 10.46 (s, I H), 11.01 (s, I H), 13.25 (s, I
H).
EXAMPLE 60: Preparation of 5-(4-(2-(1H-indazol-4-yl)-6-((4-
(methylsulfonyl)piperazin-1-yl)methyl)thieno [3,2-d] pyrimidin-4-yl)piperazin-
l-yl)-N-
hydroxypentanamide (Compound 30)
Step 60a: Tert-butyl 4-(2-chlorothieno[3,2-d]pyrimidin-4-yl)piperazine-l-
carboxylate
(Compound 0801-30)
A mixture of compound 0110 (20 g, 98 mmol), tert-butyl 1-piperazinecarboxylate
(21.9 g, 118 mmol), and triethylamine (14.8 g, 147 mmol) in methanol (400 mL)
was
stirred at room temperature for 4 h. The reaction mixture was filtered to give
a solid as first
batch of crude product. The mother liquid was concentrated and diluted with
water (500
mL) and the solid was collected as second batch of crude product. The combined
products
were dried to give compound 0801-30 (22 g, 63%) as a yellow solid. LCMS: 355
[M+1]+.
iH NMR (400 MHz, DMSO-d6): 6 1.43 (s, 9H), 3.53 (m, 4H), 3.95 (m, 4H), 7.41(d,
J= 5.6
Hz, 1H), 8.32 (d, J= 5.6 Hz , 1H).
Step 60b: Tert-butyl 4-(2-chloro-6-formylthieno[3,2-d]pyrimidin-4-
yl)piperazine-l-
carboxylate (Compound 0802-30)
Compound 0801-30 (10 g, 28.2 mmol) was suspended in THE (200 mL) and cooled
to -70 C followed by the addition of LDA solution (2M, 71 mL) dropwise and
kept the
reaction temperature below -65 C. The mixture was then stirred for 1 h. DMF
(15 mL) was
added dropwise to the mixture while the reaction temperature was kept below -
65 C. The
resulting mixture was stirred for 30 min. and quenched with saturated aqueous
NH4C1(50
mL) at -50- -60 C. The mixture was diluted with ethyl acetate (500 mL),
washed with
saturated aqueous NH4HCO3 (2 x 300 mL), water (2 x 500 mL), brine (200 mL),
dried over
Na2SO4, concentrated and the residue was purified by column chromatography on
silica gel
(methanol in dichloromethane, 2% v/v) to give compound 0802-30 (4.9 g, 45%) as
a light
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yellow solid. LCMS: 383 [M+1]+.'H NMR (400 MHz, DMSO-d6): 6 1.43 (s, 9H), 3.55
(m,
4H), 3.99 (m, 4H), 8.29 (s, I H), 10.21 (s, I H).
Step 60c: Tert-butyl 4-(2-chloro-6-(hydroxymethyl)thieno[3,2-d]pyrimidin-4-
yl)piperazine-l-carboxylate (Compound 0803-30)
To a suspension of compound 0802-30 (3.73 g, 9.764 mmol) in MeOH (20 mL) was
added NaBH4 (1.11 g, 29.293 mmol) slowly at 0 C within 10 minutes. The formed
clear
solution was stirred for 10 min., evaporated. The residue was washed with
water and
filtered to obtain compound 0803-30 (3.10 g, 83%) as a white solid. LCMS: 385
[M+1]+;
iH NMR (400 MHz, DMSO-d6): 6 1.43 (s, 9H), 3.52 (br s, 4H), 3.92 (br s, 4H),
4.81 (d, J
4.8 Hz, 2H), 5.96 (t, J = 5.2 Hz, 1 H), 7.22 (s, 1 H).
Step 60d: Tert-butyl 4-(6-(bromomethyl)-2-chlorothieno[3,2-d]pyrimidin-4-
yl)piperazine-
1-carboxylate (Compound 0804-30)
To a suspension of compound 0803-30 (3.10 g, 8.073 mmol) and PPh3 (2.54 g,
9.687 mmol) in dichloromethane (30 mL) was added NBS (1.72 g, 9.687 mmol) at
room
temperature. The mixture was stirred at room temperature for 2 h and
concentrated. The
residue was purified by column chromatography on silica gel (dichloromethane
in
petroleum ether, 10% to 50% v/v) to obtain compound 0804-30 (2.60 g, 72%) as a
white
solid. LCMS: 447 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 6 1.42 (s, 9H), 3.52 (br
s, 4H),
3.90 (br s, 4H), 5.10 (s, 2H), 7.49 (s, I H).
Step 60e: Tert-butyl 4-(2-chloro-6-((4-(methylsulfonyl)piperazin-1-
yl)methyl)thieno[3,2-
d]pyrimidin-4-yl)piperazine-l-carboxylate (Compound 0805-30)
A mixture of compound 0804-30 (3.5 g, crude product), compound 0103 (1.14 g,
4.1 mmol), and K2C03 (1.58 g, 11.4 mmol) in acetonitrile (35 mL) and DMF (17
mL) was
stirred at room temperature overnight. The reaction mixture was diluted with
water (200
mL) and extracted with dichloromethane (200 mL). The organic layer was washed
with
water (5 x 200 mL), brine, dried over Na2SO4, concentrated to give crude
product,
compound 0805-30 (3.9 g with some Ph3PO) as a yellow solid which was used in
next step
reaction without further purification. LCMS: 531 [M+1]+. 1H NMR (400 MHz, DMSO-
d6)
6 1.42 (s, 9H), 2.59 (m, 4H), 2.90 (s, 3H), 3.15 (m, 4H), 3.52 (m, 4H), 3.91
(m, 6H), 7.31
(s, 1H).
Step 60f: 2-Chloro-6-((4-(methylsulfonyl)piperazin-l-yl)methyl)-4-(piperazin-l-
yl)thieno[3,2-d]pyrimidine (Compound 0806-30)
To a solution of compound 0805-30 (5 g) in THE (50 mL) was added concentrated
hydrochloric acid (10 mL) and stirred at room temperature overnight. The
reaction mixture
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was diluted with water (200 mL) and extracted with dichloromethane (100 mL).
The orange
layer was discarded after washed with 0.1 M aqueous HC1(100 mL). The aqueous
layer
was adjusted to pH 7 with solid NaHCO3, extracted with dichloromethane (200
mL).
During extraction, small amount of methanol was added until the color of the
solution
turned to colorless from orange. The extracts was dried over Na2SO4 and
concentrated to
give compound 0806-30 (1.9 g, 76% in two steps) as a yellow solid. LCMS: 431
[M+1]+.
iH NMR (400 MHz, DMSO-d6) 6 2.57 (t, J= 4.4 Hz, 4H), 2.83 (t, J= 5.2 Hz, 4H),
2.90 (s,
3H), 3.14 (t, J = 4.4 Hz, 4H), 3.82 (t, J = 4.4 Hz, 4H), 3.90 (s, 2H), 7.28
(s, 1 H).
Step 60g: Ethyl 5-(4-(2-chloro-6-((4-(methylsulfonyl)piperazin-1-
yl)methyl)thieno[3,2-
d]pyrimidin-4-yl)piperazin-1-yl)pentanoate (Compound 0807-30)
A mixture of compound 0806-30 (450 mg, 1.04 mmol), ethyl 5-bromopentanoate
(327 mg, 1.6 mmol), and triethylamine (211 mg, 2.1 mmol) in DMF (8 mL) was
stirred for
2 h at 80 C. The mixture was poured into water (100 mL) with stirring. The
solid was
collected by filtration and purified by column chromatography on silica gel
(methanol in
dichloromethane, 2% v/v) to give compound 0807-30 (500 mg, 86 %) as a light
yellow
solid. LCMS: 560 [M+1]+. 1H NMR (400 MHz, DMSO-d6) 6 1.18 (t, J= 7.2 Hz, 3H),
1.46
(m, 2H), 1.56 (m, 2H), 2.32 (m, 4H), 2.46 (m, 4H), 2.57 (m, 4H), 2.90 (s, 3H),
3.14 (m,
4H), 3.88 (m, 6H), 4.05 (q, J=7.6 Hz, 2H), 7.29 (s, 1H).
Step 60h: Ethyl 5-(4-(2-(1H-indazol-4-yl)-6-((4-(methylsulfonyl)piperazin-l-
yl)methyl)thieno[3,2-d]pyrimidin-4-yl)piperazin-1-yl)pentanoate (Compound 0808-
30)
The title compound, 0808-30 was prepared (250 mg, 73%) as a yellow solid from
0807-30 (300 mg, 0.54 mmol), compound 0107-3 (157 mg, 0.64 mmol), NaHCO3 (135
mg,
1.61 mmol), (Ph3P)2PdC12 (19 mg, 0.03 mmol) in toluene (5 mL), ethanol (3 mL),
and
water (1.3 mL) using a procedure similar to that described for compound 0603-
107
(Example 30). LCMS: 641 [M+1]+. 1H NMR (400 MHz, DMSO-d6) 6 1.19 (t, J= 7.2
Hz,
3H), 1.51(m, 2H), 1.56(m, 2H), 2.33 (m, 4H), 2.58 (m, 8H), 2.91 (s, 3H), 3.17
(m, 4H),
3.94 (m, 2H), 4.06 (m, 6H), 7.47 (m, 2H), 7.66 (d, J = 8.4 Hz, 1 H). 8.21 (d,
J = 7.2 Hz,
I H), 8.88 (s, I H), 13.20 (s, I H).
Step 60i: 5-(4-(2-(1H-indazol-4-yl)-6-((4-(methylsulfonyl)piperazin-l-
yl)methyl)thieno[3,2-d]pyrimidin-4-yl)piperazin-1-yl)-N-hydroxypentanamide
(Compound
30)
The title compound 30 was prepared (95 mg, 39%) as a white solid from 0808-30
(250 mg, 0.39 mmol) and freshly prepared hydroxylamine methanol solution (6
mL) using
a procedure similar to that described for compound 3 (Example 1). mp 142-145
C. LCMS:
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628 [M+1]. 'H NMR (400 MHz, DMSO-d6) 6 1.53 (m, 4H), 1.99 (t, J= 6.8 Hz, 2H)
2.35
(t, J= 6.8 Hz, 2H), 2.59 (m, 8H), 2.91 (s, 3H), 3.17 (m, 4H), 3.94 (s, 2H),
4.02(m, 4H),
7.47 (m, 2H), 7.66 (d, J= 8.0 Hz, I H), 8.16 (s, I H), 8.22(d, J= 6.8 Hz, I
H), 8.88 (s, I H),
10.36 (s, 1H), 13.20 (s, 1H).
EXAMPLE 61: Preparation of 6-(4-(2-(1H-indazol-4-yl)-6-((4-
(methylsulfonyl)piperazin-1-yl)methyl)thieno [3,2-d] pyrimidin-4-yl)piperazin-
l-yl)-N-
hydroxyhexanamide (Compound 31)
Step 61a: Ethyl 6-(4-(2-chloro-6-((4-(methylsulfonyl)piperazin-1-
yl)methyl)thieno[3,2-
d]pyrimidin-4-yl)piperazin-l-yl)hexanoate (Compound 0807-31)
The title compound, 0807-31 was prepared (290 mg, 44%) as a yellow solid from
0806 (400 mg, 0.928 mmol), ethyl 6-bromohexanoate (310 mg, 1.39 mmol), and
triethylamine (187 mg, 1.86 mmol) using a procedure similar to that described
for
compound 0807-30 (Example 60). LCMS: 573 [M+1 ]+. 'H NMR (400 MHz, DMSO-d6) 65
1.18 (t, J= 7.2 Hz, 3H), 1.29 (m, 2H), 1.45 (m, 2H), 1.54 (m, 2H), 2.29 (m,
4H), 2.50 (m,
4H), 2.57 (m, 4H), 2.90 (s, 3H), 3.14 (m, 4H), 3.88 (m, 6H), 4.05 (q, J=7.2
Hz, 2H), 7.29
(s, 1H).
Step 61b: Ethyl6-(4-(2-(1H-indazol-4-yl)-6-((4-(methylsulfonyl)piperazin-l-
yl)methyl)thieno[3,2-d]pyrimidin-4-yl)piperazin-1-yl)hexanoate (Compound 0808-
31)
The title compound, 0808-31 was prepared (300 mg, 91%) as a yellow solid from
0807-31 (290 mg, 0.506 mmol), compound 0107-3 (185 mg, 0.607 mmol), NaHCO3
(128
mg, 1.52 mmol), (Ph3P)2PdC12 (18 mg, 0.025 mmol) in toluene (5 mL), ethanol (3
mL) and
water (1.3 mL) using a procedure similar to that described for compound 0603-
107
(Example 30). LCMS: 655 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.17 (t, J= 6.8
Hz,
3H), 1.20 (m, 2H), 1.52 (m, 4H), 2.32 (m, 4H), 2.58 (m, 8H), 2.91 (s, 3H),
3.17 (m, 4H),
3.95 (m, 2H), 4.01 (m, 4H), 4.05 (q, J= 6.8 Hz, 2H), 7.48 (m, 2H), 7.66 (d, J=
8.0 Hz,
1H). 8.21 (d, J= 6.8 Hz, 1H), 8.88 (s, 1H), 13.20 (s, 1H).
Step 61c: 6-(4-(2-(1H-indazol-4-yl)-6-((4-(methylsulfonyl)piperazin-l-
yl)methyl)thieno[3,2-d]pyrimidin-4-yl)piperazin-1-yl)-N-hydroxyhexanamide
(Compound
31)
The title compound 31 was prepared (41 mg, 14%) as a white solid from 0808-31
(300 mg, 0.46 mmol) and freshly prepared hydroxylamine methanol solution (5
mL) using
a procedure similar to that described for compound 3 (Example 1). m.p. 140-145
C.
LCMS: 642 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 61.28 (m, 2H), 1.51 (m, 4H), 1.96
(t,
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J= 7.2 Hz, 2H) 2.33 (t, J= 7.2 Hz, 2H), 2.60 (m, 8H), 2.91 (s, 3H), 3.17 (m,
4H), 3.94 (s,
2H), 4.01 (m, 4H), 7.47 (m, 2H), 7.66 (d, J = 8.0 Hz, 1 H). 8.21 (d, J = 7.2
Hz, 1 H), 8.68 (s,
I H), 8.88 (s, I H), 10.35 (s, I H), 13.20 (s, I H).
EXAMPLE 62: Preparation of 7-(4-(2-(1H-indazol-4-yl)-6-((4-
(methylsulfonyl)piperazin-1-yl)methyl)thieno [3,2-d] pyrimidin-4-yl)piperazin-
l-yl)-N-
hydroxyheptanamide (compound 32)
Step 62a: Ethyl 7-(4-(2-chloro-6-((4-(methylsulfonyl)piperazin-1-
yl)methyl)thieno[3,2-
d]pyrimidin-4-yl)piperazin-1-yl)heptanoate (Compound 0807-32)
The title compound, 0807-32 was prepared (550 mg, 81 %) as a yellow solid from
0806 (500 mg, 1.16 mmol), ethyl 7-bromoheptanoate (412 mg, 1.74 mmol),
triethylamine
(235 mg, 2.3 mmol) using a procedure similar to that described for compound
0807-30
(Example 60). LCMS: 587 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.18 (t, J= 7.2
Hz,
3H), 1.28(m, 4H), 1.45 (m, 2H), 1.52 (m, 2H), 2.29(m, 4H), 2.46 (m, 4H), 2.57
(m, 4H),
2.90 (s, 3H), 3.14 (m, 4H), 3.89 (m, 6H), 4.05 (q, J=7.2 Hz, 2H), 7.29 (s,
1H).
Step 62b: Ethyl7-(4-(2-(1H-indazol-4-yl)-6-((4-(methylsulfonyl)piperazin-l-
yl)methyl)thieno[3,2-d]pyrimidin-4-yl)piperazin-1-yl)heptanoate (Compound 0808-
32)
The title compound, 0808-32 was prepared (220 mg, 63%) as a yellow solid from
0807-32 (300 mg, 0.51 mmol), compound 0107-3 (150 mg, 0.61 mmol), NaHCO3 (129
mg,
1.53 mmol), (Ph3P)2PdC12 (18 mg, 0.026 mmol) in toluene (5 mL), ethanol (3
mL), and
water (1.3 mL) using a procedure similar to that described for compound 0603-
107
(Example 30). LCMS: 669 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.18 (t, J= 7.2
Hz,
3H),1.30(m, 4H), 1.52(m, 2H), 1.55(m, 2H), 2.30 (m, 4H), 2.57 (m, 8H), 2.91
(s, 3H), 3.17
(m, 4H), 3.94 (m, 2H), 4.06 (m, 6H), 7.47 (m, 2H), 7.66 (d, J = 8.4 Hz, 1 H).
8.21 (d, J = 7.2
Hz, 1 H), 8.88 (s, 1 H), 13.19 (s, 1 H).
Step 62c: 7-(4-(2-(1H-indazol-4-yl)-6-((4-(methylsulfonyl)piperazin-l-
yl)methyl)thieno[3,2-d]pyrimidin-4-yl)piperazin-1-yl)-N-hydroxyheptanamide
(Compound
32)
The title compound 32 was prepared (80 mg, 37%) as a white solid from 0808-32
(220 mg, 0.33 mmol) and freshly prepared hydroxylamine methanol solution (6
mL) using
a procedure similar to that described for compound 3 (Example 1). mp 131-134
C. LCMS:
656 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.29 (m, 4H), 1.49 (m, 4H), 1.95 (t,
J= 6.8
Hz, 2H), 2.35 (t, J= 6.4 Hz, 2H), 2.60 (m, 8H), 2.91 (s, 3H), 3.17(m, 4H),
3.95 (m, 6H),
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7.47 (m, 2H), 7.66 (d, J= 8.0 Hz, I H), 8.16 (s, I H), 8.22(d, J= 7.2 Hz, I
H), 8.88 (s, I H),
10.35 (s, 1H), 13.20 (s, 1H).
Example 63: Preparation of 2-(((2-(6-acetamidopyridin-3-yl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 112)
Step 63a: N-(5-bromopyridin-2-yl)acetamide (Compound 0601-112)
To a solution of 2-amino-5-bromopyridine (0.50 g, 2.9 mmol) in THE (10 mL) was
added pyridine (343 mg, 4.3 mmol) and acetic anhydride (295 mg, 2.9 mmol) at
room
temperature and stirred overnight. To the mixture water (30 mL) was added,
extracted with
ethyl acetate (3 x 30 mL). The combined organic layers were washed with
saturated
aqueous NaHCO3 and brine, dried over Na2SO4, concentrated to give the title
compound
(0.58 g, 92%) as a white solid LCMS: 215 [M+2]+; 'HNMR (400 MHz, DMSO-d6) 6
2.09
(s, 3H), 7.97 (dd, J= 8.8, 2.4 Hz, 1H), 8.06 (d, J= 8.8 Hz, 1H), 8.41 (d, J=
1.2 Hz, 1H),
10.64 (s, 1H).
Step 63b: N-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-
yl)acetamide
(compound 0602-112)
The title compound 0602-112 was prepared (400 mg, 57%) as a yellow solid from
0601-112 (0.57 g, 2.7 mmol), bis(pinacolato)diboron (1.00 g, 4.0 mmol),
potassium acetate
(0.78 g, 8.0 mmol), and PdC12(dppf)2 (217 mg, 0.3 mmol) using a procedure
similar to that
described for compound 0602-107 (Example 34). LCMS: 263 [M+1]+; 'HNMR (400
MHz,
DMSO-d6) 6 1.30 (s, 12H), 2.10 (s, 3H), 7.96 (dd, J= 8.4, 1.6 Hz, 1H), 8.09
(d, J= 8.4 Hz,
1 H), 8.51(d, J = 0.4 Hz, 1 H), 10.65 (s, 1 H).
Step 63c: Ethyl 2-(((2-(6-acetamidopyridin-3-yl)-4- morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (compound 0603-112)
The title compound 0603-112 was prepared (120 mg, 73%) as a off-white solid
from 0504-54 (135 mg, 0.30 mmol), 0602-112 (157 mg, 0.60 mmol), NaHCO3 (76 mg,
0.90 mmol), (Ph3P)2PdC12 (11 mg, 0.015 mmol) in toluene (2.5 mL), ethanol (1.6
mL) and
water (0.7 mL) using a procedure similar to that described for compound 0603-
107
(Example 34). LCMS: 549 [M+1]+; 'HNMR (400 MHz, CDC13) 6 1.38 (t, J= 7.2 Hz,
3H),
2.25 (s, 3H), 3.31 (s, 3H), 3.87 (t, J= 4.4 Hz, 4H), 4.01 (t, J= 4.4 Hz, 4H),
4.36 (q, J= 7.2
Hz, 2H), 5.20 (s, 2H), 7.38 (s, 1H), 8.28 (m, 2H), 8.70 (dd, J= 8.8, 2.0 Hz,
1H), 9.28 (d, J
= 2.0 Hz, 1H).
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Step 63d: 2-(((2-(6-Acetamidopyridin-3-yl)-4-morpholinothieno [3,2-d]pyrimidin-
6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (compound 112)
The title compound 112 was prepared (20 mg, 19%) as a yellow solid from 0603-
112 (108 mg, 0.20 mmol) and freshly prepared hydroxylamine methanol solution
(10 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.:
197-201 C.
LCMS: 536 [M+l]+; 'HNMR (400 MHz, DMSO-d6) 6 2.13 (s, 3H), 3.24 (s, 3H), 3.76
(t, J
= 4.4 Hz, 4H), 3.93 (t, J = 4.4 Hz, 4H), 5.21 (s, 2H), 7.46 (s, 1 H), 8.18 (d,
J = 8.8 Hz, 1 H),
8.64 (d, J = 1.2 Hz, 1 H), 8.75 (s, 2H), 9.24 (d, J = 2.0 Hz, 1 H), 10.72 (s,
1 H).
Example 64: preparation of 2-(((2-(6-(dimethylamino)pyridin-3-yl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)-N-
hydroxypyrimidine-5-carboxamide (Compound 114)
Step 64a: 5-Bromo-N,N-dimethylpyridin-2-amine (cCompound 0601-114)
To a solution of 5-bromopyridin-2-amine (1.0 g, 5.8 mmol) in THE (25 mL) was
added NaH (0.92 g, 23.1 mmol) at 0 C and stirred for 10 min. followed by the
addition of
CH3I (1 mL, 16 mmol) and stirred forl h. Water (30 mL) was added and extracted
with
ethyl acetate (3 x 30 mL). The combined organic layers were washed with brine,
dried over
Na2SO4, concentrated and purified by column chromatography on silica gel
(ethyl acetate
in petroleum ether, 10% v/v) to give the title compound (1.1 g, 94%) as a
white solid
LCMS: 203 [M+2]+; 'HNMR (400 MHz, DMSO-d6) 6 2.99 (s, 6H), 6.61 (d, J= 9.6 Hz,
1 H), 7.62 (dd, J = 9.2, 2.8 Hz, 1 H), 8.12 (d, J = 2.4 Hz, 1 H).
Step 64b: N,N-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-
2-amine
(Compound 0602-114)
The title compound 0602-114 was prepared (0.81 g, 67%) as a yellow solid from
0601-114 (1.0 g, 5.0 mmol), bis(pinacolato)diboron (1.90 g, 7.5 mmol),
potassium acetate
(1.46 g, 14.9 mmol), and PdC12(dppf)2 (1.90 g, 7.5 mmol) using a procedure
similar to that
described for compound 0602-107 (Example 34). LCMS: 167 [M-81]+; 'HNMR (400
MHz,
DMSO-d6) 6 1.27 (s, 12H), 3.05 (s, 6H), 6.58 (d, J= 8.8 Hz, 1H), 7.67 (dd, J=
8.4, 1.6 Hz,
1H), 8.32 (d, J= 1.2 Hz, 1H).
Step 64c: Ethyl 2-(((2-(6-(dimethylamino)pyridin-3-yl)-4-morpholinothieno [3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-
114)
The title compound 0603-114 was prepared (98 mg, 61%) as a off-white solid
from
0504-54 (135 mg, 0.30 mmol), 0602-114 (149 mg, 0.60 mmol), NaHCO3 (76 mg, 0.90
mmol), (Ph3P)2PdC12 (11 mg, 0.015 mmol) in toluene (2.5 mL), ethanol (1.6 mL)
and water
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(0.7 mL) using a procedure similar to that described for compound 0603-107
(Example 34).
LCMS: 535 [M+1]+; 'HNMR (400 MHz, CDC13) 6 1.38 (t, J= 7.2 Hz, 3H), 3.18 (s,
6H),
3.30 (s, 3H), 3.84 (t, J= 4.8 Hz, 4H), 3.99 (t, J= 4.8 Hz, 4H), 4.36 (q, J=
7.2 Hz, 2H),
5.18 (s, 2H), 6.58 (d, J= 9.2, 1H), 7.37 (s, 1H), 8.49 (d, J= 7.6 Hz, 1H),
8.93 (s, 2H), 9.25
(d, J = 2.0 Hz, I H).
Step 64d: 2-(((2-(6-(Dimethylamino)pyridin-3-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (compound 114)
The title compound 114 was prepared (75 mg, 82%) as a yellow solid from 0603-
114 (93 mg, 0.17 mmol) and freshly prepared hydroxylamine methanol solution (5
mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.:
190-195 C.
LCMS: 522 [M+1]+; 'HNMR (400 MHz, DMSO-d6) 6 3.10 (s, 6H), 3.23 (s, 3H), 3.74
(s,
4H), 3.89 (s, 4H), 5.19 (s, 2H), 6.89 (d, J= 7.6 Hz, I H), 7.40 (s, I H), 8.37
(d, J= 7.2 Hz,
1 H), 8.75 (s, 2H), 9.09 (s, 2H), 11.14 (d, J = 1.2 Hz, 1 H).
Example 65: preparation of 2-(((2-(2-(aminomethyl)pyrimidin-5-yl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)-N-
hydroxypyrimidine-5-carboxamide (Compound 124)
Step 65a: Tert-butyl (5-bromopyrimidin-2-yl)methylcarbamate (Compound 0601-
124)
A mixture of 5-bromo-2-methylpyrimidine (100 mg, 0.58 mmol), NBS (103 mg,
0.58 mmol), dibenzoyl peroxide (10 mg, 0.04 mmol) in tetrachloromethane (10
mL) was
refluxed for 36 h. The solvent was removed in vacuo and the residue was
purified by
column chromatography on silica gel (ethyl acetate in petroleum ether, I% v/v)
to give 5-
Bromo-2-(bromomethyl)pyrimidine (70 mg, 48%) as a white solid. 'HNMR (400 MHz,
CDC13) 6 4.49 (s, 2H), 8.72 (s, 2H).
A solution of 5-Bromo-2-(bromomethyl)pyrimidine (350 mg, 1.4 mmol) in
methanol (5 mL) was added to aqueous ammonia solution (25-28%, 10 mL) and
stirred for
2 hours at room temperature. The solvent was removed followed by addition of
ethanol (20
mL). The solvent was evaporated and to the residue was added dichloromethane
(10 mL),
triethylamine (379 mg, 3.72 mmol), Boc2O (603 mg, 2.79 mmol), DMAP (22 mg,
0.19
nmmol). The resulting mixture was stirred for 2 hours at room temperature. The
solvent
was removed and purified by column chromatography on silica gel to give 0601-
124 (380
mg, 82%) as a white solid. LCMS: 232 [M-56]+; 'HNMR (400 MHz, DMSO-d6) 6 1.38
(s,
9H), 4.28 (d, J= 5.6 Hz, 2H), 7.30 (t, J= 6.4 Hz, 1H), 8.95 (s, 2H).
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Step 65b: Tert-butyl (5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)
pyrimidin-2-
yl)methylcarbamate (Compound 0602-124)
The title compound 0602-124 was prepared (410 mg, 93%) as a yellow oil from
0601-124 (380 mg, 1.3 mmol), bis(pinacolato)diboron (0.50 g, 2.0 mmol),
potassium
acetate (388 mg, 4.0 mmol), and PdC12(dppf)2 (108 mg, 0.1 mmol) using a
procedure
similar to that described for compound 0602-107 (Example 34). LCMS: 198 [M-
137]+;
IHNMR (400 MHz, DMSO-d6) 6 1.32 (s, 12H), 1.39 (s, 9H), 4.33 (d, J= 6.0 Hz,
2H), 7.30
(t, J= 6.0 Hz, 1H), 8.87 (s, 1H).
Step 65c: Ethyl 2-(((2-(2-(aminomethyl)pyrimidin-5-yl)-4-morpholinothieno [3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-
124)
The compound ethyl 2-(((2-(2-((tert-butoxycarbonylamino)methyl) pyrimidin-5-
yl)-
4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxylate
was prepared (248 mg, 66%) as a white solid from 0504-54 (270 mg, 0.6 mmol),
0602-124
(395 mg, 1.2 mmol), NaHCO3 (152 mg, 1.8 mmol), (Ph3P)2PdC12 (21 mg, 0.03 mmol)
in
toluene (5 mL), ethanol (3.2 mL) and water (1.4 mL) using a procedure similar
to that
described for compound 0603-107 (Example 34). LCMS: 622 [M+1]+; 'HNMR (400
MHz,
CDC13) 6 1.39 (t, J= 7.2 Hz, 3H), 1.49 (s, 9H), 3.33 (s, 3H), 3.87 (t, J= 4.4
Hz, 4H), 4.01
(t, J = 4.4 Hz, 4H), 4.37 (q, J = 7.2 Hz, 2H), 4.68 (d, J = 4.4 Hz, 2H), 5.21
(s, 2H), 5.79 (s,
I H), 7.40 (s, I H), 8.94 (s, 2H), 9.62 (s, 2H).
To a solution of above compound (247 mg, 0.4 mmol) in CH2C12 (3 mL) was added
TFA (1.5 mL) and stirred at room temperature for 4 h. Water (50 mL) was added
and
extracted with CH2C12 (200 mL). The organic layer was washed with saturated
aqueous
NaHCO3, brine, dried and concentrated. The residue was recrystallized from
ethyl acetate
and petroleum ether (50% v/v) to give 0603-124 (300 mg, 100%) as an off-white
solid.
LCMS: 522 [M+1]+; 'HNMR (400 MHz, CDC13) 6 1.28 (t, J= 6.8 Hz, 3H), 3.26 (s,
3H),
3.75 (d, J = 4.4 Hz, 4H), 3.94 (d, J = 4.4 Hz, 6H), 4.26 (q, J = 7.2 Hz, 2H),
5.23 (s, 2H),
7.51 (s, 1H), 8.86 (s, 2H), 9.54 (s, 2H).
Step 65d: 2-(((2-(2-(Aminomethyl)pyrimidin-5-yl)-4-morpholinothieno[3,2-d]
pyrimidin-
6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 124)
The title compound 124 was prepared (110 mg, 54%) as a yellow solid from 0603-
124 (300 mg, 0.6 mmol) and freshly prepared hydroxylamine methanol solution
(16 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.:
>300 C.
LCMS: 509 [M+1]+;'HNMR (400 MHz, DMSO-d6) 6 3.17 (s, 6H), 3.17 (s, 3H), 3.74
(s,
4H), 3.92 (s, 4H), 3.95 (s, 2H), 5.19 (s, 2H), 7.48 (s, 1H), 8.73 (s, 2H),
9.53 (s, 2H).
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Example 66: Preparation of N-hydroxy-2-(methyl((4-morpholino-2-
phenylthieno[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 129)
Step 66a: Ethyl 2-(methyl((4-morpholino-2-phenylthieno[3,2-d]pyrimidin-6-yl)
methyl)amino)pyrimidine-5-carboxylate (Compound 0603-129)
The title compound 0603-129 was prepared (250 mg, 64%) as a yellow solid from
0504-54 (358 mg, 0.80 mmol), phenylboronic acid (195 mg, 1.60 mmol), Cs2CO3
(520 mg,
1.60 mmol) and Pd(dppf)2C12 (65 mg, 0.08 mmol) in 1,4-dioxane (6 mL) and water
(0.2
mL) using a procedure similar to that described for compound 0603-107 (Example
34).
LCMS: 491 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 1.30 (t, J= 7.2 Hz, 3H), 3.28
(s,
3H), 3.76 (t, J= 4.8 Hz, 4H), 3.94 (t, J= 4.8 Hz, 4H), 4.29 (q, J= 7.2 Hz,
2H), 5.24 (s, 2H),
7.47- 7.50 (m, 4H), 8.38-8.41 (m, 2H), 8.89 (s, 2H).
Step 66b: N-hydroxy-2-(methyl((4-morpholino-2-phenylthieno[3,2-d]pyrimidin-6-
y1)
methyl)amino)pyrimidine-5-carboxamide (Compound 129)
The title compound 129 was prepared (140 mg, 72%) as a yellow solid from 0603-
129 (200 mg, 0.41 mmol) and freshly prepared hydroxylamine methanol solution
(15 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
199 - 200 C.
LCMS: 478 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 3.23 (s, 3H), 3.76 (s, 4H), 3.93
(s,
4H), 5.20 (s, 2H), 7.47 (m, 4H), 8.39 (m, 2H), 8.75 (s, 2H).
Example 67: Preparation of N-hydroxy-2-(((2-(4-methoxyphenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide (compound 131)
Step 67a: Ethyl 2-(((2-(4-methoxyphenyl)-4-morpholinothieno[3,2-d] pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5 -carboxylate (Compound 0603-131)
The title compound 0603-131 was prepared (230 mg, 66%) as a yellow solid from
0504-54 (300 mg, 0.67 mmol), 4-methoxylphenylboronic acid (204 mg, 1.34 mmol),
Pd(PPh3)2C12 (24 mg, 0.03 mmol), NaHCO3 (202 mg, 2.01 mmol) in toluene (5 mL
),
ethanol (3 mL), and water (1.3 mL) using a procedure similar to that described
for
compound 0603-107 (Example 34). LCMS: 521 [M+1]+. 1H NMR (400 M Hz, DMSO-d6)
6 1.30 (t, J= 7.2 Hz, 3H), 3.27 (s, 1H), 3.75 (t, J= 4.4 Hz, 4H), 3.82 (s,
3H), 3.91 (t, J= 4.6
Hz, 4H), 4.29 (dd, J = 7.2, 7.2 Hz, 2H), 5.22 (s, 2H), 7.02 (d, J = 9.2 Hz,
2H), 7.45 (s, 1 H),
8.33 (d, J= 8.8 Hz, 2H), 8.88 (s, 2H).
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Step 67b: N-hydroxy-2-(((2-(4-methoxyphenyl)-4-morpholinothieno [3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 131)
The title compound 131 was prepared (60 mg, 23%) as a yellow solid from 0603-
131 (230 mg, 0.44 mmol) and freshly prepared hydroxylamine methanol solution
(8 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
247 C
(Decomposed); LCMS: 509 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 3.24 (s, 3H), 3.75-
3.77 (m, 4H), 3.82 (s, 3H), 3.90-3.92 (m, 4H), 5.20 (s, 2H), 7.02 (d, J= 8.4
Hz, 2H), 7.43
(s, 1 H), 8.34 (d, J = 8.4 Hz, 2H), 8.75 (s, 2H), 9.08 (br, 1 H), 11.12 (br, 1
H).
EXAMPLE 68: Preparation of N-hydroxy-2-(((2-(4-hydroxyphenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide (Compound 133)
Step 68a: Ethyl 2-(((2-(4-hydroxyphenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)
methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-133)
The title compound 0603-133 was prepared (170 mg, 50%) as a yellow solid from
0504-54 (300 mg, 0.67 mmol), 4-hydroxylphenylboronic acid (111 mg, 0.802
mmol),
NaHCO3 (168 mg, 2.00 mmol), (Ph3P)2PdC12 (23 mg, 0.0334 mmol) in ethanol (2.3
mL),
toluene (4 mL), and water (1 mL) using a procedure similar to that described
for compound
0603-107 (Example 34). LCMS: 507 [M+1]+. 1H NMR (400 MHz, DMSO-d6) 6 1.30 (t,
J=
6.8 Hz, 3H), 3.27 (s, 3H), 3.75 (m, 4H), 3.90 (m, 4H), 4.29 (q, J= 7.2 Hz,
2H), 5.22 (s,
2H), 6.84 (d, J= 8.8 Hz, 2H), 7.43 (s, 1H), 8.23 (d, J= 8.4 Hz, 2H), 8.88 (s,
2H), 9.81 (s,
I H).
Step 68b: N-hydroxy-2-(((2-(4-hydroxyphenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 133)
The title compound 133 was prepared (69 mg, 42%) as a white solid from 0603-
133
(170 mg, 0.336 mmol) and freshly prepared hydroxylamine methanol solution (10
mL)
using a procedure similar to that described for compound 3 (Example 1). mp.
185-195 C.
LCMS: 494 [M+1]+. 1H NMR (400 MHz, DMSO-d6) 6 3.23 (s, 3H), 3.75 (m, 4H), 3.90
(m,
4H), 5.19 (s, 2H), 6.84 (d, J = 8.4 Hz, 2H), 7.40 (s, 1 H), 8.23 (d, J = 8.4
Hz, 2H), 8.75 (s,
2H), 9.90 (s, 2H).
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Example 69: Preparation of 2-(((2-(4-(acetamidomethyl)phenyl)-4-
morpholinothieno
[3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 136)
Step 69a: 2-(((2-(4-(Acetamidomethyl)phenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 136)
The title compound 136 was prepared (75 mg, 31%) as a white solid from ethyl 2-
(((2-(4-(acetamidomethyl)phenyl)-4-morpholinothieno [3,2-d]pyrimidin-6-yl)
methyl)(methyl)amino)pyrimidine-5-carbo- xylate (250 mg, 0.45 mmol, example
48) and
freshly prepared hydroxylamine methanol solution (20 mL) using a procedure
similar to
that described for compound 3 (Example 1). m.p. 178-180 C. LCMS: 549 [M+1]+;
1H
NMR (400 MHz, DMSO-d6) 6 1.89 (s, 3H), 3.23 (s, 3H), 3.76 (m, 4H), 3.91 (m,
4H),
4.31 (d, J= 5.6 Hz, 2H), 5.20 (s, 2H), 7.34 (d, J=8.0 Hz, 2H), 7.45 (s, 1H),
8.33 (d, J= 8.4
Hz, 2H), 8.41 (t, J= 5.6 Hz, 1H), 8.75 (s, 2H).
EXAMPLE 70: Preparation of 2-(((2-(4-carbamoylphenyl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 143)
Step 70a: 4-Bromobenzamide (Compound 0601-143)
To a solution of 4-bromobenzonitrile (2 g, 10 mol) in DMSO (6 mL) was added
30% H202 (5 g, 13 mmol) and K2CO3 at 0 C and the mixture was stirred at room
temperature for 30 min. The reaction mixture was poured into water and
filtered. The
collected solid was washed with water and dried to get the compound 0601-143
(2.1 g,
96%) as a white solid. LCMS: 200 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 7.48 (s,
1H),
7.67 (d, J= 8.4 Hz, 2H), 7.81 (d, J= 8.4 Hz, 2H), 8.06 (s, 1H).
Step 70b: 4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (Compound
0602-
143)
The title compound 0602-143 was prepared (570 mg, 92%) from 0601-143 (500
mg, 2.5 mmol), bis(pinacolato)diboron (952 mg, 3.75 mmol), potassium acetate
(735 mg,
7.5 mmol), and PdC12(dppf)2 (61 mg, 0.075 mmol) using a procedure similar to
that
described for compound 0602-107 (Example 34). LCMS: 248 [M+1]+; 1H NMR (400
MHz,
DMSO-d6) 6 1.31 (s, 12H), 7.43 (s, 1 H), 7.73 (d, J = 8.4 Hz, 2H), 7.87 (d, J
= 7.6 Hz, 2H),
8.03 (s, 1H).
Step 70c: Ethyl 2-(((2-(4-carbamoylphenyl)-4-morpholinothieno[3,2-d] pyrimidin-
6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-143)
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The title compound 0603-143 was prepared (300 mg, 84%) as a yellow solid from
0504-54 (300 mg, 0.67 mmol), 0602-143 (330 mg, 1.34 mmol), NaHCO3 (168 mg, 2.0
mmol), (Ph3P)2PdC12 (23 mg, 0.03 mmol) in toluene (5 mL), ethanol (3 mL) and
water (1.3
mL) using a procedure similar to that described for compound 0603-107 (Example
34).
LCMS: 534 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.30 (t, J= 6.8Hz, 3H), 3.28 (s,
3H), 3.77 (m, 4H), 3.93 (m, 4H), 4.28 (q, J= 7.2 Hz, 2H), 5.24 (s, 2H), 7.45
(s, 1H), 7.51
(s, 1H), 7.97 (d, J= 8.4 Hz, 2H),8.07 (s, 1H), 8.43 (d, J= 8.4 Hz, 2H), 8.88
(s, 2H).
Step 70d: 2-(((2-(4-Carbamoylphenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 143)
The title compound 143 was prepared (183 mg, 63%) as a yellow solid from 0603-
143 (300 mg, 0.47 mmol) and freshly prepared hydroxylamine methanol solution
(8 mL)
using a procedure similar to that described for compound 3 (Example 1). mp 200-
202 C.
LCMS: 521 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 3.25 (s, 3H), 3.77 (m, 4H), 3.94
(m,
4H), 5.21 (s, 2H), 7.46 (s, 1 H), 7.49 (s, 1 H), 7.97 (d, J = 8.4 Hz, 2H),
8.08 (s, 1 H), 8.44 (d,
J= 8.4 Hz, 2H), 8.76 (s,1 H).
EXAMPLE 71: Preparation of 2-(((2-(4-cyanophenyl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 146)
Step 71a: Ethyl 2-(((2-(4-cyanophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)
methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-146)
The title compound 0603-146 was prepared (400 mg, 70%) as a off-white solid
from 0504-54 (500 mg, 1.1 mmol), 4-cyanophenylboronic acid (245 mg, 1.67
mmol),
NaHCO3 (280 mg, 3.34 mmol), (Ph3P)2PdC12 (39 mg, 0.05 mmol) in toluene (5 mL),
ethanol (3 mL) and water (1.3 mL) using a procedure similar to that described
for
compound 0603-107 (Example 34). LCMS: 516 [M+l]+. 'H NMR (400 MHz, CDC13) 6
1.39 (t, J= 7.6 Hz, 3H), 3.32 (s, 3H), 3.87 (m, 4H), 4.02 (m, 4H), 4.36 (q, J=
7.2 Hz, 2H),
5.21 (s, 2H), 7.39 (s, 1 H), 7.74 (d, J = 8.0 Hz, 2H), 8.53 (d, J = 8.8 Hz,
2H), 8.93 (s, 2H).
Step 71b: 2-(((2-(4-Cyanophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 146)
The title compound 146 was prepared (40 mg, 10%) as an off-white solid from
0603-146 (400 mg, 0.77 mmol) and freshly prepared hydroxylamine methanol
solution (12
mL) using a procedure similar to that described for compound 3 (Example 1). mp
214-216
C. LCMS: 503 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 3.24 (s, 3H), 3.77 (m, 4H),
3.95
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(m, 4H), 5.22 (s, 2H), 7.51 (s, 1H), 7.95 (d, J= 8.4 Hz, 2H), 8.54 (d, J= 8.2
Hz, 2H), 8.75
(s,2H), 9.13 (s, 1H), 11.11 (s, 1H).
Example 72: Preparation of 2-(((2-(4-chlorophenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(compound 147)
Step 72a: Ethyl 2-(((2-(4-chlorophenyl)-4-morpholinothieno [3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-147)
The title compound 0603-147 was prepared (250 mg, 73%) as a yellow solid from
0504-54 (300 mg, 0.67 mmol), 4-chlorophenylboronic acid (209 mg, 1.34 mmol),
Pd(PPh3)2Cl2 (24 mg, 0.03 mmol), NaHCO3 (202 mg, 2.01 mmol) in toluene (5 mL),
ethanol (3mL), and water (1.3 mL) using a procedure similar to that described
for
compound 0603-107 (Example 34). LCMS: 525 [M+1]+. 'H NMR (400 M Hz, DMSO-d6)
6 1.30 (t, J= 7.0 Hz, 3H), 3.27 (s, 3H), 3.76 (t, J= 4.8 Hz, 4H), 3.92 (t, J=
4.8 Hz, 4H),
4.29 (dd, J = 6.8, 7.2 Hz, 2H), 5.24 (s, 2H), 7.50 (s, 1 H), 7.54 (d, J = 8.8
Hz, 2H), 8.40 (d, J
= 8.4 Hz, 2H), 8.88 (s, 2H).
Step 72b: 2-(((2-(4-chlorophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 147)
The title compound 147 was prepared (69 mg, 29%) as a white solid from 0603-
147
(250 mg, 0.48 mmol) and freshly prepared hydroxylamine methanol solution (8
mL) using
a procedure similar to that described for compound 3 (Example 1). mp. 153-153
C; LCMS:
512 [M+1]+; 'H NMR (400 MHz, DMSO-d6) 6 3.24 (s, 3H), 3.75-3.77 (m, 4H), 3.91-
3.93
(m, 4H), 5.21 (s, 2H), 7.47 (s, 1H), 7.54 (d, J= 7.6 Hz, 2H), 8.39 (d, J = 8.0
Hz, 2H), 8.76
(s, 2H), 9.07 (br, 1 H), 11.14 (br, 1 H).
Example 73: Preparation of N-hydroxy-2-(((2-(4-isopropylphenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide (compound 148)
Step 73a: Ethyl 2-(((2-(4-isopropylphenyl)-4-morpholinothieno[3,2-d] pyrimidin-
6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-148)
The title compound 0603-148 was prepared (250 mg, 73%) as a yellow solid from
0504-54 (300 mg, 0.67 mmol), 4-isopropylphenylboronic acid (220 mg, 1.34
mmol),
Pd(PPh3)2C12 (24 mg, 0.03 mmol), NaHCO3 (202 mg, 2.01 mmol) in toluene (5 mL),
ethanol (3mL), and water (1.3 mL) using a procedure similar to that described
for
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compound 0603-107 (Example 34). LCMS: 533 [M+l]+. 'H NMR (400 M Hz, DMSO-d6)
6 1.24 (d, J= 7.2 Hz, 6H), 1.30 (t, J= 7.2 Hz, 3H), 2.90-2.98 (m, 1H), 3.27
(s, 3H), 3.76 (t,
J = 4.6 Hz, 4H), 3.92 (t, J = 4.4 Hz, 4H), 4.29 (dd, J = 7.2, 7.2 Hz, 2H),
5.23 (s, 2H), 7.34
(d, J= 8.0 Hz, 2H), 7.48 (s, 1H), 8.30 (d, J= 8.4 Hz, 2H), 8.88 (s, 2H).
Step 73b: N-hydroxy-2-(((2-(4-isopropylphenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 148)
The title compound 148 was prepared (58 mg, 27%) as a white solid from 0603-
148
(250 mg, 0.47 mmol) and freshly prepared hydroxylamine methanol solution (8
mL) using
a procedure similar to that described for compound 3 (Example 1). mp. 155 C
(decomposed); LCMS: 520 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 1.24 (d, J= 6.4
Hz,
6H), 2.92-2.98 (m, 1H), 3.24 (s, 3H), 3.75-3.77 (m, 4H), 3.91-3.93 (m, 4H),
5.20 (s, 2H),
7.34 (d, J= 8.0 Hz, 2H), 7.46 (s, 1H), 8.30 (d, J = 8.0 Hz, 2H), 8.76 (s, 2H),
9.08 (br, 1H),
11.10 (br, 1H).
Example 74: Preparation of N-hydroxy-2-(methyl((2-(4-(methylsulfonyl)phenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound 149)
Step 74a: Ethyl 2-(methyl((2-(4-(methylsulfonyl)phenyl)-4-morpholinothieno
[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-149)
The title compound 0603-149 was prepared (250 mg, 77%) as a yellow solid from
0504-54 (300 mg, 0.67 mmol), 4-(methanesulfonyl)phenylboronic acid (268 mg,
1.34
mmol), Pd(PPh3)2C12 (24 mg, 0.03 mmol), NaHCO3 (202 mg, 2.01 mmol) in toluene
(5
mL), ethanol (3mL), and water (1.3 mL) using a procedure similar to that
described for
compound 0603-107 (Example 34). LCMS: 569 [M+1]+. 1H NMR (400 M Hz, DMSO-d6)
6 1.30 (t, J= 7.2 Hz, 3H), 3.27 (s, 3H), 3.28 (s, 3H), 3.76-3.78 (m, 4H), 3.95-
3.97 (m, 4H),
4.29 (q, J= 7.2 Hz, 2H), 5.25 (s, 2H), 7.54 (s, 1H), 8.03 (d, J= 8.4 Hz, 2H),
8.61 (d, J= 8.8
Hz, 2H), 8.88 (s, 2H).
Step 74b: N-hydroxy-2-(methyl((2-(4-(methylsulfonyl)phenyl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 149)
The title compound 149 was prepared (160 mg, 65%) as a white solid from 0603-
149 (250 mg, 0.44 mmol) and freshly prepared hydroxylamine methanol solution
(8 mL)
using a procedure similar to that described for compound 3 (Example 1). mp.
206-208 C;
LCMS: 556 [M+1]+; 1H NMR (400 MHz, DMSO-d6) 6 3.24 (s, 3H), 3.26 (s, 3H), 3.77
(t, J
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= 4.4 Hz, 4H), 3.96 (t, J= 4.4 Hz, 4H), 5.22 (s, 2H), 7.52 (s, 1H), 8.03 (d,
J= 8.4 Hz, 2H),
8.61 (d, J = 8.8 Hz, 2H), 8.75 (s, 2H), 9.06 (br, 1 H), 11.13 (br, 1 H).
Example 75: Preparation of 2-(((2-(4-fluorophenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 200)
Step 75a: Ethyl 2-(((2-(4-fluorophenyl)-4-morpholinothieno [3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-200)
The title compound 0603-200 was prepared (300 mg, 76%) as a white solid from
0504-54 (350 mg, 0.78 mmol), 4-fluorophenylboronic acid (164 mg, 1.17 mmol),
Pd(PPh3)2C12 (27 mg, 0.039 mmol), NaHCO3 (196 mg, 2.34 mmol) in toluene (8
mL),
ethanol (5 mL), and water (2 mL) using a procedure similar to that described
for compound
0603-107 (Example 34). LCMS: 509 [M+1]+; 1H NMR (400 MHz, CDC13): 6 1.31 (t,
J=
7.2 Hz, 3H), 3.24 (s, 3H) 3.80 (t, J= 4.8 Hz, 4H), 3.94 (t, J= 4.6 Hz, 4H),
4.30 (q, J= 7.1
Hz, 2H), 5.13 (s, 2H), 7.04-7.08 (m, 2H), 7.30 (s, 1H), 8.34-8.37 (m, 2H),
8.86 (s, 2H).
Step 75b: 2-(((2-(4-fluorophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 200)
The title compound 200 was prepared (240 mg, 82%) as an off-white solid from
0603-200 (300 mg, 0.59 mmol) and freshly prepared hydroxylamine methanol
solution (16
mL) using a procedure similar to that described for compound 3 (Example 1).
m.p. 168-170
C. LCMS: 496 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 5 3.23 (s, 3H), 3.76 (t, J=
4.6
Hz, 4H), 3.92 (t, J= 4.4 Hz, 4H), 5.20 (s, 2H), 7.28-7.32 (m, 2H), 7.46 (s,
1H), 8.41-8.45
(m, 2H), 8.74 (s, 2H), 9.05 (s, 1 H), 11.12 (s, 1 H).
Example 76: Preparation of N-hydroxy-2-(methyl((4-morpholino-2-p-
tolylthieno[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 201)
Step 76a: Ethyl 2-(methyl((4-morpholino-2-p-tolylthieno[3,2-d] pyrimidin-6-
yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-201)
The title compound 0603-201 was prepared (306 mg, 78%) as a white solid from
0504-54 (350 mg, 0.78 mmol), 4-methylphenylboronic acid (212 mg, 1.56 mmol),
Pd(PPh3)2C12 (27 mg, 0.039 mmol), NaHCO3 (196 mg, 2.34 mmol) in toluene (8
mL),
ethanol (5 mL), and water (2 mL) using a procedure similar to that described
for compound
0603-107 (Example 34). LCMS: 505 [M+1]+; 1H NMR (400 MHz, CDC13): 6 1.31 (t,
J=
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7.0 Hz, 3H), 2.34 (s, 3H), 3.23 (s, 3H), 3.79 (t, J= 4.8 Hz, 4H), 3.93 (t, J=
4.8 Hz, 4H),
4.29 (q, J = 7.2 Hz, 2H), 5.12 (s, 2H), 7.19 (d, J = 8.0 Hz, 2H), 7.31 (s, 1
H), 8.24 (d, J = 8.0
Hz, 2H), 8.86 (s, 2H).
Step 76b: N-Hydroxy-2-(methyl((4-morpholino-2-p-tolylthieno[3,2-d] pyrimidin-6-
yl)methyl)amino)pyrimidine-5-carboxamide (Compound 201)
The title compound 201 was prepared (27 mg, 9%) as a off-white solid from 0603-
201 (306 mg, 0.61 mmol) and freshly prepared hydroxylamine methanol solution
(16 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.:
170-172 C.
LCMS: 492 [M+1]; 'H NMR (400 MHz, DMSO-d6): 5 2.35 (s, 3H), 3.22 (s, 3H), 3.75
(m, 4H), 3.90 (m, 4H), 5.19 (s, 2H), 7.27 (d, J = 7.6 Hz, 2H), 7.43 (s, 1 H),
8.27 (d, J = 8.4
Hz, 2H), 8.73 (s, 2H).
Example 77: Preparation of N-hydroxy-2-(methyl((4-morpholino-2-(4-
(trifluoromethyl)phenyl)thieno [3,2-d] pyrimidin-6-yl)methyl)amino)pyrimidine-
5-
carboxamide (Compound 202)
Step 77a: Ethyl 2-(methyl((4-morpholino-2-(4-(trifluoromethyl)phenyl)thieno
[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-202)
The title compound 0603-202 was prepared (527 mg, crude) as a white solid from
0504-54 (350 mg, 0.78 mmol), 4-trifluoromethylphenylboronic acid (296 mg, 1.56
mmol),
Pd(PPh3)2C12 (27 mg, 0.039 mmol), NaHCO3 (196 mg, 2.34 mmol) in toluene (8
mL),
ethanol (5 mL), and water (2 mL) using a procedure similar to that described
for compound
0603-107 (Example 34). LCMS: 559 [M+1]+; 1H NMR (400 MHz, CDC13): 6 1.32 (t,
J=
7.0 Hz, 3H), 3.24 (s, 3H), 3.81 (t, J= 4.8 Hz, 4H), 3.96 (t, J= 4.6 Hz, 4H),
4.30 (q, J= 7.1
Hz, 2H), 5.12 (s, 2H), 7.34 (s, 1 H), 7.63 (d, J = 8.4 Hz, 2H), 8.45 (d, J =
8.0 Hz, 2H), 8.86
(s, 2H).
Step 77b: N-Hydroxy-2-(methyl((4-morpholino-2-(4-(trifluoromethyl)phenyl)
thieno[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 202)
The title compound 202 was prepared (162 mg, 38%) as anoff-white solid from
0603-202 (527 mg, crude) and freshly prepared hydroxylamine methanol solution
(16 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
222-223 C.
LCMS: 546 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 63.23 (s, 3H), 3.76 (t, J= 4.4
Hz,
4H), 3.94 (t, J = 4.2 Hz, 4H), 5.21 (s, 2H), 7.50 (s, 1 H), 7.84 (d, J = 8.8
Hz, 2H), 8.57 (d, J
= 8.4 Hz, 2H), 8.74 (s, 2H), 9.06 (s, 1 H), 11.13 (s, 1 H).
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Example 78: Preparation of N-hydroxy-2-(methyl((2-(4-(methylamino) phenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound 151)
Step 78a: 4-Bromo-N-methylbenzenamine (Compound 0601-15 1) and 4-bromo-N,N-
dimethylbenzenamine (Compound 0601-152)
To a solution of 4-bromobenzenamine (3 g, 17.4 mmol) and K2C03 (3.62 g, 26.2
mmol) in THE (30 mL) was added methyl iodide (2.2 ml, 34.8 mmol) at room
temperature
and stirred overnight. Water (6 mL) was added to the reaction mixture and the
mixture was
extracted with ethyl acetate (50 mL X 3). The organic layer was dried,
concentrated and
purified by column chromatography on silica gel (ethyl acetate in petroleum
ether, 10%
v/v) to give 0601-151 (840 mg, 26%) as a white solid LCMS: 187 [M+1]+. 'H NMR
(400
MHz, DMSO-d6) 6 2.63 (s, 3H), 5.85 (br, 1H), 6.48 (d, J= 9.2 Hz, 2H), 7.20 (d,
J= 8.4 Hz,
2H) and 0601-152 (680 mg, 20%) as a white solid. LCMS: 201 [M+1]+. 'H NMR (400
MHz, DMSO-d6) 6 2.87 (s, 6H), 6.65 (d, J= 9.2 Hz, 2H), 7.29 (d, J= 8.8 Hz,
2H).
Step 78b: N-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenamine
(Compound 0602-151)
The title compound 0602-151 was prepared (301 mg, 60%) as a white solid from
0601-151 (400 mg, 2.15 mmol), bis(pinacolato)diboron (819 mg, 3.23 mmol),
potassium
acetate (632 mg, 6.4 mmol) and Pd(dppf)2C12 (351 mg, 0.43 mmol) using a
procedure
similar to that described for compound 0602-107 (Example 34). LCMS: 234
[M+1]+. 'H
NMR (400 MHz, DMSO-d6) 6 1.24 (s, 12H), 2.67 (d, J= 5.2 Hz, 3H), 6.04 (br,
1H), 6.48
(d, J= 8.4 Hz, 2H), 7.39 (d, J= 8.4 Hz, 2H).
Step 78c: Ethyl 2-(methyl((2-(4-(methylamino)phenyl)-4- morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5 -carboxylate (Compound 0603-151)
The title compound 0603-151 was prepared (100 mg, 64%) as a white solid from
0504-54 (135 mg, 0.3 mmol), 0602-151 (100 mg, 0.42 mmol), Pd(PPh3)2C12 (10.5
mg,
0.015 mmol), NaHCO3 (76 mg, 0.9 mmol) in toluene (2.5 mL), ethanol (1.6 mL),
and water
(0.7 mL) using a procedure similar to that described for compound 0603-107
(Example 34).
LCMS: 520 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.30 (t, J= 7.2 Hz, 3H), 2.73
(d, J=
4.8 Hz, 3H), 3.26 (s, 3H), 3.75 (m, 4H), 3.88 (m, 4H), 4.29 (q, J= 7.2 Hz,
2H), 5.21 (s,
2H), 6.10 (m, 1 H), 6.5 8 (d, J = 8.8 Hz, 2H), 7.3 9 (s, 1 H), 8.15 (d, J =
8.4 Hz, 2H), 8.8 8 (d,
J = 4.8 Hz, 2H).
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Step 78d: N-hydroxy-2-(methyl((2-(4-(methylamino)phenyl)-4-morpholinothieno
[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 151)
The title compound 151 was prepared (40 mg, 42%) as a yellow solid from 0603-
151 (100 mg, 0.19 mmol) and freshly prepared hydroxylamine methanol solution
(6 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.:
218-220 C.
LCMS: 507 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 2.73 (d, J= 4.4 Hz, 3H), 3.75
(d, J
= 4.0 Hz, 4H), 3.87 (d, J = 4.0 Hz, 4H), 5.18 (s, 2H), 6.11 (d, J = 4.8 Hz, 1
H), 6.5 8 (d, J =
8.0 Hz, 2H), 7.37 (s, 1H), 8.15 (d, J= 8.4 Hz, 2H), 8.75 (s, 2H), 9.06 (s,
1H), 11.13 (br,
I H).
Example 79: Preparation of 2-(((2-(4-(dimethylamino)phenyl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)-N-
hydroxypyrimidine-5-carboxamide (Compound 152)
Step 79a: N,N-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
yl)benzenamine
(Compound 0602-152)
The title compound 0602-152 was prepared (360 mg, 61%) as a white solid from
0601-152 (480 mg, 2.4 mmol), bis(pinacolato)diboron (914 mg, 3.6 mmol),
potassium
acetate (706 mg, 7.2 mmol) and Pd(dppf)2C12 (390 mg, 0.48 mmol) using a
procedure
similar to that described for compound 0602-107 (Example 34). LCMS: 248
[M+1]+. 'H
NMR (400 MHz, DMSO-d6) 6 1.25 (s, 12H), 2.93 (s, 6H), 6.66 (d, J= 8.8 Hz, 2H),
7.48 (d,
J = 8.8 Hz, 2H).
Step 79b: Ethyl 2-(((2-(4-(dimethylamino)phenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-152)
The title compound 0603-152 was prepared (100 mg, 53%) as a white solid from
0504-54 (161 mg, 0.36 mmol), 0602-152 (150 mg, 0.61 mmol), Pd(PPh3)2C12 (13
mg,
0.018 mmol), NaHCO3 (91 mg, 1.08 mmol) in toluene (2.5 mL), ethanol (1.6 mL),
and
water (0.7 mL) using a procedure similar to that described for compound 0603-
107
(Example 34). LCMS: 534 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.22 (m, 3H), 2.97
(s, 6H), 3.22 (s, 3H), 3.74 (m, 4H), 3.87 (m, 4H), 4.14 (m, 2H), 5.17 (s, 2H),
6.75 (d, J=
8.8 Hz, 2H), 7.37 (s, 1H), 8.20 (d, J= 9.2 Hz, 2H), 8.73 (s, 2H).
Step 79c: 2-(((2-(4-(dimethylamino)phenyl)-4-morpholinothieno[3,2-d] pyrimidin-
6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 152)
The title compound 152 was prepared (30 mg, 30%) as a yellow solid from 0603-
152 (100 mg, 0.19 mmol) and freshly prepared hydroxylamine methanol solution
(6 mL)
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using a procedure similar to that described for compound 3 (Example 1). m.p.:
208-2 10 C.
LCMS: 521 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 2.98 (s, 6H), 3.23 (s, 3H), 3.76
(d, J
= 4.0 Hz, 4H), 3.89 (s, 4H), 5.18 (s, 2H), 6.76 (d, J = 8.8 Hz, 2H), 7.39 (s,
1 H), 8.22 (d, J =
8.4 Hz, 2H), 8.75 (s, 2H), 9.07 (br, 1H).
Example 80: Preparation of 2-(((2-(4-(ethylamino)phenyl)-4-
morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 153)
Step 80a: 4-Bromo-N-ethylbenzenamine (Compound 0601-153)
To a solution of 4-bromobenzenamine (2.00 g, 11.67 mmol) and iodoethane (5.50
g,
35.28 mmol) in CH3CN (50 mL) was added K2C03 (6.48 g, 47.04 mmol). The mixture
was
stirred at 60 C overnight. The reaction mixture was concentrated in vacuo and
the residue
was purified by column chromatography on silica gel (ethyl acetate in
petroleum ether, 0%-
10% v/v) to give compound 0601-153 (800 mg, 38%) as a light yellow oil. LCMS:
200
[M+1]+;'H NMR (400 MHz, DMSO-d6): 6S 1.13 (t, J= 6.8 Hz, 3H), 2.97 (m, 2H),
5.73 (br
s, 1H), 6.48(d, J= 8.8 Hz, 2H), 7.17 (d, J= 8.8 Hz, 2H).
Step 80b: N-ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenamine
(Compound 0602-153)
The title compound 0602-153 was prepared (640 mg, 72%) as a white solid from
0601-153 (884 mg, 4.420 mmol), bis(pinacolato)diboron (1.68 g, 6.630 mmol),
potassium
acetate (1.30 g, 13.3mmol) and Pd(dppf)2C12 (362 mg, 0.44 mmol) using a
procedure
similar to that described for compound 0602-107 (Example 34). LCMS: 248
[M+1]+; 'H
NMR (400 MHz, DMSO-d6): 6 1.13 (m, 15H), 3.03 (m, 2H), 5.94 (br s, 1H), 6.49
(d, J=
8.4 Hz, 2H), 7.37 (d, J= 8.8 Hz, 2H).
Step 80c: Ethyl 2-(((2-(4-(ethylamino)phenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-153)
The title compound 0603-153 was prepared (200 mg, 71%) as a white solid from
0504-54 (240 mg, 0.53 mmol), 0602-153 (172 mg, 0.69 mmol), Pd(PPh3)2C12 (37
mg,
0.053 mmol), NaHCO3 (134 mg, 1.6 mmol) in toluene (8 mL), ethanol (5 mL), and
water (2
mL) using a procedure similar to that described for compound 0603-107 (Example
34).
LCMS: 534 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 1.17 (t, J= 7.2 Hz, 3H), 2.29
(t, J =
7.6 Hz, 3H), 3.09 (m, 2H), 3.25 (s, 3H), 3.73 (br s, 4H), 3.87 (br s, 4H),
4.27 (m, 2H), 5.20
(s, 2H), 6.02 (t, J = 4.8 Hz, 1 H), 6.5 8 (d, J = 8.8 Hz, 2H), 7.3 7(s, 1 H),
8.12 (d, J = 8.8 Hz,
2H), 8.87 (s, 2H). P49
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Step 80d: 2-(((2-(4-(Ethylamino)phenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
y1)
methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 153)
The title compound 153 was prepared (110 mg, 56%) as an off-white solid from
0603-153 (200 mg, 0.38 mmol) and freshly prepared hydroxylamine methanol
solution (8
mL) using a procedure similar to that described for compound 3 (Example 1).
m.p.: 185-
187 C; LCMS: 521 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 1.18 (t, J= 6.8 Hz,
3H),
3.09 (m, 2H), 3.23 (s, 3H), 3.75 (br s, 4H), 3.87 (br s, 4H), 5.18 (s, 2H),
6.02 (t, J= 4.8 Hz,
1H), 6.59 (d, J= 8.8 Hz, 2H), 7.36 (s, 1H), 8.13 (d, J= 8.4 Hz, 2H), 8.74 (s,
2H), 9.06 (s,
1 H), 11.13 (s, 1 H).
Example 81: Preparation of 2-(((2-(4-(Diethylamino)phenyl)-4-
morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 154)
Step 81a: 4-Bromo-N,N-diethylbenzenamine (Compound 0601-154)
The title compound 0601-154 (1.10 g, 40%) was synthesized according to the
synthetic procedure of making compound 0601-153 using 4-bromobenzenamine (2.00
g,
11.67 mmol) and iodoethane (5.50 g, 35.28 mmol) as starting material. LCMS:
228
[M+1]+;'H NMR (400 MHz, DMSO-d6): 6S 1.05 (t, J= 7.2 Hz, 6H), 3.30 (m, 4H),
6.58 (d,
J = 8.8 Hz, 2H), 7.22 (d, J = 8.8 Hz, 2H).
Step 81b: N,N-diethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
yl)benzenamine
(Compound 0602-154)
The title compound 0602-154 was prepared (860 mg, 63%) as a white solid from
0601-154 (1.21 g, 5.33 mmol), bis(pinacolato)diboron (2.03 g, 8.00 mmol),
potassium
acetate and Pd(dppf)2C12 using a procedure similar to that described for
compound 0602-
107 (Example 34). LCMS: 276 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 1.06 (t, J=
6.8
Hz, 6H), 1.24 (s, 12H), 3.34 (m, 4H), 6.60 (d, J= 8.4 Hz, 2H), 7.44 (d, J= 8.8
Hz, 2H).
Step 81c: Ethyl 2-(((2-(4-(diethylamino)phenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-154)
The title compound 0603-154 was prepared (260 mg, 85 %) as a white solid from
0504-54 (240 mg, 0.53 mmol), 0602-154 (220 mg, 0.8 mmol), Pd(PPh3)2C12, NaHCO3
in
toluene (8 mL), ethanol (5 mL), and water (2 mL) using a procedure similar to
that
described for compound 0603-107 (Example 34). LCMS: 562 [M+1]+; 1H NMR (400
MHz,
DMSO-d6): 6 1.12 (t, J= 7.2 Hz, 6H), 1.29 (t, J= 6.8 Hz, 3H), 3.25 (s, 3H),
3.40 (m, 4H),
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3.74 (br s, 4H), 3.87 (br s, 4H), 4.27 (m, 2H), 5.20 (s, 2H), 6.69 (d, J= 8.8
Hz, 2H), 7.39 (s,
1H), 8.18 (d, J= 8.8 Hz, 2H), 8.87 (s, 2H).
Step 81d: 2-(((2-(4-(Diethylamino)phenyl)-4-morpholinothieno[3,2-d] pyrimidin-
6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 154)
The title compound 154 was prepared (180 mg, 71%) as an off-white solid from
0603-154 (260 g, 0.463 mmol) and freshly prepared hydroxylamine methanol
solution (8
mL) using a procedure similar to that described for compound 3 (Example 1).
m.p.: 192-
196 C; LCMS: 549 [M+1]+; 'H NMR (400 MHz, DMSO-d6): 6 1.12 (t, J= 7.2 Hz,
6H),
3.23 (s, 3H), 3.41 (m, 4H), 3.75 (br s, 4H), 3.87 (br s, 4H), 5.18 (s, 2H),
6.70 (d, J= 8.8 Hz,
2H), 7.3 7 (s, 1 H), 8.19 (d, J = 8.8 Hz, 2H), 8.74 (s, 2H), 9.06 (s, 1 H),
11.11 (s, 1 H).
EXAMPLE 82: Preparation of N-hydroxy-2-(methyl((4-morpholino-2-(4-(pyrrolidin-
1-yl)phenyl)thieno [3,2-d] pyrimidin-6-yl)methyl)amino)pyrimidine-5-
carboxamide
(Compound 155)
Step 82a: 1-(4-Bromophenyl)pyrrolidine (Compound 0601-155)
A mixture of 4-bromoaniline (1 g, 5.81 mmol), Cs2CO3 (5.68 g, 17.44 mmol), 1,4-
dibromobutane (1.88 g, 8.72 mmol) in DMF (20 mL) was stirred at 60 C
overnight. After
cooled to room temperature, the mixture was diluted with water (200 mL) and
extracted
with ethyl acetate (2 x 100 mL). The organic layer was washed with water (3 x
100 mL)
and brine (100 mL), dried over Na2SO4, concentrated and purified by column
chromatography on silica gel (petroleum ether) to give compound 0601-155 (720
mg, 46%)
as a colorless oil. LCMS: 226 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.94 (t, J=
6.4
Hz, 4H), 3.18 (t, J= 6.4 Hz, 4H), 6.47 (d, J= 9.2 Hz, 1H), 7.27 (d, J= 9.2 Hz,
1H).
Step 82b: 1-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-
yl)phenyl)pyrrolidineate
(Compound 0602-155)
The title compound 0602-155 was prepared (500 mg, 57%) as a yellow solid from
0601-155 (720 mg, 3.18 mmol), bis(pinacolato)diboron (1.21 g, 4.78 mmol), AcOK
(938
mg, 9.56 mmol), PdC12(dppf)2 (78 mg, 0.0956 mmol) using a procedure similar to
that
described for compound 0602-107 (Example 34). LCMS: 274 [M+1]+. 'H NMR (400
MHz,
DMSO-d6) 6 1.25 (s, 12H), 1.95 (t, J = 6.8 Hz, 4H), 3.23 (t, J = 6.4 Hz, 4H),
6.49 (d, J =
8.8 Hz, I H), 7.46 (d, J= 8.4 Hz, 1 H).
Step 82c: Ethyl 2-(methyl((4-morpholino-2-(4-(pyrrolidin-1-yl)phenyl)thieno
[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-155)
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The title compound 0603-155 was prepared (320 mg, 86%) as a yellow solid from
0504-54 (300 mg, 0.668 mmol), 0602-155 (219 mg, 0.80 mmol), NaHCO3 (168 mg,
2.00
mmol), (Ph3P)2PdC12 (23 mg, 0.033 mmol) in ethanol (2.3 mL), toluene (4 mL)
and water
(1 mL) using a procedure similar to that described for compound 0603-107
(Example 34).
LCMS: 560 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.30 (t, J= 7.6 Hz, 3H), 1.98
(m,
4H), 3.26 (s, 3H), 3.30 (m, 4H), 3.75 (m, 4H), 3.90 (m, 4H), 4.29 (q, J= 6.8
Hz, 2H), 5.22
(s, 2H), 6.59 (d, J= 8.8 Hz, 2H), 7.40 (s, 1H), 8.21 (d, J= 8.8 Hz, 2H), 8.88
(s, 2H).
Step 82d: N-Hydroxy-2-(methyl((4-morpholino-2-(4-(pyrrolidin-1-yl)phenyl)
thieno[3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 155)
The title compound 155 was prepared (55 mg, 28%) as a yellow solid from 0603-
155 (200 mg, 0.49 mmol) and freshly prepared hydroxylamine methanol solution
(10 mL)
using a procedure similar to that described for compound 3 (Example 1). mp.
187-192 C.
LCMS: 547 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.97 (m, 4H), 3.23 (s, 3H), 3.30
(m,
4H), 3.76 (m, 4H), 3.88 (m, 4H), 5.18 (s, 2H), 6.59 (d, J= 8.8 Hz, 2H), 7.37
(s, 1H), 8.21
(d, J = 8.4 Hz, 2H), 8.75 (s, 2H), 9.06 (s, 1 H), 11.12 (s, 1 H).
Example 83: Preparation of 2-(((2-(3,4-diaminophenyl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 156)
Step 83a: 4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)benzene-1,2-diamine
(Compound 0602-156)
The title compound 0602-156 was prepared (1.0 g, 43%) as a yellow solid from 4-
bromobenzene- 1,2-diamine (1.87 g, 10 mmol), bis(pinacolato)diboron (3.9 g, 15
mmol),
Pd(dppf)2C12 (817 mg, 1 mmol) and AcOK (2.9 g, 30 mmol) using a procedure
similar to
that described for compound 0602-107 (Example 34). LCMS: 235 [M+1]+, 'H NMR
(400
MHz, DMSO-d6) 6 1.23 (s, 12H), 4.38 (s, 2H), 4.82 (s, 2H), 6.46 (d, J= 7.6 Hz,
1H), 6.77
(dd, J = 7.6, 0.8 Hz, 1 H), 6.8 8 (d, J = 0.8 Hz, 1 H).
Step 83b: Ethyl 2-(((2-(3,4-diaminophenyl)-4-morpholinothieno[3,2-d]pyrimidin-
6-yl)
methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-156)
The title compound 0603-156 was prepared (300 mg, 86%) as a white solid from
0504-54 (300 mg, 0.67 mmol), 0602-156 (190 mg, 0.80 mmol), NaHCO3 (169 mg, 2.0
mmol), CsF (203 mg, 1.34 mmol), (Ph3P)2PdC12 (47 mg, 0.067 mmol) in toluene (4
mL),
ethanol (2 mL) and water (0.5 mL) using a procedure similar to that described
for
compound 0603-107 (Example 34). LCMS: 521 [M+l]+;'H NMR (400 MHz, DMSO-d6): 6
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1.30 (t, J= 7.2 Hz, 3H), 3.26 (s, 3H), 3.75 (m, 4H), 3.87 (m, 4H), 4.29 (q, J=
7.2 Hz, 2H),
4.55 (s, 2H), 4.89 (s, 2H), 5.21 (s, 2H), 6.54 (d, J= 8.4 Hz, 1H), 7.36 (s,
1H), 7.52 (dd, J=
8.4, 2.0 Hz, I H), 7.63 (d, J= 2.0 Hz, I H), 8.88 (s, 2H).
Step 83c: 2-(((2-(3,4-diaminophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 156)
The title compound 156 was prepared (128 mg, 44 %) as a yellow solid from 0603-
156 (300 mg, 0.58 mmol) and freshly prepared hydroxylamine methanol solution
(20 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
222-225 C.
LCMS: 508 [M+1]; 'H NMR (400 MHz, DMSO-d6) 6 3.21 (s, 3H), 3.75 (m, 4H), 3.87
(m, 4H), 4.54 (s, 2H), 4.88 (s, 2H), 5.14 (s, 2H), 6.53 (d, J= 8.4 Hz, 1H),
7.33 (s, 1H), 7.52
(d, J= 7.6 Hz, I H), 7.62 (s, I H), 8.73 (s, 2H).
Example 84: Preparation of 2-(((2-(1H-indol-4-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 176)
Step 84a: Tert-butyl 4-bromo-1H-indole-l-carboxylate (Compound 0601-176)
The solution of 4-bromoindole (394 mg, 2.00 mmol), (Boc)20 (523 mg, 2.40
mmol), DMAP (293 mg, 2.4 mmol) and Et3N (0.4 mL) in MeCN (6 mL) was stirred at
room temperature for 2 hours. The solvent was removed and the residue was
dissolved in
ethyl acetate (40 mL), washed with water (3 x 20 mL) and brine (1 x 20 mL),
the organic
layer was concentrated and purified by column chromatography on silica gel
(petroleum
ether) to afford compound 0601-176 (508 mg, 85%) as a colorless oil. 'H-NMR
(400 MHz.
DMSO-d6) 6 1.64 (s, 9H), 6.67 (d, J = 3.2 Hz, 1 H), 7.28 (t, J = 8.0 Hz, 1 H),
7.48 (m, 1 H),
7.80 (d, J= 3.2 Hz, 1H), 8.08 (m, 1H).
Step 84b: Tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole-
l-
carboxylate (Compound 0602-176)
The title compound 0602-176 was prepared (448 mg, 77%) as a white solid from
0601-176 (503 mg, 1.69 mmol), bis(pinacolato)diboron (644 mg, 2.54 mmol),
Pd(dppf)2C12
(138 mg, 0.17 mmol) and AcOK (497 mg, 5.07 mmol) using a procedure similar to
that
described for compound 0602-107 (Example 34). 'H-NMR (400 MHz. DMSO-d6) 6 1.34
(s, 12H), 1.64 (s, 9H), 6.98 (d, J = 2.8 Hz, 1 H), 7.3 3 (t, J = 8.0 Hz, 1 H),
7.5 8 (m, 1 H), 7.72
(d, J= 2.8 Hz, I H), 8.20 (m, 1 H).
Step 84c: Ethyl 2-(((2-(1H-indol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)
methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-176)
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The compound tent-butyl 4-(6-(((5-(ethoxycarbonyl)pyrimidin-2-yl)(methyl)
amino)methyl)-4-morpholinothieno[3,2-d]pyrimidin-2-yl)-1H-indole-l-carboxylate
was
prepared (368 mg, 59%) as a white solid from 0504-54 (448 mg, 1.00 mmol), 0602-
176
(343 mg, 1.00 mmol), Cs2CO3 (652 mg, 2.00 mmol) and Pd(dppf)2C12 (82 mg, 0.10
mmol)
in 1,4-dioxane (6 mL) and water (0.2 mL) using a procedure similar to that
described for
compound 0603-107 (Example 34). LCMS: 630 [M+1]+.'H-NMR (400 MHz. DMSO-d6) 6
1.30 (t, J= 7.2 Hz, 3H), 1.65 (s, 9H), 3.28 (s, 3H), 3.78 (m, 4H), 3.93 (m,
4H), 4.29 (q, J=
7.2 Hz, 2H), 5.25 (s, 2H), 7.44 (m, I H), 7.55 (s, I H), 7.72 (m, I H), 7.77
(m, I H), 8.25 (m,
2H), 8.88 (s, 2H).
A mixture of above solid (368 mg, 0.59 mmol) trifluoroacetic acid (4 mL) was
stirred at room temperature for 1 hour. Adjusted to pH7 with 10% aqueous NaOH,
and
extracted with CH2C12 (60 mL). The organic layer was washed with brine (1 x 30
mL),
dried and concentrated to get compound 0603-176 (312 mg, 100%) as a yellow
solid.
LCMS: 530 [M+1]+.'H-NMR (400 MHz. DMSO-d6) 6 1.31 (t, J= 6.8 Hz, 3H), 3.29 (s,
3H), 3.79 (m, 4H), 3.94 (m, 4H), 4.30 (q, J= 7.2 Hz, 2H), 5.26 (s, 2H), 7.19
(t, J= 8.0 Hz,
I H), 7.43 (m, 2H), 7.53 (s, I H), 8.11 (m, I H), 8.89 (s, 2H), 11.23 (s, I
H).
Step 84d: 2-(((2-(1H-indol-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 176)
The title compound 176 was prepared (78 mg, 25%) as a white solid from 0603-
176
(318 mg, 0.60 mmol) and freshly prepared hydroxylamine methanol solution (10
mL) using
a procedure similar to that described for compound 3 (Example 1). m.p. 200-212
C.
LCMS: 517 [M+1]+. 'H-NMR (400 MHz. DMSO-d6) 6 3.26 (s, 3H), 3.79 (m, 4H), 3.94
(m, 4H), 5.22 (s, 2H), 7.19 (t, J= 8.0 Hz, 1H), 7.43 (m, 2H), 7.52 (m, 2H),
8.12 (d, J= 8.0
Hz, I H), 8.77 (s, 2H), 9.07 (s, I H), 11.14 (s, I H), 11.24 (s, I H).
Example 85: Preparation of N-hydroxy-2-(((2-(indolin-4-yl)-4-morpholinothieno
[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound
177)
Step 85a: Tert-butyl 4-bromoindoline-l-carboxylate (Compound 0601-177)
A mixture of 4-bromooxindole (2.77 g, 0.01 mol) and a solution of BH3 in THE
(2
M, 40 mL) was stirred at room temperature overnight. The mixture was cooled to
0 C and
diluted with 30 mL of methanol, followed by addition of 12 N HC1(7.5 mL). The
resulting
mixture was stirred at room temperature for 1 hour, adjusted to pH 8-9 with
10% aqueous
NaOH. Water was added to the mixture and extracted with ethyl acetate (3 x 100
mL). The
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organic layer was dried and concentrated to get the crude product which was
washed
through a silica gel column (ethyl acetate in petroleum ether (10%). The crude
product was
dissolved in 10% HC1(3 x 10 mL). The aqueous layer was adjusted to pH7 with
NaHCO3,
extracted with ethyl acetate (3 x 20 mL). The organic layer was dried and
concentrated to
get 4-bromoindoline (1.16 g, 45%) as an oil. LCMS: 200 [M+1]+.'H-NMR (400 MHz.
DMSO-d6) 6 2.90 (t, J= 8.8 Hz, 2H), 3.46 (t, J= 8.8 Hz, 2H), 5.86 (s, 1H),
6.43 (m, 1H),
6.64 (m, 1H), 6.83 (t, J= 8.0 Hz, 1H).
A mixture of above obtained 4-bromoindoline (759 mg, 3.81 mmol), and (Boc)20
(976 mg, 4.48 mmol) in MeCN (8 mL) was stirred at room temperature overnight.
After
evaporated, the residue was dissolved in ethyl acetate (40 mL), washed with
water (3 x 20
mL) and brine (1 x 20 mL). The organic layer was concentrated and purified by
column
chromatography on silica gel (petroleum ether) to give 0601-177(840 mg, 74%)
as a white
solid. 'H-NMR (400 MHz. DMSO-d6) 6 1.50 (s, 9H), 3.02 (t, J= 8.8 Hz, 2H), 3.94
(t, J=
8.8 Hz, 2H), 7.12 (m, 2H), 7.56 (m, 1H).
Step 85b: Tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indoline -
1-
carboxylate (Compound 0602-177)
The title compound 0602-177 was prepared (814 mg, 84%) as a white solid from
0601-177 (840 mg, 2.81 mmol), bis(pinacolato)diboron (1.07 g, 4.21 mmol),
Pd(dppf)2C12
(229 mg, 0.28 mmol) and AcOK (826 mg, 8.43 mmol) using a procedure similar to
that
described for compound 0602-107 (Example 34). LCMS: 290 [M-55]+; 'H-NMR (400
MHz. DMSO-d6) 6 1.35 (s, 12H), 1.57 (s, 9H), 3.24 (t, J= 8.4 Hz, 2H), 3.94 (t,
J= 8.4 Hz,
2H), 7.20 (m, 1H), 7.28 (m, 1H), 7.88 (m, 1H).
Step 85c: Ethyl 2-(((2-(indolin-4-yl)-4-morpholinothieno[3,2-d] pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-177)
Compound tent-butyl 4-(6-(((5-(ethoxycarbonyl)pyrimidin-2-yl)(methyl)amino)
methyl)-4-morpholinothieno[3,2-d]pyrimidin-2-yl)indoline-l-carboxylate was
prepared
(463 mg, 73%) as a white solid from 0504-54 (448 mg, 1.00 mmol), 0602-177 (345
mg,
1.00 mmol), Cs2CO3 (652 mg, 2.00 mmol) and Pd(dppf)2C12 (82 mg, 0.10 mmol) in
1,4-
dioxane (6 mL) and water (0.2 mL) using a procedure similar to that described
for
compound 0603-107 (Example 34). LCMS: 632 [M+1]+. 'H-NMR (400 MHz. DMSO-d6) 5
1.30 (t, J= 7.2 Hz, 3H), 1.52 (s, 9H), 3.27 (s, 3H), 3.55 (m, 2H), 3.87 (m,
4H), 3.94 (m,
6H), 4.29 (q, J= 7.2 Hz, 2H), 5.23 (s, 2H), 7.26 (m, I H), 7.46 (s, I H), 7.81
(m, I H), 7.89
(m, 1H), 8.87 (s, 2H).
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A mixture of above product (463 mg, 0.73 mmol) and trifluoroacetic acid (4 mL)
was stirred at room temperature for 1 hour. The mixture was adjusted to pH 7
with 10%
aqueous NaOH, extracted with CH2C12 (60 mL). The organic layer was washed with
brine
(1 x 30 mL), dried and concentrated to give 0603-177 (283 mg, 73%) as an off-
white solid.
LCMS: 532 [M+1]+.'H-NMR (400 MHz. DMSO-d6) 6 1.30 (t, J= 6.8 Hz, 3H), 3.27 (s,
3H), 3.41 (m, 4H), 3.75 (m, 4H), 3.86 (m, 4H), 4.29 (q, J= 6.8 Hz, 2H), 5.23
(s, 2H), 5.58
(s, I H), 6.58 (m, I H), 7.01 (t, J= 8.0 Hz, I H), 7.47 (m, 2H), 8.88 (s, 2H).
Step 85d: N-Hydroxy-2-(((2-(indolin-4-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)
methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 177)
The title compound 177 was prepared (130 mg, 49%) as a white solid from 0603-
177 (273 mg, 0.51 mmol) and freshly prepared hydroxylamine methanol solution
(8 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.:
146-156 C.
LCMS: 519 [M+1]. 'H-NMR (400 MHz. DMSO-d6) 6 3.24 (s, 3H), 3.42 (m, 4H), 3.75
(m,
4H), 3.87 (m, 4H), 5.20 (s, 2H), 5.57 (s, 1H), 6.58 (m, 1H), 7.01 (t, J= 8.0
Hz, 1H), 7.43 (s,
I H), 7.48 (m, I H), 8.76 (s, 2H), 9.07 (s, I H), 11.13 (s, I H).
EXAMPLE 86: Preparation of 2-(((2-(3,4-dihydro-2H-benzo[b] [1,4]oxazin-6-yl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)-N-
hydroxypyrimidine-5-carboxamide (Compound 182)
Step 86a: Tert-butyl 6-bromo-2,3-dihydrobenzo[b][1,4]oxazine-4-carboxylate
(Compound
0601-182)
A mixture of compound 4-bromo-2-nitrophenol (1 g, 4.59 mmol) and SnC12 (5.2 g,
22.9 mmol) in ethanol (10 mL) was stirred at 70 C for 2 h. After cooled down,
the mixture
was diluted with water (100 mL), adjusted to pH 7 with saturated aqueous
NaHCO3 and
extracted with ethyl acetate (100 mL). The organic layer was washed with brine
(50 mL),
dried over Na2SO4, concentrated to give 2-amino-4-bromophenol (770 mg, 89%) as
a grey
solid. LCMS: 188 [M+1 ]+. 'H NMR (400 MHz, DMSO-d6) 6 4.79 (s, 2H), 6.48 (dd,
JI =
2.4 Hz, J2 = 8.4 Hz, 1 H), 6.55 (d, J = 8.4 Hz, 1 H), 6.71 (d, J = 2.4 Hz, 1
H), 9.26 (s, 1 H).
A mixture of the product 2-amino-4-bromophenol (500 mg, 2.66 mmol), 1,2-
dibromoethane (2.5 g, 13.3 mmol) and K2CO3 (1.84 g, 13.3 mmol) in DMF (10 mL)
was
stirred at room temperature for 4 h. The mixture was diluted with water (100
mL) and
extracted with ethyl acetate (100 mL). The organic layer was washed with water
(3 x 50
mL) and brine (50 mL), concentrated and purified by column chromatography on
silica gel
(ethyl acetate in petroleum, 10% v/v) to give 5-bromo-2-(2-
bromoethoxy)benzenamine
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(250 mg, 37%) as a yellow solid. LCMS: 294 [M+1]+. 'H NMR (400 MHz, DMSO-d6)
65
3.79 (t, J= 6.0 Hz, 2H), 4.25 (t, J= 5.6 Hz, 2H), 5.06 (s, 2H), 6.63 (dd, J1=
2.4 Hz, J2 =
8.0 Hz, 1 H), 6.77 (d, J = 8.4 Hz, 1 H), 6.82 (d, J = 2.4 Hz, 1 H).
A mixture of the product 5-bromo-2-(2-bromoethoxy)benzenamine (250 mg, 0.848
mmol) and K2C03 (234 mg, 1.695 mmol) in DMF (5 mL) was stirred at 60 C for 4
h. The
mixture was diluted with water (100 mL) and extracted with ethyl acetate (100
mL). The
organic layer was washed with water (3 x 50 mL) and brine (50 mL),
concentrated to give
6-bromo-3,4-dihydro-2H-benzo[b][1,4]oxazine (170 mg, 94%) as a yellow oil.
LCMS: 214
[M+1]+. 'H NMR (400 MHz, DMSO-d6) 65 3.26 (m, 2H), 4.08 (t, J= 4.8 Hz, 2H),
6.06 (s,
1 H), 6.56 (m, 2H), 6.69 (d, J = 1.6 Hz, 1 H).
A mixture of the product 6-bromo-3,4-dihydro-2H-benzo[b][1,4]oxazine (1.37 g,
6.4 mmol), Boc2O (1.676 g, 7.68 mmol), Et3N (970 mg, 9.6 mmol), DMAP (78 mg,
0.64
mmol) in THE (27 mL) was stirred at room temperature overnight. The reaction
mixture
was diluted with water (200 mL) and extracted with ethyl acetate (100 mL). The
organic
layer was washed with water (50 mL) and brine, dried over Na2SO4 and
concentrated to
give compound 0601-182 (1.3 g, 65%) as a yellow oil. LCMS: 258 [M-55]+. 'H NMR
(400
MHz, DMSO-d6) 65 1.49 (s, 9H), 3.78 (t, J= 4.8 Hz, 2H), 4.21 (t, J= 4.4 Hz,
2H), 6.83 (d, J
= 4.4 Hz, I H), 7.12 (dd, J1= 2.0 Hz, J2 = 8.4 Hz, 2H), 8.01 (s, I H).
Step 86b: Tert-butyl 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) -2,3-
dihydrobenzo[b][1,4]oxazine-4-carboxylate (Compound 0602-182)
The title compound 0602-182 was prepared (1.4 g, 98%) as a oil from 0601-182
(1.16 g, 3.69 mmol), bis(pinacolato)diboron (1.41 g, 5.54 mmol), Pd(dppf)2C12
(90 mg,
0.111 mmol) and AcOK (1.09 g, 11.07 mmol) using a procedure similar to that
described
for compound 0602-107 (Example 34). LCMS: 306 [M-55]+. 'H NMR (400 MHz, DMSO-
d6) 651.27 (s, 12H), 1.49 (s, 9H), 3.79 (t, J = 4.4 Hz, 2H), 4.23 (t, J = 4.0
Hz, 2H), 6.88 (d, J
= 8.0 Hz, 1H), 7.25 (d, J= 9.6 Hz, 2H), 8.13 (s, 1H).
Step 86c: Ethyl 2-(((2-(3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yl)- 4-
morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxylate
(Compound 0603-182)
Compound tent-butyl 6-(6-(((5-(ethoxycarbonyl)pyrimidin-2-yl)(methyl)amino)
methyl)-4-morpholinothieo [3,2-d]pyrimidin-2-yl)-2, 3 -dihydrobenzo [b] [
1,4]oxazine-4-
carboxylate was prepared (250 mg, 58%) as a yellow solid from 0504-54 (300 mg,
0.668
mmol), 0602-182 (290 mg, 0.802 mmol), NaHCO3 (168 mg, 2.00 mmol), (Ph3P)2PdC12
(23
mg, 0.0334 mmol) in ethanol (2.3 mL), toluene (4 mL) and water (1 mL) using a
procedure
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similar to that described for compound 0603-107 (Example 34). LCMS: 648
[M+1]+. 'H
NMR (400 MHz, DMSO-d6) 6 1.30 (t, J= 6.8 Hz, 3H), 1.52 (s, 9H), 3.26 (s, 3H),
3.75 (m,
4H), 3.83 (m, 2H), 3.92 (m, 4H), 4.28 (m, 4H), 5.23 (s, 2H), 6.93 (d, J= 8.4
Hz, 1H), 7.45
(s, 1H), 8.02 (d, J= 8.8 Hz, 1H), 8.80 (s, 1H), 8.88 (s, 2H).
To a solution of the above prepared compound (250 mg, 0.386mmo1) in
dichloromethane (25 ml) was added CF3COOH(2.5 mL) and stirred at room
temperature
overnight. The mixture was diluted with ethyl acetate (100 mL) and washed with
saturated
aqueous NaHCO3 (50 mL), water (100 mL) and brine (50 mL), dried over Na2SO4,
concentrated to give 0603-182 (200 mg, 95%) as a yellow solid. LCMS: 548
[M+1]+. 1H
NMR (400 MHz, DMSO-d6) 6 1.30 (t, J= 7.2 Hz, 3H), 3.26 (s, 3H), 3.30 (m, 2H),
3.75 (m,
4H), 3.89 (m, 4H), 4.17 (m, 2H), 4.28 (q, J= 7.2 Hz, 2H), 5.22 (s, 2H), 5.92
(s, 1H), 6.70
(d, J = 8.4 Hz, 1 H), 7.42 (s, 1 H), 7.5 6 (d, J = 10.0 Hz, 1 H), 7.68 (s, 1
H), 8.8 8 (s, 2H).
Step 86d: 2-(((2-(3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yl)-4-morpholinothieno
[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 182)
The title compound 182 was prepared (72 mg, 37%) as a white solid from 0603-
182
(200 mg, 0.365 mmol) and freshly prepared hydroxylamine methanol solution (10
mL)
using a procedure similar to that described for compound 3 (Example 1). mp.
170-183 C.
LCMS: 535 [M+1]+. 1H NMR (400 MHz, DMSO-d6) 6 3.23 (s, 3H), 3.30 (m, 2H), 3.75
(m,
4H), 3.89 (m, 4H), 4.17 (m, 2H), 5.19 (s, 2H), 5.91 (s, 1H), 6.70 (d, J= 8.0
Hz, 1H), 7.40
(s, I H), 7.57 (d, J= 8.0 Hz, I H), 7.68 (s, I H), 8.75 (s, 2H), 9.07 (s, I
H), 11.01 (s, I H).
EXAMPLE 87: Preparation of 2-(((2-(1H-benzo[d]imidazol-5-yl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)-N-
hydroxypyrimidine-5-carboxamide (Compound 187)
Step 87a: Tert-butyl 5-bromo-lH-benzo[d]imidazole-l-carboxylate (Compound 0601-
187)
To a solution of 4-bromobenzene-1,2-diamine (3 g, 16 mmol) in DMF (22 mL)
were added trimethyl orthoformate (44 mL) and conc. HC1(1.5 mL) and the
mixture was
stirred at room temperature for 1 h. The mixture was diluted with water (200
mL) and
adjusted to pH7 with saturated aqueous NaHCO3, extract with ethyl acetate (200
mL). The
organic layer was dried over Na2SO4, concentrated to give 5-bromo-lH-
benzo[d]imidazole
(3.25 g, 100%) as an off-white solid. LCMS: 197 [M+1]+. 1H NMR (400 MHz, DMSO-
d6)
6 7.33 (t, J= 8.8 Hz, 1H), 7.55 (dd, J1= 7.6 Hz, J= 40 Hz, 1H), 7.79 (d, J=
47.2 Hz, 1H),
8.26 (s, 1H), 12.61 (d, J= 25.6 Hz, 1H).
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To a solution of above prepared 5-bromo-lH-benzo[d]imidazole (3.25 g, 22.1
mmol) in THE (65 mL) was added Boc2O (5.79 g, 26.5 mmol), Et3N (3.35, 33.15
mmol)
and DMAP (270 mg, 2.21 mmol). The mixture was stirred at room temperature for
4 h,
diluted with water (200 mL), extracted with ethyl acetate (200 mL). The
organic layer was
washed with water (2 x 100 mL) and brine (100 mL), dried over Na2SO4,
concentrated to
give 0601-187 (4.8 g, 98%) as a oil. LCMS: 241 [M-55]+. 'H NMR (400 MHz, DMSO-
d6)
6 1.65 (s, 9H), 7.5 7 (dd, J1= 8.4 Hz, J2 = 20 Hz, 1 H), 7.73 (d, J = 8.4 Hz,
1 H), 7.8 8 (d, J =
9.2 Hz, 1 H), 8.03 (d, J = 3 5.6 Hz, 1 H), 8.70 (d, J = 8.0 Hz, 1 H).
Step 87b: Tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-
benzo[d]imidazole-l-carboxylate (Compound 0602-187)
The title compound 0602-187 was prepared (0.94 g, 81%) as a colorless oil from
0601-187 (1 g, 3.37 mmol), bis(pinacolato)diboron (1.28 g, 5.05 mmol),
Pd(dppf)2C12 (82
mg, 0.101 mmol) and AcOK (991 mg, 10.1 mmol) using a procedure similar to that
described for compound 0602-107 (Example 34). LCMS: 289 [M-55]+. 'H NMR (400
MHz, DMSO-d6) 6 1.32 (s, 12H), 1.65 (s, 9H), 7.65 (d, J= 7.2 Hz, 0.5H), 7.74
(t, J= 8.4
Hz, 1H), 7.97 (d, J= 8.4 Hz, 1H), 8.37 (s, 0.5H), 8.69 (d, J= 18.8 Hz, 1H).
Step 87c: Ethyl 2-(((2-(1H-benzo[d]imidazol-5-yl)-4-morpholinothieno[3,2-d]
pyrimidin-
6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-187)
The compound 0603-187 was prepared (260 mg, 62%) as a yellow solid from 0504-
54 (300 mg, 0.668 mmol), 0602-187 (276 mg, 0.8 mmol), NaHCO3 (168 mg, 2.00
mmol),
(Ph3P)2PdC12 (23 mg, 0.0334 mmol) in ethanol (2.3 mL), toluene (4 mL) and
water (1 mL)
using a procedure similar to that described for compound 0603-107 (Example
34). LCMS:
531 [M+l]+. 'H NMR (400 MHz, DMSO-d6) 6 1.30 (t, J= 7.2 Hz, 3H), 3.28 (s, 3H),
3.78
(m, 4H), 3.95 (m, 4H), 4.29 (q, J= 6.8 Hz, 2H), 5.24 (s, 2H), 7.50 (s, 1H),
7.65 (d, J= 8.0
Hz, 1H), 8.32 (m, 2H), 8.64 (s, 1H), 8.88 (s, 2H), 12.59 (s, 1H).
Step 87d: 2-(((2-(1H-benzo[d]imidazol-5-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 187)
The title compound 187 was prepared (34 mg, 13%) as a yellow solid from 0603-
187 (260 mg, 0.49 mmol) and freshly prepared hydroxylamine methanol solution
(10 mL)
using a procedure similar to that described for compound 3 (Example 1). mp.
231-239 C.
LCMS: 518 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 3.25 (s, 3H), 3.79 (m, 4H), 3.95
(m,
4H), 5.21 (s, 2H), 7.48 (s, I H), 7.65 (dd, J1= 8.4 Hz, J2 = 47.2 Hz, I H),
8.31 (m, 2H), 8.64
(d, J= 44.8 Hz, 1H), 8.76 (s, 2H), 9.09 (s, 1H), 11.03 (s, 1H), 12.59 (s, 1H).
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Example 88: Preparation of N-hydroxy-2-(methyl((2-(2-methyl-3H-
benzo[d]imidazol -
5-yl)-4-morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)amino)pyrimidine-5-
carboxamide (Compound 199)
Step 88a: N,N'-(4-bromo-1,2-phenylene)diacetamide (Compound 0601-199)
To the solution of 4-bromobenzene-1,2-diamine (1.87 g, 10 mmol) and Et3N (10.1
g, 100 mmol) in CH2C12 (20 mL) was added CH30001(1.73 g, 22 mmol) at 0 C and
stirred for 2 hr at 30 C. The mixture was concentrated and the residue was
dissolved in
CH2C12, washed with water, dried over Na2SO4, concentrated to give 0601-199
(1.4 g,
52%) as a yellow solid. LCMS: 271 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 2.08 (d,
J=
3.2 Hz, 6H), 7.28 (m, 1H), 7.50 (d, J= 8.8Hz, 1H), 7.83 (s, 1H), 9.38 (d, J=
3.2 Hz, 2H).
Step 88b: N,N'-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2-phenylene)
diacetamide (Compound 0602-199)
The title compound 0602-189 was prepared (1.0 g, 63%) as a yellow solid from
0601-199 (1.4 g, 5.2 mmol), bis(pinacolato)diboron (2.0 g, 7.8 mmol),
Pd(dppf)2C12 (425
mg, 0.52 mmol) and AcOK (1.53 g, 15.6 mmol) using a procedure similar to that
described
for compound 0602-107 (Example 34). LCMS: 319 [M+1]+, 'H NMR (400 MHz, DMSO-
d6) 6 1.28 (s, 12H), 2.07 (d, J = 6.0 Hz, 6H), 7.41 (d, J = 8.0 Hz, 1 H), 7.70
(d, J = 8.4 Hz,
1 H), 7.79 (s, 1 H), 9.36 (d, J = 9.6 Hz, 2H).
Step 88c: Ethyl 2-(methyl((2-(2-methyl-3H-benzo[d]imidazol-5-yl)-4-
morpholinothieno
[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-
199)
Compound ethyl 2-(((2-(3,4-diacetamidophenyl)-4-morpholinothieno [3,2-
d]pyrimidin-6-yl) methyl)(methyl)amino)pyrimidine-5-carboxylate was prepared
(260 mg,
75%) as a white solid from 0504-54 (261 mg, 0.58 mmol), 0602-199 (240 mg, 0.75
mmol),
NaHCO3 (147 mg, 1.8 mmol) and bis(triphenylphosphine)palladium(II) chloride
(41 mg,
0.058 mmol) in toluene (4 mL), ethanol (2 mL) and water (0.5 mL) using a
procedure
similar to that described for compound 0603-107 (Example 34). LCMS: 605
[M+1]+; 1H
NMR (400 MHz, DMSO-d6): 6 1.30 (m, 3H), 2.11 (s, 6H), 3.27 (s, 3H), 3.76 (m,
4H), 3.92
(m, 4H), 4.30 (m, 2H), 5.24 (s, 2H), 7.48 (m, 1H), 7.76 (m, 1H), 8.15 (m, 1H),
8.50 (m,
1H), 8.88 (m, 2H), 8.45 (m, 2H).
To the solution of above prepared compound (360 mg, 0.6 mmol) in THE (8 mL)
was added 6M HC1(12 mL) and stirred for 10 hr at 40 C. The mixture was
adjusted to pH8
with saturated aqueous Na2CO3 at 0 C, extracted with ethyl acetate. The
organic layer was
dried, concentrated and purified by column chromatography on silica gel
(methanol in
dichloromethane, 2-5 v/v) to give title compound 0603-199 (160 mg, 50%) as a
white solid.
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LC-MS: 545 [M+1]+. 'H NMR (400 MHz, CDC13) 61.38 (t, J= 6.8 Hz, 3H), 2.60 (s,
3H), 3.30 (s, 3H), 3.85 (m, 4H), 4.01 (m, 4H), 4.36 (q, J= 7.2 Hz, 2H), 5.18
(s, 2H), 7.38
(s, 1H), 7.60 (d, J= 8.0 Hz, 1H), 8.36 (d, J= 8.4 Hz, 1H), 8.67 (s, 1H), 8.92
(s, 2H).
Step 88d: N-hydroxy-2-(methyl((2-(2-methyl-3H-benzo[d]imidazol-5-yl)-4-
morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound 199)
The title compound 199 was prepared (38 mg, 24%) as a white solid from 0603-
199
(160 mg, 0.30 mmol) and freshly prepared hydroxylamine methanol solution (20
mL) using
a procedure similar to that described for compound 3 (Example 1). m.p. 230-233
C.
LCMS: 532 [M+1]+; 'H NMR (400 MHz, DMSO-d6) (52.51 (s, 3H), 3.24 (s, 3H), 3.78
(m, 4H), 3.93 (m, 4H), 5.21 (s, 2H), 7.47 (s, 1H), 7.50 (dd, J= 42.8, 8.4 Hz,
1H), 8.25 (t, J
= 8.8 Hz, I H), 8.48 (d, J= 42.8 Hz, I H), 8.76 (s, 2H), 9.09 (s, I H), 11.00
(s, I H), 12.33 (s,
I H).
Example 89: Preparation of N-hydroxy-2-(methyl((4-morpholino-2-(2-oxo-2,3-
dihydro-1H- benzo[d]imidazol-5-yl)thieno[3,2-d]pyrimidin-6-
yl)methyl)amino)pyrimidine-5-carboxamide (Compound 186)
Step 89a: 5-Bromo-lH-benzo[d]imidazol-2(3H)-one (Compound 0601-186)
A mixture of 4-bromobenzene-1,2-diamine (3.74 g, 20 mmol), CDI (3.9 g, 24
mmol) in 1,4-dioxane(20 mL) was stirred for 1 hr at 40 C. The mixture was
filtered and
washed with petroleum ether and dichloromethane to get compound 0601-186 (3.0
g, 70%)
as a white solid. LCMS: 213 [M+1]+, 1H NMR (400 MHz, DMSO-d6) 6 6.86 (d, J=
8.0 Hz,
1H), 7.06(m, 1 H), 7.08 (m, 1H), 10.77 (s, 2H).
Step 89b: 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-
2(3H)-
one (Compound 0602-186)
The title compound 0602-186 was prepared (340 mg, 21%) as a yellow solid from
0601-186 (1.3 g, 6 mmol), bis(pinacolato)diboron (2.3 g, 9 mmol), Pd(dppf)2C12
(490 mg,
0.6 mmol) and AcOK (1.8 g, 18 mmol) using a procedure similar to that
described for
compound 0602-107 (Example 34). LCMS: 261 [M+1]+, 1H NMR (400 MHz, DMSO-d6) (S
1.27 (s, 12H), 6.91 (d, J= 7.6 Hz, 1H), 7.17 (s, 1 H), 7.28 (d, J= 7.6 Hz,
1H), 10.65 (s,
1H), 10.77 (s, 1H).
Step 89c: Ethyl2-(methyl((4-morpholino-2-(2-oxo-2,3-dihydro-lH-benzo[d]
imidazol-5-
yl) thieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxylate
(Compound
0603-186)
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The title compound 0603-186 was prepared (214 mg, 68%) as a white solid from
0504-54 (261 mg, 0.58 mmol), 0602-186 (197 mg, 0.75 mmol), NaHCO3 (147 mg, 1.8
mmol) and bis(triphenylphosphine)palladium(II) chloride (41 mg, 0.058 mmol) in
toluene
(4 mL), ethanol (2 mL) and water (0.5 mL) using a procedure similar to that
described for
compound 0603-107 (Example 34). LCMS: 547 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 6
1.30 (t, J= 6.8 Hz, 3H), 3.27 (s, 3H), 3.77 (m, 4H), 3.91 (m, 4H), 4.28 (q, J=
6.8 Hz, 2H),
5.23 (s, 2H), 6.99 (d, J = 8.0 Hz, 1 H), 7.46 (s, 1 H), 7.97 (s, 1 H), 8.08
(d, J = 9.2 Hz, 1 H),
8.88 (s, 2H), 10.71 (s, 1H), 10.80 (s, 1H).
Step 89d: N-hydroxy-2-(methyl((4-morpholino-2-(2-oxo-2,3-dihydro-lH-benzo
[d]imidazol-5-yl)thieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-
carboxamide
(Compound 186)
The title compound 186 was prepared (75 mg, 36 %) as a white solid from 0603-
186 (214 mg, 0.40 mmol) and freshly prepared hydroxylamine methanol solution
(20 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.
272-275 C.
LCMS: 534 [M+1]+; 'H NMR (400 MHz, DMSO-d6) 5 3.23 (s, 3H), 3.77 (m, 4H), 3.91
(m, 4H), 5.20 (s, 2H), 7.00 (d, J= 8.0 Hz, I H), 7.44 (s, I H), 7.98 (s, I H),
8.09 (d, J= 8.4
Hz, I H), 8.75 (s, 2H), 10.71 (s, I H), 10.82 (s, I H).
Example 90: Preparation of N-hydroxy-2-(methyl((4-morpholino-2-(2-oxoindolin-
5-yl)thieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound 194)
Step 90a: 5-Bromoindolin-2-one (Compound 0601-194)
To a mixture of 5-bromoindole-2,3-dione (2.25 g, 10 mmol), ethyleneglycol (45
mL) and hydrazine hydrate (1.06 g, 21.10 mmol) was added KOH (1.68 g, 30
mmol). The
reaction mixture was stirred at 80 C for 4 hours. The mixture was cooled to
room
temperature and poured into ice cold water and the mixture was adjusted to pH
1-2 with
12N hydrochloric acid and stirred at room temperature for 12 hours. The
mixture was
filtered and solid was washed with water (5 mL) and dried to get the crude
product which
was purified by column chromatography on silica gel (methanol in
dichloromethane, 0.5%
v/v) to give 0601-194 (785 mg, 37%) as a yellow solid. LCMS: 214 [M+1]+. 'H-
NMR (400
MHz. DMSO-d6) 6 3.51 (s, 2H), 6.76 (d, J= 8.0 Hz, 1H), 7.34 (dd, J= 8.0, 2.0
Hz, 1H),
7.38 (m, 1H), 10.49 (s, 1H).
Step 90b: 5-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-2-one
(Compound 0602-
194)
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The title compound 0602-194 was prepared (323 mg, 83%) as a yellow solid from
0601-194 (317 mg, 1.5 mmol), bis(pinacolato)diboron (572 mg, 2.25 mmol),
Pd(dppf)2C12
(126 mg, 0.15 mmol) and AcOK (441 mg, 4.5 mmol) using a procedure similar to
that
described for compound 0602-107 (Example 34). LCMS: 260 [M+1]+.'H-NMR (400
MHz.
DMSO-d6) 6 1.27 (s, 12H), 3.46 (s, 2H), 6.81 (d, J= 8.0 Hz, 1H), 7.50 (m, 2H),
10.54 (s,
I H).
Step 90c: Ethyl 2-(methyl((4-morpholino-2-(2-oxoindolin-5-yl)thieno[3,2-d]
pyrimidin-6-
yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-194)
The title compound 0603-194 was prepared (350 mg, 80%) as a yellow solid from
0504-54 (358 mg, 0.80 mmol), 0602-194 (207 mg, 0.80 mmol), Cs2CO3 (522 mg,
1.60
mmol) and Pd(dppf)2C12 (65 mg, 0.08 mmol) in 1,4-dioxane (6 mL) and water (0.2
mL)
using a procedure similar to that described for compound 0603-107 (Example
34). LCMS:
546 [M+1]+.'H-NMR (400 MHz. DMSO-d6) 6 1.30 (t, J= 7.2 Hz, 3H), 3.27 (s, 3H),
3.57
(m, 2H), 3.76 (m, 4H), 3.91 (m, 4H), 4.29 (q, J= 7.2 Hz, 2H), 5.23 (s, 2H),
6.90 (d, J= 8.4
Hz, 1H), 7.44 (s, 1H), 8.28 (m, 2H), 8.88 (s, 2H), 10.59 (s, 1H).
Step 90d: N-hydroxy-2-(methyl((4-morpholino-2-(2-oxoindolin-5-yl)thieno [3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 194)
The title compound 194 was prepared (85 mg, 25%) as a white solid from 0603-
194
(350 mg, 0.64 mmol) and freshly prepared hydroxylamine methanol solution (7.5
mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.:
270 C
(decomposed). LCMS: 533 [M+1]+. 'H-NMR (400 MHz, DMSO-d6) 6 3.24 (s, 3H), 3.57
(s,
2H), 3.77 (m, 4H), 3.91 (m, 4H), 5.20 (s, 2H), 6.90 (m, I H), 7.42 (m, I H),
8.26 (m, 2H),
8.76 (s, 2H), 10.52 (s, 1H).
EXAMPLE 91: Preparation of N-hydroxy-2-(methyl((4-morpholino-2-(1H-pyrazol-4-
yl)thieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound
196)
Step 91a: 4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(Compound 0602-
196)
The title compound 0602-196 was prepared (400 mg, 30%) as a yellow solid from
4-bromopyrazole (1 g, 6.8 mmol), bis(pinacolato)diboron (2.6 g, 10.2 mmol),
Pd(dppf)2C12
(166 mg, 0.2 mmol) and AcOK (g, 20.4 mmol) using a procedure similar to that
described
for compound 0602-107 (Example 34). LCMS: 195 [M+1]+; 1H NMR (400 MHz, DMSO-
d6) 6 1.25 (s, 12H), 7.93 (s, 2H), 13.09 (s, I H).
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Step 91b: Ethyl2-(methyl((4-morpholino-2-(1H-pyrazol-4-yl)thieno[3,2-d]
pyrimidin-6-
yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-196)
The title compound 0603-196 was prepared (150 mg, 47%) as a yellow solid from
0504-54 (300 mg, 0.67 mmol), 0602-196 (259 mg, 1.33 mmol), NaHCO3 (168 mg, 2.0
mmol), (Ph3P)2PdC12 (23 mg, 0.03 mmol) in toluene (5 mL), ethanol (3 mL) and
water (1.3
mL) using a procedure similar to that described for compound 0603-107 (Example
34).
LCMS: 481 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.30 (t, J= 6.8 Hz, 3H), 3.26
(s,
3H), 3.73 (m, 4H), 3.88 (m, 4H), 4.28 (q, J= 7.6 Hz, 2H), 5.21 (s, 2H), 7.38
(s, 1H), 8.04
(s, I H), 8.30 (s, I H), 8.88 (s, 2H), 13.07(s,1 H).
Step 91c: N-hydroxy-2-(methyl((4-morpholino-2-(1H-pyrazol-4-yl)thieno[3,2-d]
pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 196)The title
compound 196 was prepared (59 mg, 41%) as a yellow solid from 0603-196 (150
mg, 0.31
mmol) and freshly prepared hydroxylamine methanol solution (8 mL) using a
procedure
similar to that described for compound 3 (Example 1). m.p.: 214-217 C. LCMS:
468
[M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 3.23 (s, 3H), 3.73 (m, 4H), 3.87 (m, 4H),
5.18
(s, 2H), 7.3 5 (s, 1 H), 8.05 (s, 1 H), 8.3 0 (s, 1 H), 8.74 (s, 2H), 9.07 (s,
1 H), 11.11 (s, 1 H),
13.07 (s,1 H).
EXAMPLE 92: Preparation of N-hydroxy-2-(methyl((4-morpholino-2-(1H-pyrrol-3-
yl)thieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound
197)
Step 92a: 3-Bromo-l-(triisopropylsilyl)-1H-pyrrole (Compound 0601-197)
A solution n-BuLi in THE (2.5 M, 19.6 mL, 49 mmol) was added to a stirred
solution of pyrrole (3 g, 44.7 mmol) in anhydrous THE (20 mL) at -78 C in an
N2
atmosphere. Then the mixture was warmed to room temperature and stirred at
this
temperature for 10 min. The mixture was cooled again to -78 C, and
chlorotriisopropylsilane (10.5 g, 44.7mmol) was added dropwisely with
stirring. Then the
mixture was warmed to room temperature and stirred for additional 30 min.,
diluted with
water (200 mL), extracted with ether (200 mL). The organic layer was washed
with water
(2 x 100 mL) and brine (100 mL), dried over Na2SO4, concentrated to give crude
1-
(triisopropylsilyl)-1H-pyrrole (11 g, 100%) as a oil. LCMS: 224 [M+1]+. 'H NMR
(400
MHz, DMSO-d6) 6 0.98 (m, 18H), 1.40 (m 3H), 6.20 (m, 2H), 6.80 (m, 2H).
To a solution of the above prepared 1-(triisopropylsilyl)-1H-pyrrole (5.85 g,
26.2
mmol) in THE (50 mL) was added NBS (4.66 g, 26.2 mmol) at -78 C and the
resulting
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mixture was stirred at -78 C for 2 h. The mixture was warmed to room
temperature and
stirred for additional 1 h. The mixture was concentrated and purified by
column
chromatography on silica gel (petroleum) to give compound 0601-197 (6.8 g,
63%) as a
colorless oil. LCMS: 302 [M+l]+. 'H NMR (400 MHz, DMSO-d6) 6 0.98 (m, 18H),
1.47
(m 3H), 6.26 (d, J= 6.0 Hz, 1H), 6.82 (m, 1H), 6.89 (m, 1H).
Step 92b: 1-(Triisopropylsilyl)-1H-pyrrol-3-ylboronic acid (Compound 0602-197)
A solution n-BuLi in THE (2.5 M, 1.58 mL, 3.96 mmol) was added to a stirred
solution of 0601-197 (1 g, 3.31 mmol) in anhydrous THE (20 mL) at -78 C in an
N2
atmosphere. The resulting mixture was stirred at this temperature for 30 min.
To the
mixture was added trimethyl borate (687 mg, 6.6mmol) dropwise. Then the
mixture was
warmed to room temperature and stirred for additional 1 h. The mixture was
diluted with
water (200 mL), extracted with ethyl acetate (200 mL). The organic layer was
washed with
water (2 x 100 mL) and brine (100 mL), dried over Na2SO4, concentrated to give
crude
compound 0602-197 (280 mg, 32%) as a oil which was used in next step directly
without
further purification. LCMS: 268 [M+1]+.
Step 92c: Ethyl 2-(methyl((4-morpholino-2-(1H-pyrrol-3-yl)thieno[3,2-d]
pyrimidin-6-
yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-197)
The title compound 0603-197 was prepared (260 mg, 81%) as a yellow solid from
0504-54 (300 mg, 0.67 mmol), 0602-197 (0.8 g), NaHCO3 (168 mg, 2.0 mmol),
(Ph3P)2PdC12 (23 mg, 0.03 mmol) in toluene (5 mL), ethanol (3 mL) and water (1
mL)
using a procedure similar to that described for compound 0603-107 (Example
34). LCMS:
480 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 1.28 (t, J= 7.2 Hz, 3H), 3.24 (s, 3H),
3.71
(m, 4H), 3.84 (m, 4H), 4.27 (q, J= 7.2 Hz, 2H), 5.18 (s, 2H), 6.65 (s, I H),
6.77 (s, I H),
7.33 (s, I H), 7.47 (s, I H), 8.86 (s, 2H), 11.07 (s, I H).
Step 92d: N-Hydroxy-2-(methyl((4-morpholino-2-(1H-pyrrol-3-yl)thieno [3,2-
d]pyrimidin-
6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 197)
The title compound 197 was prepared (63 mg, 25%) as a yellow solid from 0603-
197 (260 mg, 0.54 mmol) and freshly prepared hydroxylamine methanol solution
(10 mL)
using a procedure similar to that described for compound 3 (Example 1). m.p.:
175-189 C.
LCMS: 467 [M+1]+. 'H NMR (400 MHz, DMSO-d6) 6 3.21 (s, 3H), 3.72 (m, 4H), 3.83
(m,
4H), 5.15 (s, 2H), 6.65 (s, I H), 6.75 (s, I H), 7.30 (s,1 H), 7.46 (s, I H),
8.73 (m, 2H), 9.05 (s,
I H), 11.05 (s, I H), 11.11 (s, I H).
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Example 93: Preparation of 2-((2-(4-aminophenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methylamino)-N-hydroxypyrimidine-5-carboxamide (Compound 211)
Step 93a: Ethyl 2-((2-(4-aminophenyl)-4-morpholinothieno [3,2-d]pyrimidin-6-
yl)methylamino)pyrimidine-5-carboxylate (Compound 0603-211)
The title compound 0603-211 was prepared (65 mg, 22%) as a yellow solid from
0504-53
(256 mg, 0.59 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2y1)phenylamine
(155 mg,
0.71 mmol), Cs2CO3 (577 mg, 1.77 mmol) and Pd(dppf)2C12 (48 mg, 0.06 mmol) in
1,4-
dioxane (6 mL) and water (0.2 mL) using a procedure similar to that described
for
compound 0603-107 (Example 34). LCMS: 492 [M+1]+.'H-NMR (400 MHz. DMSO-d6) 6
1.29 (t, J= 7.2 Hz, 3H), 3.75 (m, 4H), 3.89 (m, 4H), 4.27 (q, J= 7.2 Hz, 2H),
4.87 (d, J=
6.0 Hz, 2H), 5.53 (s, 2H), 6.60 (m, 2H), 7.28 (s, 1H), 8.09 (m, 2H), 8.83 (m,
3H).
Step 93b: 2-((2-(4-aminophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
methylamino)-
N-hydroxypyrimidine-5-carboxamide (Compound 211)
The title compound 211 was prepared (28 mg, 45%) as a yellow solid from 0603-
211 (65
mg, 0.13 mmol) and freshly prepared hydroxylamine methanol solution (6 mL)
using a
procedure similar to that described for compound 3 (Example 1). m.p.: 217-223
C. LCMS:
479 [M+1]+. 'H-NMR (400 MHz. DMSO-d6) 6 3.76 (m, 4H), 3.89 (m, 4H), 4.84 (d,
J= 5.6
Hz, 2H), 5.53 (s, 2H), 6.60 (m, 2H), 7.27 (s, 1 H), 8.90 (m, 2H), 8.51 (t, J =
5.6 Hz, 1 H),
8.66 (s, 2H), 9.05 (s, I H), 11.09 (s, I H).
Example 94: (E)-3-(4-(N-(2-(2-aminopyrimidin-5-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)sulfamoyl)phenyl)-N-hydroxyacrylamide (Compound 217)
Step 94a: 7-Bromothieno[3,2-d]pyrimidine-2,4(1H,3H)-dione (Compound 1001)
Bromine (9.1 mL, 178 mmol) was added to a solution of compound 0109 (10 g,
0.059 mol) in AcOH (166 mL) at room temperature. After the addition, the
reaction was
heated to 70 C and stirred overnight. The reaction mixture was cooled to room
temperature
and poured into ice-water (1 L). The resulting mixture was filtered and washed
with water.
The solid was suspended in saturated Na2S2O3 solution and stirred for 30 min
followed by
filtration. The solid was washed with water and saturated NaHCO3 solution,
dried in vacuo
to afford 1001 as a yellow solid (14.6 g, 87%). LCMS: 247.0 [M+1]+. 'H NMR
(400 MHz,
DMSO-d6): 6 8.22 (s, 1H), 11.40 (s, 1H), 11.51 (s, 1H).
Step 94b: 7-Bromo-2,4-dichlorothieno [3,2-d] pyrimidine (Compound 1002)
A mixture of compound 1001 (12.8 g, 51 mmol) in POC13 (150 mL) was heated to
reflux for 12 h. The excess POC13 was removed in vacuo and the residue was
poured into
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crush ice and filtered to afford the titled compound 1002 as a yellow solid.
(11.8 g, 80.2%).
LCMS: 284.8 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 8.86 (s, 1H).
Step 94c: 4-(7-Bromo-2chlorothieno[3,2-d]pyrimidin-4-yl)morpholine (Compound
1003)
Compound 1002 (11.8 g, 41.5 mmol) was suspended in methanol (150 mL) at room
temperature. Morpholine (11.2 mL, 124.5 mmol) was added. The resulting mixture
was
stirred at room temperature for 2 h before it was filtered. The solid was
washed with water,
methanol and dried in vacuo to afford compound 1003 as a yellow solid. (11.5
g, 82.7%).
LCMS: 336.0 [M+1]+. 'H NMR (400 MHz, DMSO- d6): 6 3.77 (t, J= 5.2 Hz, 4H),
3.92 (t,
J = 5.2 Hz, 4H), 8.52 (s, 1 H).
Step 94d: 4-(7-Bromo-2chloro-6-nitrothieno[3,2-d]pyrimidin- 4-yl)- morpholine
(Compound 1004)
Compound 1003 (11.5 g, 34.3 mmol) was added portions to concentrated H2SO4 (35
mL) at 0 C. Then fuming HNO3 (9 mL, 206 mmol) was added dropwise to above
solution
at 0 C over 1530 min. The resulting solution was stirred at 0 C for 2 h before
it was
poured into crush ice. The resulting mixture was filtered, washed with water
and dried in
vacuo to afford 1004 as a yellow solid (12.0 g, 90.0%). LCMS: 371.0 [M+1]+. 'H
NMR
(400 MHz, DMSO-d6): 6 3.78 (t, J= 5.2 Hz, 4H), 3.94 (t, J= 5.2 Hz, 4H).
Step 94e: 2-Chloro-4-morpholinothieno[3,2-d]pyrimidin-6-amine (Compound 1005)
A mixture of compound 1004 (12.0 g, 31.5 mmol), Tin powder (11.0 g, 94.7 mmol)
and concentrated HC1(31.6 mL) in methanol (350 mL) was stirred at 50 C for 1
h until it
became clear solution. More tin powder (7.6 g, 65.4 mmol) and concentrated
hydrochloric
acid (25 mL) were added to the solution and the resulting mixture was stirred
at 50 C
overnight. The reaction mixture was cooled to room temperature and filtered to
afford
compound 1005 as a pale yellow solid (4.0 g, 47%). LCMS: 271.0 [M+1]+. 'H NMR
(400
MHz, DMSO-d6): 6 3.69 (s, 8H), 5.94 (s, 1H), 7.15 (s, 2H).
Step 94f: (E)-Methyl 3-(4-(N-(2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)
sulfamoyl)phenyl)acrylate (Compound 1007-217)
To a mixture of 1006-217 (300 mg, 1.11 mmol) and 1005 (400 mg, 1.56 mmol) in
THE (30 mL) was added NaH (300 mg, 12.5 mmol). The reaction mixture was
stirred at
room temperature for 3 h. The reaction mixture was concentrated. Water was
added to the
mixture and stirred for 30 min. The resulting mixture was neutralized with
HOAc and
filtered. The collected solid was dissolved in MeOH (20 mL), H2SO4 (5 drops)
was added.
The reaction mixture was refluxed for 3h and concentrated. The residue was
treated with
water (20 mL) and filtered. The solid was washed with water and dried to
afford the titled
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compound 1007-217 (250 mg, 46%). It was used for the next step reaction
without further
purification.
Step 94g: (E)-Methyl 3-(4-(N-(2-(2-aminopyrimidin-5-yl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)sulfamoyl)phenyl)acrylate (Compound 1008-217)
A mixture of 1007-217 (250 mg, 0.516 mol), 5-(4,4,5,5-tetramethyl- 1,3,2-
dioxaborolan-2-yl)pyrimidin-2-amine (0602-217) (200 mg, 1.43 mmol), sat.
NaHCO3 (2
mL) and Pd(PPh3)4 (50 mg) in DMSO (15 mL) was stirred at 130 C for 5 h under
N2. To
the reaction mixture was added water and neutralized with HOAc. The
precipitate was
collected by filtration. The crude product was purified by column
chromatography to afford
the titled compound 1008-217 as a yellow solid (160 mg, 57.3%). 'HNMR (400
MHz,
DMSO-d6): 6 3.73 (s, 3H), 3.76 (s, 4H), 3.95 (s, 4H), 6.71 (d, J= 16.0 Hz,
1H), 7.4 (br,
2H), 7.66 (d, J= 16.0 Hz, 1H), 7.82 (br, 4H), 8.99 (s, 2H).
Step 94h: (E)-3-(4-(N-(2-(2-aminopyrimidin-5-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl)sulfamoyl)phenyl)-N-hydroxyacrylamide (Compound 217)
A mixture of 1008-217 (150 mg) and freshly prepard NH2OH (8 mL, 1.79 M in
MeOH)
in CH2Cl2 (5 mL) was stirred at 0 C for 2h. The reaction mixture was adjusted
to pH = 6-7
with 1.2 M aquous HC1 and filtered. The cake was purified by prep-HPLC to
afford the
titled compound 94 as a yellow solid (70 mg). M.p. >300 C. LCMS: 555 [M+1]+.
'HNMR
(400 MHz, DMSO-d6): 6 3.75 (s, 3H), 3.92 (s, 4H), 6.30 (br, 1H), 6.51 (d, J=
16.0 Hz,
1H), 7.30 (br, 1H), 7.45 (d, J= 16.0 Hz, 2H), 7.66 (d, J= 7.6 Hz, 2H), 7.81
(d, J= 8.0 Hz,
2H), 8.99 (s, 2H), 9.08 (s, 1H), 10.82 (s, 1H).
EXAMPLE 95: (E)-3-(3-(N-(2-(2-aminopyrimidin-5-yl)-4-morpholinothieno [3,2-
d]pyrimidin-6-yl)sulfamoyl)phenyl)-N-hydroxyacrylamide (Compound 218)
Step 95a: (E)-Methyl 3-(3-(chlorosulfonyl)phenyl)acrylate (1006-218)
To a solution of 3-nitro-benzaldehyde (15 g, 0.1 mol), (dimethoxy- phosphoryl)-
acetic
acid methyl ester (27 g, 0.148 mol) in DMF (100 ml) was added NaOMe (10.7 g
0.198
mol). The reaction was mornitored by TLC. After reaction was completed, the
reaction
mixture was adjusted to pH = 1 with aquous HC1 solution and evaporated. The
resulting
solid was washed with water to afford (E)-methyl 3-(3-nitrophenyl)acrylate as
a yellow
solid (20.14 g, 98%).
A solution of (E)-methyl 3-(3-nitrophenyl)acrylate (20.14 g, 0.097 mol), Fe
(32.5 g,
0.58 mol), concentrated HC1(3.5 ml) in 50% EtOH (100 ml) was refluxed for 2h.
Upon
completion, the solution was filtered through Celite and washed with EtOH. The
filtrate
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was concentrated and extracted with dichloromethane. The combined organic
phase was
dried over MgSO4 and concentrated to afford (E)-methyl 3-(3-
aminophenyl)acrylate as a
yellow solid (9.73 g, 57%).
To a solution of (E)-methyl 3-(3-aminophenyl)acrylate (11.55 g, 0.065 mol) in
AcOH
(13 mL) was added conc. HC1(45 mL). The resulting mixture was cooled to -10 C
by
EtOH/dry ice bath. NaNO2 solution (5 g in 7.2 mL water) was added dropwise at
such a
rate that internal temperature did not exceed -5 C to form diazotization
reaction solution.
SO2 (g) was introduced below the surface of HOAc (65 mL) until saturation was
evident.
CuC1(1.7 g) was added to the solution and continued the introduction of SO2
until the
yellow green suspension became blue-green and cooled to 0 C. To the reaction
mixture
was added the diazotization solution in portions prepared above and continued
reaction for
30 min. The reaction mixture was poured into ice and extracted with
dichloromethane. The
crude product was purified by column chromatography to afford 1006-218 as a
yellow
solid. 'HNMR (400 MHz, CDC13): 6 3.84 (s, 3H), 6.57 (d, J= 16 Hz, 1H), 7.66
(t, J= 8.0
Hz, 1 H), 7.72 (d, J = 16 Hz, 1 H), 7.85 (d, J = 7.6 Hz, 1 H), 8.04 (d, J =
8.0 Hz, 1 H), 8.16 (s,
I H).
Step 95b: (E)-Methyl 3-(3-(N-(2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)
sulfamoyl)phenyl)acrylate (Compound 1007-218)
To a mixture of 1006-218 (300 mg, 1.11 mmol) and 1005 (400 mg, 1.56 mmol) in
THE
(30 mL) was added NaH (300 mg, 12.5 mmol). The reaction mixture was stirred at
room
temperature for 3h. The reaction mixture was concentrated. Water was added to
the mixture
and stirred for 30 min. The resulting mixture was neutralized with HOAc and
filtered. The
collected solid was dissolved in MeOH (20 mL), H2SO4 (5 drops) was added. The
reaction
mixture was refluxed for 3h and concentrated. The residue was treated with
water (20 mL)
and filtered. The solid was washed with water and dried to afford the titled
compound
1007-218 as a yellow solid (320 mg, 58%).
'HNMR (400 MHz, DMSO-d6): 6 3.70-3.79 (m, 11H), 6.67 (s, 1H), 6.72 (d, 16.0
Hz, 1H),
7.66 (t, J = 6.0 Hz, 1 H), 7.74 (d, J = 16.0 Hz, 1 H), 7.88 (d, J = 8.4 Hz, 1
H), 8.04 (d, J = 8.0
Hz, I H), 8.17 (s, I H).
Step 95c: (E)-Methyl 3-(3-(N-(2-(2-aminopyrimidin-5-yl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)sulfamoyl)phenyl)acrylate (Compound 1008-218)
The title compound 1008-218 was prepared as a yellow solid (200 mg, 60%) from
1007-218 (300 mg, 0.6 mol), 5-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-
yl)pyrimidin-2-
amine (0602-217) (300 mg, 2.16 mmol), sat. NaHCO3 (2 mL) and Pd(PPh3)4 (50 mg)
in
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DMSO (10 mL) using a procedure similar to that described for compound 1008-217
(Example 94): 'HNMR (400 MHz, DMSO-d6): 6 3.79-3.83 (m, 7H), 4.01 (s, 4H),
6.40 (br,
1 H), 6.72 (d, J = 16.0 Hz, 1 H), 7.42 (br, 2H), 7.62 (t, J = 7.6Hz, 1 H),
7.78 (d, J = 16.0 Hz,
1H), 7.88-7.98 (m, 3H), 8.13 (s, 1H), 9.03 (s, 2H).
Step 95d: (E)-3-(3-(N-(2-(2-aminopyrimidin-5-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl)sulfamoyl)phenyl)-N-hydroxyacrylamide (Compound 218)
The title compound 218 was prepared as a light yellow solid (83 mg) from 1008-
218
(150 mg) and freshly prepared hydroxylamine methanol solution (10 mL) using a
procedure
similar to that described for compound 217 (Example 94): M.p.: >300 C. LCMS:
555[M+1]+.'HNMR: (400 MHz, DMSO-d6): 6 3.75 (s, 4H), 3.95 (s, 4H), 6.13 (br,
1H),
6.55 (d, J= 16.0 Hz, 1H), 7.47-7.55 (m, 3H), 7.70-7.79 (m, 2H), 7.98 (s, 1H),
9.00 (s, 2H),
9.15 (s, 1 H), 10.15 (br, 1 H), 10.81 (s, 1 H), 13,4 (br, 1 H).
EXAMPLE 96: (E)-N-hydroxy-3-(3-(N-(2-(6-methoxypyridin-3-yl)-4-morpho-
linothieno[3,2-d]pyrimidin-6-yl)sulfamoyl)phenyl)acrylamide (Compound 221)
Step 96a: (E)-Methyl 3-(3-(N-(2-(6-methoxypyridin-3-yl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)sulfamoyl)phenyl)acrylate (Compound 1008-221)
To a stirred mixture of 1007-218 (300 mg, 0.61 mmol) and 0602-221 (287 mg,
1.22
mmol) in DMSO (20 mL) was added Pd(PPh3)4 (36.7 mg, 0.032 mmol) and saturated
aq.
NaHCO3 (2 mL). The resulting mixture was heated at 120 C for 4h. To the
reaction
mixture was added water and adjusted to pH = 6-7 with acetic acid. The
precipitate was
collected by filtration. The crude product was purified by column
chromatography to afford
the titled compound 1008-221 as a yellow solid (250 mg, yield 72%).
1H NMR (400 MHz, DMSO-d6): 6 3.71(s, 3H), 3.76 (t, J= 4.4 Hz, 4H), 3.92-3.98
(m, 7H),
6.68 (d, J= 16.0 Hz, 1H), 6.97 (d, J= 8.8 Hz, 1H), 7.52-7.60 (m, 2H), 7.73 (d,
J= 16.0 Hz,
I H), 7.85 (d, J= 8.0 Hz, I H), 7.94 (d, J= 5.4 Hz, I H), 8.10 (s, I H), 8.43-
8.46 (m, 1H),9.02
(s, 1H).
Step 96b: (E)-N-hydroxy-3-(3-(N-(2-(6-methoxypyridin-3-yl)-4-morpholinothieno
[3,2-
d]pyrimidin-6-yl)sulfamoyl)phenyl)acrylamide (Compound 221)
The title compound was prepared as a white solid (70 mg, yield 46%) from 1008-
221
(150 mg, 0.264 mmol) and freshly prepared hydroxylamine methanol solution (10
mL, 1.79
M in MeOH) using a procedure similar to that described for compound 217
(Example 94):
M.p.: >300 C. LCMS: 569.2 [M+1]+. iH NMR (400 MHz, DMSO-d6): 6 3.77 (s, 4H),
3.94
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(s, 7H), 6.55 (d, J= 16.0 Hz, 1H), 6.97 (d, J= 8.4 Hz, 1H), 7.47-7.57 (m, 2H),
7.72-7.80
(m, 2H), 7.99 (s, I H), 8.45 (d, J= 8.0 Hz, I H), 9.02 (s, I H), 9.06 (br, I
H), 10.78 (s, I H).
EXAMPLE 97: N-Hydroxy-3-{4-[2-(6-methoxy-pyridin-3-yl)-4-morpholin-4-yl-
thieno[3,2-d]pyrimidin-6-ylsulfamoyl]-phenyl}-acrylamide (Compound 222)
Step 97a: 3 - {4- [2-(6-Methoxy-pyridin-3 -yl)-4-morpholin-4-yl-thieno [3,2-d]
pyrimidin-6-
ylsulfamoyl]-phenyl}-acrylic acid methyl ester (Compound 1008-222)
The title compound 1008-222 was prepared as a yellow solid (200 mg, 58%) from
1007-217 (300 mg, 0.61 mol), 0602-221 (287 mg, 1.22 mmol), Pd(PPh3)4 (36.7 mg,
0.032
mmol) and sat. NaHCO3 (2 mL) in DMSO (20 mL) using a procedure similar to that
described for compound 1008-221 (Example 96): LCMS: 568.2 [M+1]+. 'H NMR(400
MHz, DMSO-d6): 6 3.72 (s, 3H), 3.76 (t, J= 4.4 Hz, 4H), 3.93 (s, 7H), 5.75 (s,
1H), 6.71
(d, J = 16 Hz, 1 H), 6.96 (br, 1 H), 7.67 (d, J = 16 Hz, 1 H), 7.81-7.85 (m,
4H), 8.45 (br, 1 H),
9.02 (s, 1H).
Step 97b: N-Hydroxy-3 - {4- [2-(6-methoxy-pyridin-3 -yl)-4-morpholin-4-yl-
thieno [3,2-
d]pyrimidin-6-ylsulfamoyl] -phenyl}-acrylamide (Compound 222)
The title compound 222 was prepared as a white solid (32 mg, 21.%) from 1008-
222 (150 mg, 0.264 mmol) and freshly prepared hydroxylamine methanol solution
(5 mL,
1.79 M) using a procedure similar to that described for compound 217 (Example
94): M.p.:
>300 C. LCMS: 569.2 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 3.72 (s, 4H), 3.75
(s,
4H), 3.87 (s, 3H), 6.04 (s, 1 H), 6.44 (d, J = 16 Hz, 1 H), 6.83 (d, J = 8.8
Hz, 1 H), 7.40 (d, J
= 16 Hz, 1H), 7.56 (d, J = 8 Hz, 2H), 7.73(d, J = 7.6 Hz, 2H), 8.47(d, J = 8.4
Hz, 1H), 9.03
(m, 2H), 10.79 (s, 1H).
EXAMPLE 98: 4-(2-(2-aminopyrimidin-5-yl)-4-morpholinothieno[3,2-d] pyrimidin-6-
ylamino)-N-hydroxybutanamide (Compound 225)
Step 98a: tert-Butyl 2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-ylcarbamate
(Compound 1101)
Compound 1005 (2.0 g, 7.4 mmol) and (Boc)20 (1.95 g, 8.9 mmol) were taken into
DMF (10 mL) followed by the addition of NaH (0.90 g, 22.2 mmol). The resulting
mixture
was stirred at 0 C for 3 h. The reaction was mornitored by TLC. After
reaction
completion, the mixture was partitioned between dichloromethane and water. The
organic
layer was washed with brine, dried over Na2SO4 and evaporated to give a crude
product
which was purified by column chromatography eluted with hexanes/ethyl acetate
to afford
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the titled compound as a light yellow solid (1.3 g, 47.5%). LCMS: 371.0
[M+1]+. 'H NMR
(400 MHz, DMSO-d6): 6 1.51 (s, 9H), 3.75 (t, J= 4.8 Hz, 4H), 3.84 (t, J= 4.8
Hz, 4H),
6.64 (s, I H), 11.26 (s, I H).
Step 98b: Ethyl 4-((2-(2-aminopyrimidin-5-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)(tert-butoxycarbonyl)amino)butanoate (Compound 1103-225)
A mixture of 1101 (300 mg, 0.81 mmol), ethyl 4-bromobutanoate (236 mg, 1.22
mmol) and Cs2CO3 (528 mg, 1.62 mmol) was taken into DMF (10 mL) and heated at
100
C for 2 h. To the reaction mixture was added water and extracted with ethyl
acetate. The
ethyl acetate extract was concentrated to give a crude product 1102-225 which
was used for
the next step without further purification.
A mixture of compound 1102-225 (280 mg, 0.58 mmol), 5-(4,4,5,5-tetramethyl-
1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (0602-217) (640 mg, 2.9 mmol),
NaHCO3 (150
mg, 1.5 mmol), and bis(triphenylphosphine)palladium(II) chloride (100 mg, 0.14
mmol) in
a mixed solvents of toluene (23 mL), ethanol (14 mL) and water (6 mL) was
flushed with
nitrogen and heated at 130 C for 3.5h. The reaction mixture was concentrated
and residue
was partitioned between EtOAc and H20. The organic layer was washed with
brine, dried
over Na2SO4, and evaporated. The crude product was purified by column
chromatography
eluted with DCM / EtOAc to afford titled compound 1103-225 as a white solid
(160 mg,
51%).
HNMR: (400 MHz, CDC13): 6 1.28 (t, J= 7.2 Hz, 3H), 1.60 (s, 9H), 2.08-2.11 (m,
2H),
2.40-2.44 (m, 2H), 3.87 (t, J= 4.4 Hz, 4H), 3.98-4.02 (m, 6H), 4.17 (q, J= 7.2
Hz, 2H),
5.26 (s, 2H), 6.85 (s, 1H), 9.27 (s, 2H).
Step 98c: 4-(2-(2-Aminopyrimidin-5-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-
ylamino)-
N-hydroxybutanamide (Compound 225)
A solution of compound 1103-225 (160 mg, 0.3 mmol) in TFA (2 mL) was stirred
at
room temperature for 5h. Then the mixture was neutralized with sat. NaHCO3 and
extract
with dichloromethane. Crude 1104-225 was obtained as white solid after
concentration and
used for next step reaction without further purification.
Crude 1104-225 (160 mg, 0.36 mmol) was taken into NH2OH methanol solution (5
mL, 1.79 M) and DCM (2 mL) and stirred at room temperature overnight. The
resulting
mixture was adjusted to pH = 6-7 with acetic acid and collected the solid. The
crude
product was purified by prep-HPLC to afford the title compound 225 as an off-
white solid
(40 mg). M.p.: 220-230 C. LCMS: 431.0[M+1]+. 1H NMR (400 MHz, DMSO-d6): 6
1.81-
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1.83 (m, 2H), 2.05-2.07 (m, 2H), 3.15-3.17 (m, 2H), 3.75-3.76 (m, 8H), 6.02
(s, 1H), 6.99
(s, 2H), 7.49 (t, J= 5.2 Hz, I H), 8.72 (s, I H), 9.05 (s, 2H), 10.40 (s, I
H).
EXAMPLE 99: 4-(2-(2-Aminopyrimidin-5-yl)-4-morpholinothieno [3,2-d] pyrimidin-
6-
yl amino)-N-hydroxybutanamide (Compound 226)
Step 99a: 5-[tert-Butoxycarbonyl-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]
pyrimidin-6-
yl)-amino]-pentanoic acid ethyl ester(Compound 1102-226)
A mixture of compound 1101 (300 mg, 0.81 mmol), ethyl 5-bromopentanoate (314
mg, 1.62 mmol) and Cs2CO3 (0.53 g, 1.62 mmol) in DMF (23 mL) was stirred at
100 C for
2h. The reaction mixture was cooled to room temperature and partitioned
between EtOAc
(25 mL) and H2O (25 mL). The organic layer was washed with brine, dried over
Na2SO4,
filtered, and evaporated. The crude product 1102-226 was obtained as a yellow
solid after
concentration and used for next step without further purification. 'H NMR (400
MHz,
CDC13): 6 1.25 (J= 7.2 Hz, t, 3H), 1.59 (s, 9H), 1.60-1.80 (m, 4H), 2.34 (t,
J= 7.2 Hz,
2H), 3.86(t, J= 4.4 Hz, 4H), 3.88(t, J= 7.6 Hz, 2H), 3.95(t, J= 4.4 Hz, 4H),
4.14 (t, J
7.2 Hz, q, 2H), 6.65 (s, 1H).
Step 99b: 5 - {[2-(2-Amino-pyrimidin-5 -yl)-4-morpholin-4-yl-thieno [3,2-d]
pyrimidin-6-
yl] -t ert-butoxycarbonyl-amino}-pentanoic acid ethyl ester (Compound 1103-
226)
A mixture of 1102-226 (280 mg, 0.56 mmol), 5-(4,4,5,5-tetramethyl-1,3,2-
dioxaborolan-2-yl)pyrimidin-2-amine (0602-217) (620 mg, 2.8 mmol), NaHCO3 (150
mg,
1.8 mmol), and bis(triphenylphosphine)palladium(II) chloride (100 mg, 0.14
mmol) in a
mixed solvents of toluene (23 mL), ethanol (14 mL) and water (6 mL) was
flushed with N2
and heated at 130 C for 3.5h. Reaction mixture was concentrated and the
residue was
partitioned between EtOAc and water. The organic layer was washed with brine,
dried over
Na2SO4, and evaporated. The crude product was purified by column
chromatography eluted
with dichloromethane / EtOAc to afford 1103-226 as a white solid (160 mg,
51%). LCMS:
558 [M+1]+. 1H NMR (400 MHz, CDC13): 6 1.25 (t, J= 7.2 Hz, 3H), 1.59 (s, 9H),
1.76-
1.90 (m, 4H), 2.36 (t, J = 7.2 Hz, 2H), 3.86 (t, J = 4.4 Hz, 4H), 3.92 (t, J =
7.2 Hz, 2H),
4.00 (t, J= 4.4 Hz, 4H), 4.14 (q, J= 7.2 Hz, 2H), 5.30 (s, 2H), 6.76 (s, 1H),
9.26 (s, 2H).
Step 99c: 4-(2-(2-Aminopyrimidin-5-yl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl
amino)-
N-hydroxybutanamide (Compound 226)
The title compound 226 was prepared as an off-white solid (25 mg) from 1103-
226
(160 mg, 0.28 mmol) and TFA (2 mL), and followed by treatment with freshly
prepared
hydroxylamine methanol solution (5 mL, 1.79 M) using a procedure similar to
that
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described for compound 1103-225 (Example 98): m.p: 175-180 C. LCMS: 445.0
[M+1]+.
iH NMR (400 MHz, DMSO-d6): 6 1.57 (br, 4H), 1.99 (t, J= 6.4 Hz, 2H), 3.17 (s,
2H), 3.75
(s, 4H), 3.77 (s, 4H), 6.01 (s, I H), 6.99 (br, 2H), 7.50 (br, I H), 8.67 (s,
I H), 9.04 (s, 2H),
10.35 (s, 1H).
EXAMPLE 100: N-Hydroxy-5-(2-(2-(methylamino)pyrimidin-5-yl)-4-morpho-
linothieno[3,2-d]pyrimidin-6-ylamino)pentanamide (Compound 227)
Step 100a: Ethyl 5-(tert-butoxycarbonyl(2-(2-(methylamino)pyrimidin-5-yl)-4-
morpholinothieno[3,2-d]pyrimidin-6-yl)amino)pentanoate (Compound 1103-227)
A mixture of 1102-226 (300 mg, 0.6 mmol), N-methyl-5-(4,4,5,5-tetra methyl-
1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (0602-227) (645 mg, 4.22 mmol) and
Pd(PPh3)2C12( 12.7 mg) in a mixed solvents of toluene (6 mL), ethanol (4 mL)
and water (1
mL) was stirred at 90 C for 3h under N2 atmosphere. After the completion of
the reaction
(mornitored by TLC), the reaction mixture was concentrated and the residue was
purified
by column chromatography eluted with methanol in dichloromethane to afford
title product
as a colorless oil (150 mg, 43%).'HNMR (400 MHz, DMSO-d6): 6 1.16 (t, J= 7.2
Hz,
3H), 1.53 (s, 9H), 1.60-1.80 (m, 4H), 2.36 (t, J= 6.4Hz, 2H), 2.87 (d, J= 4.8
Hz, 3H), 3.78
(t, J = 4.4 Hz, 4H), 3.90-3.96 (m, 6H), 4.04 (q, J = 7.2 Hz, 2H), 6.99 (s, 1
H), 7.52 (d, J =
4.8 Hz, 1H), 9.14 (br, 2H).
Step 100b: Ethyl 5-(2-(2-(methylamino)pyrimidin-5-yl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-ylamino)pentanoate (Compound 1104-227)
A mixture of 1103-227 (150 mg, 0.26 mmol) and TFA (7 ml) was stirred at room
temperature for 1.5h. Upon completion of reaction, water was added to the
mixture at 10
C. The mixture was then adjusted to pH = 10-12 with 30% NaOH and extracted
with
dichloromethane. The crude product was purified by column chromatography
(dichloromethane/MeOH) to afford the title compound as a light-yellow solid
(75 mg,
61%).'HNMR (400 MHz, DMSO-d6): 6 1.16 (t, J= 7.2 Hz, 3H), 1.60-1.65 (m, 4H),
2.34
(m, 2H), 2.87 (d, J= 4.8 Hz, 3H), 3.18-3.20 (m, 2H), 3.75-3.76 (m, 8H), 4.06
(q, J= 7.2
Hz, 2H), 6.02 (s, I H), 7.41-7.47 (m, I H), 9.09 (br, 2H).
Step 100c: N-Hydroxy-5-(2-(2-(methylamino)pyrimidin-5-yl)-4-morpholinothieno
[3,2-
d]pyrimidin-6-ylamino)pentanamide (Compound 227)
The title compound 227 was prepared as a white solid (45 mg, 60%) from 1104-
227
(75 mg, 0.16 mmol) and freshly prepared hydroxylamine methanol solution (1.79
M, 20
mL) using a procedure similar to that described for compound 217 (Example 94):
m.p.:
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206-210 C. LCMS: 459.0 [M+1]+.'HNMR (400 MHz, DMSO-d6): 6 1.57-1.59 (m, 4H),
1.00 (t, J= 6.4 Hz, 2H), 2.86 (d, J= 4.8 Hz, 3H), 3.14-3.18 (m, 2H), 3.75-3.76
(m, 8H),
6.01 (s, 1H), 7.41-7.47 (m, 1H), 8.68 (s, 1H), 9.09 (br, 2H), 10.36 (s, 1H).
EXAMPLE 101: N-Hydroxy-5-(2-(6-(methylamino)pyridin-3-yl)-4-morpholino-
thieno[3,2-d]pyrimidin-6-ylamino)pentanamide (Compound 228)
Step 101a: Ethyl 5-(tert-butoxycarbonyl(2-(6-(methylamino)pyridin-3-yl)-4-
morpholino
thieno[3,2-d]pyrimidin-6-yl)amino)pentanoate (Compound 1103-228)
A mixture of 1102-226 (500 mg, 1.11 mmole), methyl-[5-(4,4,5,5-tetramethyl -
[1,3,2]dioxaborolan-2-yl)-pyridin-2-yl]-amine (0602-228) (920 mg, 6.01 mmole),
NaHCO3
(253 mg, 3.01 mmol) and Pd(PPh3)2C12 (35 mg) in toluene/EtOH/H20 (16 mL/10
mL/4
mL) was degassed with N2 and heated at 110 C overnight. TLC showed that the
reaction
was complete. The reaction mixture was concentrated. The residue was purified
by column
chromatography to afford 1103-228 as an off-white solid (370 mg, 65%). LCMS:
[M+1]+=571.
Step 101b: Ethyl 5-(2-(6-(methylamino)pyridin-3-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-y lamino)pentanoate (Compound 1104-228)
The title compound 1104-228 was prepared as yellow solid (300 mg, 98%) from
1103-228 (370 mg, 0.65 mmole) and TFA (4 ml) using a procedure similar to that
described for compound 1104-227 (Example 100): LCMS: [M+1]+=471
Step 101c: N-hydroxy-5-(2-(6-(methylamino)pyridin-3-yl)-4-morpholinothieno
[3,2-
d]pyrimidin-6-ylamino)pentanamide (Compound 228)
The title compound 228 was prepared as a light yellow solid (80 mg, 51 %) from
1104-228 (160 mg, 0.34 mmol) and a freshly prepared hydroxylamine methanol
solution
(10 mL, 1.79 M) using a procedure similar to that described for compound 217
(Example
94): M.p.:199-201 C. LCMS: 438 [M+1]+.'HNMR: (400 MHz, DMSO-d6): 6 1.58 (s,
4H),
2.00 (s, 2H), 2.82 (d, J= 4.4 Hz, 2H), 3.16 (d, J= 4.0 Hz, 2H), 3.75 (s, 8H),
5.99 (s, 1H),
6.47 (d, J= 8.8 Hz, 1H), 6.83 (d, J= 4.0 Hz, 1H), 7.43 (s, 1H), 8.23 (d, J=
8.4 Hz, 1H),
8.72 (s, I H), 8.96 (s, I H), 10.39 (s, I H).
EXAMPLE 102: N-Hydroxy-5-(2-(2-methylpyrimidin-5-yl)-4-morpholinothieno [3,2-
d]pyrimidin-6-ylamino)pentanamide (Compound 229)
Step 102a: Ethyl 5-(tert-butoxycarbonyl(2-(2-methylpyrimidin-5-yl)-4-
morpholino- thieno
[3,2-d]pyrimidin-6-yl)amino)pentanoate (Compound 1103-229)
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A mixture of 1102-226 (300 mg, 0.619 mmol), 2-methyl-5-(4,4,5,5-tetra methyl-
1,3,2-dioxaborolan-2-yl)pyrimidine (0602-229) (681 mg, 3.10 mmol), NaHCO3 (155
mg,
1.854 mmol) and Pd(PPh3)2C12 (50 mg) in toluene/EtOH/H20 (5.0 mL/3.2 mL/1.4
mL) was
stirred at 120 C for 5 h under N2. The reaction mixture was concentrated. The
residue was
purified by column chromatography to afford the titled compound as an off-
white solid
(300 mg, yield 89.8%).
Step 102b: Ethyl 5-(2-(2-methylpyrimidin-5-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
ylamino)pentanoate (Compound 1104-229)
The title compound 1104-229 was prepared as a white solid (130 mg) from 1103-
229 (300 mg) and TFA (3 mL) using a procedure similar to that described for
compound
1104-227 (Example 100).
Step 102c: N-Hydroxy-5-(2-(2-methylpyrimidin-5-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-ylamino)pentanamide (Compound 229)
The title compound 229 was prepared as a white solid (55 mg) from 1104-229
(130
mg) and a freshly prepared hydroxylamine methanol solution (10 mL, 1.79 M)
using a
procedure similar to that described for compound 217 (Example 94): M.p.: 248-
252 C.
LCMS: 444.0 [M+1]+.'HNMR (400 MHz, DMSO-d6): 6 1.58 (s, 4H), 2.00-2.01 (m,
2H),
2.69 (s, 3H), 3.18 (d, J= 5.2 Hz, 2H), 3.76 (d, J= 4.4 Hz, 4H), 3.80 (d, J=
4.4 Hz, 4H),
6.09 (s, I H), 7.60 (t, J= 5.2 Hz, I H), 8.71 (s, I H), 9.44 (s, 2H), 10.28
(s, I H).
EXAMPLE 103: 6-[2-(2-Amino-pyrimidin-5-yl)-4-morpholin-4-yl-thieno [3,2-d]
pyrimidin-6-ylamino]-hexanoic acid hydroxyamide(Compound 232)
Step 103a: 6-[tert-Butoxycarbonyl-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]
pyrimidin-6-
yl)-amino]-hexanoic acid ethyl ester (Compound 1102-232)
The title compound 1102-232 was prepared as a yellow solid (480 mg, yield
100%)
from 1101 (300 mg, 0.81 mmol), ethyl 6-bromohexanoate (360 mg, 1.62 mmol) and
Cs2CO3 (0.53 g, 1.62 mmol) in DMF (23 mL) using a procedure similar to that
described
for compound 1102-226 (Example 99): 'HNMR (400 MHz, CDC13): 6 1.16 (t, J= 7.2
Hz,
3H), 1.30-1.37 (m, 2H), 1.52 (s, 9H), 1.55-1.65 (m, 4H), 2.24-2.30 (m, 2H),
3.26 (t, 2H),
3.74 (t, J= 4.8 Hz, 4H), 3.84(t, J= 4.4 Hz, 4H), 3.90 (t, J= 7.2 Hz, 2H), 4.04
(q, J= 7.2 Hz,
2H), 6.90 (s,1 H).
Step 103b: 6-{[2-(2-Amino-pyrimidin-5-yl)-4-morpholin-4-yl-thieno[3,2-d]
pyrimidin-6-
yl]-tert-butoxycarbonyl-amino}-hexanoic acid ethyl ester (Compound 1103-232)
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The title compound 1103-232 was prepared as a white solid (300 mg, 67%) from
1102-232 (400 mg, 0.78 mmol), 5-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-
yl)pyrimidin-
2-amine (0602-217) (730 mg, 3.3 mmol), NaHCO3 (200 mg, 2.4 mmol) and
bis(triphenylphosphine)palladium(II) chloride (140 mg, 0.39 mmol) in toluene
(32 mL),
ethanol (20mL) and water (8 mL) using a procedure similar to that described
for compound
1103-226 (Example 99): LCMS: 572.2[M+1]+. 1H NMR (400 MHz, CDC13): 6 1.25 (t,
J=
7.2 Hz, 3H), 1.39-1.45 (m, 2H), 1.59(s, 9H), 6 1.67-1.80 (m, 4H), 2.32 (t, J=
7.2 Hz, 2H),
3.85-3.91 (m, 6H), 4.00 (t, J = 5.2 Hz, 4H), 4.14 (q, J = 7.2 Hz, 2H), 6.75
(s, 1 H), 9.27 (s,
2H).
Step 103c: 6-[2-(2-Amino-pyrimidin-5-yl)-4-morpholin-4-yl-thieno[3,2-
d]pyrimidin- 6-
ylamino]-hexanoic acid ethyl ester (Compound 1104-232)
The title compound 1104-232 was prepared as a white solid (200 mg) from 1103-
232 (300 mg, 0.5 mmol) and TFA (3 mL) using a procedure similar to that
described for
compound 1104-227 (Example 100): LCMS: 472.2[M+1]+. 'H NMR (400 MHz, CDC13): 6
1.17 (t, J= 7.2 Hz, 3H), 1.35-1.40 (m, 2H), 1.55-1.61 (m, 4H), 2.29 (t, J= 7.2
Hz, 2H),
3.13-3.18 (m, 2H), 3.70-3.75 (m, 8H), 6 4.04 (q, J= 7.2 Hz, 2H), 5.99 (s, 1H),
6 6.91 (s,
2H), 6 7.39 (t, J= 5.6 Hz, 1H), 9.04 (s, 2H).
Step 103d: 6-[2-(2-Amino-pyrimidin-5-yl)-4-morpholin-4-yl-thieno[3,2-
d]pyrimidin- 6-
ylamino]-hexanoic acid hydroxyamide (Compound 232)
The title compound 232 was prepared as an off-white solid (20 mg) from 1104-
232
(100 mg, 0.21 mmol) and a freshly prepared hydroxylamine methanol solution (5
mL, 1.79
M) using a procedure similar to that described for compound 217 (Example 94):
m.p: 220-
225 C. LCMS: 459.0 [M+1]+. 'HNMR (400 MHz, DMSO-d6): 6 1.30-1.37 (m, 2H), 1.49-
1.62 (m, 4H), 1.96 (t, J = 7.2 Hz, 2H), 3.15 -3.17 (m, 2H), 3.76-3.91 (m, 8H),
6.11 (s, 1 H),
7.2 (br, 2H), 8.6 (br, I H), 9.09 (s, 2H), 10.39 (s, I H), 11.8 (br, I H).
EXAMPLE 104: 6-[2-(6-Methoxy-pyridin-3-yl)-4-morpholin-4-yl-thieno [3,2-d]
pyrimidin-6-ylamino]-hexanoic acid hydroxyamide (Compound 233)
Step 104a: 6-{tert-Butoxycarbonyl-[2-(6-methoxy-pyridin-3-yl)-4-morpholin-4-yl-
thieno [3,2-d]pyrimidin-6-yl] -amino }-hexanoic acid ethyl ester(Compound 1103-
233)
A mixture of compound 1102-232 (200 mg, 0.39 mmol), 0602-221 (183 mg, 0.78
mmol), NaHCO3 (98 mg, 1.17 mmol) and bis(triphenylphosphine)palladium(II)
chloride
(14 mg, 0.039 mmol) in toluene (14 mL), ethanol (8.8 mL) and water (3.5 mL)
was flushed
with nitrogen and heated at 120 C overnight. The reaction mixture was
concentrated and
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partitioned between EtOAc and water. The organic layer was washed with brine,
dried over
Na2SO4. The crude product was purified by column chromatography eluted with
dichloromethan / EtOAc to afford the titled compound 1103-233 as a white solid
(200 mg,
88%). LCMS: 586.2[M+1]+.
Step 104b: Ethyl 6-(2-(6-methoxypyridin-3-yl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
ylamino)hexanoate (Compound 1104-233)
The title compound 1104-233 was prepared as a white solid (160 mg, 96%) from
1103-233 (200 mg, 0.34 mmol) and TFA (3 mL) using a procedure similar to that
described
for compound 1104-227 (Example 100): LCMS: 486.5 [M+1]+. 'H NMR (400 MHz,
CDC13): 6 1.26 (t, J= 7.2 Hz, 3H), 1.30-1.45 (m, 2H), 1.46-1.80 (m, 4H), 2.30-
2.34 (m,
2H), 3.30-3.40 (m, 2H), 3.84-3.90 (m, 8H), 4.00 (s, 3H), 4.14 (m, 2H), 4.62
(br, 1H), 6.22
(s, 1 H), 6.79 (d, J = 8.8 Hz, 1 H), 8.5 5 (dd, J = 8.8 Hz, 2.4 Hz, 1 H), 9.10
(s, 1 H).
Step 104c: 6-[2-(6-Methoxy-pyridin-3-yl)-4-morpholin-4-yl-thieno[3,2-
d]pyrimidin-6-
ylamino]-hexanoic acid hydroxyamide (Compound 233)
The title compound 233 was prepared as an off-white solid (50 mg) from 1104-
233
(160 mg, 0.33 mmol) and a freshly prepared hydroxylamine methanol solution (10
mL,
1.79 M) using a procedure similar to that described for compound 217 (Example
94): m.p:
125-130 C. LCMS: 473 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.32-1.34 (m, 2H),
1.51-1.60 (m, 4H), 1.94-1.98 (m, 2H), 3.14-3.16 (m, 2H), 3.74-3.79 (m, 8H),
3.91(s, 3H),
6.03 (s, I H), 6.88 (d, J= 8.8 Hz, I H), 7.48 (t, J= 5.2 Hz, I H), 8.51 (dd,
J= 8.4 Hz, 2.4
Hz, I H), 8.67 (s, I H), 9.08 (s, I H), 10.34 (s, I H).
EXAMPLE 105: 7-[2-(2-Amino-pyrimidin-5-yl)-4-morpholin-4-yl-thieno [3,2-d]
pyrimidin-6-ylamino]-heptanoic acid hydroxyamide(Compound 234)
Step 105a: 7-[tert-Butoxycarbonyl-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]
pyrimidin-6-
yl)-amino]-heptanoic acid ethyl ester(Compound 1102-234)
The title compound 1102-234 was prepared as a yellow solid (480 mg, 100%) from
1101 (300 mg, 0.81 mmol), ethyl 7-bromoheptanoate (383 mg, 1.62 mmol) and
Cs2CO3
(0.53 g, 1.62 mmol) in DMF (23 mL) using a procedure similar to that described
for
compound 1102-226 (Example 99): LCMS: 527.2 [M+1]+.
Step 105b: 7- {[2-(2-Amino-pyrimidin-5-yl)-4-morpholin-4-yl-thieno[3,2-
d]pyrimidin- 6-
yl]-tert-butoxycarbonyl-amino}-heptanoic acid ethyl ester(Compound 1103-234)
The title compound 1103-234 was prepared as a white solid (220 mg, 49%) from
1102-234 (400 mg, 0.76 mmol), 5-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-
yl)pyrimidin-
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2-amine (0602-217) (840 mg, 3.8 mmol), NaHCO3 (200 mg, 2.4 mmol) and
bis(triphenylphosphine)palladium(II) chloride (140 mg,0.39mmol) in toluene (32
mL),
ethanol (20 mL) and water (8 mL) using a procedure similar to that described
for
compound 1103-226 (Example 99): LCMS: 586[M+1]+. 1H NMR (400 MHz, CDC13): 6
1.25 (t, J= 7.2 Hz, 3H), 1.40-1.41 (m, 2H), 1.59 (s, 9H), 1.63-1.76 (m, 2H),
2.30 (t, J= 7.6
Hz, 2H), 3.85-3.90 (m, 6H), 4.00 (t, J= 4.4 Hz, 4H), 4.13 (q, J= 7.2 Hz, 2H),
5.38 (s, 2H),
6.76 (s, 1H), 9.26 (s, 2H).
Step 105c: 7-[2-(2-Amino-pyrimidin-5-yl)-4-morpholin-4-yl-thieno[3,2-
d]pyrimidin-6-
ylamino]-heptanoic acid ethyl ester(Compound 1104-234)
The title compound 1104-234 was prepared as a white solid (200 mg) from 1103-
234 (220 mg, 0.38 mmol) and TFA (3 mL) using a procedure similar to that
described for
compound 1104-227 (Example 100): LCMS: 486.3[M+1]+.
Step 105d: 7-[2-(2-Amino-pyrimidin-5-yl)-4-morpholin-4-yl-thieno[3,2-d]
pyrimidin-6-
ylamino]-heptanoic acid hydroxyamide(Compound 234)
The title compound 234 was prepared as an off-white solid (100 mg) from 1104-
234
(100 mg, 0.21 mmol) and a freshly prepared hydroxylamine methanol solution (5
mL, 1.79
M) using a procedure similar to that described for compound 217 (Example 94):
m.p: 235-
240 C. LCMS: 473.0[M+1]+. 1H NMR (400 MHz, DMSO-d6): 6 1.23-1.35 (m, 4H),
1.48-
1.59 (m, 4H), 1.95 (t, J= 7.6 Hz, 2H), 3.12-3.19 (m, 2H), 3.74-3.75 (m, 8H),
5.99 (s, 1H),
6.97 (s, 2H), 7.43 (t, J= 5.2 Hz, 1H), 8.64 (s, 1H), 9.04 (s, 2H), 10.32(s,
1H)
EXAMPLE 106: N-Hydroxy-2-(methyl((2-(6-(methylamino)pyridin-3-yl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound 111)
Step 106a: (2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-
methyl-amine
(Compound 0503)
To the solution of 0112 (20.0 g, 70.4 mmol) in methanol (125 mL) was added
methylamine solution in methanol (27% v/v, 75 mL, 563.2 mmol) under nitrogen
atmosphere. The reaction mixture was stirred at room temperature overnight and
the
solvent was removed in vacuo to give a crude solid product, which was
dissolved in
methanol (550 mL) and THE (220 mL) under nitrogen. Sodium borohydride (8 g,
211.2
mmol) was added in portions and reaction mixture was stirred at room
temperature
overnight. The reaction mixture was evaporated in vacuo and water (300 mL) was
added.
The aqueous mixture was extracted with methylene chloride and the combined
extracts
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were dried over Na2SO4 and concentrated. The residue was dissolved in 6M
HC1(230 mL)
and stirred for 30 min. The aqueous solution was washed with methylene
chloride for
several times, and adjusted to pH = 9-10 with NaOH (4N). The precipitated
solid was
collected by filtration and dried (60 C, 6h) to give a light yellow solid (18
g, 85%).
LCMS: 299 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 2.32 (s, 3H), 3.74 (t, J= 5.2
Hz,
4H), 3.88 (t, J = 5.2 Hz, 4H), 3.96 (s, 2H), 7.24 (s, 1 H).
Step 106b: 2-[(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-
methyl-
amino]-pyrimidine-5-carboxylic acid ethyl ester (Compound 0504)
The mixture of 0503 (10 g, 33.6 mmol), CH3CN (400 mL) and 0305 (6.8 g, 36.4
mmol) was stirred at room temperature. Diisopropylethylamine (DIPEA) (220 mL,
1.26
mol) was then added and the solution was stirred overnight and evaporated.
After
methylene chloride (300 mL) was added, the organic phase was washed with
water, dried
over Na2SO4 and concentrated in vacuo to leave a residue. To the residue was
added ethyl
acetate and the mixture was stirred in ice/water bath for 50 min. The titled
product 0504
was collected as a white solid (10.6 g, 70%). LCMS: 449 [M+1]+; 'HNMR (400
MHz,
DMSO-d6): 6 1.30 (t, J= 7.2 Hz, 3H), 3.25 (s, 3H), 3.71 (t, J= 5.2 Hz, 4H),
3.83 (t, J= 4.8
Hz, 4H), 4.29 (m, 2H), 5.21 (s, 2H), 7.39 (s, 1H), 8.87 (s, 2H).
Step 106c: Ethyl 2-(methyl((2-(6-(methylamino)pyridin-3-yl)-4-
morpholinothieno[3,2-
d]pyri -midin-6-yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-111)
A mixture of N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-
amine (0602-227) (351 mg, 1.5 mmol), 0504 (314 mg, 0.7 mmol), NaHCO3 (176 mg,
2.1
mmol) and Pd(PPh3)2C12 (24.6 mg, 0.035 mmol) were dissolved in Toluene/ EtOH/
H2O
(2.5 mL/ 1.6 mL/ 0.7 mL) Then the reaction was stirred at 120 C in microwave
for 2 h.
Water (8 mL) was added to the mixture and extracted with ethyl acetate (15 mL
x 3). The
organic layer was dried, concentrated, purified by column chromatography
(methanol in
dichloromethane, 5% v/v) to give the title compound 0603-111 (150 mg, 41%) as
a white
solid. LCMS: 521 [M+1]+. 1H NMR (400 MHz, DMSO-d6): 6 1.28 (t, J= 7.2 Hz, 3H),
2.81
(d, J= 4.4 Hz, 3H), 3.24 (s, 3H), 3.73 (d, J= 4.4 Hz, 4H), 3.86 (d, J= 4.4 Hz,
4H), 4.27 (q,
J = 7.2 Hz, 2H), 5.20 (s, 2H), 6.48 (d, J = 8.4 Hz, 1 H), 6.91 (d, J = 4.4 Hz,
1 H), 7.39 (s,
1H), 8.25 (d, J= 8.4 Hz, 1H), 8.86 (s, 2H), 8.90 (s, 1H).
Step 106d: N-Hydroxy-2-(methyl((2-(6-(methylamino)pyridin-3-yl)-4-
morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound 111)
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The title compound 111 was prepared as a brown solid (21 mg, 14%) from 0603-
236 (150 mg, 0.29 mmol) and a freshly prepared hydroxylamine methanol solution
(6 mL)
using a procedure similar to that described for compound 217 (Example 94): mp:
193-195
C. LCMS: 508 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 2.83 (d, J= 4.8 Hz, 3H),
3.23
(s, 3H), 3.74 (m, 4H), 3.89 (m, 4H), 5.20 (s, 2H), 6.50 (d, J= 8.8 Hz, 1H),
6.92 (d, J= 5.2
Hz, 1 H), 7.3 9 (s, 1 H), 8.27 (dd, J = 8.8, 2.0 Hz, 1 H), 8.75 (s, 2H), 9.01
(d, J = 2.0 Hz, 1 H),
9.07 (br, 1H).
EXAMPLE 107: N-Hydroxy-2-(methyl(1-(2-(6-(methylamino)pyridin-3-yl)-4-
morpholinothieno [3,2-d] pyrimidin-6-yl)ethyl)amino)pyrimidine-5-carboxamide
(Compound 237)
Step 107a: 1-(2-Chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)ethanone
(Compound
1201)
To a solution of compound 0111 (5 g, 20 mmol) in THE (100 mL) at -78 C was
dropwise added a solution of nBuLi/THF (2.5 M, 11 mL). The reaction mixture
was stirred
at room temperature for about 30 minutes. DMA (7 g, 80 mmol) was added and the
reaction
mixture was allowed to slowly warm up to room temperature and stirred for 2 h.
Reaction
mixture was poured into a cold aqueous HC1 solution. The crude product was
precipitated
from solution and collected by filtration to afford the title compound as a
yellow solid (5 g,
86%).
Step 107b: 1-(2-Chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)-N-
methylethanamine
(Compound 1202)
To a suspension of 1201 (2.36 g, 7.9 mmol) in MeOH (120 ml) was added
MeNH2/MeOH (7.3 g, 27%).The reaction mixture was stirred at 45 C overnight.
The
raction mixture was concentrated and the reidue was dissolved in MeOH/THF (58
ml/23
ml). To the resulting solution was added MgSO4 (2.4 g, 0.02 mol), NaBH4 (897
mg, 23.7
mmol) and stirred at room temperature overnight. To the reaction mixture was
added water.
The mixture was adjusted to pH = 5-6 with HC1 and washed with dichloromethane.
The
aqueous layer was adjusted to pH = 8-9 with aqueous NaOH and extracted with
dichloromethane. The combine organic layers were dried and concentrated under
reduced
pressure to afford the crude product as a yellow solid (1.5 g, 60%). It was
used for next
step without further purification.
Step 107c: Ethyl 2-((I-(2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)ethyl)(methyl)
amino)pyrimidine-5-carboxylate (Compound 1203)
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To a solution of 1202 (1.29 g, 4.12 mmol) in dioxane (40 mL) were added 0305
(962 mg, 5.15 mmol) and DIPEA (1.28 g, 9.9 mmol). The reaction mixture was
refluxed
overnight. The reaction mixture was concentrated and residue was purified by
column
chromatograph to afford titled compound as a white solid (1.9 g, 99%).
1H NMR (400 MHz, DMSO-d6): 6 1.31 (t, J= 7.2 Hz, 3H), 1.71 (d, J= 7.2 Hz, 3H),
3.01
(s, 3H), 3.70 (t, J= 4.4 Hz, 4H), 3.82 (t, J= 4.0 Hz, 4H), 4.29 (q, J= 7.2 Hz,
2H), 6.50 (q, J
= 7.2 Hz, 1H), 7.38 (s, 1H), 8.88 (s, 2H).
Step 107d: Ethyl2-(methyl(l-(2-(6-(methylamino)pyridin-3-yl)-4-
morpholinothieno [3,2-
d]pyrimidin-6-yl)ethyl)amino)pyrimidine-5-carboxylate (Compound 1204-237)
To a stirred mixture of compound 1203 (350 mg, 0.76 mmol) and compound 0602-
237 (533 mg, 2.3 mmol) in toluene/EtOH/H20 (20mL/l OmL/2m1) was added
Pd(PPh3)2C12
(28.7 mg, 0.038 mmol) and NaHCO3 (191.52 mg, 2.28 mmol). The resulting mixture
was
heated to 120 C and stirred for 5 h. The resulting mixture was concentrated
to remove
most of solvent. The residue was purified by column chromatography to afford
the tiled
compound 111-327-1 as a yellow solid (260 mg, 64%), 1H NMR (400 MHz, CDC13): 6
1.39
(t, J= 7.2 Hz, 3H), 1.74 (s, 3H), 2.99 (d, J= 5.2 Hz, 3H), 3.05 (s, 3H), 3.84
(t, J= 4.8 Hz,
4H), 3.97 (t, J= 4.4 Hz, 4H), 4.36 (q, J= 7.2 Hz, 2H), 4.89 (d, J= 4.0 Hz, I
H), 6.45 (d, J=
8.8 Hz, I H), 6.62 (q, J= 6.4 Hz, I H), 7.33 (s, I H), 8.46 (dd, J= 8.4 Hz,
2.0 Hz, I H), 8.93
(s, 2H), 9.17 (s, 1H).
Step 107e: N-Hydroxy-2-(methyl(l-(2-(6-(methylamino)pyridin-3-yl)-4-morpholino-
thieno[3,2-d]pyrimidin-6-yl)ethyl)amino)pyrimidine-5-carboxamide (Compound 111-
327)
A mixture of 1204-237 (150 mg, 0.28 mmol) and a freshly prepared NH2OH in
methanol (10 mL, 1.79 M in MeOH) was stirred at room temperature for 0.5 h.
The mixture
was adjusted to pH = 5-6 with 2M aq. HC1 and concentrated. The residue was
purified by
prep-HPLC to afford the titled compound 111-237 as a white solid (60 mg, 40%).
M.p.:
270 - 275 C. LCMS: 522 [M+1]+. 'HNMR (400 MHz, DMSO-d6): 6 1.71 (s, 3H), 2.84
(d,
J= 4.0 Hz, 3H), 2.98 (s, 3H), 3.74 (s, 4H), 3.88 (s 4H), 6.45-6.25 (m, 2H),
6.90 (s, 1H),
7.3 9 (s, 1 H), 8.27 (d, J = 8.4 Hz, 1 H), 8.76 (s, 2H), 9.02 (s, 1 H), 9.06
(s, 1 H), 11.13 (s, 1 H).
EXAMPLE 108: N-Hydroxy-2-(2-(2-(6-(methylamino)pyridin-3-yl)-4-morpho-
linothieno [3,2-d] pyrimidin-6-yl)propan-2-ylamino)pyrimidine-5-carboxamide
(Compound 240)
Step 108a: 2-(2-Chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)propan-2-ol
(Compound
1205)
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To a suspension of 0111 (5.10 g, 20.00 mmol) in THE (120 mL) at -70 C was
dropwise added a solution of n-BuLi/hexane (2.5 M, 22 mL, 55 mmol). The
suspension was
stirred at -20 C for 45 min. The resulting red solution was re-cooled and dry
acetone (5.8
g, 100 mmol) was then added at -70 C and stirred at -20 C for 5 h. The
reaction mixture
was quenched with ice water (125 mL) and extracted with ethyl acetate. The
combined
organic layers were dried over Na2SO4 and concentrated in vacuo to afford the
crude
product, which was purified by column chromatography (ethyl acetate in hexane)
to afford
the titled compound 1205 as a white solid (3.79 g, 60%). LC-MS: 314.1 [M+1]+.
'H NMR
(400 MHz, DMSO-d6): 6 1.56 (s, 6H), 3.74 (m, 4H), 3.89 (m, 4H), 5.94 (s, 1H),
7.22 (s,
1H).
Step 108b: N-(2-(2-Chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)propan-2-yl)
formamide (Compound 1206)
To a suspension of 1205 (3.79 g, 12 mmol) in TMSCN (6 g, 60 mmol) was added
conc. H2SO4 (13 g, 132 mmol) slowly at room temperature. The reaction mixture
was
stirred at room temperature for 5 h. The reaction mixture was quenched with
ice water,
basified with NH3 = H2O and extracted with dichloromethane.
The crude product was purified by column chromatography (ethyl acetate/hexane)
to afford the titled compound 1206 as an off-white solid (3.56 g, 87%). LC-MS:
341.1
[M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.69 (s, 6H), 3.74 (m, 4H), 3.86 (m, 4H),
7.23
(s, I H), 7.98 (s, I H), 8.71 (s, I H).
Step 108c: 2-(2-Chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)propan-2-amine
(Compound 1207)
A mixture of 1206 (3.55 g, 10.40 mmol) in MeOH (18 mL), H2O (12 ml) and conc.
HC1(47 mL) was stirred at room temperature overnight. Reaction mixture was
concentrated and re-dissolved in water, neutralized with sat. NaHCO3 and
extracted with
dichloromethane. The combined organic layers were dried over Na2SO4 and
concentrated in
vacuo to afford the crude product, which was purified by column chromatography
(ethyl
acetate/hexane) to afford the titled compound 1207 as a white solid (2.93 g,
90 %).
LC-MS: 313 [M+l]+. 'H NMR (400 MHz, DMSO-d6): 6 1.49 (s, 6H), 2.40 (s, 2H),
3.74
(m, 4H), 3.89 (m, 4H), 7.23 (s, 1H).
Step 108d: Ethyl 2-(2-(2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-yl)propan-
2-yl-
amino)pyrimidine-5-carboxylate (Compound 1208)
A mixture of 1207 (1.50 g, 5 mmol), 0305 (1.85 g, 10 mmol) and DIPEA (1.92 g,
15 mmol) in dioxane (30 mL) was heated at 120 C for 48 h. The reaction was
then
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quenched with water and extracted with ethyl acetate. The combined organic
layers were
dried over Na2SO4 and concentrated in vacuo to afford the crude product, which
was
purified by column chromatography (ethyl acetate / hexane) to afford the title
compound
1208 as an off-white solid (976 mg, 42 %). LC-MS: 463.3 [M+1]+. 'H NMR (400
MHz,
DMSO-d6): 6 1.25 (t, J= 6.8 Hz, 3H), 1.82 (s, 6H), 3.70 (m, 4H), 3.81 (m, 4H),
4.23 (q, J-
6.8 Hz, 2H), 7.20 (s, 1H), 8.71 (m, 3H).
Step 108e: Ethyl 2-(2-(2-(6-(methylamino)pyridin-3-yl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)propan-2-ylamino)pyrimidine-5-carboxylate (Compound 1210-240)
A mixture of 1208 (185 mg, 0.40 mmol), 0602-237 (187 mg, 0.80 mmol), NaHCO3
(101 mg, 1.20 mmol) and PdC12(PPh3)2 (28 mg, 0.04 mmol) in toluene (8 mL),
EtOH (5
mL) and water (2 mL). The suspension was heated at 120 C overnight. The
reaction
mixture was concentrated and the residue was purified by column chromatography
(ethyl
acetate/hexane) to afford the titled compound 1210-240 as a white solid (71
mg, 33%). LC-
MS: 535 [M+1]+. 'H NMR: (400 MHz, DMSO-d6): 6 1.25 (t, J= 7.2 Hz, 3H), 1.85
(s, 6H),
2.83 (d, J= 0.8 Hz), 3.74 (m, 4H), 3.87 (m, 4H), 4.22 (q, J= 7.2 Hz, 2H), 6.50
(d, J= 8.8
Hz, I H), 6.88 (d, J= 4.8 Hz, I H), 7.24 (s, I H), 8.27 (dd, J= 2.0 and 8.8
Hz, I H), 8.69 (m,
3H), 9.01 (d, J= 2.4 Hz, 1H).
Step 108f: N-Hydroxy-2-(2-(2-(6-(methylamino)pyridin-3-yl)-4-morpholinothieno
[3,2-
d]pyrimidin-6-yl)propan-2-ylamino)pyrimidine-5-carboxamide (Compound 240)
A mixture of 1210-240 (100 mg, 0.19 mmol) and a freshly prepared NH2OH/MeOH
solution (25 mL, 1.79M) in CH2C12 (4 mL) and stirred at room temperature for
lh. The
solution was filtrated and adjusted to pH = 7 with acetic acid. To the mixture
was added
water (30 mL) and compound 240 (96 mg) was collected by filtration. m.p. 200-
205 C.
LC-MS: 522 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.83 (s, 6H), 2.83 (d, J= 4.8
Hz,
3H), 3.74 (m, 4H), 3.86 (m, 4H), 6.49 (d, J= 8.8 Hz, I H), 6.88 (s, I H), 7.23
(s, I H), 8.27
(dd, J = 8.8 Hz , 2.4 Hz, 1 H), 8.34 (s, 1 H), 8.5 3 (br, 2H), 8.97 (s, 1 H),
9.01 (d, J = 2.4 Hz,
I H), 10.95 (s, I H).
EXAMPLE 109: N-Hydroxy-2-(methyl((2-(1-methyl-lH-pyrazol-4-yl)-4-morpho-
linothieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound
246)
Step 109a: Ethyl 2-(methyl((2-(I-methyl-IH-pyrazol-4-yl)-4-
morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxylate (Compound 0603-246)
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The title compound 0603-246 was prepared as a yellow solid (196mg, 35%) from
0504 (500 mg, 1.11 mmol), 1-methyl-4-(4,4,5,5-tetramethyl -1,3,2-dioxaborolan-
2-yl)-1H-
pyrazole (579 mg, 2.78 mmol) and Pd(PPh3)2C12 (23 mg,Ø033 mmol) in toluene
(11.2
mL), ethanol (7 mL) and water (2.3 mL) using a procedure similar to that
described for
compound 1103-226 (Example 99).
Step 109b: N-Hydroxy-2-(methyl((2-(1-methyl-lH-pyrazol-4-yl)-4-
morpholinothieno [3,2-
d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide (Compound 246)
The title compound 246 was prepared as a white solid (80 mg, 45%) from 0603-
246
(190 mg, 0.38 mmol) and a freshly prepared hydroxylamine methanol solution (20
mL,
1.79 M in methanol) in dichloromethane (2 mL) using a procedure similar to
that described
for compound 217 (Example 94). m.p.:164-169 C. LCMS :482[M+1]+. 'HNMR (400
MHz, DMSO-d6): 6 3.22 (s, 3H), 3.73 (d, J= 4.4 Hz, 4H), 3.86-3.90 (m, 7H),
5.18 (s, 2H),
7.35 (s, I H), 7.97 (s, I H), 8.31 (s, I H), 8.75 (s, 2H), 9.09 (s, I H),
11.15 (s, I H).
EXAMPLE 110: N-hydroxy-2-((1-(2-(6-methoxypyridin-3-yl)-4-morpholino-
thieno [3,2-d] pyrimidin-6-yl)ethyl)(methyl)amino)pyrimidine-5-carboxamide
(Compound 247)
Step 110a: Ethyl2-((1-(2-(6-methoxypyridin-3-yl)-4-morpholinothieno[3,2-d]
pyrimidin-
6- yl)ethyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 1210-247)
The title compound 1210-247 was prepared as a yellow solid (240 mg, 95%) from
1208 (200 mg, 0.43 mmol), 2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-
2-
yl)pyridine (153 mg, 0.65 mmol), NaHCO3 (109 mg, 1.29 mmol) and
bis(triphenylphosphine)palladium(II) chloride (15 mg, 0.02 mmol) in toluene (8
ml),
ethanol (5 ml) and water (1 ml) using a procedure similar to that described
for compound
1204-237 (Example 107): LCMS: 536.0 [M+1]+. 1H NMR (400 MHz, DMSO-d6): 6 1.31
(t,
J = 7.2 Hz, 3H), 1.74 (d, J = 7.2 Hz, 3H), 3.03 (s, 3H), 3.75 (t, J = 4.4 Hz,
4H), 3.91-3.93
(m, 7H), 4.30 (q, J = 7.2 Hz, 2H), 6.50 (q, J = 7.2 Hz, 1 H), 6.92 (d, J = 8.4
Hz, 1 H), 7.48 (s,
1 H), 7.5 7 (dd, J = 8.4 Hz, 2.4 Hz, 1 H), 8.89 (s, 2H), 9.15 (d, J = 2.4 Hz,
1 H).
Step 110b: N-Hydroxy-2-((1-(2-(6-methoxypyridin-3-yl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)ethyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 247)
The title compound 247 was prepared as an light pink solid (120 mg, 56%) from
1210-247 (220 mg, 0.41 mmol) and a freshly prepared hydroxylamine methanol
solution
(30 mL, 1.79 M) using a procedure similar to that described for compound 237
(Example
107): LCMS: 523.0 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.70 (d, J= 6.8 Hz,
3H),
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2.97 (s, 3H), 3.32 (m, 1H), 3.73-3.75 (m, 4H), 3.91-3.93 (m, 7H), 6.46 (d, J=
6.8 Hz, 1H),
6.91 (d, J= 8.8 Hz, I H), 7.45 (s, I H), 8.57 (dd, J= 8.8 Hz, 2.0 Hz, I H),
8.76 (s, 2H), 9.15
(d, J = 2.4 Hz, 1 H).
EXAMPLE 111: N-Hydroxy-2-(2-(2-(6-methoxypyridin-3-yl)-4-morpholino-
thieno[3,2-d]pyr- imidin-6-yl)propan-2-ylamino)pyrimidine-5-carboxamide
(Compound 250)
Step 111a: Ethyl 2-(2-(2-(6-methoxypyridin-3 -yl)-4-morpholinothieno [3,2-
d]pyrimidin-6-
yl)propan-2-ylamino)pyrimidine-5-carboxylate (Compound 1210-250)
The title compound 1210-250 was prepared as a white solid (160 mg, 75%) from
1208 (185 mg, 0.40 mmol), 0602-222 (188 mg, 0.80 mmol), NaHCO3 (101 mg, 1.20
mmol) and PdC12(PPh3)2 (28 mg, 0.04 mmol) was taken into toluene (8 mL), EtOH
(5 mL)
and water (2 mL) using a procedure similar to that described for compound 1204-
237
(Example 107): LC-MS: 536.3 [M+1]+. 'H NMR: (400 MHz, DMSO-d6): 6 1.30 (t, J=
6.8
Hz, 3H), 1.91 (s, 6H), 3.81 (m, 4H), 3.96 (m, 4H), 3.99 (s, 3H), 4.28 (q, J=
6.8 Hz, 2H),
6.97 (d, J= 8.8 Hz, I H), 7.36 (s, I H), 8.62 (dd, J= 8.4 Hz, 2.4 Hz, I H),
8.74 (m, 3H), 9.20
(d, J = 2.0 Hz, I H).
Step 111b: N-Hydroxy-2-(2-(2-(6-methoxypyridin-3-yl)-4-morpholinothieno [3,2-
d]pyr-
imidin-6-yl)propan-2-ylamino)pyrimidine-5-carboxamide (Compound 250)
The title compound 247 was prepared as a white solid (140 mg) from 1210-250
(160 mg, 0.30 mmol) and a freshly prepared hydroxylamine methanol solution (30
mL,
1.79 M) in CH2C12 (4 mL) using a procedure similar to that described for
compound 237
(Example 107): LC-MS: 523 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.90 (s, 6H),
3.81
(m, 4H), 3.96 (m, 4H), 3.99 (s, 3H), 6.97 (d, J= 8.4 Hz, I H), 7.36 (s, I H),
8.43 (s, I H),
8.60 (m, 3H), 9.04 (s, I H), 9.20 (d, J= 2.0 Hz, I H), 11.10 (s, I H).
EXAMPLE 112: N-Hydroxy-2-((2-(2-(6-methoxypyridin-3-yl)-4-morpho-
linothieno [3,2-d] pyrimidin-6-yl)propan-2-yl)(methyl)amino)pyrimidine-5-
carboxamide (Compound 251)
Step 112a: Ethyl 2-((2-(2-chloro-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)propan-2-
yl)(methyl)amino)pyrimidine-5-carboxylate (Compound 1209)
NaHDMS (2.0 M in THF, 1.5 mL) was added to a solution of compound 1208 (278
mg, 0.60 mmol) in dry THE (25 mL) at -70 C followed by the addition of MeOTf
(0.34
mL, 3 mmol). The resulting mixture was stirred at -70 C for 0.5 h. The
reaction was
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quenched with water, and extracted with EtOAc. The combined organic layers
were dried
with Na2SO4. The crude product was purified by column chromatography
(Hexanes/EtOAc) to afford the titled compound 1209 as a white solid (52 mg, 18
%).
LCMS: 477.3[M+l]+. 1H NMR: (400 MHz, DMSO-d6): 6 1.24 (t, J= 7.2 Hz, 3H), 1.89
(s,
6H), 3.48 (s, 3H), 3.69-3.71 (m, 4H), 3.78-3.80 (m, 4H), 4.21 (q, J= 7.2 Hz,
2H), 7.16 (s,
I H), 8.65 (s, 2H).
Step 112b: Ethyl 2-((2-(2-(6-methoxypyridin-3-yl)-4-morpholinothieno[3,2-d]
pyrimidin-
6- yl)propan-2-yl)(methyl)amino)pyrimidine-5-carboxylate (Compound 1211-251)
A mixture of 1209 (52 mg, 0.11 mmol), 6-methoxypyridin-3-ylboronic acid (68
mg,
0.44 mmol), NaHCO3 (27 mg, 0.32 mmol) and PdC12(PPh3)2 (7.8 mg, 0.011 mmol) in
toluene (2.6 mL), EtOH (1.6 mL) and water (0.6 mL) was heated at 120 C
overnight. The
reaction mixture was concentrated and purified by column chromatography
(Hexanes/EtOAc) to afford the titled compound 1211-251 as a white solid (53
mg, 88%).
LCMS: 550.4 [M+1]+. 1H NMR: (400 MHz, DMSO-d6): 6 1.23 (t, J= 6.8 Hz, 3H),
1.93 (s,
6H), 3.50 (s, 3H), 3.74-3.76 (m, 4H), 3.88-3.90 (m, 4H), 3.92 (s, 3H), 4.20
(q, J= 7.2 Hz,
2H), 6.90 (d, J= 8.4 Hz, 1H), 7.25 (s, 1H), 8.55 (dd, J= 2.0 Hz, 8.8 Hz, 1H),
8.65 (s, 2H),
9.12 (d, J= 2.0 Hz, 1H).
Step 112c: N-Hydroxy-2-((2-(2-(6-methoxypyridin-3-yl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)propan-2-yl)(methyl)amino)pyrimidine-5-carboxamide (Compound
251)
The title compound 251 was prepared as a white solid (32 mg, 35%) from 1210-
250
(95 mg, 0.17 mmol) and a freshly prepared hydroxylamine methanol solution (30
mL, 1.79
M) in CH2C12 (4 mL) using a procedure similar to that described for compound
237
(Example 107): m.p. 175-178 C. LCMS: 537.4 [M+1]+. 1H NMR (400 MHz, DMSO-d6):
6
1.91 (s, 6H), 3.46 (s, 3H), 3.73-3.74 (m, 4H), 3.86-3.90 (m, 4H), 3.92 (s,
3H), 6.90 (d, J=
8.8 Hz, 1H), 7.24 (s, 1H), 8.53-8.57 (m, 3H), 8.91 (br, 1H), 9.12 (d, J= 2.0
Hz, 1H), 10.91
(br, 1 H).
EXAMPLE 113: 2-[2-(6-Methoxy-pyridin-3-yl)-4-morpholin-4-yl-thieno[3,2-d]
pyrimidine-6-sulfonylamino]-pyrimidine-5-carboxylic acid hydroxyamide
(Compound
252)
Step 113a: 2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidine-6-sulfonyl
chloride
(Compound 1301)
To a suspension of 0 111 (10 g, 39.2 mmol) in THE (220 mL) at -78 C was added
slowly 2.5 M solution of n-BuLi in hexane (18.8 mL, 47.06 mmol) under
nitrogen. The
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resulting slurry was allowed to warm up to -40 C and a clear brown solution
was observed.
The solution was cooled to -50 C and S02C12 (6.3 mL, 78.43 mmol) was added
slowly.
The resulting solution was stirred at -40 C for 2 h. The reaction mixture was
concentrated
and purified by column chromatography eluted with hexanes/ethyl acetate to
afford
compound 1301 as a yellow solid (3.7 g, 26.8 %). LCMS: 353.8[M+1]+. 'HNMR (400
MHz, DMSO-d6): 6 3.75 (t, J= 4.4 Hz, 4H), 3.89 (t, J= 4.4 Hz, 4H), 7.27 (s,
1H).
Step 113b: 2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidine-6-sulfonic acid
amide
(Compound 1302)
1301 (2 g, 5.67 mmol) was taken into a solution of NH3 in methanol and stirred
at
room temperature overnight. The reaction mixture was concentrated and purified
by
column chromatography eluted with 2% methanol in dichloromethane to afford
compound
1302 (0.97 g, 51 %). LCMS: 335.0[M+1]+. 'HNMR (400 MHz, DMSO-d6): 6 3.77 (t, J
4.4 Hz, 4H), 3.92 (t, J= 4.4 Hz, 4H), 7.75 (s, 1H), 8.15 (s, 2H).
Step 113c: 2-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidine-6-
sulfonylamino)-
pyrimidine-5-carboxylic acid ethyl ester (Compound 1303)
A mixture of 1302 (400 mg, 1.2 mmol), 0305 (335 mg, 1.8 mmol) and TEA in
dichloromethane was stirred at reflux for 3 days. The reaction mixture was
diluted by
dichloromethane and washed with water. The crude product was purified by
column
chromatography to afford compound 1303 (210 mg, 36 %).
LCMS: 485.0[M+1]+. 'HNMR (400 MHz, DMSO-d6): 6 1.27 (t, J= 7.2 Hz, 3H), 3.79
(s,
4H), 3.94 (s, 4H), 4.27 (q, J = 7.2 Hz, 2H), 7.64 (s, 1 H), 8.71 (s, 2H).
Step 113d: 2-[2-(6-Methoxy-pyridin-3-yl)-4-morpholin-4-yl-thieno[3,2-
d]pyrimidine- 6-
sulfonylamino]-pyrimidine-5-carboxylic acid ethyl ester (Compound 1304-252)
The title compound 1304-252 was prepared as a gray solid (100 mg, 38 %) from
1303 (260 mg, 0.54 mmol), 6-methoxypyridin-3-ylboronic acid (246 mg, 1.61
mmol) and
Pd(PPh3)4 (10 mg) in dioxane (9 mL) and sat. NaHCO3 (3 mL) using a procedure
similar to
that described for compound 1211-251 (Example 112): 'HNMR (400 MHz, DMSO-d6):
6
1.28 (t, J= 7.2 Hz, 3H), 3.80 (t, J= 4.4 Hz, 4H), 3.92 (s, 3H), 4.00 (t, J=
4.4 Hz, 4H), 4.29
(q, J = 7.2 Hz, 2H), 6.93 (d, J = 8.4 Hz, 2H), 8.02 (s, 1 H), 8.5 8 (dd, J =
8.8 Hz, 2.4 Hz,
I H), 8.99 (s, 2H), 9.16 (s, I H).
Step 113e: 2-[2-(6-Methoxy-pyridin-3-yl)-4-morpholin-4-yl-thieno[3,2-
d]pyrimidine-
sulfonylamino]-pyrimidine-5-carboxylic acid hydroxyamide (Compound 252)
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The title compound 252 was prepared as a white solid (21 mg, 23%) from 1304-
252
(100 mg) and a freshly prepared hydroxylamine methanol solution (16 mL, 1.79
M) using a
procedure similar to that described for compound 237 (Example 107): m.p.: 259-
265 C.
LCMS: 545.3[M+1]+. 'HNMR (400 MHz, DMSO-d6): 6 3.81 (s, 4H), 3.96 (s, 3H),
4.00 (s,
4H), 6.93 (d, J= 8.4 Hz, 2H), 7.99 (s, 1H), 8.58 (d, J= 8.0 Hz, 1H), 8.84 (s,
2H), 9.16 (s,
I H), 9.18 (s, I H), 11.23 (s, I H).
EXAMPLE 114: 2-(((2-(6-ethoxypyridin-3-yl)-4-morpholinothieno [3,2-d]
pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 255)
Step 114a: 5-Bromo-2-ethoxypyridine (Compound 0601-255)
A solution of freshly prepared sodium ethoxide in ethanol was treated with 2,5-
Dibromo-pyridine and heated at reflux for 28 h. The mixture was concentrated
and
partitioned between dichloromethane and water, organic layer was washed with
brine, dried
over MgSO4, filtered and evaporated in vacuum to get compound 0601-255 (3 g,
70%).
LCMS: 202.0 [M+1]+
Step 114b: 2-Ethoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(Compound
0602-255)
A slurry of compound 0601-255 (3.0 g, 15.0 mmol), bis(pinacolatio)diboran (5.7
g,
22.0 mmol), PdC12(dppf) (327 mg, 0.45 mmol) and potassium acetate (4.4 g, 45.0
mmol) in
anhydrous 1,4-dioxane (30 ml) was heated at 110 C overnight. The reaction
mixture was
filtered and concentrated. The crude product was purified by column
chromatography to
afford titled compound (2.7 g, 73%) as a white solid. 'HNMR (400 MHz, DMSO-
d6): 6
1.29 (s, 12H), 1.29 (t, J= 7.2 Hz, 3H), 4.34 (q, J= 7.2 Hz, 2H), 6.76 (d, J=
8.4 Hz, 1H),
7.85 (dd, J = 8.4 Hz, 1.6 Hz, 1 H), 8.3 8 (d, J = 1.6 Hz, 1 H).
Step 114c: Ethyl 2-(((2-(6-ethoxypyridin-3-yl)-4-morpholinothieno [3,2-
d]pyrimidin-6-yl)
methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-255)
The title compound 0603-255 was prepared as a white solid (335 mg, 94%) from
0504 (300 mg, 0.67 mmol), 0602-255 (333 mg, 1.34 mmol), NaHCO3 (168 mg, 2
mmol)
and bis(triphenylphosphine)palladium(II) chloride (23 mg, 0.03 mmol) in
toluene (8 ml),
ethanol (5 ml) and water (1 ml) using a procedure similar to that described
for compound
1103-226 (Example 99): 'HNMR (400 MHz, DMSO-d6): 6 1.31-1.37 (m, 6H), 3.76 (t,
J=
4.4 Hz, 4H), 3.87 (3, 3H), 3.92 (t, J= 4.4 Hz, 4H), 4.30 (q, J= 7.2 Hz, 2H),
4.40 (q, J= 7.2
Hz, 2H), 5.24 (s, 2H), 6.87 (d, J= 8.8 Hz, I H), 7.46 (s, I H), 8.56 (dd, J=
8.4 Hz, 1.2 Hz,
1H), 8.88 (s, 2H)., 9.13 (s, 1H).
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Step 114d: 2-(((2-(6-Ethoxypyridin-3-yl)-4-morpholinothieno[3,2-d]pyrimidin- 6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 255)
The title compound 255 was prepared as a white solid (90 mg, 46%) from 0603-
255
(200 mg, 0.37 mmol) and a freshly prepared hydroxylamine methanol solution (50
mL,
1.79 M) using a procedure similar to that described for compound 237 (Example
107):
m.p.:162-165 C. LCMS: 523.0 [M+1]+.'HNMR (400 MHz, DMSO-d6): 6 1.34 (t, J=
7.2
Hz, 3H), 3.24 (s, 3H), 3.75 (t, J= 4.0 Hz, 4H), 3.92 (t, J = 4.0 Hz, 4H), 4.37
(q, J = 7.2 Hz,
2H), 5.20 (s, 2H), 6.8 8 (d, J = 8.4 Hz, 1 H), 7.44 (s, 1 H), 8.5 6 (dd, J =
8.4 Hz, 1.2 Hz, 1 H),
8.75 (s, 2H)., 9.04 (s, 1 H), 9.12 (s, 1 H), 11.12 (s, 1 H).
EXAMPLE 115: N-Hydroxy-2-(((2-(6-(2-hydroxyethoxy)pyridin-3-yl)-4-morpho-
linothieno[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5- carboxamide
(Compound 256)
Step 115a: 2-(5-Bromopyridin-2-yloxy)ethanol (Compound 0601-256)
To s stirred solution of 2, 5-dibromopyridine (20 g, 85 mmol) in NMP (100 mL)
was added NaH (17 g, 0.425 mol) portionwise at 0 C. The resulting mixture was
stirred at
0 C for 1 h. Ethylene glycol (26.4 g, 0.425 mol) was added. The resulting
mixture was
heated at 110 C for 4h. The reaction mixture was cooled to room temperature
and
partitioned between EtOAc and water. The organic layer was washed with water,
dried over
Na2SO4 and concentrated. The residue was purified by column chromatography
eluted with
hexanes/EtOAc to afford 0601-256 (7.2 g, yield 39%) as an off-white solid.
LCMS: 218.0 [M+1]+. 'H NMR (400 MHz, CDC13): 6 3.45-3.48 (t, 1H, J= 5.6 Hz),
3.92-
3.95 (m, 2H), 4.39-4.42 (m, 2 H), 6.70 (d, J = 8.8 Hz, 1 H), 7.66 (dd, J = 8.8
Hz, 2.4 Hz,
I H), 8.16 (d, J= 2.4 Hz, I H).
Step 115b: 2-(5-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-
yloxy)ethanol
(Compound 0602-256)
A mixture of 0601-256 (5 g, 0.023 mol), bis(pinacolato)diboron (7 g, 0.028
mol),
KOAc (6.8 g, 0.069 mol) and Pd(dppf)C12 (336.7 mg, 0.46 mmol) in dry dioxane
(50 mL)
was degassed with N2 and heated at 90 C for 4h. The reaction mixture was
concentrated.
The residue was suspended in CH2C12 and stirred for 30 min and filtered. The
filtrate was
concentrated and purified by column chromatography eluted with hexanes / EtOAc
to
afford 0602-256 (9 g, containing Bis(pinacolato)diboron) as a yellow oil.
LCMS: 266.0
[M+1]+. 'H NMR (400 MHz, CDC13): 6 1.334 (s, 12 H), 3.87 (m, 1H), 3.94 (m,
2H), 4.48-
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4.51 (m, 2 H), 6.77 (d, J = 8.4 Hz, 1 H), 7.95 (dd, J = 8.4 Hz, 2.0 Hz, 1 H),
8.50 (d, 1 H, J =
1.2 Hz, 1H).
Step 115c: Ethyl2-(((2-(6-(2-hydroxyethoxy)pyridin-3-yl)-4-
morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-
256)
The title compound 0603-256 was prepared as an off-white solid (150 mg, 30%)
from 0602-256 (708 mg, 2.67 mmole), 0504 (400 mg, 0.89 mmole), NaHCO3 (224 mg,
2.67 mmole) and Pd(PPh3)2C12 (31 mg,0.05 mmole) in toluene /EtOH/H20 (16 mL/10
mL/4 mL) using a procedure similar to that described for compound 1103-226
(Example
99): LCMS: 552 [M+1]+.
Step 115d: N-Hydroxy-2-(((2-(6-(2-hydroxyethoxy)pyridin-3-yl)-4-
morpholinothieno [3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 256)
The title compound 256 was prepared as a light yellow solid (55 mg, 36%) from
0603-256 (150 mg, 0.27 mmol) and a freshly prepared hydroxylamine methanol
solution
(20 mL, 1.79 M) using a procedure similar to that described for compound 237
(Example
107): m.p.: 143-145 C. LCMS: 539.0 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 3.24
(s,
3H), 3.75 (s, 6H), 3.92 (s, 4H), 4.35 (s, 2H), 4.84-4.87 (br,1 H), 5.20 (s,
2H), 6.90 (d, J=
6.8 Hz,1 H), 7.44 (s,1 H), 8.5 7 (d, J = 6.4 Hz, 1 H), 8.76(s, 2H), 9.06 (s, 1
H), 9.12(s, 1 H),
11.26 (s, 1H).
EXAMPLE 116: 2-(((2-(6-(2-(Dimethylamino)ethoxy)pyridin-3-yl)-4-morpho-
linothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-
carboxamide (Compound 257)
Step 116a: 2-(2-Bromoethoxy)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
yl)pyridine
(Compound 0601-257)
A mixture of 0602-256 (2.9 g, 0.01 mol) and PBr3 (3 mL) in toluene (30 mL) was
heated at 80 C for 3 h. The reaction mixture was cooled to 0 C and aq.
NaHCO3 was
added slowly until pH = 7-8 and extracted with EtOAc. The combined organic
layers were
washed with brine, dried over Na2SO4 and concentrated to afford crude 0601-257
(2 g,
61.2%) as a pink solid. LCMS: 328.1 [M+1]+. 'H NMR (400 MHz, CDC13): 6 1.24
(s, 12
H), 3.65 (t, J= 6.4 Hz, 2H), 4.25 (t, J= 6.4 Hz, 2 H), 6.56 (d, J= 8.8 Hz,
1H), 7.60 (dd, J
8.8 Hz, 1.6 Hz, I H), 7.70 (d, J = 1.6 Hz, I H).
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Step 116b: Ethyl 2-(((2-(6-(2-(dimethylamino)ethoxy)pyridin-3-yl)-4-mor-
pholinothieno[3, 2-d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxylate
(Compound 0603-257)
0601-257 (600 mg, 1.83 mmol) was taken into Me2NH (20 mL in CH3OH) and
stirred at room temperature for 3 h. The reaction mixture was concentrated to
give crude
0602-257 as a yellow solid (650 mg) which was used in the next step without
further
purification.
A mixture of 0602-257 (600 mg), 0504 (350 mg, 0.78 mmol), NaHCO3 (196 mg,
2.33 mmol) and Pd(PPh3)2C12 (27 mg) in toluene/EtOH/H20 (16 mL/10 mL/4 mL) was
degassed with N2 and heated at 110 C for 3 h. The reaction mixture was
concentrated. The
residue was suspended in CH2C12 (50 mL) and filtered. The filtrate was
concentrated. The
crude product was purified by column chromatography to afford 0603-257 as a
white solid
(200 mg, 44%).
Step 116c: 2-(((2-(6-(2-(Dimethylamino)ethoxy)pyridin-3-yl)-4-
morpholinothieno[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 257)
The title compound 257 was prepared as a light yellow solid (30 mg, 30%) from
0603-257 (100 mg, 0.28 mmol) and a freshly prepared hydroxylamine methanol
solution
(10 mL, 1.79 M) in CH2C12 (2 mL) using a procedure similar to that described
for
compound 237 (Example 107). m.p.: 140-143 C. LCMS: 566 [M+1]+. 'H NMR (400
MHz, DMSO-d6): 6 2.21(s, 6H), 2.56 (t, J= 5.2 Hz, 2H), 3.23 (s, 3H), 3.75 (s,
4H), 3.90 (s,
4H), 4.12 (s, 2H), 5.16 (s, 2H), 6.46 (dd, J = 8.8 Hz, 1.2 Hz, 1 H), 7.40 (s,
1 H), 8.17 (s, 1 H),
8.30 (dd, J = 8.4 Hz, 4.4 Hz, 1 H), 8.63 (d, J = 12 Hz, 1 H), 8.80 (s, 2H),
11.11 (br, 1 H).
EXAMPLE 117: N-Hydroxy-2-(methyl((2-(6-(2-(methylamino)ethoxy)pyridin-3-yl)- 4-
morpholinothieno [3,2-d] pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound 258)
Step 117a: N-Methyl-2-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-
pyridin-2-yloxy)
ethanamine (Compound 0602-258)
A mixture of 0601-257 (300 mg, 0.917 mmol) and MeNH2 (20 mL, in alchohol)
was stirred at room temperature for 3 h. The reaction mixture was concentrated
to afford
crude 0602-25 8 which was used in the next step without further purification.
LCMS: 279.1
[M+1]+. 'H NMR (400 MHz, CDC13): 6 1.24 (s, 12 H), 3.13 (s, 3H), 4.06-4.08 (m,
2H),
4.60-4.64 (m, 2 H), 6.84 (d, J = 9.2 Hz, 1 H), 7.92 (s, 1 H), 8.04 (d, J = 8.8
Hz, 1 H).
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Step 117b: Ethyl 2-(methyl((2-(6-(2-(methylamino)ethoxy)pyridin-3-yl)-4-
morpholinothieno [3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxylate
(Compound 0603-258)
The title compound 0603-258 was prepared as an grey solid (120 mg, 24%) from
0602-258 (prepared above), 0504 (400 mg, 0.89 mmol), NaHCO3 (230 mg, 2.75
mmol) and
Pd(PPh3)2C12 (10 mg) in toluene/EtOH/H20 (16 mL/10 mL/4 mL) using a procedure
similar to that described for compound 1103-226 (Example 99): LCMS: 565
[M+1]+. 1H
NMR (400 MHz, CDC13): 6 1.38 (t, J= 7.2 Hz, 3H), 3.19 (s, 3H), 3.32 (s, 3H),
3.87 (t, J=
5.2Hz, 4H), 3.99 (t, J= 5.2Hz, 4H), 4.15 (t, J= 6.4 Hz, 2H), 4.37 (q, 2H, J=
7.2 Hz), 4.76
(t, J = 5.6Hz, 2H), 5.20 (s, 2H), 6.93 (d, J = 8.8 Hz, 1 H), 7.32 (s, 1 H),
8.71 (d, J = 2.4 Hz,
1H), 8.78 (dd, J= 9.2 Hz, 1.6 Hz, 1H), 8.93 (s, 2H).
Step 117c: N-Hydroxy-2-(methyl((2-(6-(2-(methylamino)ethoxy)pyridin-3-yl)-4-
mor-
pholinothieno[3,2-d]pyrimidin-6-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound
258)
The title compound 258 was prepared as a light yellow solid (55 mg, 47%) from
0603-258 (120 mg, 0.21 mmol) and a freshly prepared hydroxylamine methanol
solution
(10 mL, 1.79 M) in CH2C12 (2 mL) using a procedure similar to that described
for
compound 237 (Example 107). m.p.: 170-175 C. LCMS: 552.3 [M+1]+. 1H NMR (400
MHz, DMSO-d6): 6 2.98 (s, 3H), 3.23 (s, 3H), 3.71-3.77 (m, 6H), 3.90-3.92 (m,
4H), 4.32
(br, 2H), 5.19 (s, 2H), 6.94 (d, 1H, J= 9.6 Hz), 7.39 (s, 1H), 8.42-8.45 (d,
2H), 8.68 (s, 1H),
8.71 (s, 2H).
EXAMPLE 118: 2-(((2-(6-(2-Aminoethoxy)pyridin-3-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 259)
Step 118a: 2-(2-Azidoethoxy)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
yl)pyridine
(Compound 0602-259)
A mixture of 0601-257 (260 mg, 0.8 mmol) and NaN3 (103.4 mg, 1.59 mmol) in
DMF was heated at 60 C for 2 h. The reaction mixture was partitioned between
CH2C12 and
water. The organic layer was washed with brine, dried over Na2SO4. The crude
product was
dissolved in toluene and was used in the next step directly.
LCMS: 291.1 [M+1]+.
Step 118b: Ethyl 2-(((2-(6-(2-aminoethoxy)pyridin-3-yl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-
259)
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A mixture of 0602-259, 0504 (350 mg, 0.78 mmol), NaHCO3 (200 mg, 2.4 mmol)
and Pd(PPh3)2C12 (10 mg) in toluene/EtOH/H20 (16 mL/10 mL/4 mL) was degassed
with
N2 and heated at 110 C for 3 h. The reaction mixture was concentrated. The
residue was
purified by chromatography eluted with CH2Cl2/MeOH to afford the titled
compound, ethyl
2-(((2-(6-(2-azidoethoxy)pyridin-3-yl)-4-morpholinothieno[3,2-d] pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate. It was dissolved in methanol
and can
be used in the next step directly. LCMS: 577.3 [M+1]+
A mixture of ethyl 2-(((2-(6-(2-azidoethoxy)pyridin-3-yl)-4-
morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (prepared
above) and
Pd/C (100 mg) in MeOH (10 mL) was stirred at room temperature under H2 (1 atm)
overnight. The reaction mixture was diluted with CH2Cl2 (10 mL) and filtered
through
celite. The filtrate was concentrated to afford the tiled compound 0603-259 as
a light
yellow solid (130 mg, 15.3%, 2 steps). LCMS: 551 [M+1]+. 1H NMR (400 MHz,
CDC13):
6 1.38 (t, J= 7.2 Hz, 3H), 3.15 (q, J= 5.6 Hz, 2H), 3.31 (s, 3H), 3.84-3.87
(m, 4H), 3.94-
3.97 (m, 4H), 4.12 (t, J= 6 Hz, 2H), 4.36 (q, 2H, J= 7.2 Hz), 5.18 (s, 2H),
6.63 (d, J= 9.2
Hz, 1 H), 7.27 (s, 1 H), 8.3 7 (dd, J = 9.6 Hz, 2.4 Hz, 1 H), 8.51 (d, J = 2.4
Hz, 1 H), 8.93 (s,
2H).
Step 118c: 2-(((2-(6-(2-Aminoethoxy)pyridin-3-yl)-4-morpholinothieno[3,2-d]
pyrimidin-
6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 259)
The title compound 259 was prepared as a light yellow solid (40 mg, 31%) from
0603-259 (130 mg, 0.24 mmol) and a freshly prepared hydroxylamine methanol
solution
(10 mL, 1.79 M) in CH2Cl2 (2 mL) using a procedure similar to that described
for
compound 237 (Example 107). m.p.: 110-115 C. LCMS: 538.3 [M+1]+. 1H NMR (400
MHz, DMSO-d6): 6 2.89 (q, J= 6 Hz, 2H), 3.23 (s, 3H), 3.75 (t, J= 4.8Hz, 4H),
3.90 (t, J
4.4 Hz, 4H), 4.01 (t, J = 6 Hz, 2H), 5.19 (s, 2H), 6.47 (d, J = 9.2 Hz, 1 H),
7.3 7 (s, 1 H), 8.32
(dd, J = 9.6 Hz, 2.4 Hz, 1 H), 8.63 (d, J = 2.4 Hz, 1 H), 8.75 (s, 2H).
EXAMPLE 119: N-Hydroxy-2-(((2-(2-methoxypyrimidin-5-yl)-4-morpho-
linothieno[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carbo-
xamide
(Compound 261)
Step 119a: 5-Bromo-2-methoxypyrimidine (Compound 0601-261)
5-Bromo-2-chloropyrimidine (10g, 52mmol) and sodium methoxide (8.42 g, 156
mmol) were taken in methanol (50 mL) and stirred at room temperature
overnight. Reaction
mixture was concentrated followed by the addition of water. The resulting
mixture was
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extracted with ethyl acetate and dried over MgSO4. The titled compound was
obtained after
concentration (8.07 g, 82.5%).
Step 119b: 2-Methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine
(Compound 0602-261)
A mixture of 0601-261(0.5 g, 2.65 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-
bi(1,3,2-dioxaborolane) (1.35 g, 5.3 mmol), KOAc (780 mg, 7.95 mmol),
Pd(dppf)C12 (100
mg, 0.133 mmol) was taken into 1,4-dioxane (10 mL) in a pressure vessel was
heated at
100 C for 5 h. The reaction mixture was filtered through silica pad eluted
with
dichloromethane (50 mL). The titled compound was obtained as a white solid
(0.5 g, 90%)
after concentration.
Step 119c: Ethyl 2-(((2-(2-methoxypyrimidin-5-yl)-4-morpholinothieno[3,2-d]
pyrimidin -
6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-261)
The title compound 0603-261 was prepared as an grey solid (120 mg, 24%) from
0504 (450mg, 1.0 mmol), 0602-261 (472 mg, 2.Ommol), NaHCO3 (252 mg, 3.0 mmol),
bis(triphenylphosphine)palladium(II)chloride (120 mg, 0.15 mmol) was taken
into ethanol
(10 mL), water(6 mL), toluene(8 mL) using a procedure similar to that
described for
compound 1103-226 (Example 99).
Step 119d: N-Hydroxy-2-(((2-(2-methoxypyrimidin-5-yl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 261)
The title compound 261 was prepared as a white solid (30 mg, 19.4%) from 0603-
261 (160 mg, 0.31 mmol) and a freshly prepared hydroxylamine methanol solution
(20 mL,
1.79 M) using a procedure similar to that described for compound 237 (Example
107).
m.p.: 215-220 C. LCMS: 510.0[M+1]+.'HNMR (400 MHz, DMSO-d6): 6 3.24 (s, 3H),
3.74-3.76 (m, 4H), 3.92-3.95 (m, 4H), 4.00 (s, 3H), 5.20 (s, 2H), 7.46 (s,
1H), 8.75 (s, 2H),
9.04 (s, 1 H), 9.42 (s, 2H), 11.12 (s, 1 H).
EXAMPLE 120: N-Hydroxy-2-((1-(2-(2-methoxypyrimidin-5-yl)-4-morpho-
linothieno [3,2-d] pyrimidin-6-yl)ethyl)(methyl)amino)pyrimidine-5-carboxamide
(Compound 262)
Step 120a: Ethyl 2-((I-(2-(2-methoxypyrimidin-5-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6- yl)ethyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 1204-
262)
The title compound 1204-262 was prepared as an grey solid (150 mg, 72%) from
1203 (180 mg, 0.388 mmol), 602 (183mg, 0777mmo1), NaHCO3(98 mg, 1.17 mmol),
bis(triphenylphosphine)Pdlladium(II)chloride (50 mg, 0.02 mmol), ethanol (5
mL) ,
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water(2 mL), toluene(8 mL) using a procedure similar to that described for
compound
1204-237 (Example 107): LCMS: 537.2 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.31
(t, J= 7.2 Hz, 3H), 1.74 (d, J= 6.8 Hz, 3H), 3.03 (s, 3H), 3.74 (s, 4H), 3.94
(s, 4H), 4.00 (s,
3H), 4.30 (q, J= 7.2 Hz, 2H), 6.52 (s, 1H), 7.51 (s, 1H), 8.89 (s, 2H), 9.42
(s, 2H).
Step 120b: N-Hydroxy-2-((1-(2-(2-methoxypyrimidin-5-yl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)ethyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 262)
The title compound 262 was prepared as a white solid (80 mg, 55%) from 1204-
262
(150 mg, 0.28 mmol) and a freshly prepared hydroxylamine methanol solution (20
mL,
1.79 M) using a procedure similar to that described for compound 237 (Example
107).
m.p.: 210-215 C. LCMS: 524.1 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.72 (d, J=
6.8 Hz, 3H), 2.98 (s, 3H), 3.74 (s, 4H), 3.92 (s, 4H), 4.00 (s, 3H), 6.48 (q,
J= 5.2 Hz, 1H),
7.48 (s, 1H), 8.76 (s, 2H), 9.41 (s, 2H).
EXAMPLE 121: N-Hydroxy-2-(2-(2-(2-methoxypyrimidin-5-yl)-4-morpho-
linothieno [3,2-d] pyr- imidin-6-yl)propan-2-ylamino)pyrimidine-5-carboxamide
(Compound 263)
Step 121a: Ethyl 2-(2-(2-(2-methoxypyrimidin-5-yl)-4-morpholinothieno[3,2-d]
pyrimidin-
6-yl)propan-2-ylamino)pyrimidine-5-carboxylate (Compound 1210-263)
The title compound 1210-263 was prepared as a white solid (167 mg, 78%) from
1208 (185 mg, 0.40 mmol), boronate ester (189 mg, 0.80 mmol), NaHCO3 (101 mg,
1.20
mmol) and PdC12(PPh3)2 (28 mg, 0.04 mmol) in toluene (8 mL), EtOH (5 mL) and
water (2
mL) using a procedure similar to that described for compound 1204-237 (Example
107):
LC-MS: 537.3 [M+1]+.
Step 121b: N-Hydroxy-2-(2-(2-(2-methoxypyrimidin-5-yl)-4-morpholinothieno [3,2-
d]pyr-
imidin-6-yl)propan-2-ylamino)pyrimidine-5-carboxamide (Compound 263)
The title compound 263 was prepared as a white solid (30 mg) from 1210-263
(167
mg, 0.31 mmol) and a freshly prepared hydroxylamine methanol solution (20 mL,
1.79 M)
in CH2C12 (4 mL) using a procedure similar to that described for compound 237
(Example
107). m.p. 202-207 C. LC-MS: 524.3 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.84
(s,
6H), 3.74 (m, 4H), 3.92 (m, 4H), 4.00 (s, 3H), 7.31 (s, 1H), 8.38 (s, 1H),
8.43 (s, 1H), 8.52
(br, 2H), 8.98 (s, I H), 9.41 (s, 2H), 10.95 (s, I H).
EXAMPLE 122: 2-(2-(2-(4-Aminophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)propan-2-ylamino)-N-hydroxypyrimidine-5-carboxamide (Compound 269)
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Step 122a: Ethyl 2-(2-(2-(4-aminophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)pro-
pan-2-ylamino)pyrimidine-5-carboxylate (Compound 1210-269)
The title compound 1210-269 was prepared as a white solid (100 mg, 48%) from
1208 (185 mg, 0.40 mmol), boronate ester (0602-217) (175 mg, 0.80 mmol),
NaHCO3 (101
mg, 1.20 mmol) and PdC12(PPh3)2 (28 mg, 0.04 mmol) in toluene (8 mL), EtOH (5
mL) and
water (2 mL) using a procedure similar to that described for compound 1204-237
(Example
107): LC-MS: 520.4 [M+1]+.
Step 122b: 2-(2-(2-(4-Aminophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)
propan-2-
ylamino)-N-hydroxypyrimidine-5-carboxamide (Compound 269)
The title compound 269 was prepared as a white solid (77 mg, 75%) from 1210-
269
(100 mg, 0.19 mmol) and a freshly prepared hydroxylamine methanol solution (20
mL,
1.79 M) in CH2C12 (4 mL) using a procedure similar to that described for
compound 237
(Example 107). LC-MS: 507.3 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.82 (s, 6H),
3.28 (s, 1H), 3.74 (m, 4H), 3.85 (m, 4H), 5.48 (s, 2H), 6.60 (d, J= 8.8 Hz,
2H), 7.19 (s,
1H), 8.07 (d, J= 8.8 Hz, 2H), 8.33 (s, 1H), 8.53 (br, 2H), 8.96 (s, 1H), 10.94
(s, 1H).
EXAMPLE 123: 2-[2-(4-Amino-phenyl)-4-morpholin-4-yl-thieno[3,2-d] pyrimidine-6-
sulfonylamino]-pyrimidine-5-carboxylic acid hydroxyamide (Compound 271)
Step 123a: 2-[2-(4-Amino-phenyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine- 6-
sulfonylamino]-pyrimidine-5-carboxylic acid ethyl ester ( Compound 1304-271)
A mixture of 1303 (270 mg, 0.56 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-
dioxaborolan-2-yl)aniline (368 mg, 1.67 mmol) and Pd(PPh3)4(200 mg,Ø3 mmol)
in
dioxane (9 mL) and sat. NaHCO3 (3 mL) was stirred at 110 C for 3h under N2
atmosphere.
Upon completion, the reaction was concentrated and purified by column
chromatography to
afford compound 1304-271 (170 mg, 56 %). LCMS: 542.3[M+1]+. 'HNMR (400 MHz,
DMSO-d6): 6 1.28 (t, J= 7.2 Hz, 3H), 3.79 (t, J= 4.4 Hz, 4H), 3.96 (t, J= 4.4
Hz, 4H),
4.27 (q, J= 7.2 Hz, 2H), 6.62 (d, J= 8.4 Hz, 2H), 7.85 (s, 1H), 8.08 (d, J=
8.4 Hz, 2H),
8.89 (s, 2H).
Step 123b: 2-[2-(4-Amino-phenyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine-6-
sulfonylamino]-pyrimidine-5-carboxylic acid hydroxyamide (Compound 271)
The title compound 271 was prepared as a white solid (26 mg, 17%) from 1304-
271
(160 mg) and a freshly prepared hydroxylamine methanol solution (20 mL, 1.79
M) using a
procedure similar to that described for compound 237 (Example 107). m.p.: 197-
205 C.
LCMS: 529.2[M+1]+.'HNMR (400 MHz, DMSO-d6): 6 3.79 (s, 4H), 3.94 (s, 4H), 6.60
(d,
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J= 8.4 Hz, 2H), 7.83 (s, I H), 8.08 (d, J= 8.4 Hz, 2H), 8.76 (s, 2H), 9.16 (s,
I H), 11.18(s,
I H).
EXAMPLE 124: 2-(((2-(4-Amino-3-chlorophenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl) methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 275)
Step 124a: Ethyl 2-(((2-(4-amino-3-chlorophenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-275)
The mixture of 4-bromo-2-chloroaniline (516 mg, 2.5 mmol),
bis(pinacolato)diboron (952 mg, 3.75 mmol), PdC12(dppf) (102 mg, 0.125 mmol)
and
KOAc (735 mg, 7.5 mmol) in anhydrous dioxane (10 mL) was heated at 100 C for
3h in a
pressure vessel. The resulting solution was filtered and the solid was washed
with CH2C12
(20 mL). The filtrate was concentrated in vacuo. The obtained deep brown solid
was mixed
with 0504 (505 mg, 1.125 mmol), NaHCO3 (280 mg, 3.375 mmol) and PdC12(PPh3)2
(39
mg, 0.056 mmol) in toluene (11 mL), EtOH (7 mL) and water (2.8 mL). The
suspension
was heated at 130 C overnight in a pressure vessel. After cooling to room
temperature,
CH2C12 (10 mL) and water (10 mL) were added and the mixture was filtered. The
organic
layer was separated, washed with brine, dried over MgSO4, and evaporated. The
crude
compound was purified by column chromatography (Hexanes/ethyl acetate) to
afford the
titled compound as a light yellow solid. (478 mg, 78%). LC-MS: 540.3 [M+1 ]+.
'H NMR:
(400 MHz, DMSO-d6): 6 1.31 (t, J= 6.8 Hz, 3H), 3.24 (s, 3H), 3.74 (t, J= 4.4
Hz, 4H),
3.89 (t, J= 4.0 Hz, 4H), 4.29 (q, J= 6.8 Hz, 2H), 5.22 (s, 2H), 5.75 (s, 2H),
6.85 (d, 1H),
7.40 (s, 1H), 8.07 (dd, J= 8.4 Hz, 1.6 Hz, 1H), 8.18 (d, J= 2.0 Hz, 1H), 8.87
(s, 3H).
Step 124b: 2-(((2-(4-Amino-3-chlorophenyl)-4-morpholinothieno[3,2-d]pyrimidin-
6-yl)
methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 275)
The title compound 275 was prepared as a light yellow solid (20 mg, 21 %) from
0603-275 (100 mg, 0.185 mmol) and a freshly prepared hydroxylamine methanol
solution
(2.1 mL, 1.79 M)) in CH2C12 (2 mL) using a procedure similar to that described
for
compound 237 (Example 107). m.p. 241-246 C. LC-MS: 527.4 [M+1]+. 'H NMR (300
MHz, DMSO-d6): 6 3.21 (s, 3H), 3.74 (s, 4H), 3.87 (s, 4H), 5.16 (s, 2H), 5.74
(s, 2H), 6.85
(m, I H), 7.36 (s, I H), 8.07 (m, I H), 8.17 (s, I H), 8.72 (s, 2H).
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EXAMPLE 125: 2-(((2-(4-Amino-3-tuorophenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl) methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 276)
Step 125a: Ethyl 2-(((2-(4-amino-3-fluorophenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-276)
The title compound 0603-276 was prepared as a light yellow solid (503 mg, 85%)
from 4-bromo-2-fluoroaniline (475 mg, 2.5 mmol), bis(pinacolato) diboron (952
mg, 3.75
mmol), PdC12(dppf) (102 mg, 0.125 mmol) and KOAc (735 mg, 7.5 mmol) in
anhydrous
dioxane (10 mL) followed by 0504 (505 mg, 1.125 mmol), NaHCO3 (280 mg, 3.375
mmol)
and PdC12(PPh3)2 (39 mg, 0.056 mmol) in toluene (11 mL), EtOH (7 mL) and water
(2.8
mL) using a procedure similar to that described for compound 0603-275 (Example
124):
LC-MS: 524.3 [M+1]+. 1H NMR: (400 MHz, DMSO-d6): 6 1.31 (t, J= 7.2 Hz, 3H),
3.27 (s,
3H), 3.75 (t, J= 4.8 Hz, 4H), 3.89 (t, J= 4.8 Hz, 4H), 4.30 (q, J= 7.2 Hz,
2H), 5.22 (s, 2H),
5.55 (s, 2H), 6.84-6.79 (m, 1H), 7.40 (s, 1H), 7.97-7.91 (m, 2H), 8.88 (s,
2H).
Step 125b: 2-(((2-(4-Amino-3-fluorophenyl)-4-morpholinothieno[3,2-d]pyrimidin-
6-yl)
methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 276)
The title compound 276 was prepared as a light yellow solid (55 mg, 38%) from
0603-276 (150 mg, 0.286 mmol) and a freshly prepared hydroxylamine methanol
solution
(3.3 mL, 1.79 M in methanol) in CH2C12 (3 mL) using a procedure similar to
that described
for compound 237 (Example 107). m.p. 203-206 C. LC-MS: 511 [M+1]+ . 1H NMR
(300
MHz, DMSO-d6): 6 3.23 (s, 3H), 3.82 (m, 4H), 3.88 (m, 4H), 5.18 (s, 2H), 5.53
(s, 2H),
6.79-6.83 (m, I H), 7.37 (s, I H), 7.91-7.96 (m, 2H), 8.74 (s, 2H), 9.02 (br,
I H), 11.08 (br,
I H).
EXAMPLE 126: 2-(((2-(4-Amino-3-nitrophenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 278)
Step 126a: Ethyl 2-(((2-(4-amino-3-nitrophenyl)-4-morpholinothieno[3,2-
d]pyrimidin -6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-278)
The title compound 0603-278 was prepared as an orange solid (538 mg, 86%) from
4-bromo-2-nitroaniline (543 mg, 2.5 mmol), bis(pinacolato) diboron (952 mg,
3.75 mmol),
PdC12(dppf) (102 mg, 0.125 mmol) and KOAc (735 mg, 7.5 mmol) in anhydrous
dioxane
(10 mL) followed by 0504 (505 mg, 1.125 mmol), NaHCO3 (280 mg, 3.375 mmol) and
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PdC12(PPh3)2 (39 mg, 0.056 mmol) in toluene (11 mL), EtOH (7 mL) and water
(2.8 mL)
using a procedure similar to that described for compound 0603-275 (Example
124).
Step 126b: 2-(((2-(4-Amino-3-nitrophenyl)-4-morpholinothieno[3,2-d]pyrimidin-
6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 278)
The title compound 278 was prepared as a light pink solid (35 mg, 23%) from
111-
243-2 (150 mg, 0.272 mmol) and a freshly prepared hydroxylamine methanol
solution (3.3
mL, 1.79 M in methanol) in CH2C12 (3 mL) using a procedure similar to that
described for
compound 237 (Example 107). m.p. 202-206 C. LCMS: 538.5 [M+1]+. 'H NMR (400
MHz, DMSO-d6): 6 3.24 (s, 3H), 3.77 (m, 4H), 3.91 (m, 4H), 5.19 (s, 2H), 7.10
(d, J= 6.6
Hz, 1 H), 7.44 (s, 1 H), 7.69 (s, 2H), 8.3 7 (dd, J = 6.6, 1.5 Hz, 1 H), 8.74
(s, 2H), 9.00 (d, J =
1.5 Hz, 2H).
EXAMPLE 127: 2-(((2-(4-Amino-3-hydroxyphenyl)-4-morpholinothieno [3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 279)
Step 127a: Ethyl 2-(((2-(4-amino-3-hydroxyphenyl)-4-morpholinothieno[3,2-d]
pyrimidin-
6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-279)
The title compound 0603-279 was prepared as a yellow solid (200 mg) from 2-
amino-5-bromophenol (1 g, 5.3 mmol), 4,4,4',4',5,5,5',5'- octamethyl- 2,2'-
bi(1,3,2-
dioxaborolane) (2.7 g, 10.6 mmol), KOAc (1.5 g, 15.9 mmol) and Pd(dppf)C12
(200 mg) in
DMSO (15 mL) followed by 0504 (500 mg, 2.12 mmol), 2-amino-5-(4,4,5,5-
tetramethyl-
1,3,2-dioxaborolan-2-yl)phenol (a solution in DMSO prepared in previous step,
sat.
NaHCO3 (2 mL) and Pd(PPh3)4 (100 mg) in DMSO (50 mL) using a procedure similar
to
that described for compound 0603-275 (Example 124).
Step 127b: 2-(((2-(4-Amino-3-hydroxyphenyl)-4-morpholinothieno[3,2-d]pyrimidin-
6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 279)
The title compound 279 was prepared as a yellow solid (50 mg, 25%) from 111-
247-3 (200 mg) and a freshly prepared hydroxylamine methanol solution (8 mL,
1.79 M in
methanol) in CH2C12 (3 mL) using a procedure similar to that described for
compound 237
(Example 107). m.p.: 214-218 C. LCMS: 509 [M+1]+.'HNMR (400 MHz, DMSO-d6): 6
3.20 (s, 3H), 3.76 (t, J= 4.4 Hz, 4H), 3.88 (t, J= 4.0 Hz, 4H), 5.17 (s, 2H),
6.49 (s, 2H),
6.58-6.64 (m, 2H), 7.44 (s, 1H), 7.73 (s, 1H), 8.69 (s, 2H).
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EXAMPLE 128: 2-(((2-(4-Amino-3-cyanophenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 280)
Step 128a: 2-Amino-5-bromobenzonitrile (Compound 0601-280)
2-Aminobenzonitrile (5.0 g, 42.3 mmol) was taken into dichloromethane (60 mL).
NBS (7.54 g, 42.3 mmol) was added and stirred at 0 C for 3h. The reaction
mixture was
diluted with dichloromethane and washed with brine, dried over MgSO4. The
crude product
was purified by column chromatography (hexane/ethyl acetate = 10/1) to afford
the titled
compound (7.20 g, 87%).'HNMR (400 MHz, DMSO-d6): 6 6.22 (s, 2H), 6.75 (d, J=
9.2
Hz, 1 H), 7.42 (dd, J = 8.8 Hz, 2.4 Hz, 1 H), 7.5 8 (s, 1 H).
Step 128b: Ethyl 2-(((2-(4-amino-3-cyanophenyl)-4-morpholinothieno[3,2-d]
pyrimidin- 6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-280)
A mixture of 0601-280 (500 mg, 2.54 mmol), 4,4,4',4',5,5,5',5'-octamethyl -
2,2'-
bi(1,3,2-dioxaborolane) (775 mg, 3.04 mmol), KOAc( 742 mg, 7.62 mmol),
PdC12(dppf)
(66 mg) in anhydrous 1,4-dioxane (20 mL) was heated to 90 C for 12h. The
reaction
mixture was filtered and concentrated to afford crude product 0602-280 as a
brown solid
(550 mg) which was used for the next step reaction without further
purification.
A mixture of 0602-280 (500 mg, prepared above), 0504 (500 mg, 1.11 mmol),
NaHCO3 (269 mg, 3.2 mmol), PdC12(Ph3P)2 (60 mg), in toluene/ethanol/water(17.1
mL/10.5 mL/ 4.5 mL) was stirred at 140 C overnight. The reaction mixture was
filtered
and concentrated in vacuo. The crude product was purified by column
chromatography to
afford 0603-280 (140 mg, 24%) as a white solid. LCMS: 531.4 [M+1]+. 'HNMR,
(400
MHz, DMSO-d6): 6 1.31 (t, J= 7.2 Hz, 3H), 3.27(s, 3H), 3.75(s, 4H), 3.91(s,
4H), 4.29
(q, J = 7.2 Hz, 2H), 5.22 (s, 2H), 6.44 (s, 2H), 6.87 (d, J = 8.8 Hz, 1 H),
7.41 (s, 1 H), 8.29-
8.34 (m, 2H), 8.88 (s, 1H).
Step 128c: 2-(((2-(4-Amino-3-cyanophenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 280)
The title compound 280 was prepared as a yellow solid (20 mg, 14%) from 0603-
280 (150 mg, 0.28 mmol) and a freshly prepared hydroxylamine methanol solution
(6 mL,
1.79 M in methanol) in CH2C12 (2 mL) using a procedure similar to that
described for
compound 237 (Example 107). m.p.:237-240 C. LCMS: 518.0 [M+1]+. 'HNMR, (400
MHz, DMSO-d6): 6 3.23 (s, 3H), 3.75 (s, 4H), 3.89 (s, 4H), 5.18 (s, 2H), 6.86
(s, 2H), 6.87
(d, J = 8.4 Hz, 1 H), 7.3 8 (s, 1 H), 8.29-8.33 (m, 2H), 8.74 (s, 1 H).
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EXAMPLE 129: 2-{[2-(3-Acetyl-4-amino-phenyl)-4-morpholin-4-yl-thieno [3, 2-d]
pyrimidin -6-yl methyl]-methyl-amino}-pyrimidine-5-carboxylic acid
hydroxyamide
(Compound 281)
Step 129a: 1-(2-Amino-5-bromo-phenyl)-ethanone (Compound 0601-281)
To a mixture of 1-(2-aminophenyl)ethanone (5.0 g, 0.037 mol) in
dichloromethane
(60 mL) was added NBS (7.3 g, 0.04 mol). The resulting mixture was cooled to 0
C
followed by the addition of H2SO4 (0.05 mL). It was allowed to warm to room
temperature
and stirred overnight. To the reaction mixture was added water and extracted
with
dichloromethane, dried over MgSO4. The crude product was purified by column
chromatography eluted with ethyl acetate/hexanes to afford the titled compound
(4.6 g,
58%) as a yellow solid. m.p 87-89 C. 1H NMR (400 MHz, CDC13): 6 2.56 (s, 3H),
6.31 (s,
2H), 7.5 6 (d, J = 8.8 Hz, 1 H), 7.3 3 (dd, J = 2.4 Hz, 1 H), 7.80 (d, J = 2.4
Hz, 1 H).
Step 129b: 1-[2-Amino-5-(4, 4, 5, 5-tetramethyl-[1, 3, 2] dioxaborolan-2-yl)-
phenyl]-
ethanone (Compound 0602-28 1)
A mixture of 0601-281(1 g, 4.7 mmol), bis(pinacolato) diboron(1.66 g, 6.5
mmol),
PdC12(dppf) (230 mg, 0.28 mmol) and KOAc (1.48 g, 15 mmol) in anhydrous
dioxane (30
mL) was heated at 100 C for 4h in a pressure vessel. The reaction mixture was
filtered and
the solid was washed with CH2Cl2. The filtrate was concentrated in vacuo and
purified by
column chromatography (3% ethyl acetate/hexanes) to afford 0602-281 (1 g ,
83%) as an
off-white solid. m.p 45-48 C. 1H NMR (400 MHz, CDC13): 6 1.33 (s, 12H), 2.62
(s, 3H),
6.49 (s, 2H), 6.60 (d, J= 8.4 Hz, 1H), 7.66 (d, J= 8.0 Hz, 1H), 8.18 (s, 1H).
Step 129c: 2- {[2-(3 -Acetyl-4-amino-phenyl)-4-morpholin-4-yl-thieno [3, 2-d]
pyrimidin-6-
ylmethyl]-methyl-amino}-pyrimidine-5-carboxylic acid ethyl ester (Compound
0603-281)
The title compound 0603-281 was prepared as a light yellow solid (450 mg, 92%)
from 0602-281 (720 mg, 2.7mmol), 0504 (412 mg, 0.9 mmol), NaHCO3 (232 mg, 2.7
mmol) and PdC12(PPh3)2 (107 mg) in toluene (7.5 mL), EtOH (4.8 mL) and water
(2.1 mL)
using a procedure similar to that described for compound 1103-226 (Example
99): LCMS:
548.4 [M+1 ]+.
Step 129d: 2- {[2-(3 -Acetyl-4-amino-phenyl)-4-morpholin-4-yl-thieno [3, 2-d]
pyrimidin -
6-yl methyl]-methyl-amino}-pyrimidine-5-carboxylic acid hydroxyamide (Compound
281)
The title compound 281 was prepared as a pale yellow solid (40 mg) from 0603-
281
(500 mg) and a freshly prepared hydroxylamine methanol solution (12 mL, 1.79 M
in
methanol) in CH2Cl2 (5 mL) using a procedure similar to that described for
compound 237
(Example 107). m.p.:156-158 C. LC-MS: 535 [M+1]+. 1H NMR (400 MHz, DMSO-d6):
6
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2.61 (s, 3H), 3.23 (s, 3H), 3.76 (s, 4H), 3.90 (s, 4H), 5.19 (s, 2H), 6.84 (m,
1H), 7.40 (s,
1H), 7.53 (s, 2H), 8.27 (m, 1H), 8.78 (d, J= 23.2 Hz, 3H), 9.02 (s, 1H), 11.09
(s, 1H).
EXAMPLE 130: 2-(((2-(4-Amino-3,5-difluorophenyl)-4-morpholinothieno [3,2-d]
pyrimidin-6-yl) methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 282)
Step 130a: 2,6-Difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline
(Compound 0602-282)
The title compound 0602-282 was prepared as an off-white solid (3 g) from 4-
bromo-2,6-difluoroaniline (2.08 g, 10 mmol), 4,4,4',4',5,5,5',5'-octamethyl-
2,2'-bi(1,3,2-
dioxaborolane (3.79 g, 15 mmol), KOAc (3 g, 30 mmol) and Pd(dppf)C12 (400 mg,
0.5
mmol) in dioxane (50 mL) using a procedure similar to that described for
compound 0602-
281 (Example 129). 1H NMR (400 MHz, CDC13): 6 1.31 (s, 12H), 3.91 (br, 2H),
7.22-7.25
(m, 2H).
Step 130b: Ethyl 2-(((2-(4-amino-3,5-difluorophenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-
282)
The title compound 0603-282 was prepared as a yellow solid (200 mg, 55%) from
0602-282 (500 mg, 2 mmol), 0504 (300 mg, 0.669 mmol), NaHCO3 (168 mg, 2 mmol)
and
Pd(PPh3)2C12 (50 mg) in toluene/EtOH/H20 (5 mL/3.2 mL/1.4 mL) using a
procedure
similar to that described for compound 1103-226 (Example 99): 'HNMR: (400 MHz,
CDC13): 6 1.31 (t, J= 7.2 Hz, 3H), 3.28 (s, 3H), 3.76-3.77 (m, 4H), 3.89-3.91
(m, 4H), 4.30
(q, J= 7.2 Hz, 2H), 5.23 (s, 2H), 5.59 (s, 2H), 7.42 (s, 1H), 7.86(d, J= 9.6
Hz, 2H), 8.88 (s,
2H).
Step 130c: 2-(((2-(4-Amino-3,5-difluorophenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-
yl) methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 282)
The title compound 282 was prepared as a white solid (95 mg, 63%) from 0603-
282
(150 mg, 0.277 mmol) and a freshly prepared hydroxylamine methanol solution (4
mL,
1.79 M in methanol) in CH2C12 (2 mL) using a procedure similar to that
described for
compound 237 (Example 107). m.p.: 190-193 C. LCMS: 529 [M+1]+. 'HNMR: (400
MHz,
CDC13): 6 3.23 (s, 3H), 3.75 (br, 4H), 3.89 (br, 4H), 5.18 (s, 2H), 5.62 (s,
2H), 7.39 (s, 1H),
7.85 (d, J= 8.4 Hz, 2H), 8.74 (s, 2H), 9.02 (br, 1H), 11.08 (s, 1H).
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EXAMPLE 131: 2-{[2-(4-Amino-2-fluoro-phenyl)-4-morpholin-4-yl-thieno[3,2-d]
pyrimidin-6-ylmethyl]-methyl-amino}-pyrimidine-5-carboxylic acid hydroxyamide
(Compound 283)
Step 131a: 4-Bromo-3-fluoro-phenylamine (Compound 0601-283)
3-fuloroaniline (4.44 g, 40 mmol), NBS (7.11 g, 40 mmol) were taken into
dichloromethane (30 mL). Cone. H2SO4 (2 drops) was added. The resulting
mixture was
stirred at 0 C for 4 h. The reaction mixture was diluted with water and
extracted with ethyl
acetate. The combined extract was washed with aq. NaHCO3, dried over Na2SO4.
The
crude product was purified by column chromatography (hexanes/ethyl acetate =
10/1) to
afford the titled compound (5.00 g, 66%) as a light yellow solid. LCMS: 190
[M+1]+.
iH NMR (400 MHz, DMSO-d6): 6 5.53 (s, 2H), 6.34 (dd, J= 8.8 Hz, 3.2 Hz, 1H),
6.45 (dd,
J = 11.6 Hz, 2.4 Hz, 1 H), 7.18 (t, 1 H).
Step 131b: 2-{[2-(4-Amino-2-fluoro-phenyl)-4-morpholin-4-yl-thieno[3,2-d]
pyrimidin-6-
ylmethyl]-methyl-amino}-pyrimidine-5-carboxylic acid ethyl ester (Compound
0603-283)
The title compound 0603-283 was prepared as a light yellow solid (250 mg, 28%)
from 0601-283 (500 mg, 3.44 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-
bi(1,3,2-
dioxaborolane) (1.74 g, 6.87 mmol), KOAc ( 842 mg, 8.60 mmol), PdC12(dppf) (50
mg) in
anhydrous 1,4-dioxane (5 mL) followed by 3-Fluoro-4-(4,4,5,5-tetramethyl-
[1,3,2]dioxaborolan-2-yl)- phenylamine, 0504 (772 mg, 1.72 mmol), NaHCO3 (433
mg,
5.16 mmol), PdC12(Ph3P)2 (75 mg) in toluene/ethanol/water(8 mL/4 mL/2 mL)
using a
procedure similar to that described for compound 0603-275 (Example 124). LCMS:
524
[M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.31 (t, J= 6.8 Hz, 3H), 3.25(s, 3H),
3.72 (m,
4H), 3.85 (m, 4H), 3.88 (m, 2H), 4.28 (q, J= 7.2 Hz, 2H), 5.20 (s, 2H), 6.34
(d, J= 12.8
Hz, 1H), 6.43 (d, J= 8.4 Hz, 1H), 7.38 (s, 1H), 7.83-7.90 (m, 1H), 8.86 (s,
2H).
Step 131c: 2-{[2-(4-Amino-2-fluoro-phenyl)-4-morpholin-4-yl-thieno[3,2-d]
pyrimidin-6-
ylmethyl]-methyl-amino }-pyrimidine-5-carboxylic acid hydroxyamide (Compound
283)
The title compound 283 was prepared as a white solid (50 mg, 20%) from 0603-
283
(250 mg, 0.4 8mmol) and a freshly prepared hydroxylamine methanol solution (8
mL, 1.79
M in methanol) in CH2C12 (2 mL) using a procedure similar to that described
for compound
237 (Example 107). m.p:195-197 C. LCMS: 511 [M+1]+. 1H NMR (400 MHz, DMSO-
d6): 6 3.22 (s, 3H), 3.71-3.74 (m, 4H), 3.84-3.87 (m, 4Hm), 5.18 (s, 2H), 5.74
(s, 2H), 6.34
(dd, J= 13.6 Hz, 1.6 Hz, I H), 6.43 (dd, J= 8.4 Hz, 1.6 Hz, I H), 7.36 (s, I
H), 7.85 (t, J=
8.4 Hz, 1 H), 8.74 (2H, s), 9.02 (br, 1 H), 11.01 (br, 1 H).
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EXAMPLE 132: 2-(((2-(4-Amino-2-chlorophenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 284)
Step 132a: 3-Chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline
(Compound
0602-284)
The title compound 0602-284 was prepared as a light yellow solid (1 g, 66.7%)
from 4-bromo-3-chloroaniline (1.23 g, 5.9 mmol), 4,4,4',4',5,5,5',5'-
octamethyl-2,2'-
bi(1,3,2-dioxaborolane) (2.2 g, 8.9 mmol), KOAc (1.7 g, 17.7 mmol) and
Pd(dppf)C12 (200
mg, 0.24 mmol) in dioxane using a procedure similar to that described for
compound 0602-
281 (Example 129).'HNMR (400 MHz, CDC13): 6 3.84 (br, 2H), 1.33 (s, 12H), 6.49
(d, J=
8.0 Hz, 1 H), 6.44 (s, 1 H), 7.51 (d, J = 8.0 Hz, 1 H).
Step 132b: Ethyl 2-(((2-(4-amino-2-chlorophenyl)-4-morpholinothieno[3,2-d]
pyrimidin -
6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-284)
The title compound 0603-284 was prepared as a yellow solid (200 mg, 55%) from
0602-284 (220 mg, 0.869 mmol), 0504 (300 mg, 0.66 mmol), NaHCO3 (166 mg, 2
mmol)
and Pd(PPh3)2C12 (50 mg) in toluene/EtOH/H20 (5 mL/3.2 mL/1.4 mL) using a
procedure
similar to that described for compound 1103-226 (Example 99): 'HNMR (400 MHz,
DMSO-d6): 6 1.31 (t, J= 7.2 Hz, 2H), 3.72 (t, J= 4.0 Hz, 4H), 3.87 (t, J= 4.0
Hz, 4H),
4.29 (q, J= 7.2 Hz, 2H), 5.23 (s, 2H), 5.61 (br, 2H), 6.57 (d, J= 8.4 Hz, 1H),
6.67 (s, 1H),
7.41 (s, 1H), 7.60 (d, J= 8.0 Hz, 2H), 8.88 (s, 2H).
Step 132c: 2-(((2-(4-Amino-2-chlorophenyl)-4-morpholinothieno[3,2-d]pyrimidin-
6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 284)
The title compound 284 was prepared as a light tan solid (31 mg, 15%) from
0603-
284 (200 mg, 0.37 mmol) and a freshly prepared hydroxylamine methanol solution
(4 mL,
1.79 M in methanol) in CH2C12 (2 mL) using a procedure similar to that
described for
compound 237 (Example 107). m.p.:179-183 C. LCMS: 527.0[M+l]+. 'HNMR (400 MHz,
DMSO-d6): 6 3.22 (s, 3H), 3.72 (t, J= 4.8 Hz, 4H), 3.87 (t, J= 4.0 Hz, 4H),
5.19 (s, 2H),
5.61 (s, 2H), 6.5 7 (dd, J = 8.4 Hz, 2 Hz, 1 H), 6.67 (s, 1 H), 7.3 9 (s, 1
H), 7.60 (d, J = 8.4 Hz,
2H), 8.73 (s, 2H).
EXAMPLE 133: 2-(((2-(4-Amino-3-(hydroxymethyl)phenyl)-4-morpholinothieno [3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 285)
Step 133a: (2-Amino-5-bromophenyl)methanol (Compound 0601-285)
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A solution of 2-Amino-5-bromo-benzoic acid (5.0 g, 23.15 mmol) in anhydrous
THE
was added dropwise to the mixture of LiA1H4 (2.2 g, 58 mmol) in THE under N2
protection
at 0 C. The resulting mixture was warmed to room temperature and stirred for
2h. The
reaction was quenched by adding Na2SO4 10H20 to the reaction mixture at 0 C.
The
resulting mixture was filtered and the filtrate was concentrated to afford the
titled
compound 0601-285 (2.05 g, 43%) as an off-white solid, m.p.103-106 C. The
crude
product was used for next step reaction without further purification. 1H NMR
(400 MHz,
CDC13): 6 4.61 (s, 2H); 6.56 (d, J= 8.0 Hz, 2H,); 7.19-7.26 (m, 2H).
Step 133b: (2-Amino-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)
methanol
(Compound 0602-285)
The title compound 0602-285 was prepared as a yellow solid (900 mg, 77%) from
0601-285 (950 mg, 4.7 mmol), bis(pinacolato)diboron (1.8 g, 7.08 mmol),
AcOK(1.38 g,
14 mmol) PdC12(dppf)2 (390 mg, 0.478 mmol) in anhydrous dioxane using a
procedure
similar to that described for compound 0602-281 (Example 129). LCMS: 250.3
[M+1]+.
'HNMR (400 MHz, CDC13): 6 1.31 (s, 12H); 4.40 (br, 2H); 4.66 (s, 2H); 6.66 (d,
J= 8.0
Hz, 1 H); 7.51 (s, 1 H); 7.5 8 (dd, J = 8.0 Hz, 1.2 Hz, 1 H).
Step 133c: Ethyl 2-(((2-(4-amino-3-(hydroxymethyl)phenyl)-4-
morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-
285)
The title compound 0603-285 was prepared as a light yellow solid (239 mg,
55.8%)
from 0602-285 (450 mg, 1.8 mmol), 0504 (360 mg 0.8 mmol) NaHCO3 (202 mg, 2.4
mmol), Pd(PPh3)2C12 (60 mg, 0.085 mmol) in toluene (12 mL), ethanol (7 ml) and
water (3
ml) using a procedure similar to that described for compound 1103-226 (Example
99): mp
215-217 C. LCMS, 536.3 [M+1]+; 'HNMR (400 MHz, DMSO-d6): 6 1.33, (m, 3H), 3,77
(m, 4H), 3,92 (m, 4H), 3.30 (m, 3H), 4.32 (m, 2H), 4.50 (m, 2H), 5.04 (m, 1H),
5.22 (m,
2H), 5.33 (m, 2H), 6.70 (m, I H), 7.40 (m, I H), 8.07 (m, I H), 8.19 (m, I H),
8.92 (s, 2H).
Step 133d: 2-(((2-(4-Amino-3-(hydroxymethyl)phenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound
285)
The title compound 285 was prepared as a light yellow solid (97 mg) from 0603-
285 (115 mg, 0.2147 mmol) and a freshly prepared hydroxylamine methanol
solution (30
mL, 1.79 M in methanol) in CH2C12 (2 mL) using a procedure similar to that
described for
compound 237 (Example 107). m.p.: 195-200 C. LCMS: 523.0 [M+1]+. 'HNMR (400
MHz, DMSO-d6): 6 3.17 (s, 3H); 3.76 (s, 4H); 3.89 (s, 4H); 4.46 (s, 2H); 5.18
(s, 2H); 5.33
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(s, 2H); 8.67 (t, J = 3.6 Hz, 1 H); 7.37 (s, 1 H); 8.04 (d, J = 7.2 Hz, 1 H);
8.16 (s, 1 H), 8.74 (s,
2H).
EXAMPLE 134: 2-({2-[3-(2-Hydroxy-ethoxy)-phenyl]-4-morpholin-4-yl-thieno [3,2-
d]pyrimidin-6-ylmethyl}-methyl-amino)-pyrimidine-5-carboxylic acid
hydroxyamide
(Compound 289)
Step 134a: [2-(3-Bromo-phenoxy)-ethoxy]-t-butyl-dimethyl-silane (Compound 0601-
289)
To a solution of 2-(3-Bromo-phenoxy)-ethanol (2.0 g, 9.2 mol) in
dichloromethane
(15 mL) at ice water bath temperature was added (1.25 g, 1.84 mmol) imidazole
followed
by the addition of TBSC1(1.67 g, 11 mmol) under nitrogen protection. TLC
showed
reaction was complete in 1 h. The reaction mixture was filtered. The filtrate
was washed
with water and dried over Na2SO4. The crude product was purified by column
chromatography eluted with hexanes/ethyl acetate to afford titled product as a
light color
oil (2.65 g, 87%).
Step 134b: 2-{3-[2-(tert-Butyl-dimethyl-silanyloxy)-ethoxy]-phenyl}-4,4,5,5-
tetramethyl-
[1,3,2]dioxaborolane (Compound 0602-289)
The title compound 0602-289 was prepared as an off-white solid (1.94 g, 64%)
from 0601-289 (2.65 g, 8 mmol), bis (pinacolato) diboron, KOAc (2.36 g, 24
mmol) and
Pd(dppf)C12 (0.117 g, 2 mmol) in anhydrous dioxane (30 mL) using a procedure
similar to
that described for compound 0602-281 (Example 129).
Step 134c: 2-({2-[3-(2-Hydroxy-ethoxy)-phenyl]-4-morpholin-4-yl-thieno[3,2-d]
pyrimidin-6-ylmethyl}-methyl-amino)-pyrimidine-5-carboxylic acid ethyl ester
(Compound 0603-289)
A mixture of 0602-289 (1.94 g, 5.1 mmol), 0504 (0.5079 g, 1.3 mmol) and NaHCO3
(0.323 g, 3.8 mmol) were taken into l Oml toluene (10 mL), ethanol (6.4 mL)
and water (2.8
mL) followed the addition of Pd(PPh3)2C12 (54 mg). The resulting mixture was
heated at
120 C overnight. TLC showed the reaction complete. The reaction mixture was
filtered
through celite and washed with dichloromethane. The crude product was purified
by
column chromatography eluted with hexanes/ethyl acetate to afford 2-[(2-{3-[2-
(tert-Butyl-
dimethyl-silanyloxy)-ethoxy]-phenyl}-4-morpholin- 4-yl-thieno[3,2-d]pyrimidin-
6-
ylmethyl)-methyl-amino]-pyrimidine-5-carboxylic acid ethyl ester (0.71 g, 83%)
as an off-
white solid. LC-MS: 665[M+1]+.
2-[(2-{3-[2-(tert-Butyl-dimethyl-silanyloxy)-ethoxy]-phenyl}-4-morpholin- 4-yl-
thieno[3,2-d]pyrimidin-6-ylmethyl)-methyl-amino]-pyrimidine-5-carboxylic acid
ethyl
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ester (680 mg, 1.02 mmol) was taken into THE (20 mL) followed by the addition
of
tetrabutyl ammonium fluoride (533 mg, 2.04 mmol). The resulting mixture was
stirred at
room temperature for 30 min. TLC showed reaction complete. Reaction mixture
was
diluted with ethyl acetate, washed with water and dried with Na2SO4. The crude
product
was purified by column chromatography eluted with hexanes/ethyl acetate to
afford titled
compound as an off-white solid (540 mg, 96%). m.p.: 216-220 C. LC-MS: 551.3
[M+1] +.
iH NMR (400 MHz, CDC13): 6 1.38 (t, J= 6.8 Hz, 3H), 3.31 (s, 3H), 3.86 (t, J=
5.2 Hz,
4H), 3.99-4.02 (m, 6H), 4.20 (t, J = 4.4 Hz, 2H), 4.37 (q, J = 7.2 Hz, 2H),
5.19 (s, 2H). 7.01
(dd, J= 8 Hz, 2 Hz, 1H), 7.35-7.39 (m, 2H). 8.01-8.05 (m, 2H), 8.92 (s, 2H).
Step 134e: 2-({2-[3-(2-Hydroxy-ethoxy)-phenyl]-4-morpholin-4-yl-thieno[3,2-d]
pyrimidin-6-ylmethyl }-methyl-amino)-pyrimidine-5 -carboxylic acid
hydroxyamide
(Compound 289)
The title compound 289 was prepared as a white solid solid (70 mg, 71.7%) from
0603-289 (100 mg) and a freshly prepared hydroxylamine methanol solution (3
mL, 1.79 M
in methanol) in CH2C12 (1 mL) using a procedure similar to that described for
compound
237 (Example 107). m.p.: 148-152 C. LC-MS: 538.0 [M+1] +. 1H NMR (400 MHz,
DMSO-d6): 6 3.24 (S, 3H), 3.75-3.78 (m, 6H), 3.92 (t, J= 4.8 Hz, 4H), 4.06 (t,
J= 5.2 Hz,
2H). 4.85 (t, J = 4.4 Hz, 1 H), 5.20 (s, 2H), 7.05 (dd, J = 8.0 Hz, 2 Hz, 1
H). 7.3 8 (t, J = 8
Hz, 1 H), 7.46 (s, 1 H). 7.92 (s, 1 H), 7.97(d, J = 3.6 Hz, 1 H), 8.75 (s,
2H), 9.02 (s, 1 H),
11.09 (s, 1H).
EXAMPLE 135: N-Hydroxy-2-(((2-(3-(2-hydroxyethylthio)phenyl)-4-morpho-
linothieno[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5- carboxamide
(Compound 290)
Step 135a: 2-(3-Bromophenylthio)ethanol (Compound 0601-290)
A mixture of 3-bromobenzenethiol (1.5 g, 7.93 mmol), 2-bromoethanol (1.19 g,
9.52 mmol) and Cs2CO3 (5.16 g, 15.8 mmol) in DMF (l Oml) was heated to 50 C
overnight. The reaction mixture was diluted with ethyl acetate and filtered.
The filtrate was
washed with water and dried. The crude product was purified by column
chromatography
to afford the titled compound (1.5 g, 81%). 'HNMR (400 MHz, CDC13): 6 3.12 (t,
J= 6.0
Hz, 2H), 3.77 (t, J= 6.4 Hz, 2H), 7.14 (t, J= 8.0 Hz, 1H), 7.26-7.34 (m, 2H),
7.51 (s, 1H).
Step 135b: Ethyl 2-(((2-(3-(2-hydroxyethylthio)phenyl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-
290)
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The title compound 0603-290 was prepared as a yellow solid (525 mg, 70%) from
0601-290 (487 mg, 2.1 mmol), bis(pinacolato)diboron (0.8g, 3.l5mmol), AcOK
(720mg
7.35mmol) and PdC12(dppf)2 (190 mg, 0.26 mmol) in dioxane followed by 0504
(600 mg,
1.33 mmol), 0602-290, NaHCO3 (400 mg, 4.76 mmol), Pd(PPh3)2C12 (60 mg, 0.086
mmol)
in toluene (12 ml), ethanol (8 ml), water (6 ml) using a procedure similar to
that described
for compound 0603-275 (Example 124). m.p.: 144-147 C. LCMS: 567.2[M+l]+.
'HNMR
(400 MHz, DMSO-d6): 6 1.30 (t, J= 6.8 Hz, 3H), 3.10 (t, J= 6.8 Hz, 2H), 3.59-
3.64 (m,
2H), 3.76 (br, 4H), 3.93 (br, 4H), 4.29 (q, J = 6.8 Hz, 2H), 4.91 (t, J = 4.2
Hz, 1 H), 5.24 (s,
2H), 7.43-7.45 (br, 2H); 8.19 (d, J= 7.2 Hz, 1H), 8.33 (s, 1H); 8.87 (s, 2H).
Step 135c: N-Hydroxy-2-(((2-(3-(2-hydroxyethylthio)phenyl)-4-morpholinothieno
[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 290)
The title compound 290 was prepared as a white solid solid (97 mg) from 0603-
290
(260 mg, 0.46 mmol) and a freshly prepared hydroxylamine methanol solution (30
mL,
1.79 M in methanol) using a procedure similar to that described for compound
237
(Example 107). m.p.: 188-192 C. LCMS: 554.0[M+1]+. 'H NMR (400 MHz, DMSO-d6):
6
3.10 (t, J= 6.8 Hz, 2H), 3.24 (s, 3H), 3.58-3.63 (m, 2H), 3.76 (br, 4H), 3.92
(br, 4H), 4.99
(t, J= 5.2 Hz, I H), 5.20 (s, 2H), 7.43-7.49 (br, 3H), 8.18 (d, J= 6.8 Hz, I
H), 8.32 (s, I H),
8.75 (s, 2H), 9.08 (br, I H), 11.15 (s, I H).
EXAMPLE 136: 2-(((2-(2-Fluoro-5-hydroxyphenyl)-4-morpholinothieno[3,2-d]
pyrimidin-6- yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 292)
Step 136a: Ethyl 2-(((2-(2-fluoro-5-hydroxyphenyl)-4-morpholinothieno[3,2-d]
pyrimidin-
6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-292)
The title compound 0603-292 was prepared as a grey solid (1.32 g 74.9%) from 3-
bromo-4-fluorophenol (1.425 g, 7.5 mmol), bis(pinacolato) diboron (2.86 g,
11.25 mmol),
PdC12(dppf) (306 mg, 0.375 mmol), and KOAc (2.205 g, 22.5 mmol) in anhydrous
dioxane
(20 mL) followed by 0504 (1.5 g, 3.375 mmol), NaHCO3 (850 mg, 10.13 mmol) and
PdC12(PPh3)2 (118 mg, 0.168 mmol) in toluene (11 mL), EtOH (7 mL) and water
(2.8 mL)
using a procedure similar to that described for compound 0603-275 (Example
124).
LCMS: 525.2 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.30 (m, 3H), 3.27 (s, 3H),
3.74
(m, 4H), 3.88 (m, 4H), 4.29 (m, 2H), 5.24 (s, 2H), 6.83 (m, 1H), 7.07 (m, 1H),
7.42 (m,
2H), 8.88 (s, 2H), 9.48 (s, 1H).
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Step 136b: 2-(((2-(2-Fluoro-5-hydroxyphenyl)-4-morpholinothieno[3,2-
d]pyrimidin-6-
yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide (Compound 292)
The title compound 292 was prepared as a tan solid (380 mg) from 0603-292 (560
mg, 1.07 mmol) and a freshly prepared hydroxylamine methanol solution (30 mL,
1.79 M
in methanol) using a procedure similar to that described for compound 237
(Example 107).
m.p. 250-255 C. LCMS: 512 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 3.24 (s, 3H),
3.73 (m, 4H), 3.88 (m, 4H), 5.21 (s, 2H), 6.83 (m, I H), 7.06 (m, I H), 7.42
(m, I H), 7.45 (s,
1 H), 8.75 (s, 2H), 9.04 (s, 1 H), 9.45 (s, 1 H), 11.11 (s, 1 H).
EXAMPLE 137: N-Hydroxy-2-(((2-(3-hydroxy-4-nitrophenyl)-4-morpholinothieno
[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound
293)
Step 137a: 2-Nitro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol
(Compound
0602-293)
The title compound 0602-293 was prepared as yellow solid (2,9 g, 80%) from 5-
bromo-2-nitrophenol (3 g, 13.76 mmol), 4,4,4',4',5,5,5',5'-octamethyl -2,2'-
bi(1,3,2-
dioxaborolane) (5.25 g, 20.69 mmol), KOAc (3.75 g, 41.28 mmol) and Pd(dppf)C12
(300
mg) in dioxane (100 mL) using a procedure similar to that described for
compound 0602-
281 (Example 129). LCMS: 266.2[M+1]+.'HNMR (400 MHz, CDC13): 6 1.44 (s, 12H),
6.80 (d, J = 8.8 Hz, 1 H), 6.84 (s, 1 H), 8.04 (d, J = 8.8 Hz, 1 H).
Step 137b: Ethyl 2-(((2-(3-hydroxy-4-nitrophenyl)-4-morpholinothieno[3,2-d]
pyrimidin -
6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-293)
The title compound 0603-293 was prepared as a yellow solid (100 mg, 27%) from
0602-293 (350 mg, 1.337 mmol), 0504 (300 g, 0.668 mmol), sat. NaHCO3 (4 mL)
and
Pd(PPh3)4 (100 mg) in dioxane (12 mL) using a procedure similar to that
described for
compound 1103-226 (Example 99): LCMS: 552.3[M+1]+. 'HNMR: (400 MHz, DMSO-d6):
6 1.36 (t, J= 6.8 Hz, 3H), 3.76 (s, 4H), 3.86 (s, 4H), 4.35 (d, J= 6.4 Hz,
2H), 5.30 (s, 2H),
7.00 (d, J= 7.6 Hz, 1H), 7.33 (s, 1H), 7.52 (s, 1H), 7.85 (d, J= 8.8 Hz, 1H),
8.94 (s, 2H).
Step 137c: N-Hydroxy-2-(((2-(3-hydroxy-4-nitrophenyl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 293)
The title compound 293 was prepared as a yellow solid (40 mg, 30%) from 0603-
293 (130 mg) and a freshly prepared hydroxylamine methanol solution (3 mL,
1.79 M in
methanol) using a procedure similar to that described for compound 237
(Example 107).
m.p.: 258-261 C. LCMS: 559[M+1]+.'HNMR: (400 MHz, DMSO-d6): 6 3.23 (s, 3H),
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3.69(s, 4H), 3.79 (s, 4H), 5.20 (s, 2H), 6.90 (s, I H), 7.20 (s, I H), 7.44
(s, I H), 7.79 (s, I H),
8.74 (s, 2H).
EXAMPLE 138: N-Hydroxy-2-(((2-(3-(1-hydroxyethyl)phenyl)-4-morpho-
linothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide
(Compound 294)
Step 138a: Ethyl2-(((2-(3-(1-hydroxyethyl)phenyl)-4-morpholinothieno[3,2-d]
pyrimidin-
6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-294)
1-(3-bromophenyl) ethanone (1.00 g, 5.02 mmol), 4,4,4',4',5,5,5',5'-
octamethyl-
2,2'-bi(1,3,2-dioxaborolane) (2.55 g, 10.05 mmol), KOAc ( 1.23 g, 12.55 mmol),
PdC12(dppf) (100 mg) were taken into anhydrous 1,4-dioxane (lOmL) and heated
to 100 C
for 3h. After cooling to room temperature the reaction mixture was filtered
and
concentrated. The crude product was isolated as brown solid (1.2 g) and was
used for the
next step reaction without further purification.
A mixture of 0602-294 (1.2 g, prepared above), 0504 (563 mg, 1.26 mmol),
NaHCO3 (318 mg, 3.78 mmol), PdC12(Ph3P)2 (55 mg), in toluene/ethanol/water(4
mL/2
mL/l mL) was stirred at 130 C overnight. The reaction mixture was filtered
and
concentrated. The crude product was purified by column chromatography to
afford ethyl 2-
(((2-(3-acetylphenyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl) methyl)
(methyl)amino)pyrimidine-5-carboxylate as an off-white solid (350 mg, 52%).
LCMS
533.3 [M+1]+.'HNMR (400 MHz, DMSO-d6): 6 1.31 (t, J= 7.2 Hz, 3H), 2.66 (s,
3H), 3.30
(s, 3H), 3.76-3.80 (m, 4H), 3.92-3.97 (m, 4H), 4.29 (q, J= 7.2 Hz, 2H), 4.79-
4.84 (m, 1H),
5.24 (s, 2H), 7.40-7.49 (m, 2H), 7.49 (s, 1 H), 8.07 (dd, J = 6.4 Hz, 1.6 Hz,
1 H), 8.63 (d, J =
7.6 Hz, 1H), 8.88 (s, 2H), 8.90 (s, 1H).
A mixture of ethyl 2-(((2-(3-acetylphenyl)-4-morpholinothieno[3,2-d]pyrimidin-
6-
yl) methyl) (methyl)amino)pyrimidine-5-carboxylate (350 mg, 0.66 mmol), NaBH4
(124
mg, 3.29 mmol) in anhydrous THE was stirred at room temperature for 0.5 h. The
mixture
was quenched with water and extracted with dichloromethane. The combined
organic
layers were washed brine and dried over MgSO4. Crude product 0603-294 was
obtained as
a white solid (200 mg, 57%) after filtration and was used directly for next
step reaction
without further purification. LCMS: 535.4 [M+l+.'HNMR (400 MHz, DMSO-d6): 6
1.31
(t, J= 7.2 Hz, 3H), 1.38 (d, J= 6.4Hz, 3H), 3.28 (s, 3H), 3.76-3.80 (m, 4H),
3.92-3.97 (m,
4H), 4.29 (q, J= 7.2 Hz, 2H), 4.79-4.84 (m, 1H), 5.24 (s, 2H), 7.40-7.49 (m,
2H), 7.49 (s,
1H), 8.23-8.26 (m, 1H), 8.39 (s, 1H), 8.88 (s, 2H).
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Step 138b: N-Hydroxy-2-(((2-(3-(1-hydroxyethyl)phenyl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 294)
The title compound 294 was prepared as a light yellow solid (70 mg, 36% yield)
from 0603-294 (200mg, 0.37mmol) and a freshly prepared hydroxylamine methanol
solution (8 mL, 1.79 M in methanol) in dichloromethane (2mL) using a procedure
similar
to that described for compound 237 (Example 107). m.p.:168-171 C. LCMS: 522.0
[M+1]. 'H-NMR, (400 MHz, DMSO-d6): 6 (ppm) 1.37 (d, J= 6.0 Hz, 3H), 3.24 (s,
3H),
3.77 (s, 4H), 3.93 (s, 4H,), 4.81 (br, 1H), 5.20 (s, 3H),7.39-7.47 (3H, m),
8.25 (d, J= 6.0
Hz, I H), 8.38 (s, I H), 8.75 (s, 2H), 9.02 (s, I H), 11.09 (s, I H).
EXAMPLE 139: 2-(((2-(2-Fluoro-5-(1-hydroxyethyl)phenyl)-4-morpholinothieno
[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound 295)
Step 139a: Ethyl 2-(((2-(5-acetyl-2-fluorophenyl)-4-morpholinothieno[3,2-d]
pyrimidin- 6-
yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-295)
The title compound 0603-295 was prepared as a light yellow solid (500 mg, 79%)
from 1-(3-bromo-4-fluorophenyl)ethanone (500 mg, 2.3 mmol),
4,4,4',4',5,5,5',5'-
octamethyl-2,2'-bi(1,3,2-dioxaborolane (1.17 g, 4.6 mmol), KOAc (1.35 g, 6.9
mmol),
Pd(dppf)C12 (84 mg, 0.115 mmol) inl,4-dioxane (10 mL) followed by 0504 (516
mg, 1.15
mmol), NaHCO3(290 mg, 3.45 mmol), bis(triphenylphosphine)palladium(II)chloride
(40
mg, 0.06 mmol) in ethanol (5 mL) , water (10 mL), toluene (2.5 mL) using a
procedure
similar to that described for compound 0603-275 (Example 124).
Step 139b: 2-(((2-(2-Fluoro-5-(1-hydroxyethyl)phenyl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)methyl)(methyl)amino)-N-hydroxypyrimidine-5-carboxamide
(Compound
295)
A mixture of 0603-295 (180 mg, 0.33 mmol), NaBH4 (110 mg, 1.3 mmol), and THE
(10 mL) was stirred at room temperature for 2h. To the reaction was added
water and
extracted with dichloromethane. The combined organic layers were washed with
brine and
dried with MgSO4. The crude product was obtained as an off-white solid (180
mg) after
comncentration and was used for the next step reaction without further
purification.
The hydroxyethyl intermediate (160 mg, 0.29 mmol) was taken into NH2OH
methanol solution (20 mL, 1.79M in methanol) and stirred for 1h. The reaction
mixture was
adjusted pH to 6 with acetic acid followed by concentration. The residue was
triturated with
water and filtered. The crude product was purified by prep-HPLC to afford the
titled
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compound 295 as a white solid (45 mg, 29%). m.p.:230-235 C. LCMS: 540.0
[M+1]+.
IHNMR (400 MHz, DMSO-d6): 6 1.34 (d, J= 6.4 Hz, 3H), 3.23 (s, 3H), 3.72-3.75
(m, 4H),
3.87-3.90 (m, 4H), 4.78 (q, J = 6.4 Hz, 1 H), 5.21 (s, 2H), 7.21 (dd, J = 10.8
Hz, 8.4 Hz,1 H),
7.43 (d, J = 1.2 Hz, 1 H), 7.48 (s, 1 H), 7.96 (dd, J = 7.6 Hz, 2.4 Hz, 1 H),
8.74 (s, 2H), 9.15
(br,1 H),11.13 (s,1 H).
EXAMPLE 140: (S)-N-Hydroxy-2-(((2-(3-(1-hydroxyethyl)phenyl)-4-morpho-
linothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide
(Compound 296)
Step 140a: (S)-Ethyl 2-(((2-(3-(1-hydroxyethyl)phenyl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-
296)
The title compound 0603-296 was prepared as an off-white solid (400 mg, 82%)
from (S)-1-(3-bromophenyl)ethanol (500 mg, 2.49 mmol), 4,4,4',4',5,5,5',5'-
octamethyl-
2,2'-bi(1,3,2-dioxaborolane) (1.26 g, 4.98 mmol), KOAc (6.1 g, 6.22 mmol),
PdC12(dppf)
(50 mg) in DMSO (10 mL) followed by 0504 (400 mg, 0.89 mmol), NaHCO3(150 mg,
1.78 mmol), PdC12(Ph3P)2 (40 mg) in toluene /ethanol/water (12 mL/6 mL/3 mL)
using a
procedure similar to that described for compound 0603-275 (Example 124). LCMS:
535.0
[M+1]+. 'H-NMR, (400 MHz, DMSO-d6): 6(ppm) 1.31 (t, J= 6.8 Hz, 3H), 1.37 (d,
J= 6.4
Hz, 3H), 3.28 (s, 3H), 3.76-3.78 (m, 4H), 3.92-3.95 (m, 4H), 4.28 (q, J= 6.8
Hz, 2H), 4.78-
4.84(m, 1H), 5.17 (d, J= 4.4 Hz, 1H), 5.24 (s, 2H), 5.24 (1H, s), 7.39-7.44
(m, 2H), 7.45 (s,
I H), 8.22-8.26 (m,1 H), 8.39 (s, I H), 8.90 (s, 2H).
Step 140b: (S)-N-Hydroxy-2-(((2-(3-(1-hydroxyethyl)phenyl)-4-morpholinothieno
[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 296)
The title compound 296 was prepared as a white solid (110 mg, 57%) from 0603-
296 (200 mg, 0.37 mmol) and a freshly prepared hydroxylamine methanol solution
(10 mL,
1.79 M in methanol) in dichloromethane (4 mL) using a procedure similar to
that described
for compound 237 (Example 107). m.p.:168-172 C. LCMS: 522.0 [M+1]. 'H-NMR,
(400
MHz, DMSO-d6): 6(ppm) 1.37 (d, J= 6.4 Hz, 3H), 3.22 (s, 3H), 3.77 (m, 4H),
3.93 (m,
4H), 4.81 (br, 1 H), 5.20 (s, 2H), 5.24 (1 H, s), 7.41-7.48 (m, 3H), 8.25 (d,
J = 6.4 Hz, 1 H),
8.39 (s,1H), 8.75 (s, 2H).
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EXAMPLE 141: (R)-N-Hydroxy-2-(((2-(3-(1-hydroxyethyl)phenyl)-4-morpho-
linothieno [3,2-d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-
carboxamide
(Compound 297)
Step 141a: (R)-Ethyl 2-(((2-(3-(1-hydroxyethyl)phenyl)-4-morpholinothieno[3,2-
d]
pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxylate (Compound 0603-
297)
The title compound 0603-297 was prepared as an off-white solid (350 mg, 74%)
from (R)-1-(3-bromophenyl)ethanol (500 mg, 2.49 mmol), 4,4,4',4',5,5,5',5'-
octamethyl-
2,2'-bi(1,3,2-dioxaborolane) (1.26 g, 4.98 mmol), KOAc (6.10 g, 6.22 mmol),
PdC12(dppf)
(50 mg) in DMSO (10 mL) followed by 0504 (400 mg, 0.89 mmol), NaHCO3 (150 mg,
1.78 mmol), PdC12(Ph3P)2 (40 mg) in toluene/ ethanol/water (12 mL/6 mL/3 mL)
using a
procedure similar to that described for compound 0603-275 (Example 124). LCMS:
535.0
[M+1]+. 'H-NMR (400 MHz, DMSO-d6): 6 1.30 (t, J= 7.2 Hz, 3H), 1.37 (d, J= 6.4
Hz,
3H), 3.28 (s, 3H), 3.75-3.78 (m, 4H), 3.92-3.95 (m, 4H), 4.28 (q, J= 7.2 Hz,
2H), 4.78-4.84
(m, 1 H), 5.17 (d, J = 4.4 Hz, 1 H), 5.24 (s, 2H), 5.24 (1 H, s), 7.3 9-7.44
(m, 2H), 7.45 (s,
I H), 8.22-8.26 (m,1 H), 8.39 (s, I H), 8.90 (s, 2H).
Step 141b: (R)-N-Hydroxy-2-(((2-(3-(1-hydroxyethyl)phenyl)-4-morpholinothieno
[3,2-
d]pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 297)
The title compound 297 was prepared as a white solid (130 mg, 67%) from 0603-
297 (200 mg, 0.37 mmol) and a freshly prepared hydroxylamine methanol solution
(10 mL,
1.79 M in methanol) in dichloromethane (4 mL) using a procedure similar to
that described
for compound 237 (Example 107). m.p.:169-173 C. LCMS: 522.0 [M+l]. 'H-NMR (400
MHz, DMSO-d6): 6 1.37 (d, J= 6.4 Hz, 3H), 3.25 (s, 3H), 3.77 (m, 4H), 3.93 (m,
4H),
4.80-4.84 (m, 1H), 5.21 (s, 2H), 5.22 (1H, s), 7.40-7.48 (m, 3H), 8.25 (d, J=
6.8 Hz, 1H),
8.3 9(s,1 H), 8.75(s, 2H), 9.05 (br, I H), 11.14 (br, I H)
EXAMPLE 142: 2-(4-Aminophenyl)-N-((l-(5-(hydroxycarbamoyl)pyrimidin-2-yl)
piperidin-4-yl)methyl)-4-morpholinothieno [3,2-d] pyrimidine-6-carboxamide
(Compound 307)
Step 142a: 2-Chloro-4-morpholinothieno[3,2-d]pyrimidine-6-carboxylic acid
(Compound
1401)
0112 (5 g, 17.67 mmol) was suspended in dichloromethane (100 mL) followed the
addition of mCPBA (75%, 4g, 22.97 mmol). The resulting mixture was heated to
reflux for
24 h. Upon completion, the mixture was cooled to room temperature and
concentrated to 1/2
volume. The mixture was filtered to afford a crude yellow solid which was re-
crystallized
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from Hexanes/Ethyl acetate = 1/1 to give the titled compound as a light yellow
solid (3.2 g,
61%).'HNMR (400 MHz, DMSO-d6): 6 3.74-3.3.78 (m, 4H), 3.92-3.95 (m, 4H), 7.86
(s,
I H).
Step 142b: Ethyl 2-(4-((2-chloro-4-morpholinothieno[3,2-d]pyrimidine-6-
carboxamido)
methyl)piperidin-1-yl)pyrimidine-5-carboxylate (Compound 1402)
To a suspension of 1401 (400 mg, 1.34 mmol), 0401 (384 mg, 1.47 mmol) and
TBTU (644 mg, 2.07 mmol) in dichloromethane (80 mL) was added DIPEA (518 mg,
4.02
mmol) at 0 C. The resulting mixture was stirred at room temperature for 1 h.
The reaction
mixture was diluted with dichloromethane and washed with water. The crude
product was
triturated with water and filtered to afford titled compound as a white solid
(473 mg, 65%).
1HNMR (400 MHz, DMSO-d6): 6 1.15-1.43 (m, 7H), 1.78-1.82 (m, 2H), 1.82-1.84
(m,
1H), 2.98-3.15 (m, 2H), 3.77 (br, 4H); 3.93 (br, 4H), 4.25-4.28 (m, 2H), 4.75-
4.78 (m, 2H),
8.02 (s, 1H), 8.78 (s, 2H), 9.02-9.06 (m, 1H).
Step 142c: Ethyl 2-(4-((2-(4-aminophenyl)-4-morpholinothieno[3,2-d]pyrimidine-
6-
carboxamido)methyl)piperidin-l-yl)pyrimidine-5-carboxylate.(Compound 1403-307)
1402 (300 mg, 0.55 mmol), 4-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)
aniline
(180 mg, 0.82 mmol) and Pd(PPh3)2C12 (12 mg) were taken into toluene (6 mL),
ethanol
(3.6 mL) and water (1.2 mL) and stirred at 120 C for 2h under N2 atmosphere.
Upon
completion the reaction was cooled with ice-water bath, followed by the
addition of water
(50 mL). After stirring for lh, the solid was collected by filtration and
purified by column
chromatography to afford titled compound as a white solid (200 mg, 60%). 'HNMR
(400
MHz, DMSO-d6): 6 1.10-1.40 (m, 7H), 1.80-2.00 (m, 3H), 2.98-3.10 (m, 2H), 3.21-
3.25
(m, 2H), 3.80 (br, 4H); 3.99 (br, 4H), 4.26 (t, J= 6.8 Hz, 2H), 4.75-4.79 (m,
2H), 5.55 (s,
2H), 6.62 (d, J= 8.4 Hz, 2H), 8.06 (s, 1H), 8.10 (d, J = 8.8 Hz, 2H), 8.77 (s,
2H), 8.95 (br,
1H).
Step 142d: 2-(4-Aminophenyl)-N-((1-(5-(hydroxycarbamoyl)pyrimidin-2-
yl)piperidin- 4-
yl) methyl)-4-morpholinothieno[3,2-d]pyrimidine-6-carboxamide (Compound 307)
1403-307 (190 mg, 0.32 mmol) was taken into freshly prepared NH2OH methanol
solution (20 mL, 1.79 M in methanol) and dichloromethane (3 mL). The resulting
mixture
was sealed and stirred at room temperature for 4h. Upon completion, the
reaction solution
was adjusted to pH 7-8 with HC1 aqueous solution (1.2 M). Water was added
followed by
filtration to afford product as a yellow solid (130 mg, 70%). m.p:232-238 C.
LCMS:590.0[M+1]+. 'HNMR (400 MHz, DMSO-d6): 81.09 (m, 2H),1.79 (d, J= 11.6 Hz,
2H), 1.93 (m, 1H), 2.93-2.99 (m, 2H), 3.21-3.31 (m, 2H), 3.80 (br, 4H), 3.96
(br, 4H), 4.72
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(d, J= 13.2 Hz, 2H), 5.55 (s, 2H), 6.62 (d, J= 8.4 Hz, 2H), 8.07-8.12 (m, 3H),
8.66 (s, 2H),
8.95 (d, J= 10.0 Hz, 2H), 11.03(s,1H).
EXAMPLE 143: N-((1-(5-(Hydroxycarbamoyl)pyrimidin-2-yl)piperidin-4-yl) methyl)-
2-(6-(methylamino)pyridin-3-yl)-4-morpholinothieno[3,2-d]pyrimidine-6-
carboxamide
(Compound 308)
Step 143a: Ethyl 2-(4-((2-(6-(methylamino)pyridin-3-yl)-4-morpholinothieno[3,2-
d]
pyrimidine-6-carboxamido)methyl)piperidin-1-yl)pyrimidine-5-carboxylate
(Compound
1403-308)
The title compound 0403-308 was prepared as a yellow solid (110 mg, 41%) from
1402 (240 mg, 0.44 mmol), N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
yl)pyridin-2-amine (154 mg, 0.66 mmol), NaHCO3(l 11 mg, 1.3 mmol) and
Pd(PPh3)2C12
(9.6 mg) in toluene (8 mL), ethanol (4.8 mL) and water (1.6 mL) using a
procedure similar
to that described for compound 1403-307 (Example 142). 'HNMR (400 MHz, DMSO-
d6) :
6 1.16-1.20 (m, 2H), 1.29 (t, J= 7.2 Hz, 3H), 1.78-1.82 (m, 2H), 1.95-2.00 (m,
1H), 2.84
(d, J= 4.4 Hz, 3H), 3.01 (t, J= 11.6 Hz, 2H), 3.21-3.24 (m, 2H), 3.79 (t, J=
5.2 Hz, 4H),
3.98 (t, J= 4.8 Hz, 4H), 4.26 (q, J= 6.8 Hz, 2H), 4.76 (d, J= 12.6 Hz, 2H),
6.52 (d, J= 8.8
Hz, 2H), 6.91 (d, J = 4.8 Hz, 1 H), 8.11 (s, 1 H), 8.27 (dd, J = 8.8 Hz, 2 Hz,
1 H), 8.76 (s,
2H), 8.91 (t, J = 4.2 Hz, 1 H), 9.04 (s, 1 H).
Step 143b: N-((1-(5-(Hydroxycarbamoyl)pyrimidin-2-yl)piperidin-4-yl)methyl)-2-
(6-
(methylamino)pyridin-3-yl)-4-morpholinothieno[3,2-d]pyrimidine-6-carboxamide
(Compound 308)
The title compound 308 was prepared as a white solid (55 mg, 51%) from 1403-
308
(110 mg, 0.18 mmol) and a freshly prepared hydroxylamine methanol solution (20
mL,
1.79 M in methanol) in dichloromethane (3 mL) using a procedure similar to
that described
for compound 307 (Example 142). m.p:215-218 C. LCMS:605.0[M+1]+.
IHNMR (400 MHz, DMSO-d6): 6 1.16-1.23 (m, 2H), 1.78-1.82 (m, 2H), 1.90-1.94
(m,
1H), 2.84 (d, J= 4.8 Hz, 3H), 2.93 (t, J= 4.8 Hz, 2H), 3.21-3.24 (m, 2H), 3.80
(t, J= 5.2
Hz, 4H), 3.98 (t, J = 4.8Hz, 4H), 4.72 (d, J = 13.2 Hz, 2H), 6.51 (d, J = 8.8
Hz, 2H), 6.96
(d, J = 4.8 Hz, 1 H), 8.11 (s, 1 H), 8.27 (dd, J = 8.8 Hz, 2 Hz, 1 H), 8.66
(s, 2H), 8.96 (d, J =
13.2 Hz, 2H), 9.04 (s, I H), 11.05 (s, I H).
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EXAMPLE 144: N-Hydroxy-2-(4-((2-(6-(methylamino)pyridin-3-yl)-4-morpho-
linothieno [3,2-d] pyrimidine-6-sulfonamido)methyl)piperidin-1-yl)pyrimidine-5-
carboxamide (Compound 310)
Step 144a: Ethyl 2-(4-((2-(6-(methylamino)pyridin-3-yl)-4-morpholinothieno[3,2-
d]
pyrimidine-6-sulfonamido)methyl)piperidin-1-yl)pyrimidine-5-carboxylate
(Compound
1405-310)
The title compound 0405-3 10 was prepared as a yellow solid (350 mg, 78%) from
1404 (400 mg, 0.704 mmol), N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-
2-
yl)pyridin-2-amine (247 mg, 1.06 mmol), NaHCO3 (177 mg, 2.11 mmol) and
(PPh3)4Pd (40
mg, 0.035 mmol) were taken into toluene (8 mL), ethanol (16 mL) and water (4
mL) using
a procedure similar to that described for compound 1403-307 (Example 142).
LCMS: 654.0
[M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 1.071.13 (m, 2H),1.28 (t, J= 7.2 Hz, 3H),
1.761.79 (m, 3H), 2.78-3.00 (m, 7H), 3.80 (t, J= 4.4 Hz, 4H), 3.97 (t, J= 4.4
Hz, 4H),
4.25 (q, J = 6.8 Hz, 2H), 4.72 (d, J = 12.4 Hz, 2H), 6.52 (d, J = 8.8 Hz, 1
H), 6.97 (d, J = 4.4
Hz, 1 H), 7.60 (d, J = 6.4 Hz, 1 H), 7.82 (s, 1 H), 8.29 (d, J = 4.4 Hz, 1 H),
8.34 (s, 1 H), 8.75
(s, 2H), 9.04 (s, 1H).
Step 144b: N-Hydroxy-2-(4-((2-(6-(methylamino)pyridin-3-yl)-4-morpholinothieno
[3,2-
d]pyrimidine-6-sulfonamido)methyl)piperidin-1-yl)pyrimidine-5-carboxamide
(Compound
310)
The title compound 310 was prepared as a white solid (183 mg, 94%) from 1405-
310 (200 mg, 0.306 mmol) and a freshly prepared hydroxylamine methanol
solution (25
mL, 1.79 M in methanol) in dichloromethane (3 mL) using a procedure similar to
that
described for compound 307 (Example 142). M.p.:190-197 C. LCMS: 641.0 [M+1]+.
iH NMR (400 MHz, DMSO-d6): 6 1.021.11 (m, 2H), 1.74-1.77 (m, 3H), 2.83-2.94
(m,
7H), 3.79 (t, J= 4.4 Hz, 4H), 3.97 (t, J= 4.4 Hz, 4H), 4.67 (d, J= 12.8 Hz,
2H), 6.52 (d, J
= 8.8 Hz, 1 H), 6.99 (d, J = 4.8 Hz, 1 H),7.82 (s, 1 H), 8.28 (dd, J = 8.8 Hz,
2 Hz, 1 H), 8.63
(s, 2H), 9.03 (d, J= 2.4 Hz, 1H).
EXAMPLE 145: N-Hydroxy-2-(methyl((7-morpholino-5-(2-(trifluoromethyl)-1H-
benzo [d] imidazol-5-yl)benzo [b] thiophen-2-yl)methyl)amino)pyrimidine-5-
carboxamide (Compound 318)
Step 145a: 5-Bromo-2-(trifluoromethyl)-1H-benzo[d]imidazole (Compound 0601-
318)
A mixture of 4-bromobenzene-1, 2-diamine (3.0 g, 16.0 mmol), TFA (9.5 mL,
128.3 mmol) and aq. HC1(3 M, 32 mL) was heated to reflux overnight. The
reaction
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mixture was concentrated and purified by column chromatography to afford the
titled
compound (2.60 g, 60%). LCMS: 265.0[M+1]+. 'HNMR (400 MHz, DMSO-d6): 6 7.52
(dd, J = 8.8 Hz, 1.2 Hz, 1 H), 7.69 (d, J = 8.4 Hz, 1 H), 7.95 (s, 1 H), 14.20
(br, 1 H).
Step 145b: Ethyl2-(methyl((7-morpholino-5-(2-(trifluoromethyl)-1H-benzo[d]
imidazol-
5-yl)benzo[b]thiophen-2-yl)methyl)amino)pyrimidine-5-carboxylate (Compound
0603-
318)
A mixture of 0601-318 (100 mg, 0.38 mmol), 4,4,4',4',5,5,5',5'-octamethyl -
2,2'-
bi(1,3,2-dioxaborolane)( 107 mg, 0.42 mmol), PdC12(dppf) (10 mg), KOAc (74 mg,
0.76
mmol) in anhydrous 1,4-dioxane (2 mL) was stirred at 100 C overnight. The
reaction
mixture was filtered and concentrated. The crude product was obtained as a
brown solid
(150 mg) which was used for next step without further purification.
A mixture of above crude boronate, 0504 (119 mg, 0.27 mmol), PdC12(Ph3P)2 (10
mg), NaHCO3 (68 mg, 0.81 mmol) were taken into toluene/ethanol/water (4 mL/2
mL /1
mL) and stirred at 130 C for 3 h. The reaction mixture was filtered and
concentrated. The
residue was purified by column chromatography to afford titled compound as an
off-white
solid (129 mg, 80%). 'HNMR, (400 MHz, DMSO-d6): 6 1.30 (t, J= 6.8 Hz, 3H),
3.28 (s,
3H), 3.77-3.81 (m, 4H), 3.95-3.97 (m, 4H), 4.28 (q, J= 6.4 Hz, 2H), 5.24 (s,
2H), 7.50 (s,
1H), 7.75 (br, 1H), 8.48 (br, 1H), 8.48 (d, J= 7.6 Hz, 1H), 8.73 (s, 1H), 8.87
(s, 2H).
Step 145c: N-Hydroxy-2-(methyl((7-morpholino-5-(2-(trifluoromethyl)-1H-
benzo[d]
imidazol-5-yl)benzo[b]thiophen-2-yl)methyl)amino)pyrimidine-5-carboxamide
(Compound
318)
The title compound 318 was prepared as an off-white solid (15 mg, 15%) from
0603-318 (100 mg, 0.167mmol) and a freshly prepared hydroxylamine methanol
solution
(2 mL, 1.79 M in methanol) using a procedure similar to that described for
compound 307
(Example 142). m.p.>300 C. LCMS: 586 [M+1]+. 'HNMR (400 MHz, DMSO-d6): 6 3.25
(s, 3H), 3.77-3.80 (m, 4H), 3.95-3.97 (m, 4H), 5.22 (s, 2H), 7.50 (s, 1H),
7.70 (d, J= 8.4
Hz, 0.5H), 7.88 (d, J= 8.4 Hz, 0.5H), 8.44 (d, J= 8.4 Hz, 0.5H), 8.52 (d, J=
8.4 Hz, 0.5H),
8.65 (s, 0.5H), 8.81 (s, 0.5H), 8.76 (s, 2H), 9.02 (s, 1H), 11.09 (s, 1H),
14.03 (s, 1H).
EXAMPLE 146: N-Hydroxy-5-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholinothieno[3,2-d]pyrimidin-2-yl)phenoxy)pentanamide (Compound 319)
Step 146a: 5-(4-Bromo-phenoxy)-pentanoic acid ethyl ester(Compound 1502-319)
A mixture of 4-Bromo-phenol (12 g, 68.0 mmol), Cs2CO3 (44.0 g, 0.136 mmol), 5-
bromo-pentanoic acid ethyl ester (16.9 g, 82.0 mmol) in DMF was heated to 80
C for 4h.
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The reaction mixture was diluted with water and extracted ethyl acetate. The
combined
organic layers were washed with water, brine and dried with Na2SO4. The crude
product
was purified by column chromatography to afford the titled compound 1502-319
as an oil
(16.0 g). 'HNMR (400 MHz, CDC13): 6 1.25 (t, J= 7.2 Hz, 3H); 1.81 (m, 4H);
2.35-2.39
(m, 2H), 3.93 (t, J = 6.0 Hz, 2H), 4.13 (q, J = 7.2 Hz, 2H), 6.76 (d, J = 8.8
Hz, 2H), 7.35 (d,
J = 8.8 Hz, 2H).
Step 146b: Ethyl 5-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)
pentanoate
(Compound 1503-319)
A mixture of 1502-319 (5.0 g, 0.0167 mmol), bis(pinacolato)diboron (6.4 g,
0.025
mol), PdC12(dppf)2 (610 mg, 0.835 mmol), KOAc (5 g, 0.05 mol) in anhydrous
dioxane
was heated to reflux overnight. The reaction mixture was diluted with
EtOAc/hexanes and
stirred for 30 min. It was filtered to remove solid and concentrated to afford
the titled
compound 1503-319 as a brown solid (3.5 g, 60%). 'HNMR: (400 MHz, CDC13): 6
1.28 (t,
J= 7.2 Hz, 3H), 1.33 (s, 12H), 1.82 (br, 4H), 2.37 (m, 2H), 3.99 (m, 2H), 4.13
(q, J= 7.2
Hz, 2H), 6.87 (d, J= 8.8 Hz, 2H), 7.73 (d, J= 8.4 Hz, 2H).
Step 146c: Ethyl 5-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholino
thieno[3,2-d]pyrimidin-2-yl)phenoxy)pentanoate (Compound 1504-319)
A mixture of 1503-319 (35 0mg, 1 mmol), 0113 (300 mg, 0.486 mmol), NaHCO3
(126 mg, 1.5 mmol) and bis(triphenylphosphine)palladium(II) chloride (40 mg,
0.057
mmol) in toluene (12 ml), ethanol (8.0 ml) and water (3.0 ml) was heated at
108 C for 4 h
under N2 atmosphere. The reaction mixture was partitioned between water and
EtOAc and
separated. The crude product was purified by column chromatography to afford
the title
compound as a light yellow solid (180 mg, 60%). m.p.: 110-114 C. LCMS : 618.4
[M+1]+.
1H NMR: (400 MHz, CDC13): 61.26 (t, J= 7.2 Hz, 3H), 1.85 (m, 4H), 2.39 (t, J=
6.8 Hz,
2H), 2.67 (t, J= 4.8 Hz, 4H), 2.80 (s, 3H), 3.29 (t, J= 4.8 Hz, 4H), 3.86-3.90
(m, 6H),
4.03-4.05 (m, 6H), 4.14 (q, J = 6.8 Hz, 2H), 6.95 (d, J = 8.8 Hz, 2H), 7.31(s,
I H), 8.37 (d, J
= 8.8 Hz, 2H).
Step 146d: N-Hydroxy-5-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-mor-
pholinothieno[3,2-d]pyrimidin-2-yl)phenoxy)pentanamide (Compound 319)
The title compound 319 was prepared as a white solid (100 mg, 66%) from 1504-
319 (150 mg, 0.243 mmol) and a freshly prepared hydroxylamine methanol
solution (30
mL, 1.79 M in methanol) using a procedure similar to that described for
compound 307
(Example 142). m.p. 140-144 C. LC-MS: 605.2[M+1]+. 1H NMR: (400 MHz, DMSO-
d6):
6 1.66-1.72 (br, 4H), 2.01-2.04 (m, 2H), 2.59 (m, 4H), 2.90 (s, 3H), 3.16 (d,
J= 3.6 Hz,
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4H); 3.80 (t, J= 4.0 Hz, 4H), 3.95-4.05 (m, 8H), 7.01 (d, J= 8.8 Hz, 2H), 7.38
(s, 1H), 8.33
(d, J = 8.8 Hz, 2H), 10.39 (br, 1 H).
EXAMPLE 147: N-Hydroxy-6-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)- 4-
morpholino thieno[3,2-d]pyrimidin-2-yl)phenoxy)hexanamide (Compound 320)
Step 147a: Ethyl 6-(4-bromophenoxy)hexanoate (Compound 1502-320)
The title compound 1502-320 was prepared as an oil (4.0 g. 74%) from 4-Bromo-
phenol (3.0 g, 17.34 mmol), Cs2CO3 (11.3 g, 34.68 mmol), ethyl 6-
bromohexanoate (16.9
g, 82 mmol) in DMF using a procedure similar to that described for compound
1502-319
(Example 146). 'HNMR (400 MHz, CDC13): 6 1.25 (t, J= 7.2 Hz, 3H), 1.54 (q, J=
8.4
Hz, 2H), 1.66-1.73 (m, 2H); 1.74-1.82 (m, 2H), 2.32 (t, J= 7.6 Hz, 2H), 3.91
(t, J= 6.4 Hz,
2H), 4.13 (q, J= 7.2 Hz, 2H), 6.75 (d, J= 9.2 Hz, 2H), 7.35 (d, J= 8.8 Hz,
2H).
Step 147b: Ethyl 6-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)
hexanoate
(Compound 1503-320)
The title compound 1503-320 was prepared as a brown oil (1.5 g, 67%) from 1502-
320 (2.0 g, 6.35 mmol), bis(pinacolato)diboron (2.42 g, 9.5 mmol),
PdC12(dppf)2 (300 mg,
0.41 mmol), KOAc(1.86 g,19.0 mmol) in anhydrous dioxane using a procedure
similar to
that described for compound 1503-319 (Example 146). 'HNMR: (400 MHz, CDC13): 6
1.25
(t, J= 7.2 Hz, 3H), 1.33 (s, 12H), 1.46-1.54 (m, 2H), 1.63-1.73 (m, 2H), 1.77-
1.84 (m, 2H),
2.32 (t, J= 7.2 Hz, 2H), 3.98 (t, J= 6.4Hz, 2H); 4.13 (q, J= 7.2 Hz, 2H), 6.87
(d, J= 8.8
Hz, 2H), 7.73 (d, J= 8.8 Hz, 2H).
Step 147c: Ethyl 6-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholino
thieno[3,2-d]pyrimidin-2-yl)phenoxy)hexanoate (Compound 1504-320)
The title compound 1504-320 was prepared as an off-white solid (185 mg, 50%)
from 1503-320 (420 mg, 1.16 mmol), 0113(250 mg, 0.58 mmol), NaHCO3 (150 mg,
1.74
mmol) and bis(triphenylphosphine)palladium(II) chloride (50 mg, 0.071 mmol) in
toluene
(10 mL), ethanol ( 8 mL) and water (3 mL) using a procedure similar to that
described for
compound 1504-319 (Example 146). m.p.: 125-129 C. LCMS: 632.4 [M+1]+. 'HNMR:
(400 MHz, CDC13): 6 1.26 (t, J= 7.2 Hz, 3H), 1.50-1.57 (br, 2H), 1.68-1.74 (m,
2H), 1.80-
1.87 (m, 2H), 2.34 (t, J= 7.2 Hz, 2H), 2.67 (m, 4H), 2.80 (s, 3H), 3.29 (m,
4H), 3.82-3.90
(m, 6H), 4.01-4.04 (m, 6H), 4.14 (q, J = 7.2 Hz, 2H), 6.95 (d, J = 8.8 Hz,
2H), 7.31 (s, 1 H),
8.37 (d, J= 8.8 Hz, 2H).
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Step 147d: N-Hydroxy-6-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholino
thieno[3,2-d]pyrimidin-2-yl)phenoxy)hexanamide (Compound 320)
The title compound 320 was prepared as a white solid (70 mg, 36%) from 1504-
320
(200 mg, 0.316 mmol) and a freshly prepared hydroxylamine methanol solution
(30 mL,
1.79 M in methanol) using a procedure similar to that described for compound
307
(Example 142). m.p. 140-142 C. LCMS: 619.2[M+1]+.'HNMR: (400 MHz, DMSO-d6): 6
1.42-1.44 (m, 2H), 1.54-1.58 (m, 2H), 1.72-1.754 (m, 2H), 1.98 (t, J= 7.2 Hz,
2H), 2.59
(br, 4H), 2.90 (s, 3H), 3.15 (br, 4H), 3.79 (t, J= 4.0 Hz, 4H), 3.81-4.04 (m,
8H), 7.01 (d, J
= 8.8 Hz, 2H), 7.37 (s, I H), 8.32 (d, J= 8.8 Hz, 2H), 8.67 (s, I H), 10.35
(s, I H).
EXAMPLE 148: N-Hydroxy-7-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-
morpholinothieno[3,2-d]pyrimidin-2-yl)phenoxy)heptanamide (Compound 321)
Step 148a: Ethyl 7-(4-bromophenoxy)heptanoate (Compound 1502-321)
The title compound 1502-321 was prepared as an oil (1.6 g, 80%) from 4-Bromo-
phenol (1.1 g, 6.5 mmol), Cs2CO3 (4.0 g, 13 mmol), ethyl 7-bromoheptanoate
(1.7 g, 7.2
mmol) in DMF using a procedure similar to that described for compound 1502-319
(Example 146). 'HNMR: (400 MHz, CDC13): 6 1.25 (t, J= 7.2 Hz, 3H), 1.35-1.51
(m, 4H),
1.617-1.69 (m, 2H), 1.75-1.78 (m, 2H), 2.30 (t, J= 7.2 Hz, 2H), 3.90 (t, J=
6.4 Hz, 2H),
4.12 (q, J= 7.2 Hz, 2H), 6.76 (d, J= 8.8 Hz, 2H), 7.35 (d, J= 8.8 Hz, 2H).
Step 148b: Ethyl?-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)
heptanoate
(Compound 1503-321)
The title compound 1503-321 was prepared as an oil (1.7g, 60%) from 1502-321
(1.9g, 5.8 mmol), bis(pinacolato)diboron (2.2g, 8.7 mmol), PdC12(dppf)2(
212mg, 0.29
mmol), KOAc(1.8 g, 17.4 mmol) in anhydrous dioxane using a procedure similar
to that
described for compound 1503-319 (Example 146). 'HNMR: (400MHz, CDC13): 6 1.25
(t, J
= 7.2 Hz, 3H), 1,33 (s, 12H), 1.37-1.43 (m, 2H), 1.46-1.50 (m, 2H), 1.62-1.70
(m, 2H);
1.75-1.80 (m, 2H), 2.30 (d, J= 7.6 Hz, 2H), 3.97 (t, J= 6.8 Hz, 2H), 4.12 (q,
J= 7.2 Hz,
2H), 6.87 (d, J= 8.8 Hz, 2H), 7.73 (d, J= 8.8 Hz, 2H).
Step 148c: N-Hydroxy-7-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-mor-
pholinothieno[3,2-d]pyrimidin-2-yl)phenoxy)heptanamide (Compound 1504-321)
The title compound 1504-321 was prepared as a pale yellow solid (260 mg, 50%)
from 1503-321 (600 mg, 1.60 mmol), 0113 (350 mg, 0.81 mmol), NaHCO3 (200 mg,
2.4
mmol) and bis(triphenylphosphine)palladium(II) chloride (60 mg, 0.085 mmol) in
toluene
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(12 mL), ethanol (9 mL) and water (4 mL) using a procedure similar to that
described for
compound 1504-319 (Example 146). m.p.: 154-157 C. LCMS: 646.4 [M+l]+. 'H NMR:
(400 MHz, CDC13): 6 1.25 (t, J= 7.2 Hz, 3H), 1.47-1.54 (m, 2H), 1.54-1.60 (m,
2H), 1.66-
1.68 (m, 2H), 1.78-1.85 (m, 2H), 2.31 (d, J= 7.6 Hz, 2H), 2.67 (m, 4H), 2.79
(s, 3H), 3.29
(m, 4H), 3.85-3.89 (m, 6H), 4.00-4.03 (m, 6H), 4.13 (q, J= 7.2 Hz, 2H), 6.96
(d, J= 8.8
Hz, 2H), 7.31 (s, 1H), 8.37 (d, J= 8.8 Hz, 2H).
Step 148d: N-Hydroxy-7-(4-(6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-mor-
pholinothieno[3,2-d]pyrimidin-2-yl)phenoxy)heptanamide (Compound 321)
The title compound 321 was prepared as a white solid (130 mg, 54.5%) from 1504-
321 (220 mg, 0.34 mmol) and a freshly prepared hydroxylamine methanol solution
(30 mL,
1.79 M in methanol) using a procedure similar to that described for compound
307
(Example 142). m.p. 143-145 C. LC-MS: 633.2[M+1]+. 'HNMR: (400 MHz, DMSO-d6):
6
1.30-1.56 (m, 6H), 1.73 (t, J= 6.4 Hz, 2H), 1.96 (t, J= 7.6 Hz, 2H), 2.59 (br,
4H), 2.89 (s,
3H), 3.15 (br, 4H), 3.80 (m, 4H), 3.80-4.04 (m, 8H), 7.01 (d, J= 8.8 Hz, 2H),
7.38 (s, 1H),
8.32 (d, J= 8.8 Hz, 2H), 8.66 (s, 1H), 10.33 (s, 1H).
EXAMPLE 149: N-Hydroxy-2-(((2-(6-methoxypyridin-3-yl)-4-morpholinothieno[3,2-
d] pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 236)
Step 149a: 5-Bromo-2-methoxypyridine (Compound 0601-236)
A solution of 2-methoxy-pyridine (100 g, 0.92 mole), NBS (180 g, 1.0 mole) in
acetonitrile (1.OL) was stirred at reflux for 21 h. TLC showed reaction
complete. The
reaction mixture was cooled to room temperature and concentrated. -900ml
solvent was
collected. The resulting suspension was filtered and washed with n-hexane (-
400mL). The
filtrate was concentrated again to afford crude product. The crude product was
distilled at
reduced pressure (30 C/-0.3mmHg) to afford the title compound as clear oil
(146 g, 84%).
LCMS (m/z): 190.0 [M+1]+. 'H NMR (400 MHz, CDC13): 6 3.90 (s, 3H), 6.65 (d, J=
8.8
Hz, 1 H), 7.62 (dd, J = 8.8 Hz, 2.4Hz, 1 H), 8.19 (s, 1 H).
Step 149b: 6-Methoxypyridin-3-ylboronic acid (Compound 1601-236):
To a solution of compound 0601-236 (20 g, 0.11 mole) in anhydrous THE (180 ml)
was added dropwise n-BuLi (59 mL, 2M in THF) at -78 C, the resulting mixture
was
stirred for 1 h. Triisopropyl borate (37mL) was added at -78 C and the
reaction mixture
was warmed to room temperature and continued to stir overnight. TLC
(hexanes/ethyl
acetate =5:1) showed reaction complete. The mixture was adjusted pH to 3-4
with 4N HC1
(90 ml). The precipitate was collected by filtration to afford crude compound
1601-236 (21
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g, 128%). The crude compound 1601-236 (21g) was dissolved in water (200 ml)
and the
solution was adjusted pH to 8-9 with concentrated ammonia solution, the
precipitate was
collected by filtration to afford the pure title compound 1601-236 as a white
solid.(l 1 g,
67%). LCMS (m/z): 154.1 [M+1]+. 'H NMR (400 MHz, DMSO-d6): 6 3.86 (s, 3H),
6.76
(d, J = 8.4 Hz, 1 H), 7.99 (dd, J = 8.4 Hz, 2.0 Hz, 1 H), 8.05 (br, 2H), 8.52
(d, J = 2.0 Hz,
I H).
Step 149c: 2-Methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(Compound 0602-236)
A mixture of compound 0601-236 (55 g, 0.29 mol), 4,4,4',4',5,5,5',5'-
octamethyl -
2,2'-bi(1,3,2-dioxaborolane) (90 g, 0.35 mol), potassium acetate (57 g, 0.58
mol) and
bis(triphenylphosphine)palladium(II) chloride (2.2 g, 3 mmol) in anhydrous
dioxane (500
mL) was heated at 108 C under N2 atmosphere overnight. The reaction mixture
was
concentrated and purified by column chromatography eluted with hexanes/ethyl
acetate to
afford titled compound 0602-236 (58 g, 84 %). 1H NMR (400 MHz, DMSO-d6): 6
1.30 (s,
12H), 3.88 (s, 3H), 6.81 (d, J= 8.0 Hz, 1H), 7.88 (dd, J= 8.0 Hz, 2.0Hz, 1H),
8.41 (d, J=
2.0Hz, 1H).
Step 149d: Ethyl-2-(((2-(6-methoxypyridin-3-yl)-4-morpholinothieno [3,2-
d]pyrimidin- 6-
yl) methyl) (methyl)amino)pyrimidine-5-carboxylate (Compound 0603-236)
Method A: A mixture of compound 0504 (12 g, 26.7 mmol), 1601-236 (4.9 g, 32
mmol), NaHCO3 (6.7 g, 80.1 mmol) and bis(triphenylphosphine)palladium(II)
chloride
(188 mg, 0.267 mmol) in a mixed solvents of toluene (80 ml), ethanol (50 ml)
and water
(10 ml) was heated at 108 C for 4.5 h under N2 atmosphere. TLC showed
reaction was
complete. The reaction mixture was then cooled to room temperature and water
(20 ml)
was added. The resulting solid was collected by filtration and was then
suspended in
ethanol (100 mL). The suspension was stirred at room temperature for 30
minutes and
filtered. The collected solid was washed with ethanol and dried in vacuo to
afford titled
compound 0603-236 as a white solid (10g, 72%).
Method B: A mixture of compound 0504 (1.5 g, 3.34 mmol), 0602-236 (1.6 g, 6.68
mmol), NaHCO3 (0.84 g,10.0 mmol) and bis(triphenylphosphine)palladium(II)
chloride
(118 mg, 0.167 mmol) in a mixed solvents of toluene (24 ml), ethanol (15
ml),and water (3
ml) was heated at 108 C under N2 atmosphere overnight. The reaction mixture
was
partitioned between dichloromethane and water. The organic layer was separated
and was
washed with brine, dried over Na2SO4, filtered and evaporated in vacuo to give
a residue
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which was purified by column chromatography eluted with hexanes/ethyl acetate
to afford
compound 0603-236 as a white solid (1.7 g, 98 %).
M.p.198-202 C. LCMS: 522.30 [M+1]. 'H NMR (400 MHz, DMSO-d6): 6 1.31
(t, J= 7.2 Hz, 3H), 3.28 (s, 3H), 3.76 (t, J= 4.4 Hz, 4H), 3.93 (t, J= 4.4 Hz,
4H), 3.94 (s,
3H), 4.30 (q, J= 7.2 Hz, 2H), 5.24 (s, 2H), 6.92 (d, J= 8.8 Hz, 1H), 7.47 (s,
1H), 8.57 (dd,
J= 8.8 Hz, 2.OHz, 1H), 8.88 (s, 2H), 9.15 (d, J= 2.0 Hz, 1H).
Step 149e: 2-[(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-
methyl-
amino]-pyrimidine-5-carboxylic acid methyl ester (Compound 1602-236)
A mixture of compound 0503 (25 g, 84 mmol), CH3CN (500 mL) and methyl 2-
chloropyrimidine-5 -carboxylate (16 g, 92 mmol) was stirred at room
temperature.
Diisopropylethylamine (DIPEA) (500 mL, 2.9 mol) was added. The solution was
stirred
overnight and evaporated. After methylene chloride (500 mL) was added, the
organic
phase was washed with water, dried with Na2SO4 and concentrated in vacuo. To
the
residue was added ethyl acetate (200 mL) and the mixture was stirred in
ice/water bath for
50 min. The title product was collected as a white solid (29.4 g, 81%). LCMS
(m/z): 435.2
[M+1]+. 'HNMR (400 MHz, DMSO-d6): 3.25 (s, 3H), 3.71 (t, J= 5.2 Hz, 4H), 3.82-
3.84
(m, 7H), 5.21 (s, 2H), 7.39 (s, 1H), 8.87 (s, 2H).
Step 149f: Methyl-2-(((2-(6-methoxypyridin-3-yl)-4-morpholinothieno [3,2-
d]pyrimidin-6-
yl) methyl) (methyl)amino)pyrimidine-5-carboxylate (Compound 1603-236)
A mixture of compound 1602-236 (29.4 g, 67.7 mmol), 1601-236 (12.4 g, 81.3
mmol), NaHCO3 (17.1 g, 203 mmol) and bis(triphenylphosphine)palladium(II)
chloride
(475 mg, 0.68 mmol) in toluene (480 ml), ethanol (300 ml) and water (60 ml)
was heated
at 108 C for 6.5 h under N2 atmosphere. TLC showed reaction complete. The
reaction
mixture was cooled to room temperature and filtered to remove any insoluble
solid, then
water/EtOH (150 ml/150 mL) was added. The resulting precipitate was collected
by
filtration. The solid was suspended in ethanol (200 mL) and stirred at room
temperature
for 30 min. It was filtered and washed with ethanol and dried in vacuo to
afford titled
compound 1603-236 (25 g, 73%). LCMS (m/z): 508.30 [M+1]+. 1H NMR (400 MHz,
DMSO-d6): 6 3.27 (s, 3H), 3.75 (t, J = 4.4 Hz, 4H), 3.82 (s, 3H), 3.93 (t, J=
4.4 Hz, 4H),
3.90-3.93 (m, 7H), 5.23 (s, 2H), 6.90 (d, J= 8.8 Hz, 1H), 7.47 (s, 1H), 8.57
(dd, J= 8.4
Hz, 2.0Hz, 1 H), 8.88 (s, 2H), 9.15 (d, J = 2.0 Hz, 1 H).
Step 149g: N-Hydroxy-2-(((2-(6-methoxypyridin-3-yl)-4-morpholinothieno[3,2-d]
pyrimidin-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide (Compound 236)
Preparation of hydroxylamine methanol solution
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A mixture of NH2OH.HCl (80 g, 1.12 mol) in MeOH (400 mL) was heated at 60-65
C for lh to form a clear solution. It was then cooled in an ice-water bath. To
the cold
mixture was added a solution of KOH (96 g, 1.68 mol) in MeOH (240 mL) dropwise
while
maintaining the reaction temperature at 0-10 C. The resulting mixture was
stirred at 0 C
for 30 minutes and then filtered through a constant pressure funnel filled
with anhydrous
Na2SO4 (700 g). The filtrate was collected under an ice-bath and stored in
refrigerator for
future use.
Preparation of Compound 236 from compound 0603-236
Compound 0603-236 (10 g, 19 mmol) was suspended in the above freshly prepared
hydroxylamine methanol solution (1.79M, 350 ml). To this mixture was added
dichloromethane (100 mL). The reaction flask was sealed and the mixture was
stirred at
room temperature for 5 h before it turned into clear solution. Reaction was
stirred for
additional 9 h. and was filtered to remove any insoluble solid. The filtrate
was adjusted to
pH 6-7 with the addition of acetic acid to form solid precipitate. The solid
was collected by
filtration and washed with water and minimum amount of methanol, dried in
vacuo at 60
C for 5h to afford compound 236 as a white solid (9.2g, 96%). m.p. 177-180 C.
LCMS:
509.3 [M+1]+. 1H NMR (400 MHz, DMSO-d6): 6 3.24 (s, 3H), 3.76 (t, J= 5 Hz,
4H), 3.92
(t, J= 5 Hz, 4H), 3.92 (s, 3H), 5.20 (s, 2H), 6.90 (d, J= 8.8 Hz, 1H), 7.44
(s, 1H), 8.57 (dd,
J= 8.8 Hz, 2.4Hz, I H), 8.75 (s, 2H), 9.01 (s, I H), 9.14 (d, J= 2.0 Hz, I H),
11.08 (s,1 H).
Preparation of Compound 236 from compound 1603-236
Compound 1603-236 (500 mg, 0.98 mmol) was suspended in above hydroxylamine
methanol solution (1.79M, 12 ml). Dichloromethane (5 mL) was added. The
reaction flask
was sealed and stirred at room temperature for 5 h before it turned into clear
solution.
Reaction solution was filtered to remove any insoluble solid and added water
(5 mL).
Acetic acid was added to adjust pH to 9 and added water (10 mL) dropwise. The
resulting
reaction mixture was filtered and washed with water and minimum amount of
methanol.
The white solid was collected and dried in vacuo at 60 C for 5h to afford
compound 236
as a white solid (388 mg, 77%). LCMS (m/z): 509.4 [M+1]+. 1H NMR (400 MHz,
DMSO-d6): 6 3.24 (s, 3H), 3.76 (t, J = 5 Hz, 4H), 3.92 (t, J = 5 Hz, 4H), 3.93
(s, 3H), 5.20
(s, 2H), 6.91 (d, J = 8.8 Hz, I H), 7.45 (s, I H), 8.57 (dd, J = 8.8 Hz, 2.0
Hz, I H), 8.75 (s,
2H), 9.01 (s, I H), 9.15 (d, J = 2.0 Hz, I H), 11.09 (s,1 H).
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Biological Assays:
The following assays are used to determine the IC50 of compounds of the
present
invention with respect to inhibition of P13 kinase, HDAC and mTOR.
(a) An in vitro assay which determines the ability of a test compound to
inhibit PI3Ka.
Activity of PI3Ka was measured using ADP-Glo assay from Promega. PI3Ka, a
complex
of N-terminal GST-tagged recombinant full-length human pl l Oa (GenBank,
Accession
No.U79143) and N-terminal His tagged recombinant full length human p85 a
(GenBank
Accession No.XM_043865) were co-expressed in a Baculovirus infected Sf9 cell
expression system. The ADP-Glo assay was performed to measure the amount of
ADP
generated from ATP hydrolysis coupled with PIP2 phosphorylation by the
purified
recombinant PI3Ka (pl l0a/p85a). PI3Ka was incubated with 20 M PIP2 substrate
in the
reaction buffer (50 mM HEPES, pH 7.4, 150 mM NaCl, 5 mM MgC12, 1 MM DTT, 3 uM
Naorthovanadate, 10 M ATP and 0.5% DMSO) for 30 minutes at room temperature.
The
ADP generated was then detected by ADP-Glo assay from Promega, a luminescent
ADP
detection assay. The ADP-Glo assay is performed in two steps; first, after the
kinase
reaction, ADP-Glo Reagent is added to terminate the kinase reaction and
deplete the
remaining ATP. Second, the Kinase Detection Reagent is added to simultaneously
convert
ADP to ATP and allow the newly synthesized ATP to be measured using a
luciferase/luciferin reaction.
(b) An in vitro assay which determines the ability of a test compound to
inhibit PI3K ..
Activity of PI3K(3 was measured using ADP-Glo assay from Promega. P1 3Ka, a
complex
of N-terminal His-tagged recombinant full-length human pl 100 (GenBank,
Accession
No.NM_006219) and untagged recombinant full length human p85 a (GenBank
Accession
No.XM_043865) were co-expressed in a Baculovirus infected Sf9 cell expression
system.
The ADP-Glo assay was performed to measure the amount of ADP generated from
ATP
hydrolysis coupled with PIP2 phosphorylation by the purified recombinant
PI3K(3
(pl 10(3/p85a). PI3K(3 was incubated with 20 M PIP2 substrate in the reaction
buffer (10
mM Tris-HC1, pH 7.5, 50 mM NaCl, 5 mM MgC12, 10 M ATP and 0.5% DMSO) for 90
minutes at room temperature. The ADP generated was then detected by ADP-Glo
assay
from Promega, a luminescent ADP detection assay. The ADP-Glo assay is
performed in
two steps; first, after the kinase reaction, ADP-Glo Reagent is added to
terminate the kinase
reaction and deplete the remaining ATP. Second, the Kinase Detection Reagent
is added to
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simultaneously convert ADP to ATP and allow the newly synthesized ATP to be
measured
using a luciferase/luciferin reaction.
(c) An in vitro assay which determines the ability of a test compound to
inhibit PI3K6.
Activity of PI3K6 was measured using ADP-Glo assay from Promega. PI3K6, a
complex
of N-terminal GST-tagged recombinant full-length human p1106 (GenBank,
Accession
No.NM_005026) and untagged recombinant full length human p85 a (GenBank
Accession
No.XM_043865) were co-expressed in a Baculovirus infected Sf9 cell expression
system.
The ADP-Glo assay was performed to measure the amount of ADP generated from
ATP
hydrolysis coupled with PIP2 phosphorylation by the purified recombinant PI3K6
(pl 106/p85a). PI3K6 was incubated with 20 M PIP2 substrate in the reaction
buffer (10
mM Tris-HC1, pH 7.5, 50 mM NaCl, 5 mM MgC12, 10 M ATP and 0.5% DMSO) for 90
minutes at room temperature. The ADP generated was then detected by ADP-Glo
assay
from Promega, a luminescent ADP detection assay. The ADP-Glo assay is
performed in
two steps; first, after the kinase reaction, ADP-Glo Reagent is added to
terminate the kinase
reaction and deplete the remaining ATP. Second, the Kinase Detection Reagent
is added to
simultaneously convert ADP to ATP and allow the newly synthesized ATP to be
measured
using a luciferase/luciferin reaction.
(d) An in vitro assay which determines the ability of a test compound to
inhibit PI3Ky.
Activity of PI3Ky was measured using ADP-Glo assay from Promega. PI3Ky, N-
terminal
His-tagged recombinant full-length human p120y (GenBank, Accession
No.AF327656)
was expressed in a Baculovirus infected Sf9 cell expression system. The ADP-
Glo assay
was performed to measure the amount of ADP generated from ATP hydrolysis
coupled
with PIP2 phosphorylation by the purified recombinant PI3Ky (pl 106/p85a).
PI3Ky was
incubated with 20 M PIP2 substrate in the reaction buffer (10 mM Tris-HC1, pH
7.5, 50
mM NaCl, 5 mM MgC12, 10 M ATP and 0.5% DMSO) for 90 minutes at room
temperature. The ADP generated was then detected by ADP-Glo assay from
Promega, a
luminescent ADP detection assay. The ADP-Glo assay is performed in two steps;
first, after
the kinase reaction, ADP-Glo Reagent is added to terminate the kinase reaction
and deplete
the remaining ATP. Second, the Kinase Detection Reagent is added to
simultaneously
convert ADP to ATP and allow the newly synthesized ATP to be measured using a
luciferase/luciferin reaction.
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(e) An in vitro assay which determines the ability of a test compound to
inhibit HDAC
enzymatic activity.
HDAC inhibitory activity was assessed using the Biomol Color de Lys system (AK-
500, Biomol, Plymouth Meeting, PA). Briefly, HeLa cell nuclear extracts were
used as a
source of HDACs. Different concentrations of test compounds were serially
diluted in
dimethylsulphoxide (DMSO) and added to HeLa cell nuclear extracts in the
presence of a
colorimetric artificial substrate. Final assay condition contained 50 mM
Tris/Cl, pH 8.0,
137 mM NaCl, 2.7 mM KC1 and 1 mM MgC12. Reactions were carried in room
temperature (25 C) for 1 hour before addition of developer for termination.
Relative
enzyme activity was measured in the WALLAC Victor 111420 microplate reader as
fluorescence intensity (excitation: 350- 380 nm ; emission: 440-460 nm). Data
were
analyzed using GraphPad Prism (v4.0a) with a sigmoidal dose response curve
fitting for
IC50 calculation.
(f) An in vitro assay which-determines the ability of a test compound to
inhibit mTor
serine/threonine protein kinase.
The ability of compounds to inhibit mTor activity was assayed using standard
radioisotope assay for kinase. Briefly, FLAG-tagged, recombinant full-length
human mTor
(GenBank accession No. NM_004958) was expressed using baculovirus expression
system
in Sf2l cells and purified using antibody affinity column. Purified enzyme was
incubated
with c-terminal fragment of p70S6K as it's substrate in the presence of ATP.
p33 ATP
tracers were included in the assay to monitor the enzyme activity. Final assay
condition was
with 50 mM HEPES pH 7.5, 1 mM EGTA, 0.01% Tween 20, 2 mg/ml substrate, 3 mM
Manganese Chloride and 70uM of ATP and was carried out at room temperature for
40
minutes. The reaction was then stopped by the addition of 3% phosphoric acid
solution. 10
ul of the reaction was spotted onto a P30 filtermat and washed three times for
5 minutes in
75 mM phosphoric acid and once in methanol prior to drying and scintillation
counting.
Different concentrations of compounds were added to reaction to assess the
activity of
compounds to inhibit mTor kinase. IC50 was calculated using Prism software
with
sigmoidal dose-response curve fitting.
The following TABLE B lists compounds representative of the invention and
their
activity in HDAC, P13K and m-TOR assays. In these assays, the following
grading was
used: I > 10 M, 10 M > II > 1 M, 1 M > III > 0.1 M, and IV < 0.1 M for
IC50.
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TABLE B
Compound HDAC PI3Ka PI3KO PI3Ky PI3K6 No. TOR
3 IV I
4 IV I
IV I
7 III III
8 IV I
9 IV I
11 II
12 III IV
13 III III
14 IV IV IV IV III II
IV III I
16 III IV II
18 IV III
19 IV III
IV III
III I
31 III I
32 III I
34 III I
IV I
36 IV I
41 IV III
42 IV II
43 IV II II
44 III IV
I IV
46 II III
48 IV IV
49 III IV
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50 IV IV
51 III IV
53 IV IV IV IV IV
54 IV IV IV IV IV
60 IV I
61 IV I
62 IV I
63 IV I
65 IV III
66 IV III
67 IV III
68 IV III
69 IV IV II
70 IV IV III
71 IV III
73 IV III
74 IV III
75 III IV
76 IV IV
78 IV III
79 IV IV
80 III IV
81 II IV
83 II III
84 IV IV
85 IV III
86 IV IV IV
87 IV IV II
88 III IV
89 IV IV II
90 IV IV
91 IV IV
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92 IV IV
93 III III
94 IV IV
95 III IV
96 IV IV
97 IV IV
98 IV IV
99 IV IV III
101 I III III
102 III III III
103 IV III
104 III IV
105 III I
106 II IV
107 IV II
108 IV II
109 IV IV III
110 IV IV IV
111 IV IV III III IV IV
112 IV IV
114 IV III
115 IV IV IV
116 IV IV IV
117 IV IV II
119 IV IV
120 IV IV III
121 IV IV II
122 IV IV
124 IV IV
125 IV IV
129 IV I
130 IV III
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131 IV I
132 IV IV IV
133 IV III
134 IV III
135 IV III
136 I I
137 IV II
138 IV IV IV
139 IV III
140 IV III
141 IV III
142 IV III III
143 IV III
144 IV II
146 IV I
147 IV I
148 IV I
149 IV I
150 IV IV IV
151 IV I
152 IV I
153 IV I
154 IV I
155 IV I
156 IV III
157 IV IV IV
158 IV I
159 IV II
160 IV I
161 IV III
162 IV II
163 IV III
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164 IV IV
165 IV I
166 IV I
167 IV III
168 IV II
176 IV III
177 IV I
178 IV I
181 IV II
182 IV II
183 IV I
184 IV II III
186 IV I
187 IV IV
191 IV
192 IV III
193 IV I
194 IV I
196 IV III
197 IV IV
199 IV IV
200 IV I
201 IV III
202 IV III
203 III III
204 III III
206 IV III
207 IV III
209 IV I
210 III III
211 IV III
214 IV
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215 IV III
216 IV IV
217 IV IV
218 IV IV
219 IV IV
220 IV IV
221 IV IV
222 IV III
223 IV IV
224 IV IV
225 II IV
226 IV IV
227 IV IV
228 IV IV
229 IV IV
230 IV IV
231 IV IV
232 IV IV
233 IV IV
234 IV IV
235 IV IV
236 IV IV III III IV II
237 IV IV
240 IV IV
241 IV III
243 III IV
245 IV IV
246 IV III I
247 IV IV
250 IV IV
251 IV II
252 IV IV
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253 III IV
254 IV III
255 IV IV
256 IV IV I
257 IV I
258 IV I
259 IV I
260 IV I
261 IV IV
262 IV IV
263 IV IV
264 IV III
265 IV IV I
266 IV III
269 IV II
270 IV
271 IV IV
272 IV IV
273 III IV
274 IV IV III
275 IV III
276 IV IV
277 IV IV
278 IV IV
279 IV IV
280 IV III
281 IV III
282 IV III
283 IV III
284 IV IV
285 IV IV IV
286 IV III
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287 IV I
288 IV I
289 IV IV
290 IV III
291 IV I
292 IV IV IV
293 IV IV I
294 IV IV III
295 IV III
296 IV III I
297 IV III III
298 IV III
299 IV III
300 IV III
301 IV II I
302 IV I
303 IV III
304 IV II
305 IV III
306 IV III
307 IV III
308 IV IV
309 III III
310 IV IV
311 IV III I
312 IV III
313 IV III I
314 IV III I
315 IV IV
316 IV IV
317 IV IV
318 IV IV III
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319 III III
320 III III
321 III III
322 III III
323 III IV
324 III III
325 III II
326 III III
327 III II
328 III IV
329 III IV
Cell Proliferation Assay:
Cancer cell lines were plated at 5,000 to 10,000 per well in 96-well flat
bottomed
plates with various concentration of compounds. The cells were incubated with
compounds
for 72 hours in the presence of 0.5% of fetal bovine serum. Growth inhibition
was accessed
by adenosine triphosphate (ATP) content assay using Promega CellTiter-Glo kit.
Promega
CellTiter-Glo kit is an ATP monitoring system based on firefly luciferase.
Briefly, l6 1 of
mammalian cell lysis and substrate solution was added to 84 l of culture
medium per well
to lyse the cells and stabilize the ATP. The mixture was shaken and incubated
for 30
minutes and subsequently the luminescence was measured.
The following TABLES C, D and E list compounds representative of the
invention,
reference compounds and their antiproliferative activity (IC50 ( M)) in cell-
based assays.
In these assays, the following grading was used: I > 10,000 nM, 10,000 nM > II
> 1000 nM,
1000nM>III>100nM,100nM>IV>10nM,andV<10nmforIC50=
TABLE C
Cancer Type Cell Line SAHA GDC- SAHA/ 0941 GDC-0941 150 236 111 261 70 116 120
138
WiDr II III III IV IV IV IV IV IV
HCT116 II III III V V V
Colon SW403 II II II V V V
SW620 II III III V V V
SWI-116 II III III V V V IV
T-84 II II II IV IV IV
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H358 II II II V V V
H292 II II II V V V
NSCLC H2122 II II II V V V
H460 II III II IV IV IV
A549 II III II V IV IV
Calu6 II II II V IV IV IV IV IV
MiaPacal II II III V IV V IV IV IV
CaPan2 II II II V IV IV IV III IV
Pancreas CaPan1 II II III III IV
CFPAC-1 II II II IV IV IV III III IV
PANC-1 II II II IV IV IV IV III IV
SW1990 II II III V V V IV IV IV
Breast MDA-MB- II I II V III IV IV IV IV
Prostate PC-3 11 11 11 IV
LN-Cap III III IV V
Ovarian OVCAR-5 11 III III V
Multiple OPM-2 III III III V V V V
Myeloma
TABLE D
CancerType Cell Line 125 284 289 157 294 115 110 132 54 296 308 292
Colon WiDr IV V III IV III IV III IV V III IV
HCT116 IV V V V V V III IV
H358 V V V IV III IV
H292 V IV IV III
NSCLC H2122 V V V IV III IV
H460 IV IV IV III IV III IV
A549 IV IV IV IV IV III III
Calu6 IV IV IV IV IV IV
MiaPacal IV V V IV IV III IV
CaPan2 IV
Pancreas CaPan1 IV
CFPAC-1 III
PANC-1 IV
SW1990 IV
Breast MDA-MB-231 IV IV
Multiple Myeloma OPM-2 IV V
TABLE E
Cancer Type Cell Line BEZ-235 199 187 112
Colon WiDr II IV IV III
NSCLC Calu6 III III III IV
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MiaPacal II IV IV IV
CaPan2 II II III III
Pancreas CaPan1 III
CFPAC-1 II III III
PANC-1 II III III IV
SW1990 IV IV IV IV
Breast MDA-MB-231 I IV III IV
The patent and scientific literature referred to herein establishes the
knowledge that
is available to those with skill in the art. All United States patents and
published or
unpublished United States patent applications cited herein are incorporated by
reference.
All published foreign patents and patent applications cited herein are hereby
incorporated
by reference. All other published references, documents, manuscripts and
scientific
literature cited herein are hereby incorporated by reference.
While this invention has been particularly shown and described with references
to
preferred embodiments thereof, it will be understood by those skilled in the
art that various
changes in form and details may be made therein without departing from the
scope of the
invention encompassed by the appended claims.
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