Note: Descriptions are shown in the official language in which they were submitted.
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LA PRESENTE PARTIE DE ~; I I t DENIANDE OU CE BREVET
COMPREND PLUS D'UN TOME.
CECI EST LE TOME I DE ;2
NO~: Pour les tomes additionels, veuillez c~ntacter le 8ureau canadien des
brevets
JUMBO APPLICATIONSIPATENTS
THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE
THAN ONE VOLlJME
THIS IS VOLUME -1 - OF ,~
NOTE: Fcr additicnal Yclumes please c~nlact the Canadian Patent Office
WO 94/20062 2 1 5 ~ PCT/US94/02283
BALANOIDS
~ ,
FIELD OF THE ~Nv~ ON
The present invention relates to the field of
treatments for inflammatory, cardiovascular, metabolic, nervous
system, viral infectious, neoplastic and other diseases. The
invention provides compounds which can inhibit protein kinase
C enzymes. More particularly, the present invention relates to
novel compounds which are referred to herein as "balanoids".
R~R~.~UND OF THE INVENTION
Protein kinase C (PRC) is a family of calcium- and
phospholipid-dependent serine/threonine-specific protein
kinases which play an important role in cellular growth
control, regulation, and differentiation. Protein kinase C is
activated by diacylglycerol (DAG), a neutral lipid, and when
activated will transfer the y-phosphate of MgATP to a serine or
threonine residue on a substrate protein. The mechanisms of
protein kinase C action have been described in U.S. Patent
4,816,450 issued March 28, 1989 to Bell et al., which is
incorporated herein by reference.
The activation of protein kinase C has been implicated
in several human disease processes, including cancer tumors,
inflammation and reperfusion injury. Accordingly, protein
kinase C is a target for therapeutic agents useful in treating
these conditions.
Cancer is a disease characterized in part by
uncontrolled cell growth. Protein kinase C is directly
involved in cellular growth control and is believed to be
involved in tumor formation. Protein kinase C is fundamental
W094/20062 PCT~S94/02283 _
2 1 5 l ~ 1 2 -- 2
to the processes involved in tumorigenicity, since it is the
major high-affinity receptor for endogenous cellular DAGs as
well as for several classes of tumor promoters. These tumor
promoters also stimulate protein kinase C catalysis. Castagna
et al., (1982) J. Biol. Chem. 257:7847, reported direct
activation of protein kinase C by tumor promoting phorbol
esters.
Protein kinase C is the major, if not exclusive,
intracellular receptor of phorbol esters, which are very potent
tumor promoters. Phorbol esters and other tumor promoters bind
to and activate protein kinase C. Since DAG and phorbol esters
interact at the same site, DAGs have been suggested to be the
"endogenous phorbol esters", analogous to the opiate receptor
where the conservation of a high affinity receptor implied the
existence of an endogenous analogue. DAG has been shown to
increase the affinity of protein kinase C for Ca 2 and
phospholipid and thus activates protein kinase C at cellular
levels of these essential cofactors. Extracellular signals
including hormones, growth factors, and neurotransmitters are
known to stimulate phosphatidylinositol turnover resulting in
the generation of IP3 and DAG.
Structures of 40 distinct oncogenes of viral and
cellular origin have revealed that oncogenes encode altered
forms of normal cellular proteins. Several of the gene
products appear related to growth factors or other elements
involved in transmembrane signalling. These oncogene products
appear to function by altering the level of critical second
messengers. Cells transformed with the oncogenes ras, sis,
erbB, abl, and src have been shown to contain elevated levels
of DAG which is then believed to activate protein kinase C.
Studies on ras transformed cells have shown protein kinase C
activation to be concomitant with elevation of DAG.
Phorbol esters, such as phorbol myristate acetate
(PM~A), have complex effects on cells including effects on
membrane function, mitogenesis, differentiation, and gene
expression. Synthetic DAGs mimic many of the effects of PMA in
vitro and inhibitors of protein kinase C have been shown to
215~412
W094/20062 PCT~S94/02283
-- 3
block PMA-induced effects on cells. Thus, protein kinase C may
mediate the actions of certain oncogenes, such as ras, which
cause intracellular increases in DAG and concomitant increases
in protein kinase C. In addition, activation of protein kinase
C leads to the expression of c-myc, c-fos, c-cis, c-fms,
nuclear protooncogenes which are important in cell
transformation. Overexpression of protein kinase C in NIH 3T3
cells causes altered growth regulation and enhanced tumorige-
nicity, and in rat fibroblasts leads to anchorage-independent
growth in soft agar. Overexpression of protein kinase C in
these cells resulted in tumor formation in animals receiving
transplanted cells.
Several studies have shown increased expression of
protein kinase C in certain tumor types such as breast and lung
carcinomas. Activated protein kinase C has also been detected
in human colon carcinomas although increased expression at the
gene level was not seen. Topoisomerases are directly modulated
by protein kinase C as substrates for the enzyme.
Protein kinase C inhibitors have been reported to
potentiate the antitumor activity of chemotherapeutic agents
such as cis-platin both in vitro and in vivo (Grunicke, et al.
(1989) Adv. Enzyme Regul. 28:201; and German Offenlegungs-
schrift DE 3827974). In addition, it has been suggested that
protein kinase C would be a potential target for therapeutic
design because of its central role in cell growth (Tritton,
T.R. and J.A. Hickman, (1990) Cancer Cells 2:5-102). German
Offenlegungsschrift DE 3827974 Al discloses therapeutic
preparations comprising a protein kinase C inhibitor in
combination with a lipid, a lipid analogue, a cytostatic agent
or phospholipase inhibitor which are useful for cancer therapy.
Inflammation and reperfusion injury, particularly
pertaining to cardiac injury, are common conditions for which
there exists no definitive treatment despite extensive
research. Appropriate treatments for these conditions are
needed.
Protein kinase C inhibitors have been demonstrated to
block platelet aggregation and release of neutrophil activating
W094/20062 PCT~S94/02283
~15~12 - 4 ~
agents such as platelet activating factor (PAF) (Schachtele, et
al. (1988) Biochem. Biophy. Res. Commun. 151:542; Hannun, et
al. (1987) J. Biol. Chem. 262:13620; Yamada, et al. (1988)
Biochem. Pharmacol. 37:1161). Protein kinase C inhibitors have
also been shown to inhibit neutrophil activation, and
chemotactic migration (McIntyre, et al. (1987) J. Biol Chem.
262:15730; Lambreth, et al. (1988) J. Biol. Chem. 263:3818;
Pittet, et al. (1987) J. Biol. Chem. 262:10072; and Gaudry, et
al. (1988) Immunology 63:715), as well as neutrophil
degranulation and release of proteolytic enzymes and reactive
oxygen intermediates (Wilson, et al. (1986) J. Biol. Chem.
261:12616; Fujita et al. (1986) Biochem. Pharmacol. 35:4555;
Berkow, et al. (1987) J. Leukoc. Biol. 41: 441; Salzer, et al.
(1987) Biochem. Biophys. Res. Commun. 148:747; Kramer, et al.
(1989) J. Biol. Chem. 262:5876; and Dewald, et al. (1989)
Biochem. J. 26~:879).
Thus, inhibitors of protein kinase C have the
capability of blocking all three of the most significant
mechanisms of pathogenesis associated with myocardial
reperfusion injury. Protein kinase C is, accordingly, a drug
target for therapeutic agents. Additionally, the inhibitory
effect of protein kinAse C inhibitors on keratinocytes, and on
the oxidative burst in neutrophils, provides an
anti-inflammatory effect.
Groth, T., et al., Proc. Adabori Conf: 3rd Ger.-Jap.
Symp. Pept. Chem., E. Wuensch, ed., 91 (1989) disclose
ophiocordin, an antibiotic with antifungal activity having
certain structural similarities to certain compounds of the
invention.
~UNMARY OF ~HE l~v~.,lON
The present invention relates to a novel class of
compounds referred to herein as balanoids. Compounds according
to the present invention have the following formula:
2157~12
W O 94/20062 PCTrUS94102283
-- 5 --
G F E
1'
D l~ B B Z
~ ~ 1 2
n(~ ~)m
A
wherein:
A is: CH2, NR, S, S 2 or O;
Bl is : NR, O or CH2;
B2 is: C0, CS, or SO2;
Z is: R4, aryl, heteroaryl, substituted aryl or
substituted heteroaryl;
D is: NR3, O or CH2;
E is: R5, aryl, heteroaryl, substituted aryl or
substituted heteroaryl:
F is : CO , CS , CH (OR6), CH2 , O , S or NR;
G is: R7, aryl, heteroaryl, substituted aryl,
substituted heteroaryl or substituted cycloalkyl;
K is: hydrogen or lower alkyl;
X is: CO, CS, CH2, CNR or CCR R
R , R , R , R , R , R , R , R and Rl are, independently,
hydrogen, lower alkyl, aryl or JR ;
R5 is: lower alkyl or aryl;
J is: CO, C=NR, S 2 or P(O)0 alkyl;
Rll is: hydrogen, lower alkyl, aryl, alkamino,
arylamino, aryloxy or alkoxy;
Rl2 is: straight or branched alkyl, aryl;
m is: 1-4;
n is: 1-4; and
m plus n is up to 5;
providing that if m is 3, A is NH, Bl is 0, B2 is CO,
Z is p-hydroxyphenyl, D is NH, X is CO, and E, F, and G, taken
together, are
W094/20062 PCT~S94/02283
215~12 - 6 -
OH HO
HOOC ~$ C
OH
then n is not 1.
Additionally, the present invention relates to pharma-
ceutically acceptable salts of the above compounds and to
formulations comprising the above compounds in pharmaceutically
acceptable carriers. Prodrugs such as carbonates and esters of
phenolic functional groups and other species metabolizable into
compounds of the invention are also considered to be within the
scope of the present invention.
The present invention relates to a method of
inhibiting protein kinase C activity which comprises contacting
protein kinase C with an inhibitory amount of a balanoid of the
invention.
The present invention also relates to methods of
treating an animal, preferably a mammal, that is suffering from
a PRC-related disease, especially an inflammatory,
cardiovascular and/or neoplastic diseases by administering an
effective amount of a balanoid to the animal.
DET~TT-~n ~ TPTION OF ~K~KK~ EMBODINENT8
This invention is directed to a family of novel
compounds denominated "balanoids". Members of the family have
been found to exhibit the ability to inhibit enzymes of the
family of enzymes known as protein kinase C enzymes.
Selectivity in inhibitors among the isoforms of protein kinase
C (PKC) has been shown for balanoids and it is believed that
balanoids will be useful in the treatment of disease linked to
PKC enzymes.
Methods for preparing balanoids together with
synthetic intermediates, are further objects of the invention
as are methods for testing for PKC-linked diseases.
W094/20062 215 ~ PCT~S94102283
-- 7
The present invention relates to balanoids, and their
pharmaceutically acceptable salts and formulations. Compounds
according to the present invention have been shown to inhibit
protein kinase C. PKC inhibitors are known to be useful in the
treatment of cancer, inflammatory and reperfusion injury
through their antiproliferative and anti-inflammatory
activities in human neutrophils, human keratinocytes, and human
tumor cells.
The present invention relates to methods of inhibiting
protein kinase C activity which comprises contacting said
protein kinase C with an effective amount of a balanoid or a
pharmaceutically acceptable salt thereof. Protein kinase C
inhibitors are useful as anti-inflammatory, antitumor, and
reperfusion injury agents through their antiproliferative and
anti-inflammatory activities in human neutrophils, human
keratinocytes, and human tumor cells. The present invention
relates to methods of treating animals, specifically mammals,
suffering from inflammatory, cardiovascular and/or neoplastic
diseases by administering an amount of a balanoid or a pharma-
ceutically acceptable salt thereof to the animal. Humantherapeutics are preferred.
The methods of the present invention comprise inhibit-
ing protein kinAs~ C activity by contacting protein kinase C
with an inhibitory effective amount of a balanoid. Balanoids
been discovered to inhibit the activity of protein kinase C.
Exposure of cells in vitro to balanoids results in the
inhibition of PKC activity. Inhibition of PKC activity in
cells impedes cellular activities associated with several
disease conditions. Of particular note is the selectivity
exhibited by Balanoids which permits selective inhibition of
one or more isoforms (isozymes) of PKC to a greater degree than
other isoforms. Such selectivity has long been desired and is
indicative of great therapeutic usefulness.
The methods of the present invention are useful in the
treatment of diseases which involve cellular growth, regulation
and differentiation such as inflammatory, cardiovascular and
neoplastic diseases. PKC activity is associated with disease
W094l20062 PCT~S94/02283
2157~112 - 8 -
conditions such as cancer, inflammation and reperfusion injury.
Accordingly, the present invention relates to methods of
treating a mammal suffering from cancer, inflammation such as
the type associated with arthritis, reperfusion injury or other
PKC-linked conditions. The methods comprise administering to
the mammal an effective amount of a balanoid or a phar-
maceutically acceptable salt thereof which inhibits PKC
activity connected with disease.
PKC phosphorylates certain molecules, referred to
herein as phosphorylation acceptor molecules. In order to
identify compounds that inhibit PKC activity, an appropriate
assay is performed. An exemplary and convenient assay is one
in which radio labelled ATP is combined with a phosphorylation
acceptor molecule in the presence of PKC and a balanoidal PKC
inhibitor-candidate compound (hereinafter referred to as a
"test compound"). Various amounts of test compound are used to
determine the level of inhibitory activity that a particular
test compound possesses. As a control, radio labelled ATP,
phosphorylation acceptor molecule and PKC are combined without
test compound. Assay conditions such as pH, salt and cofactor
conditions are preferably maintained to be similar to
physiological levels in order to duplicate in vivo conditions.
In the assay, if PKC is active, the phosphorylation receptor
molecule will be phosphorylated, gaining a radiolabelled
phosphorus atom. Thus, the inhibitory activity of the test
compound can be determined by incubating PKC, 3ZP-ATP,
phosphorylation receptor molecule and test compound and then
measuring the level of phosphorylation activity by measuring
the level of radioactive phosphorus present in the
phosphorylation receptor molecule.
A convenient way to determine the selectivity of PKC
inhibitory activity, test compounds are investigated for cAMP
dependent protein kinase (PKA) inhibitory activity. As in the
PKC assay, the level of inhibitory activity is determined by
measuring the level of phosphorylation of a phosphorylation
acceptor molecule incubated with radiolabelled ATP and PKA.
W094/20062 2 1 3 7 4 1 ~ PCT~S94/02283
_ g _
Preferred PKC inhibitors are selective inhibitors and do not
effect the activity of PKA.
In order to investigate the effect that balanoid PKC
inhibitors of the present invention have on cell growth and
activity, assays are performed such as to determine human tumor
cell growth inhibition, human keratinocyte inhibition and
- neutrophil superoxide anion release. Briefly, a human tumor
cell growth inhibition assay measures the growth of tumor cells
in the presence PKC inhibitors by measuring the incorporation
of radiolabelled amino acid in cells. The human keratinocyte
inhibition assay measures the proliferation of human epidermal
keratinocytes in the same manner as tumor cell growth is
measured. Hyperproliferation of keratinocytes is symptomatic
of many disease conditions associated with inflammation. The
neutrophil superoxide anion release assay measures a PKC
inhibitors ability to block the PMA-induced effects on cells.
The ability of the PKC inhibitors to affect superoxide release
by PMA stimulated neutrophils is determined by measuring
cytochrome C reduction. Cytochrome C is measured by measuring
optical density.
In vivo studies to determine the anti-inflammatory
activity of a test substance are conducted using the phorbol
12-myristate 13-acetate (PMA) induced mouse ear edema model
which is a mouse model of acute inflammation. Using this
model, the efficacy of various test compounds as anti-
inflammatory agents are determined.
The compounds of the present invention are referred
to herein as balanoids. Novel compounds according to the
present invention can be expressed by the formula:
G F E
X K
D ~/ B~B2Z
n(~ )) m
A
W094l20062 PCT~S94/02283
2 ~ o -
wherein:
A is: CH2, NR , S, SO2 or O;
Bl is: NR , O or CH2;
B2 is: CO, CS, or SO2;
Z is: R , aryl, heteroaryl, substituted aryl or
substituted heteroaryl;
D is: NR , O or CH2;
E is: R , aryl, heteroaryl, substituted aryl or
substituted heteroaryl;
F is: CO, CS, CH(OR), CH2, O, S or NR ;
G is: R7, aryl, heteroaryl, substituted aryl,
substituted heteroaryl or substituted cycloalkyl;
R is: hydrogen or lower alkyl;
X is: CO, CS, CH2, CNR or CCR R ;
R , R , R , R , R , R , R , R and R are, independently,
hydrogen, lower alkyl, aryl or JR1l;
R5 is: lower alkyl or aryl;
J is: CO, C=NRl2, SO2 or P(O)O alkyl;
R is: hydrogen, lower alkyl, aryl, alkamino,
arylamino, aryloxy or alkoxy;
Rl2 is: straight or branched alkyl, aryl;
m is: 1-4;
n is: 1-4; and
m plus n is up to 5;
providing that if m is 3, A is NH, Bl is O, B2 is CO,
Z is p-hydroxyphenyl, D is NH, X is CO, and E, F, and G, taken
together, are
OH HO
HOOC~C~
OH
W094/20062 2 1 ~ 7 ~12 PCT~S94/02283
then n is not 1. Compounds according to the present invention
include pharmaceutically acceptable salts of these compounds.
Prodrugs, such as those having carbonates and esters of
phenolic groups are also within the scope of the invention.
Lower alkyl means a straight chain, branched or cyclic
moiety having from 1 to 6 carbon atoms.
Compounds according to the present invention can have
at position A: CH2, NR1, S, SO2 or O. It is preferred that A be
NH, CH2, or NRl. When A is NR1, it is preferred that R1 be H or
lower alkyl, aryl or JR~ wherein J is CO, C=NH or SOz and R~ is
lower alkyl, aryl, alkylamino, arylamino, aryloxy or alkoxy.
Thus, compounds of the present invention can have at position
A: CH2, NH, S, SO2, O, NSO2CH3, NSO2phenyl, NCONHphenyl, NCO
phenyl, NCH2phenyl, NCH(CH3)2, NCOCH3, NCOCF3, NSO2-(5-
dimethylamino-l-naphthalene, NSO2-1-naphthalene, NSO2-2-
naphthalene, NSO2-2-methyl-5-nitrophenyl, NSO2-2-nitrophenyl,
NSO2-4-nitrophenyl, NCH=NC(CH3)3, NCONHCH3, Nco(cH2)l4cH3~
NCOOCH2phenyl, NCOOCH2CH(CH3) 2 ~ NCOOC(CH3)3, NCOCH2phenyl,
NP=O(OCH2CH3)2, NCH2CH3, NCOOCH3, NSO2CH2phenyl or N(CH2)40H.
Compounds according to the present invention can have
at position B1: NR2, O or CH2. It is preferred that B1 be NR or
O. B1 is more preferably NH or NCH3.
Compounds according to the present invention-can have
at position B2: CO, CS, or SO2. It is preferred that B2 be CO
or CS; B2 is more preferably CO.
Compounds according to the present invention can have
at position K: H, or lower alkyl, such as methyl, ethyl or
propyl. It is preferred that K be H.
- Compounds according to the present invention can have
at position Z: R , aryl, heteroaryl, substituted aryl or
substituted heteroaryl. In some embodiments, Z is preferably
hydroxy substituted aryl, ether substituted aryl, hydroxy
substituted heteroaryl or ether substituted aryl. In some
embodiments, Z is pyridine, pyrrole, oxazole, indole, purine,
furan, thiophene, pyridazine, pyrimidine, pyrazine, imidazole,
W094/20062 PCT~S94102283
21S74~
- 12 -
thiazole, isoxazole, pyrazole, isothiazole, benzene, methyl
benzene, dimethyl benzene, trimethyl benzene, tetramethyl
benzene, ethyl benzene, tetraethyl benzene, propyl benzene,
tetrapropyl benzene, butyl benzene, tetrabutyl benzene, pentyl
benzene, tetrapentyl benzene, methoxy benzene, dimethoxy
benzene, trimethoxy benzene, tetramethoxy benzene, ethoxy
benzene, diethoxy benzene, nitro benzene, dinitro benzene, halo
benzene, dihalo benzene, trihalo benzene, tetrahalo benzene,
benzene carboxylic acid, benzene dicarboxylic acid, benzamide,
benzene diamide, 3,5-dihydroxy benzene, trihydroxy benzene,
tetrahydroxy benzene, pentahydroxy benzene, triethoxy benzene,
tetraethoxy benzene, propoxy benzene, dipropoxy benzene,
tripropoxy benzene, tetra propoxy benzene, aniline, diamino
benzene, methoxy pyridine, dimethoxy pyridine, hydroxy
pyridine, dihydroxy pyridine, ethoxy pyrrole, dihydroxy
pyrrole, dimethoxy indole, hydroxy purine, dimethoxy furan,
hydroxy thiophene, methoxy pyridazine, dimethoxy pyridazine,
hydroxy pyrimidine, diamido pyrimidine, amido pyrazine,
cyanobenzene, butyloxybenzene, hydroxyindole, diethoxy
pyrazine, phenyl, quinoline, methoxy quinoline, dimethoxy
quinoline, trimethoxy quinoline, hydroxy quinoline, dihydroxy
quinoline, ethoxy quinoline, amino quinoline, diamido
quinoline, trihalo quinoline, quinoline carboxylic acid,
quinazoline, methoxy quinazoline, dimethoxy quinazoline,
trimethoxy quinazoline, hydroxy quinazoline, trihydroxy
quinazoline, tetraethoxy quinazoline, diamino quinazoline,
triamido quinazoline, tetrahalo quinazoline, quinazoline
dicarboxylic acid. Z is more preferably p-hydroxy phenyl, p-
benzyloxy phenyl, p-benzoate phenyl, p-carboxy phenyl, 4-(2-
hydroxyphenylcarbonyl)-3,5-dihydroxyphenyl,p-aminophenyl,4-
fluoro phenyl, 4-benzyloxy phenyl, p-methyl phenyl, p-
benzyloxycarbonyl phenyl, p-nitrophenyl, 5-benzyloxy-2-indole,
5-hydroxy-2-indole, 3,4-dihydroxy phenyl, 2-benzyloxy phenyl,
2-hydroxyphenyl, phenyl, p-NHSO2CH3phenyl, p-methoxymethyleneoxy
phenyl, p-acetoxy phenyl. It is more preferred that Z be
substituted phenyl. It is most preferred that Z be p-hydroxy
phenyl, p-halophenyl or 5-hydroxy indole.
W O 94/20062 2 ~ 12 PCTrJS94/02283
- 13 -
Compounds according to the present invention can have
at position D: NR , O or CH2. It is preferred that D be NR , or
o. It is more preferred that D be O or NH.
Compounds according to the present invention can have
at position E: R4, aryl, heteroaryl, substituted aryl or
substituted heteroaryl. In some embodiments, E is preferably
hydroxy substituted aryl, ether substituted aryl, hydroxy
substituted heteroaryl or ether substituted aryl. In other
embodiments, E may be pyridine, pyrrole, oxazole, indole,
purine, furan, thiophene, pyridazine, pyrimidine, pyrazine,
imidazole, thiazole, isoxazole, pyrazole, isothiazole, benzene,
methyl benzene, dimethyl benzene, trimethyl benzene,
tetramethyl benzene, ethyl benzene, tetraethyl benzene, propyl
benzene, tetrapropyl benzene, butyl benzene, tetrabutyl
benzene, pentyl benzene, tetrapentyl benzene, methoxy benzene,
dimethoxy benzene, trimethoxy benzene, tetramethoxy benzene,
ethoxy benzene, diethoxy benzene, nitro benzene, dinitro
benzene, halo benzene, dihalo benzene, trihalo benzene,
tetrahalo benzene, benzene carboxylic acid, benzene
dicarboxylic acid, benzamide, benzene diamide, 3,5-dihydroxy
benzene, trihydroxy benzene, tetrahydroxy benzene, pentahydroxy
benzene, triethoxy benzene, tetraethoxy benzene, propoxy
benzene, dipropoxy benzene, tripropoxy benzene, tetra propoxy
benzene, aniline, diamino benzene, methoxy pyridine, dimethoxy
pyridine, hydroxy pyridine, dihydroxy pyridine, ethoxy pyrrole,
dihydroxy pyrrole, dimethoxy indole, hydroxy purine, dimethoxy
furan, hydroxy thiophene, methoxy pyridazine, dimethoxy
pyridazine, hydroxy pyrimidine, diamido pyrimidine, amido
pyrazine, diethoxy pyrazine, phenyl, quinoline, methoxy
quinoline, dimethoxy quinoline, trimethoxy quinoline, hydroxy
quinoline, dihydroxy quinoline, ethoxy quinoline, amino
quinoline, diamido quinoline, trihalo quinoline, quinoline
carboxylic acid, quinazoline, methoxy quinazoline, dimethoxy
quinazoline, trimethoxy quinazoline, hydroxy quinazoline,
trihydroxy quinazoline, tetraethoxy quinazoline, diamino
quinazoline, triamido quinazoline, tetrahalo quinazoline,
quinazoline dicarboxylic acid, 2-hydroxy benzene, 3-hydroxy
W094/20062 PCT~S94/02283
21S~12 - 14 -
benzene, 3-butyloxy benzene, 3-butyloxy-5-hydroxy benzene, 3-
hexanoyloxy-5-hydroxy benzene, 3,5-dioctyloxy benzene, 3-
octyloxy-5-hydroxy benzene, 3-methoxy-5-hydroxy benzene, 3,5-
bis(acetoxy)benzene,3-(methoxycarbonyl)oxy-5-hydroxy benzene,
3,5-dihydroxy phenyl, 3-ethoxy-5-hydroxy phenyl, 3,5-
dibenzyloxy phenyl, 3,5-dimethoxy phenyl, 3-hydroxy-5-benzoate
phenyl, phenyl, 3,5-dimethoxymethyleneoxy phenyl, 3-
methoxycarbonyloxy phenyl, 3-acetoxy-5-hydroxy phenyl. It is
more preferred that E be 3,5-hydroxy benzene or 3-acyloxy-5-
hydroxy benzene. It is most preferred that E be 3,5-hydroxy
benzene.
Compounds according to the present invention can have
at position F: CO, CS, CH(OR6), CH2, O, S or NR6. It is
preferred that F be CO or CH2. It is most preferred that F be
CO.
Compounds according to the present invention can have
at position G: R~, aryl, heteroaryl, substituted aryl,
substituted heteroaryl or substituted cycloalkyl. In some
embodiments, G is preferably hydroxy substituted aryl, carboxy
substituted aryl, hydroxy substituted heteroaryl or carboxy
substituted heteroaryl. In other embodiments, G may be
pyridine, pyrrole, oxazole, indole, purine, furan, thiophene,
pyridazine, pyrimidine, pyrazine, imidazole, thiazole,
isoxazole, pyrazole, isothiazole, benzene, methyl benzene,
dimethyl benzene, trimethyl benzene, tetramethyl benzene, ethyl
benzene, tetraethyl benzene, propyl benzene, tetrapropyl
benzene, butyl benzene, tetrabutyl benzene, pentyl benzene,
tetrapentyl benzene, methoxy benzene, dimethoxy benzene,
trimethoxy benzene, tetramethoxy benzene, ethoxy benzene,
diethoxy benzene, nitro benzene, dinitro benzene, halo benzene,
dihalo benzene, trihalo benzene, tetrahalo benzene, benzene
carboxylic acid, benzene dicarboxylic acid, benzamide, benzene
diamide, 3,5-dihydroxy benzene, trihydroxy benzene,
tetrahydroxy benzene, pentahydroxy benzene, triethoxy benzene,
tetraethoxy benzene, propoxy benzene, dipropoxy benzene,
tripropoxy benzene, tetra propoxy benzene, aniline, diamino
benzene, methoxy pyridine, dimethoxy pyridine, hydroxy
W094t20062 21~ ~ ~1~ PCTtUS94/02283
- 15 -
pyridine, dihydroxy pyridine, ethoxy pyrrole, dihydroxy
pyrrole, dimethoxy indole, hydroxy purine, dimethoxy furan,
hydroxy thiophene, methoxy pyridazine, dimethoxy pyridazine,
hydroxy pyrimidine, diamido pyrimidine, amido pyrazine,
5 diethoxy pyrazine, phenyl, quinoline, methoxy quinoline,
dimethoxy quinoline, trimethoxy quinoline, hydroxy quinoline,
dihydroxy quinoline, ethoxy quinoline, amino quinoline, diamido
quinoline, trihalo quinoline, quinoline carboxylic acid,
quinazoline, methoxy quinazoline, dimethoxy quinazoline,
trimethoxy quinazoline, hydroxy quinazoline, trihydroxy
quinazoline, tetraethoxy quinazoline, diamino quinazoline,
triamido quinazoline, tetrahalo quinazoline, quinazoline
dicarboxylic acid, 2-cyano-6-hydroxy benzene,, 2-hydroxy-
5,6,7,8-tetrahydro-naphthalene,2-acetoxy-6-carboxybenzene,2-
hydroxy-6-tetrazolylbenzene,2-hydroxy-naphthalene,2-hydroxy-
6-(methoxycarbonyl) benzene, 2-carboxy-3-pyridinyl, 2-
(ethoxycarbonyl)-6-hydroxy benzene, 2,3-dihydroxy benzene, 2-
carboxy cyclohexane, 2,6-dihalo benzene, 2-acetoxy-6-
(ethoxycarbonyl) benzene. In some embodiments, G is preferably
2-carboxy-6-hydroxy phenyl, 2-ethoxycarbonyl-6-hydroxy phenyl,
2-hydroxy phenyl, 2-benzyloxycarbonyl phenyl, 2-hydroxy
naphthyl, 2,3,5,6,-tetramethyl phenyl, 2,6-dihydroxy phenyl,
2,6-dimethoxy phenyl, 2-carboxy cyclohexane, 2-hydroxy
cyclohexane, 2-hydroxy-1-naphthyl, 2,6-dichloro phenyl, 2-
methoxy-6-hydroxy phenyl, 2-carboxy-3-pyridine, 3-carboxy-2-
pyridine, phenyl, 3,4-dihydroxy phenyl, 2-methoxycarbonyl-6-
hydroxy phenyl, 2-butoxycarbonyl-6-hydroxy phenyl, 2-(2-
methylpropyloxycarbonyl)-6-hydroxyphenyl, 2-nitrilo-6-hydroxy
phenyl, 2-carboxy phenyl, 2-(4-acetoxybenzyloxycarbonyl)-6-
hydroxy phenyl, 2-benzyloxycarbonyl-6-benzyloxy phenyl, 2,6-
dibenzyloxy phenyl, 2-benzyloxycarbonyl cyclohexane, 1-
benzyloxy-2-naphthyl, 2-methoxy-6-benzyloxy phenyl, 2-
benzyloxycarbonyl-3-pyridinyl, 3-benzyloxycarbonyl-2-pyridinyl,
2-benzyloxyphenyl, 2-nitrilo-6-benzyloxy phenyl, 3,4-
dibenzyloxyphenyl, 2-benzyloxy-1-naphthyl, 6-benzyloxy-2-
tetrazolylphenyl, 6-hydroxy-2-tetrazolylphenyl, 2-
methyltetrazolylphenyl,3-methyltetrazolylphenyl,2-hydroxy-1-
W O 94/20062 PCTrUS94/02283
2~ 2 - 16 -
(5,6,7,8-tetrahydro) naphthyl,3-benzyloxycarbonyl-4-benzyloxy
phenyl, 3-carboxy-4-hydroxy phenyl, 2-methoxymethyleneoxy
phenyl, 2-ethoxycarbonyl-6-benzyloxy phenyl, 2-benzyloxy
carbonyl-l-naphthyl, 2-carboxy-1-naphthyl, 2-benzyloxy-6-methyl
phenyl, 2-methyl-6-hydroxy phenyl, 2-acetoxy-6-ethoxycarbonyl
phenyl, 2-cyclohexylmethoxycarbonyl-6-hydroxy phenyl, 2-
carboxy-6-benzyloxy phenyl, 2-methoxycarbonyl-6-benzyloxy
phenyl, 2-hexanoyloxy-6-carboxy phenyl. It is more preferred
that G be 2-carboxy-6-hydroxyphenyl, 2-hydroxy-6-(tetrazol-2-
y)phenyl, 2,6-dihydroxy phenyl, 2-hydroxy-1-naphthyl, 2-
methoxycarbonyl-6hydroxyphenyl, 2-cyano-6-hydroxy phenyl, and
2-hydroxy-6-(trifluoromethylsulfonamino)phenyl. It is most
preferred that G be 2-carboxy-6-hydroxy benzene and its ester
or acyl derivatives as well as 2-R-6-hydroxyphenyl where R is
carboxylic acid surrogate such as tetrazole or N-
sulfonylcarboxamide.
Compounds according to the present invention can have
at position X: CO, CS, CNR3 or CCR9R10. It is preferred that X
be C0 or CH2.
Compounds according to the present invention can have
2 3 4 6 R7 R3 R9 and Rl, independently: Y
lower alkyl, aryl or JRl1 wherein J is CO, CN or S02 and R11 is
lower alkyl, aryl, alkylamino, arylamino, aryloxy or alkoxy.
Compounds according to the present invention can have
at position R5 lower alkyl or aryl.
In compounds according to the present invention, m is
1-4. It is preferred that m be 1-2, preferably 1.
In compounds according to the present invention, n is
1-4. It is preferred that n be 1-3.
In compounds according to the present invention, n
plus m is less than or equal to 5. It is preferred that n plus
m is less than or equal to 4.
It will be appreciated that atoms within the moieties
defined by n and m may have substituents. Such substituents
may preferably include hydrocarbyl groups such as the lower
alkyl groups, methyl, ethyl and propyl together with larger
aliphatic and aromatic functions.
WOg4/20062 2 i ~ 7 `1~ 2 PCT~S94/02283
- 17 -
It will also be appreciated that m and n may contain
other functional species such as carbonyl, thiocarbonyl,
hydroxy, amino, halo and others. Preferred species are
carbonyl and thiocarbonyl.
Preferred compounds contain aryl groups in positions
Z, E and G. In preferred compounds, Z, E and G are hydroxy
substituted aryl, ether substituted aryl, hydroxy substituted
heteroaryl, or ether substituted aryl. E and G are preferably
substituted aryls or substituted heteroaryls whereby the
substitutions are located such that the two aryl rings are
conformationally "crowded" out of plane with each other.
Additionally, A is preferably NR or CHz.
Pharmaceutically acceptable salts of these compounds
may be used in accordance with the present invention. One
having ordinary skill in the art could readily appreciate what
salts would be appropriate. Pharmaceutically acceptable salts
include, but are not limited to sodium, trialkyl ammonium,
potassium, calcium, zinc, lithium, magnesium, aluminum,
diethanolamine, ethylenediamine, meglumine and acetate.
Preferred salts are sodium and potassium.
In certain preferred compounds of the present inven-
tion A is CH2, NR , S, or O; Bl is NR , O, or CH2; B2 is CO or
CS; Z is R4, aryl, heteroaryl, substituted aryl or substituted
heteroaryl or D is NR3, O or CH2; E is R5, aryl, heteroaryl,
substituted aryl or substituted heteroaryl; F is CO or CS; G is
R7, aryl, heteroaryl, substituted aryl or substituted
heteroaryl; X is CO or CS; R, R, R, R, R, R, R, R and R
are, independently: hydrogen, lower alkyl or aryl; R5 is lower
alkyl or aryl; m is 1-4; and n is 1-4; where n plus m is less
than or equal to 5.
In other preferred compounds of the present invention
A is CH2, NRl, S, or O; Bl is NR2 or O; B2 is CO or CS; Z is
hydroxy substituted aryl, ether substituted aryl, hydroxy
substituted heteroaryl, halo substituted aryl; D is NR3 or 0;
E is hydroxy substituted aryl, ether substituted aryl, hydroxy
substituted heteroaryl, acyloxy substituted aryl; F is CO or
CS; G is hydroxy substituted aryl, ether substituted aryl,
W094/20062 PCT~S94/02283
2 1 ~ 7 ~1~ 18 -
hydroxy substituted heteroaryl, carboxy substituted aryl; X is
CO or CS; Rl, R2 and R3 independently: hydrogen, lower alkyl or
aryl; m is 1-2; and n is 1-3; where n plus m is less than or
equal to 4.
In still other preferred compounds of the present
invention A is NH , CH2 or NR; Bl is NR or O; B2 is CO or CS;
Z is hydroxyphenyl or halophenyl; D is NR3, O or CH2; E is 3,5-
hydroxy benzene or 3,5-alkoxy benzene; F is CO or CH2; G is 2-
carboxy-benzene, 2-hydroxy benzene, 2,6-dihydroxy benzene, 2-
methoxy benzene, 2,6-dimethoxy benzene, or 6-hydroxy benzene-
2-carboxylic acid; X is CO; Rl, R2, or R3 are, independently
hydrogen, lower alkyl or aryl; m is l; and n is 3.
In certain preferred compounds of the present inven-
tion A is NH or CH2; Bl is NH; B2 is CO; Z is p-hydroxyphenyl;
15 D is O; E is 3,5-hydroxy benzene; F is CO; G is 2-carboxy-6-
hydroxyphenyl, 2-hydroxy-6-(tetrazol-2-y)phenyl, 2,6-dihydroxy
phenyl, 2-hydroxy-1-naphthyl, 2-methoxycarbonyl-6hydroxyphenyl,
2-cyano-6-hydroxy phenyl, and 2-hydroxy-6-
(trifluoromethylsulfonamino)phenyl; X is CO; m is l; and n is
3.
As will be appreciated, it is generally the case that
one stereoisomer is more biologically active than its
enantiomer. It is envisioned that preferred stereoisomerism
will be determined for active species and that such preferred
compounds will be selected for therapeutic and other uses.
Compounds of the present invention may be synthesized
from readily available starting materials by st~n~rd
~er-~n;ques such as by following the basic synthesis set out
below. One having ordinary skill in the art may employ other
well known synthetic schemes to produce compounds according to
the present invention.
Prodrugs such as carbonates and carboxy esters of
phenolic OH and NH groups can be prepared by the derivatization
of OH and NH groups with acylating agents, such as methyl
chloroformate, ethyl chloroformate, isobutyryl chloride,
methoxypropionyl chloride, methyl chlorosuccinate, ethyl
chlorosuccinate and benzoyl chloride, for example.
21~74~ 2
W094l20062 PCT/US94/02283
-- 19 --
Additionally, prodrugs of compounds which contain a carboxylic
acid can be prepared by derivitazation with alkylating agents,
such as methyl iodide or acetoxymethyl chloride.
Reaction Scheme I provides syntheses for producing
compounds according to the present invention including the use
of a cyclic carbonyl or a heterocyclic carbonyl such as the
seven membered lactam shown as a starting material. For
example, a lactam can be benzylated with base in
tetrahydrofuran to protect the nitrogen functionality. It is
then reacted with base and phenylselenyl chloride followed by
sodium periodate to yield the unsaturated lactam. Oxidation
with osmium tetroxide followed by benzoylation yields the
hydroxy benzoate shown. Further reaction with
trifluoromethanesulfonic anhydride and sodium azide provides
the anti azido ester which can be reduced with, for example,
lithium aluminum hydride. Reaction with a carboxylic acid
substituted with a Z functionality yields the amide. Further
reaction with a GFE carboxylic function followed by
deprotection provides the family of ester/amides.
WO 94120062
PCT/US9 1102283
2157~112 -20-
O
_~z = ~ Z ti
`C
T ~ T
O t_
~Z= e_~
.' T ~,, o T
m . .
~ m ~:
Q IL Q I
Z Jl-- ~ O
C~ 11
-- ' --C
W094/20062 ~i 7 11~ PCT~S94/02283
- 21 -
Scheme II provides a synthesis scheme for producing
compounds according to the present invention including the use
of an enol ether lactam such as the azepinone shown as a
starting material. For example, an enolether lactam can be
benzylated with base in tetrahydrofuran to protect the nitrogen
function. It is then hydrolyzed with acid and reacted with
sodium nitrite in acetic acid to form the oxime. Catalytic
hydrogenation in the presence of acetic anhydride gives the
acetamide, which can be reduced with, for example, sodium
borohydride and hydrolyzed to the syn (or anti) aminohydroxy
lactam. Reduction with, for example, lithium aluminum hydride
and reaction with a carboxylic acid substituted with a Z
functionality yields the amide, which can be further reacted
with GFE carboxylic function and deprotected to provide the
family of ester/amides.
WO 94noo62 rCT/US94/02283
- 22 -
2 1 5 7 ~c~jeme II
OEt OEt ~O
1)KOtBu ~ HCUH20 ~
~ N ~ o 2)PhCH28r Ph N~ ~h
Y.Tamuraetal.,
Chem.Pharm.Bull.,
~(3).523-8,529-34(1971)
1 ) NaBH4
~2)HCUH20/EtOH
NaNO2 ~ NOH H2/RaneyNi ~ NHCOCH3 3) NaOH
ACOH/Ac20 ~ N ~ O AcOHlAc20 ~ O
Ph Ph
H ~OH Z-COCI OH
NHz LAH (~ IH2 Et3N/DMAP(~;~NHCO--Z
Ph Ph Z-C-N ~N Ph
G-F-E-COCI
Et3NtDMAP lc_E-F-G H2 olC_E-F-G
~ ~NHCO - Z
G-F-E-COOH ( ~ NHCO - Z Pd(OH)2;C H
DCCIDMAP
W094/20062 21~ 2 PCT~S94/02283
- 23 -
Reaction Scheme III provides methods for producing
compounds according to the present invention including the use
as a starting material of the previously mentioned syn
aminohydroxy lactam. Oxidation of the alcohol using, for
example, oxalyl chloride, dimethylsulfoxide, and triethylamine
(Swen oxidation) provides the Keto intermediate which is
treated with HONH2-HCl followed by reduction using, for
example, RaNi (Raney nickel) catalyst and hydrogen affords the
amino-amide. Reaction with a GFE carboxylic acid function
followed by deprotection provides the family of diamides.
WO 94/20062
215 7 ~ 24 - PCTIUS9~/02283
Scheme m
~OH O
(~NHCO--Z oxalyl chloride ~NHCO Z 1) NH20H-HCI
DMSO, Et3N, -65C NJ 2) RaNi. H2
Ph Ph
G-F-E-COCI
~NH2 Rm Et3N/DMAP NHCO-E-F-G
(;;)~NHCO{~ or O~ NHCO--Z
G-F-E-co-N~g N
Ph
Ph
NHCO-E-F-G
H2
~NHCO Z
Pd(OH)2/C H
W094eO062 ~ 3 5, ~12 PCT1594/02283
Scheme IV provides a synthesis scheme for producing
compounds according to the present invention including the use
as a starting material of a cyclic olefin as shown.
Epoxidation with peracetic acid followed by reaction with
sodium azide affords the anti azido alcohol, which is O-
protected and reduced to the aminoalcohol ether. Reaction with
a carboxylic acid substituted with a Z functionality yields the
amide, which is O-deprotected and reacted with a GFE carboxylic
function to provide the family of ester/amides.
WO 94/20062 rCT/US94/02283
2 1 5 ~ 26 -
t~ T
=:E O ~
IL -
~3 ~
m
~ ~0~
~0
s
_ c ~
_
c
T
T
Z~
, E m
~
z~
~ o
T T
0~ ~
N _
T
~ o U~f~O
z 5 0
C~ ~ Z`'-
o , ~ r~l ~0
W094/2006~ 2 1 ~ 2
_ PCT/US94/02283
- 27 -
Scheme V provides a syntheses for preparing compounds
according to the present invention including a synthesis scheme
for producing 6, 7 and 8 member cyclic and heterocyclic groups
including Bl and D with stereo specific attachment, including
the use as a starting material of the unsaturated aldehyde.
For example, tin-mediated condensation and subsequent
cyclization of the aldehyde with an isothiocyanate affords the
oxazolidine thione, which can be reduced with lithium aluminum
hydride, ozonolyzed, and further reduced with, for example,
sodium borohydride to the diol. Mesylation with
methanesulfonyl chloride and base followed by ring closure with
benzylamine affords the azepine, which is hydrolyzed with an
acid, for example, hydrochloric açid. The resultant anti
aminoalcohol is reacted with a carboxylic acid substituted with
a Z functionality to give the amide, which is further reacted
with a GFE carboxylic function and deprotected to provide the
family of ester/amides with predictable stereochemistry at the
positions of attachment of B1 and D to the ring system.
WO 94/2006~
- PCTIUS9 1/02283
21~7 4~ -28
ô
CL
o~ ~ I I
Z
m
~<O~
--~--LIJ_~
~O~cn z
J Z
~ .
T I oN
> ,_ Z O ~
O=~ c ~ ~ U e~
0=( ~ 5
O ~<O~
N~ 3~ ~> O
O ~ ,~
Z N m
, N
3~ C`' ~.) C~
0~ ~ (~ _
~ Z_~m O O
0~
W094/20062 2 1 ~ 7 ~ 1 2
PCT/US94/02283
- 29 -
The reactions of Scheme VI provide a synthesis scheme
for producing 6, 7, and 8 membered cyclic and heterocyclic
groups including Bl and D with stereo specific attachment
including the use as a starting material of N-carbobenzyloxy
asparagine. For example, CBZ-asparagine is reacted with
bis(trifluoroacetoxy) iodobenzene to give the mono-protected
- diaminoacid, which is differentially protected with di-t-butyl
dicarbonate and reduced with, for example,
borane/tetrahydrofuran. Oxidation to the aldehyde and
condensation with an unsaturated organometallic affords the
diprotected diamino alcohol, which gives the terminal tosyloxy
compound after hydroboration/oxidation and treatment with
toluene sulfonyl chloride. Removal of the butoxycarbonyl is
accomplished by acid treatment, for example, formic acid or
trifluoroacetic acid, followed by removal of the amine-
protecting group, which can be removed with hydrogen.
Selective reaction with a carboxylic acid substituted with a Z
functionality followed by treatment with benzyl chloroformate
and base gives the protected amide, which is further reacted
with a GFE carboxylic function and deprotected to provide the
family of ester/amides with predictable stereochemistry at the
positions of attachment of B1 and D to the ring system.
WO 94/20062 PCT/US91/02283
2 1 5 ~ l 1 2
- 30
Scheme Vl
H~N$~ ~_NHcez DMAP NEI ~~~ ez
NHCeZ ~ ~ S wcr~
HN ~aM5CI , HN
80C n=0-2 80C 80C
l) BBN
2) H~02
3) Tsa
TsO/~ z) tase ~ NHCeZ ~ Pd~OH) ~ N
Azapine 1.
Int~..c~ with alk...~ti~ synthcsis
shown on nc:a shccet
SUBSTITUTE SHEET (RULE 263
W094/20062 21~ ~ k ~ ~ PCT/US94102283
- 31 -
Scheme VII provides a syntheses for producing
compounds according to the present invention including a
synthesis scheme for producing 6, 7, and 8 member cyclic and
heterocyclic groups including Bl and D with stereo specific
attachment including the use as a starting material of
phthalimide alkyl aldehyde. for example, the aldehyde can be
reacted with methyl isocyanoacetate in the present of gold
ferrocene catalyst to give the oxazolidine, which is hydrolyzed
to the diaminohydroxy ester salt with for example, hydrochloric
acid. Base mediated cyclization affords the lactam, which can
be reduced to an aminohydroxy compound. Reaction with a
carboxylic acid substituted with a Z functionality followed by
treatment with benzyl chloroformate and base gives the
protected amide, with is further reacted with a GFE carboxylic
function and deprotected to provide the family of ester/amides
with predictable stereo chemistry at the positions of
attachment of B1 and D to the ring system.
.'O 94/20062
PCT/US94/02283
2~7~12 -32-
Scheme Vll
O Gold F~ ucc~c O
~N~r~ C ?~'CH,CO2CH3 [~ ~; O
O CHO O ,~
H3CO2C ~N"
n=0-2
1) 6NHCI
2) MeOH, HCI
OH `~
(~ .,NH2 MeOH,K2CO3 yH2-HCI
~/ ~ HCI-H2N ~C02CH3
N ~o OH
LAH
OH 1) ZCOCI OH
~'EL3, D~ N~r z
N CBZ
H
Azapinc 1
G-F-E-COCI
~ ~Et3, DMAP
E--F-G
E--F-G O~
o H2, Pd(OH)2 _~ H
~,... ~N~rz lFA.ELOH C8Z
W094/20062 2 ~ ~ 7 ~ ~ 2 rcT~sg~/02283
- 33 -
Scheme VIII A provides a synthesis scheme for
producing compounds according to the present invention
including a synthesis scheme for producing 5 and 6 member
cyclic and heterocyclic groups including Bl and D with syn
attachment including the se of the protected cyclic ketone as
starting material. For example, the ketone is deprotonated and
the enolate aminated to afford the butoxycarbonylamino ketone
which can be stereo specifically reduced with, for example,
sodium borohydride to the syn aminoalcohol. Reactions with a
carboxylic acid substituted with a Z functionality yields the
amide. Further reaction with a GFE carboxylic function
followed by deprotection provides the family of syn
ester/amides.
WO 94/20062 PCTtUS9 1tO2283
7 ~ 1 2 34
o
I
N m
_~ ,O~ZT
Z
m O m ,~ I
._ T
O-~Z~ +' TZ
~ ~0--~Z ~ ~
T _ O
O~Z~ ~
+ + ~
,~ ~ X o X ,.
~ o U U D
-- o~Z~ o o
z
-
W094/20062 21~ PCT~S91/02283
- 35 -
Scheme VIII B provides a synthesis scheme for
producing compounds according to the present invention
including a synthesis scheme for producing 5 and 6 member
cyclic and heterocyclic groups including B1 and D with anti
attachment including the use as starting material of the syn
hydroxyamide from Scheme VIII A. For example, the syn
hydroxyamide can be inverted to anti hydroxyamide by treatment
with carboxylic acid, such as acetic acid, in the present of
triphenylphosphine and diethylazodicarboxylate followed by
treatment with sodium methoxide. Reaction with a GFE
carboxylic function followed by deprotection provides the
family of anti ester/amides.
WO 9~/20062 PCT/US9~/02283
21~7~12 -36-
~
X X IZ ~,
U U O ~ ZI Nl C
U
Q
Z
T~ a~ .,
O ~ JQ -- o
o
D D C~
X ~ T
U O
~N
~ IZ '~
o ~Z ~ 11
u _ E
C~
W094120062 21~ ~ ~12 PCT/US94/02283
- 37 -
Scheme VIII C provides methods for producing compounds
according to the present invention including a synthesis scheme
for producing 5 and 6 member cyclic and heterocyclic groups
including Bl and D with anti attachment including the use as
starting material of the syn hydroxyamide from Scheme VIII A.
For example, the syn hydroxyamide can b~ inverted to anti amino
amide by treatment with trifluoromethane sulfonic anhydride and
sodium azide followed by reduction with, for example, tin (II)
chloride. Reaction with a GFE carboxylic function followed by
deprotection provides the family of anti diamides.
WO 94/2006~
PCT/US9 1/02283
21 ~! 112 -38-
_
I
r ~
o Z.. ~ZT c~l C ¦
U U U Z"
~ ' _
o
t~l N
m .q
Z~Z Z
D O o
~ ~ _
N ~ N
_~ a ~J
W094/20062 2 ~ 3 PCT~S91/02283
- 39 -
Scheme VIII D provides a synthesis' scheme for
producing compounds according to the present invention
including a synthesis scheme for producing 5 and 6 member
cyclic and heterocyclic groups including Bl and D with syn
attachment including the use as starting material of the anti
hydroxyamide from Scheme VIII B. For example, the anti
hydroxyamide can be inverted to syn amino amide by treatment
with trifluoromethanesulfonic anhydride and sodium azide
followed by reduction with, for example, tin (II) chloride.
Reaction with a GFE carboxylic function followed by
deprotection provides the family of syn diamides.
~'O 94/20062
PCT/US9 1/02283
N
O
+ <, 3
U O X ~ Z
,, ~, U
N
T
a
I_~z~ . Z
N
O
~ o ~ E
W094/20062 21~ 7 41~ PCT/US94/02283
- 41 -
Scheme VIII E provides a synthesis scheme for
producing compounds according to the present invention
including producing 5 and 6 member cyclic and heterocyclic
groups including Bl and D with anti attachment including the
use of the protected cyclic epoxide as starting material. For
example, the epoxide is opened with ammonia to provide the anti
aminoalcohol or with azide to provide the azido-alcohol.
Reduction of the latter with, for example, triphenylphosphine
provides the amino alcohol. Reaction with a carboxylic acid
substituted with a Z functionality yields the anti
hydroxyamide. Further reaction with a GFE carboxylic function
followed by deprotection provides the family of anti
ester/amides.
WO 94/20062 rCT/US91/02283
2 1 ~ 7 4 1 2 -42-
D
X m
Ll O U --, GZ I U~l
/ e
x
Z I
~ Z~
N ~
m o
Z--,G n ~ C~L"_
e ~ ~ ~
D <~ --3 z ~
~, o ~ I o
m N ~ z
e ~J5 D
Z~~\ n +~
~
S O~z~
WO9ql20062 21~ PCT/159~;02283
Scheme VIII F provides a synthesis scheme for
producing compounds according to the present invention
including a synthesis scheme for producing 5 and 6 member
cyclic and heterocyclic groups including Bl and D with syn
attachment including the use of the anti hydroxyamide from
Scheme VIII E as starting material. For example, the anti
hydroxyamide can be inverted to syn hydroxyamide by treatment
with carboxylic acid, such as acetic acid in the presence of
triphenylphosphine and diethyl azodicarboxylate followed by
deprotection provides the family of syn ester/amides.
WO 94/20062 44 PCT/US94/02283
2~^~7~
o I ~, N
~-) XO T
~ U --_~
~~
X
U
N ~
0~ 0
~
w
O D
X tL
u
m O
'~QZ E
E
-
C~
W094/20062 2 ~ 12 PCT/US9~/02283
Scheme VIII G provides a synthesis scheme for
producing compounds according to the present invention
including a synthesis scheme for producing 5 and 6 member
cyclic and heterocyclic groups including B~ and D with syn
attachment including the use of the anti hydroxyamide from
Scheme VIII E as starting material. For example, the anti
hydroxyamide can be inverted to syn amino amide by treatment
with trifluoromethanesulfonic anhydride and sodium azide
followed by reduction with, for example, tin (II) chloride.
Reaction with a GFE carboxylic function followed by
deprotection provides the family of syn diamides.
WO 94/20062 PCT/US94/02283
- 46 -
2 1~ l 4 12
O
+ <~31 -~ZI
U ,, U
O ~ ~ _
tO ~3
o
TZ X~Z ~ Z
N ~_
n --I ~ ~ CL
c~ I ~ _
X S
r~ , U
N
E I E E
U,
W094/20062 215 ~ ~ ~ 2 PCT/US94/02283
- 47 -
Scheme VIII H provides a synthesis scheme for
producing compounds according to the present invention
including a synthesis scheme for producing 5 and 6 member
cyclic and heterocyclic groups including the use of the syn
hydroxyamide from Scheme VIII F as starting material. For
example, the syn hydroxyamide can be inverted to anti amino
amide by treatment with trifluoromethanesulfonic anhydride and
sodium azide followed by reduction with, for example, tin (II)
chloride. Reaction with a GFE carboxylic function followed by
deprotection provides the family of anti diamides.
WO 9~/20062
PCTIUS94/02283
2 1 ~ ~ 4 1 2 48
~I tL
Z .~, I
</~
~ x I .. ~
U U X
o ~ U
o
o
OI
a ~ t~
~ C D
Z'--~ Z
~ O
D _, Q~ C
U
~ a
T--~ ~ ~ N z
C~
W094/20062 21~ ~ ~1~ PCT/US9S/02283
- 49 -
Scheme IXA provides a synthesis scheme for producing
intermediate GFE carboxylic acids including the use of 4-
bromobenzoic acids as starting material. For example a 4-
bromobenzoic acid can be esterified by treatment with
carbonyldiimidazole followed by an alcohol such as tert-
butanol. The ester is treated with n-butyl lithium followed by
an addition of N, N-dimethyl formamide to give the aldehyde
ester, or by addition of carbon dioxide to give the acid ester,
which is converted to the acid chloride ester with oxalyl
chloride. The ester can also be reacted with n-butyl lithium
and a benzaldehyde to give the diphenyl carbinol, which is
oxidized with, for example, chromic acid, and deprotected to
provide GFE carboxylic acids. The ester can be reacted with n-
butyl lithium and benzoyl chloride to give the diphenyl ketone,
which is deprotected to also provide GFE carboxylic acids.
WO 94/20062 PCT/US94/02283
21~12
.so.
Scheme rx A L) n-suLiO c~c~2~
Er~CO2H ,, ~,~C02R --
2) CO2~CO2R
~ (COC~).
CIOC~CO2R
X 1) ~ ~CO R
O O
~COzR ~CO2H
Er~ 1) B .I; ~CO2R
o
~CO2H
SUBS~lTUTE SHEET (RULE 2~
2~7412
WO9~/20062 PCT/US91l02283
- 51 -
Scheme IXB provides a synthesis scheme for producing
intermediate GFE carboxylic acids including the use of either
4-bromobenzoic esters from scheme IXA or benzyl alcohols as
starting material. For example, a 4-bromobenzoic ester can be
treated with n-butyl lithium and phthalic anhydride to afford
the 2'-carboxybenzophenone ester, which is protected with, for
example, benzyl alcohol esterification at the 2' position and
deprotected at the other ester position to provide GFE
carboxylic acids. In an alternative, preferred method, the
alcohol is coupled with the acid chloride and treated with one
equivalent of nBuLi which provides, upon rearrangement, the
hydroxy ketone. Oxidation to the acid is effected in a two-
step process using first pyridinium dichromate (PDC) or TEMPO
(2,2,6,6-tetramethyl-1-piperidinyloxy) followed by
tetrabutylammonium permanganate (nBu4MnO4) or sulfamic acid
(NH2SO3H) and sodium chlorite (NaClO2). The resultant acid is
protected, for example, by conversion to the benzyl ester and
then deprotected at the other ester position to provide the
family of GFE carboxylic acids.
21~7 4~ 2
~O 94/20062 PCT/US94/02283
Scheme IX B - 52-
HO2C O
/=\ 1) BuLi ~ protect
~ 2) 5~ Iyl/J x ~CO2R
R'02C O R'02C O
~ dep~tec~, 5~CO2H
HO HO~
Br ~CO2R []
CIOC ~ CO2R ,KOtBu
X
2) nBuLi
HO2C O R'02C O
~ protect ~CO2R
R'02C O
depl~)tect ~CO2H
2~5 ~J~1~
W094/20062 rcTluss4lo2283
- 53 -
Scheme IXC provides a synthesis scheme for producing
GFE carboxylic acids including the use of bromobenzenes as
starting material. For example, a bromobenzene can be treated
with n-butyl lithium and the acid chloride ester from scheme
IXA to afford the benzophenone ester, which is deprotected to
provide GFE carboxylic acids. The bromobenzene can also be
treated with n-butyl lithium and the aldehyde ester from scheme
IXA to give the diphenylcarbinol ester, which is either
deprotected directly, oxidized with, for example, chromic acid
and deprotected, or reduced with, for example, hydrogen and
deprotected to provide the family of GFE carboxylic acids.
2157~1~?
WO 94/20062 PCT/US94/02283 _
- 54 -
Scheme ~X C
~r 1) ~'-r ~ CO R ~COzR
tCy~utc~L ~CO2H
L~.U...,L ~`CO2H
2) ~CO R ~CO2R
O ~ \
~CO2R ~CO2R
d~y~lt~ deprotect
~CO2H ~CO2H
S~I~UTE S! IEE~ (RULE 2~J
2157~12
WO9~/20062 PCT/US91102283
- 55 -
Scheme IXD provides a synthesis scheme for producing
compounds according to the present invention including the use
of halobenzenes, for example, bromobenzenes, and heterocyclic
or cyclic compounds substituted with B1-B2-Z and DH, which are
described in other schemes, as starting materials. For
example, a bromobenzene can be reacted with a 4-heterobenzoate
ester, in dimethyl formamide/triethylamine with heat or copper
catalyst to give GFE acid chloride. This can be treated with
heterocyclic compounds substituted with B1-B2-Z and DH in the
presence of tetrahydrofuran/triethylamine to provide this
family of balanoids.
WO 94/20062
PCT/US9~/02283
- 56 -
21~7~
Scheme ~X D
Arm 1 synthesis (continued):
DMF/Et3N
,Br HQ~ DMFlEt~N/Cu catalyst
Q=O,~H,S
~3,a~ Ester ~ ~3`cocl
X DMF/CH2CI2
HD~ B2--Z y ~CO
D~B, - B2--Z
Et3N/THF A
W094/20062 21~ ~ 4 ~ 2 rcTlus94lo2283
- 57 -
Scheme IXE provides a synthesis scheme f~or producing
compounds according to the present invention including the use
of aromatic or heteroaromatic halides protected, hetero-
substituted aryl or heteroaryl acids or alcohols, and
heterocyclic or cyclic compounds substituted with Bl-B2-X and
DH, which are described in other schemes, as starting
materials. For example, a bromoaromatic can be reacted with a
protected 4-hetero-substituted aryl acid or alcohol in the
presence of transition metal catalyst, for example, copper, and
base to afford protected GFE acid or alcohol, which can be
deprotected and treated with, for example, phosphorous penta
chlorides to provide GFEX chloride. This is reacted with
heterocyclic or cyclic compounds substituted with Bl-B2-Z and
DH to provide another family of balanoids.
WO 94/20062
PCT/US94/02283
2 1 ~
- s8 -
Scheme IX E
Cu catalyst
G Br+ HF E XOR ~ G F E--XOR
or Pd(0) or Ni (0)
E~3N
d~plotccL PCIS
G F E XOH
HD B~B2Z G F E--X\
~. D B1 Bz-Z
G--F--E XCI , \1~
E-,~IT~ / \
~A~)n
W094/20062 2 1 5 7 ~ ~ ~ PCTIUS9~102283
- 59 -
Scheme IX F provides a preferred method for the
preparation of the GFE-CO2H. Intermediate aldehyde is first
protected, for example, as a cyclic acetal, which is then
treated with nBuLi and DMF to afford aldehyde 1. Aryl bromide,
is treated with n-butyl lithium and then aldehyde I to afford
the alcohol. Oxidation using, for example MnO2 followed by
acidic hydrolysis of the acetal gives the ketoaldehyde.
Oxidation to the acid using, for example, sodium chlorite and
sulfamic acid, followed by the appropriate deprotection
sequences affords GFE-CO2H.
wo 94/20062 PCT/US94/02283
2 1 5 ~ 60 -
Scheme IX F
o o
X2 OH
I Y I X~
/=1=\ 1) nBuLi ~/ ~ ~/~ 1) oxidation
2) 1 C~~ c/J~ 2) acid, H20
~/~ 1) o~id~ion ~f/
2)protection /~1~
CO2R3)deprotection / CO2H
x2 x2
W094/20062 21~ 7 412 PCT~S9~/022~3
-
- 61 -
Scheme IX G describes a preferred method for the
preparation of GFE-CO2H where E is substituted with two OH or
OR groups. Protected bis-phenol is treated with n-butyl
lithium followed by aldehyde (prepared in Scheme IX F)
provides, after oxidation with, for example, MnO2 the ketone
intermediate. Following deprotection, the primary alcohol is
oxidized, using, for example MnO2 followed by sodium
chlorite/hydrogen peroxide. Hydrolysis of the acetal followed
by treatment with MnO2, KCN, acetic acid and the alcohol R'-OH
provides the desired GFE-CO2H.
A detailed scheme for benzophenone synthesis is shown
in Scheme IX H.
wo 94/2006~ - 62- PCT/US94/02283
Scheme IX G
IOCH3
O OCH3 y
L~ l)nBuLi
~ OCH3 ~/r CHO O~ OCH3
o
3) oxidation
o o OCH3
1) deprotection ~ H30+
2) oxidation ll o ! _ .
3) esterification ~ ~ / ~ CO2R
~ ~
OCH3
o OCH3 O OCH3
Y ~ 1 1) KCN, AcOH /Y
~/~ ~MnO2, R'OH
CHO O/~2) dep~otection ~ ~ C02R' ~ CO2H
OCH3 OCH3
WO9~/2006~ 2 1 ~ 7 ~ l 2
- PCT/US9~/02283
- 63 -
Scheme IX H
Synthesis of MOM-Protected Benzophenone
CO.M~ CO.Me ~OH
~3~ MOMCI ~ LAH, T~ ~
HO OH MOMO~OMOM MOMOJ~OMOM
OT~DMS
TBDMSCI r
e ~
1~/10~10~0~10~1
HO nnO BnO
K,C03,BnBr ~ 1,3~ ,l).u~ H7~-C
CHO ~ PhMe,~ ~ o
O~
HO hlOMO MOMO
f~q N.-lH, MOMCI ~ "BuLi, C6HI2 ~CHO
n ~MF ~
OTBDMS MOMlo hlOhlO~OTl~DMS
+ ~, I ''BuLi,0''C ~<
MOMO OhlOlM O J .1 ~
WO 9~/20062 PCT/US9 ~/022~3
2 1 ~
~ 64 ~
MOMO MOMO~,OMOM MOMO~OTBDMS
¢~0 OT~DMSh-- OMOM MnO~, CH,CI
o J "BuLi, 0"C. THF
O
MOMO~ OTBDMSMOMO~ OH
MOMO o ~ MOMO o ~ \\
~< TBAF, THF ~ MnO., CH.CI,
OMOM > ~ OhlOM
O O
MOMO ~_~CHO hlOhlO~CO,hle
MO~ KOH, 1, MO~
W~ OhlOlM MeOH. 0"C W~ OM()hl
O O
NaCI02. HtO~ 18% H~SO~
N.aH2PO.~, MeCN-H.O a~l;orbe~ n SiO2
CH2( 12
MOh~O CO~H hlOlMO CO~Me
MO~ MO~
~ OhlOhl W~CHO
WO 94/2006~
~1 J ~ ~ _ ~ PCT/US94/02283
-- 65 --
~,HO MOMO~,OMOM 'Blll i O"C B~ U~OTBDMS
~ ll l OMOM
OOTBDMS ~
o
MlOhlO~OTBDMS MOMO ~--OH
MnO.. CH2CI- n,~ TBAF B~
W~ OMOM W~ OMIOMI
O O
MOMO~ ~CHO M~ lO~ CO2H
BnO O \ \\ BnO ~ \\
MnO-. CH-a2 ~< NsClO2, NaH2PO1 ~ <
H,02,MeCN-H.O l~'~~ OMOM
O O
W094/20062 PCT/US94/02283
2~ 2 - 66 -
Scheme X provides means for produciAg compounds
according to the present invention including the use of
heterocyclic or cyclic compounds substituted with B1-Bz-Z and
DXEFG where F is C=o, which are described in other schemes, as
starting materials. For example, a heterocyclic or cyclic
compound substituted with B1-B2-Z and DXEFG where F is C=O can
be treated with a reducing agent such as sodium borohydride to
afford balanoids where F is CHOH, or it can be treated with a
sulfurizing agent, for example, phosphorus pentasulfide, to
provide the family of balanoids with B2 and/or F equal to C=S.
~ro 9~/20062 21 S 7 412 PCTIUS9~102283
- 67 -
Scheme X
;~ G 1l HE~ G
I E
m=0-3
P4SIOor
Lawesson's
reagent
111 E~G
DC ll3
B~ C_ z
A
X~,X2,X3 in~cpe~ ly = O or S
W094/20062 PCT/US9S/02283
2 1 J7 '~12 - 68 -
Scheme XIA provides a synthesis scheme for producing
compounds according to the present invention including the use
of heterocyclic compounds substituted with B1-B2-Z and DXEFG
where A is NH, described in other schemes, as starting
materials. For example, a heterocyclic compound substituted
with Bl-B2-Z and DXEFG where A is NH can be treated with an
alkylating agent, sulfonylating agent, or acylating agent, for
example acetyl chloride, in the presence of base to provide the
family of balanoids with a substanted nitrogen.
In an alternative approach, Scheme XI B, the R group
can be appended at an earlier stage in the synthesis using
appropriately protected intermediates. For example,
heterocyclic intermediate can be treated with an alkylating
agent, sulfonylating agent or acylating agent, in the presence
of base to provide the intermediate where Rl is not hydrogen.
WO94/20062
2 PCT/US9 ~/02283
Scheme XI A - 69 -
G--F--E--X-D~--B2--Z G--F--E--X-D 31-3z--Z
n( )m Et3N n(~ )m
H Rl
m=1-4,n=14 Rl=lower alkyl, aryl, or JR2 where
m+n~or=5 J=CO,CN, or SO2 and R2=lower
alkyl, aryl, alkylamino, or alkoxy
Scheme XI B
HO~B1 -B2 ~Z ~B1 -B2 ~Z
N Et3N N
H Rl
G--F--E--X-D B1-B2--Z
as described )--(
in Scheme I Rl
2157~1~
WO9~12006' PCT/US94102283
- 70 -
Scheme XII provides a synthesis scheme for producing
compounds according to the present invention including the use
of cyclic or heterocyclic compounds substituted with B1-B2-Z and
DXEFG where X is C=O, which are described in other schemes, as
starting materials. For example, a heterocyclic compound
substituted with B1-B2-Z and DXEFG where is C=O can be treated
with an alkylating agent, for example dimethyl sulfate and an
alcohol, such as methanol, to give the intermediate ketal.
This can be reacted with an organometallic, for example, the
lithium salt of diethyl malonate, to provide the family of
balanoids in which x is C=CR3R4. The ketal can be reacted with
a primary amine, for example, butylamine, to provide the family
of balanoids in which X is C=NR2.
WO 94/2006~ 7
- - 71 - ~ cL ~ PCT/US911022S3
X--\
E /
U~
a
X E X E
O~
G U U
C~
oN
_ o C~.~
m c
E ~X E/
O=~
~ U
~VO91/20062 PCT/US91/02283
57 41~ Scheme XIII provides syntheses for producing compounds
according to the present invention including the use of GFE
carboxylic acids and cyclic or heterocyclic compounds
substituted with B1-B2-Z and DH or B,H and DXEFG, which are
described in other schemes, as starting materials. For
example, a GFE carbinol can be treated with mesyl chloride and
an amine base, such as triethylamine, followed by treatment
with an iodide source such as sodium iodide to afford a GFE
methyl iodide. This can be reacted with a cyclic or
heterocyclic compound substituted with Bl-B2-Z and DH in the
presence of base such as sodium hydride to provide the family
of balanoids in which X is CH2.
The cyclic or heterocyclic compound substituted with
B,H and DXEFG can be reacted with a Z sulfonyl chloride, for
example, benzenesulfonyl chloride, in the presence of base to
provide the family of balanoids in which B2 is SO2.
WO 94/20062 215 7 417 PCT/US9-1/02283
N
o~ I
U
U ~ ' ~,u
z ~ m E
s z I ' X~s
X
u
~ U Z
E
U~ .
W094/20062 PCT/US94/02283
21~7~ 74 -
Scheme XIV A provides a synthesis scheme for producing
intermediates which are heterocyclic compounds substituted with
Bl=NH2 and D=OH and in which A is oxygen. For example, a cyclic
diene such as cyclopentadiene can be treated with a peracid
such as peracetic acid and the epoxide opened with an amino
species, for example, (dibenzylamino) dimethyl aluminum,
followed by butyldimethylsilyl chloride to obtain the protected
amino alcohol. This can be ozonolyzed and reduced to the diol,
which is treated with tosyl chloride to give the chloro
tosylate. Treatment with bis(trimethyltin)oxide or preferably
potassium superoxide and a crown ether, for example, 18-C-6,
provides the chloroalcohol which is ring closed on treatment
with a strong base, such as methyllithium or butyllithium to
afford the protected heterocyclic compound, which is
deprotected to heterocyclic compounds substituted with B~=NH2
and D=OH and in which A is oxygen.
~'O 91/2006' 5 2 1 5 7 4 1 ~ PCT/US9 ~/02283
Scheme XIV A
1) Bn2NAIMe2 NBn2
Ac02H ~ 2) TBDMS-CI ~
1-3 1-3 ~ 'OTBDMS
1) 03 OTBDMS OTBDMS
2) NaBH4 Ts-CI .
HO~OH ~ Cl ~OTs
NBn2 NBn2
2) BuLi, toluene, heat TBDMSOs NBn2 [H] TBDMSOs NH2
n(i~)m n(~ )m
F- HOs NH2
n(~ )m
2 1~7 ~ ~/20062 - 76 - PCT/US9~/02283
Scheme XIV B provides a synthesis scheme for producing
intermediates which are heterocyclic compounds substituted with
B1=NH2 and D=OH and in which A is sulfur. For example, a cyclic
diene such as cyclopentadiene can be treated with a peracid
such as peracetic acid and the epoxide opened with an azide
species, for example, sodium azide, followed by t-
butyldimethylsilyl chloride to obtain the protected amino
alcohol. This can be ozonolyzed and reduced to the diol, which
is treated with mesyl chloride to give the dimesylate and ring
closed with, for example, bis(trimethyltin) sulfide or
preferably, lithium sulfide and an amine base such as
triethylamine. Reduction of the azide and deprotection occurs
on treatment with lithium aluminum hydride to give heterocyclic
compounds substituted with Bl = NH2 and D = OH and in which A
is sulfur.
Compounds where A is SO2 can be prepared from
compounds where A is sulfur by treatment with an oxidizing
agent, for example, peracetic acid,followed by deprotection to
provide compounds wherein A is SO2 (Scheme XIV C).
WO 94/20062 21 S 1 ~ ~ ~ PCT/US91/02283
- 77-
Scheme XIV B
1) NaN3, NH4CI, CH30H/N3
Ac02H ~< 2) TBDMS-CI
1-3 1-3 ~ "OTBDMS
1) 03 OTBDMS OTBDMS
2) NaBH4 . Ms-CI
HO~OH ~ MsO ~OMs
N3 NBn2
1 ) Li2S, Et3N, CH30H HO NH2
2)LiAlH4
S
-'O 9~/~006'
- - PCTAJS91/02283
Scheme X~V C - 78 - `-
215~
G-F-E-X-D B1-B2-Z G-F-E-X-D B1-B2-Z
/ /
___ 1) RCO3H ~ A
S~ )m 2) De~rut~c~ion n( ~ ~ )m
O O
~ro94/20062 215 ~ ~1~ PCT/US94/02283
- 79 -
Scheme XV provides a synthesis scheme for producing
compounds according to the present invention including the use
of Z sulfonyl chlorides, such as benzenesulfonyl chloride, and
cyclic or heterocyclic compounds substituted with Bl=NH2 and DH,
which are described in other schemes, as starting materials.
For example, a Z sulfonyl chloride and a cyclic or heterocyclic
compound substituted with Bl=NH2 and DH are combined in the
presence of base to afford the sulfonamide, which is reacted
with GFEX halide to provide the family of balanoids in which B
is N and B2 is SO2.
WO 94/2006' PCT/US93/022S3
2 ~r'J - 80 -
Scheme XV
ol o
D NH2 Cl--S--Z D N--S--Z
m( ~ ) n Et~N m( ~ ) n
G F E X~ H ¦¦
G - F E--X--Cl D N--S--Z
m( ~A~ ) n
G--F E X~ R
D N_S--Z
K2CO3, R2X \~ ( o
m( g~A~ ) n
W094/20062 2 1 5 ~ ~ ~ ~? PCT/US9S/02283
- 81 -
Scheme XVI provides a synthesis scheme for producing
compounds according to the present invention including the use
of ketones, for example, acetophenone, and azepinediones, such
as 1-benzylazepin-2,4-dione, as starting materials. For
example, the ketone can be brominated with bromine in acetic
acid and protected with ethylene glycol to afford the
bromomethyl ketal, which is added to a base-treated solution of
the azepinedione to give the alkylated azepinedione. This is
reduced to the hydroxyazepine with, for example, lithium
aluminum hydride, and reacted with base and GFEX halide, which
is described in other schemes, then deprotected to provide the
family of balanoids where Bl is CH2, Bz is C=O, and D is O.
These compounds can be reacted with, for example, phosphorus
tetrasulfide, to provide the family of balanoids where Bl is
CH2, B is C=S and D is O.
WO 94/2006'
- PCT/US94/02283
2 1 5 ~ 82 -
Scheme XVI
--'6Z Br2, HOAcBr~_~Z HOCH2CH20H, Br 7~
O o pTsOH, Benzene, O J
Dean-Stark Trap
~,~ 1 ) NaH ~ LIAIH~
G--F E X
OH O
(~<Z 1.) NaH (~<Z
N \J 2-~ Cl X E--F--G N \_l
Rl R
G--F E X
1.) PPTS, Acetone
H20 , (~' P4S, o
N
G--F E X\
o
(~Z
N S
-'09~/20062 ~ 2 PCT~S9~10228
- 83 -
Scheme XVII provides a synthesis scheme for producing
compounds according to the present invention including GFE
carboxylic acids, which are described in other schemes, and
cyclic or heterocyclic groups substituted with OH and BlB2Z,
also described in other schemes, as starting materials. For
example, a GFE carboxylic acid can be treated with oxalyl
chloride and N,O-dimethylhydroxylamine to afford the
methoxymethyl amide, which is reacted with a methyl
organometallic to give the methyl ketone. This is then
deprotonated with base and reacted with the product from
treatment of a cyclic or heterocyclic group substituted with DH
and BlB2z with tosyl chloride to provide the family of balanoids
with D as CH2 and X as C=O.
WO 9~/2006~
- PCT/US9~/022~3
- 84-
2157~12
Scheme XVII
?
) (COC1~2/D~
CH2Cl2 ~'.~"
G F--E--CO2H ~ G F--E--CON(OMe)(Me)
2) MeNHOMe
CH3M
M=Li, MgBr
G F E--COCHa
HO B~ B2Z TsCI TsO B~ B2Z
(~)n ~ (~) LiN(SiMe3)2
G--F E--COC~Bl B2Z
(b~A~)n
W094/2006~ 215 i ~ CT/US94/02t~3
- 85 -
Scheme XVIII provides a synthetic scheme for compounds
of the invention where K is not equal to H. Oxidation of the
alcohol using, for example, oxalyl chloride and
dimethylsulfoxide (Swern oxidation) followed by addition of an
organometallic reagent, for example, the complex of
trimethylaluminum and methyl magnesium chloride to afford the
tertiary alcohol. This intermediate is converted using the
Scheme I to compounds of the invention.
wO 9~/2006~
- 86 - PCTIUS9 1102283
Scheme XVIII
2157~12
HO B1 - B2--z 1 ) oxalyl chloride HO ~ B1 - B2--Z
)~ DMSO, Et3N )~
A/~ 2) Me3AI, MeMgCl Q\A/Q
as described
Targets where K = CH3
in Scheme I
~094/2006~ 21 S 7 41~ PCT/US94/02283
- 87 -
Scheme XIX provides a synthetic scheme for compounds
of the invention where the group G is substituted with an
alkoxycarbonyl group. Compounds wherein G is substituted with
a carboxy group are treated with an alkylating agent, for
example, methyl iodide, and a base such as sodium carbonate to
provide the target compounds.
WO 9~/20062 - 88 - PCT/US9~1/022~3
~cheme ~lX
21S7412
C~X~
X D K B IB2Z R-halo
~A~ )m K2CO3, DMF
OH
Y I X
X~
x2 DK B IB2Z
n(~ )m
where R = alkyl, substit-1tPd alkyl
wo 9~,2006? 2 1 S 7 4 1 2 PCT/US9~/02283
- 89 -
Scheme XX provides a synthetic route for compounds of
the invention where G and/or E residues are substituted with
acyloxy groups. Target compounds, which possess one or more
hydroxyl groups on G or E is treated with an acylating agent,
for example, acetyl chloride, ethyl chloroformate, and the
like, in the presence of a base such as pyridine or
triethylamine to provide the target compounds.
WO 94/2006 2 PCTIUS9 ~/022~3
Scheme XX ~ 90 ~
lS~ 41~ o
OH HO~ X 1 ) _1l
D K B IB2Z R Cl
pyridine
~A~ 2) dep~~ L
~O~o~X~ ~
o~L O~ D~ B~B2Z
n(~A~ )m
~'O9~/20062 21~ 7 ~12 PCT/US9~/022X3
-- 91 --
Scheme XXI provides a synthetic scheme for the
preparations of compounds where Bl is N-R and B2 is C=O.
Intermediate amide, following protection of the D
functionality, is treated with a strong base such as KOtBu or
KH and an alkylating agent such as methyliodide or
dimethylsulfate to provide the intermediate where B1 is N-R.
This can be converted using the procedures outlined in Scheme
I to afford the target compound.
WO 9~/2006~
2 PCT/US9`1/02283
Scheme XXI ~ 9
2157412
HD K N~ 1) Protection HD K N~
\)~( o 2) strongbase, \~4 0
~ A/~ alkylating agent
as described
target where R = lower alkyl
in Scheme I
W094/20062 2 1 5 7 4 1 2 PCT/US91/02283
- 93 -
Scheme XXII describes the synthesis of compounds of
the invention in which group G is substituted with a tetrazole
ring. Keto aldehyde (prepared as described in Scheme IX F) is
treated with hydroxylamine hydrochloride in dimethylformamide
to provide the nitrile. Following deprotection to the acid, it
is coupled to provide target compounds wherein G is substituted
with a nitrile group. Treatment with trimethylsilylazide and
nBu2SnO followed by deprotection affords the target compounds.
WO 91/20062 PCT/US9~/02283
Scheme ~Xll 94 ~
2157412
1 HONH2-HCl ~9/l----¢~/
~CHO 0~CO2R CN /J~CO2R
t~D K B ,B2Z CN 5~ ~
~4 X~ D K B ,B2Z
n(~ )~ )m n(~)m
) TMSN3 Y ll x'
nBu2SnO ~/~
2) Deprotect ~
D K B~B2Z
n(~A~ )m
WO9~/2006' 21~ 7 ~12 PCT/US9~/02283
- 95 -
Scheme XXIII provides a synthetic scheme for the
preparation of compounds where D is N-R. Amine intermediate
(prepared as described in Scheme III) is converted to the
trifluoroacetamide. Treatment with a strong base, such as
KOtBu and an alkylating agent such as methyl iodide or
dimethylsulfate followed by cleavage of the trifluorocetamide
provides the intermediate amine wherein D is N-R. This is
converted to compounds of the invention using procedures
outlined in Scheme III.
WO 9~/20062
n~ PCT/US9~1/02283
Scheme XXIII - ~u- -
Z157412 o
1~
NH2 B1-B2--Z / CF3
\ HN Bl-B2--Z 1) KOtBu
n(~ )m 1 ) TFA, CDI )_~ (CH3)2S04
g\ A/~ 2) KOH, CH30H
~ CH3
HN B1-B2--Z
as described
, Targets where D = -NCH3
n(~ ~A/Q)m in Scheme I
W094/20062 215 7 412 PCT/US91/02283
- 97 -
Persons of ordinary skill in the art will recognize
that the foregoing schemes are exemplary only and should not be
construed to be limiting.
Pharmaceutical preparations incorporating compounds
according to the present invention can be used to block PKC
activity related to abnormal or undesirable cellular events and
activity including tumorogeneis and cellular activity related
to inflammation and reperfusion injury. Treatment of disorders
and disease conditions can be performed by administration of
effective amounts of pharmaceutical preparation that comprise
compounds according to the present invention. Compounds can be
formulated for human and animal prophylactic and therapeutic
applications by those having ordinary skill in the art. The
range of amounts of a compound to be administered to mammals,
lS particularly humans, to be effective in inflammatory, tumor or
reperfusion injury therapy can routinely be determined by those
having ordinary skill in the art.
The compounds and pharmaceutical compositions of the
invention may be administered by any method that produces
contact of the active ingredient with the agent's site of
action in the body of a mammal or in a body fluid or tissue.
These methods include but not limited to oral, topical,
hypodermal, intravenous, intramuscular and intraparenteral
methods of administration. The compounds may be administered
singly or in combination with other compounds of the invention,
other pharmaceutical compounds such as chemotherapeutic
compounds, or in conjunction with therapies such as radiation
treatment. The compounds of the invention are preferably
administered with a pharmaceutically acceptable carrier
selected on the basis of the selected route of administration
and standard pharmaceutical practice.
The compounds of the invention are administered to
mammals, preferably humans, in therapeutically effective
amounts which are effective to inhibit protein kinase C, to
inhibit tumor cell growth, inhibit inflammation of tissue,
inhibit keratinocyte cell proliferation, inhibit oxidative
burst from neutrophils or inhibit platelet aggregation. The
W094/2006' PCT/US94/02283
2157~1~ I 98 -
dosage administered in any particular instance will depend upon
factors such as the pharmacodynamic characteristics of the
compound of the invention, its mode and route of adminis-
tration; age, health, and weight of the recipient; nature and
extent of symptoms; kind of concurrent treatment, frequency of
treatment, and the effect desired.
Compounds according to the present invention inhibit
the activity of PKC in cells. The range of the amount of
inhibitory compound that is effective for inhibiting PKC
activity can be determined by one having ordinary skill in the
art. By inhibiting PKC activity, balanoids are useful in the
treatment of disease conditions in which control of cellular
growth, regulation and/or differentiation is desirable. An
effective amount of a balanoid can be administered to mammals
who are suffering from inflammatory, cardiovascular or
neoplastic diseases, particularly inflammation, reperfusion
injury and cancer, in order to counter the disease at the
cellular level.
It is contemplated that the daily dosage of a compound
of the invention will be in the range of from about 1 ~g to
about lOO mg per kg of body weight, preferably from about 1 ~g
to about 40 mg per kg body weight, more preferably from about
lO ~g to about 20 mg per kg per day.
Pharmaceutical compositions of the invention may be
administered in a single dosage, divided dosages or in
sustained release forms. Persons of ordinary skill will be
able to determine dosage forms and amounts with only routine
experimentation based upon the considerations of this inven-
tion. Isomers of the compounds and pharmaceutical
compositions, particularly optically active stereoisomers, are
also within the scope of the present invention.
The compounds of the invention may be administered as
a pharmaceutical composition orally in solid dosage forms, such
as capsules, tablets, and powders, or in liquid dosage forms,
such as elixirs, syrups, and suspensions. The compounds may
also be administered parenterally in sterile liquid dosage
forms or topically in a carrier. The compounds of the
~094/20062 2 1 5 7 ~ 1 2 PCT/US94/02283
_ 99 _
invention may be formulated into dosage forms according to
standard practices in the field of pharmaceutical preparations.
See ~emington's Pharmaceutical Sciences, A. Osol, Mack
Publishing Company, Easton, Pennsylvania.
Compounds of the invention may be mixed with powdered
carriers, such as lactose, sucrose, mannitol, starch, cellulose
derivatives, magnesium stearate, and stearic acid for insertion
into gelatin capsules, or for forming into tablets. Both
tablets and capsules may be manufactured as sustained release
products for continuous release of medication over a period of
hours. Compressed tablets can be sugar or film coated to mask
any unpleasant taste and protect the tablet from the atmosphere
or enteric coated for selective disintegration in the
gastrointestinal tract.
Liquid dosage forms for oral administration may
contain coloring and flavoring to increase patient acceptance,
in addition to a pharmaceutically acceptable diluent such as
water, buffer or saline solution.
For parenteral administration, a compound of the
invention may be mixed with a suitable carrier or diluent such
as water, an oil, saline solution, aqueous dextrose (glucose)
and related sugar solutions, glycols such as propylene glycol
or polyethylene glycols. Solutions for parenteral
administration contain preferably a water soluble salt of the
compound of the invention. Stabilizing agents, antioxidizing
agents and preservatives may also be added. Suitable
antioxidizing agents include sodium bisulfite, sodium sulfite,
and ascorbic acid, citric acid and its salts, and sodium EDTA.
Suitable preservatives include benzalkonium chloride, methyl-or
propyl-paraben, and chlorbutanol.
Animal studies have shown that perhaps 50% or more of
ischemic-related myocardial damage can be attributed to
polymorphonuclear leukocytes (neutrophils) which accumulate at
the site of occlusion. Damage from the accumulated neutrophils
may be due to the release of proteolytic enzymes from the
activated neutrophils or the release of reactive oxygen
intermediates (ROI). Much of the "no reflow" phenomenon
W094/20062 PCT/US94/02283
2,15~12 - 100 -
associated with myocardial ischemia is attributed to myocardial
capillary plugging. The plugging of capillaries has been
attributed to both aggregated platelets and aggregated
neutrophils. Although both cell types are aggregate during the
ischemic event, the relative contribution of each to capillary
plugging has not yet been established. It is accepted that the
damage by neutrophils to myocardial tissue proceeds through a
cascade of events, one of the earliest being the bonding of
activated neutrophils to damaged vascular endothelium. However
the binding of the neutrophils is significantly enhanced by
their activation. Thus, an even earlier event is the
generation of molecules (such as cytokines, and chemotactic
factors) which can function as activation stimuli. These
molecules probably originate from damaged and aggregated
platelets, from damaged vascular endothelium, or from the
oxidation of plasma proteins or lipids by endothelial-derived
oxidants.
Strategies for overcoming the deleterious effects of
reactive oxygen intermediates have centered on the development
of scavengers for the molecules. Superoxide dismutase (SOD)
has been shown to be a particularly effective scavenger of
superoxide, but suffers from a very short half-life in the
blood. Several companies have approached this problem by
creating versions of the enzyme with increased half-lives by
techniques such as liposome encapsulation or polyethylene
glycol conjugation. Reports on the effectiveness of these new
versions are mixed. Catalase, a scavenger of hydrogen
peroxide, and hydroxyl radical scavengers have also been tested
and found to be effective to varying degrees. However, none of
the strategies designed to scavenge reactive oxygen
intermediates will prevent the aggregation of platelets, the
release of chemotactic molecules, the activation and adherence
of neutrophils to vascular endothelium, or the release of
proteolytic enzymes from activated neutrophils.
One advantage of protein kinase C inhibitors as
therapeutics for reperfusion injury is that they have been
demonstrated to: 1) block platelet aggregation and release of
W094l20062 21~ 7 ~12 PCT/US94102283
-- 101 --
neutrophil activating agents such as PAF; 2) block neutrophil
activation, chemotactic migration, and adherence to activated
or damaged endothelium; and 3) block neutrophil release of
proteolytic enzymes and reactive oxygen intermediates. Thus,
these agents have the capability of blocking all three of the
most significant mechanisms of pathogenesis associated with
reperfusion injury and should thus have a decided therapeutic
advantage.
The table below contains a list of compounds according
to the present invention. For convenience, in some cases the
substituient groups described in their noun form rather than as
adjectives, e.g. pyridine rather than pyridyl. It is to be
understood that, unless specified, points of attachment of
these functional groups can be any which are typically found in
organic chemistry. Thus, for example, recitation of "pyridine"
as a substituient comprehends attachment at the 2, 3, or 4
position .
Cmpd. A 81 ~ 2 Z D E F G X K m n
NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3
phenyl
2 NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3
phenyl
3 NH NH SO2 p-methyl phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3
phenyl
4 NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO phenyl CO H 1 3
phenyl
CH2 NH CO p-hydroxy phenyl 0 2,6-dihydroxy CH2 phenyl CO H 1 3
phenyl
6 CH2 NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-hydroxy phenyl CO H 1 3
phenyl
7 CH2 NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-carboxy phenyl CO H 1 `3
phenyl
8 NH NH CO p-hydroxy phenyl 0 2,6-dihydroxy CO 2-carboxy phenyl CO H 1 3
phenyl
9 NH NH CO pyridine 0 3,5-dihydroxy CO 2-- I,UAy 6- CO H 1 3
phenyl hydroxy benzene
NH NH CO pyrrole 0 3,5-dihydroxy CO 2-~.~ b~)A~6- CO H 1 3
phenyl hydroxy phenyl
11 NH NH CO oxazole 0 3,5~" 'hoA~ CO 2-- bUAY 6- CO H 1 3
phenyl methyl phenyl
12 NH NH CO indole 0 3-hydroxy-5- CO 2-hydroxy-6- CO H 1 3 r
`h~l~ h_.,~1 methylphenyl
13 NH NH CO purine 0 3 1 `h y 5- CO 2,6-dimethyl CO H 1 3 i~
hJII h~ l phenyl
Cmpd. A 8 1 ~ 2 ¦ D E F ¦ G ¦ X K ¦ m n
14 NH NH CO ~uran O 3-hydroxy CO 2,6-~tn hvAy CO H 1 3
phenyi phenyi
NH NH CO lI,k~i-,_.,e O 3,5-dihydroxy CO 2,6-dihydroxy CO H 1 3
phenyi phenyi
16 NH NH CO PJ-i ' ' ,e O 3,5~1n 'hGA~ CO 2~ L.oAf 6- CO H 1 3
phenyl hydroxy benzene
17 NH NH CO ,~".i ,hli.,e O 3-hydroxy-5- CO 2~arboxy4- CO H 1 3
. . ._lhJ4 1 ~i hydroxy benzene
18 NH NH CO pyrazine O 3.5~' '~ Y CO 2~ IJVA~ 6- CO H 1 3
phenyi hydroxy benzene
19 NH NH CO imidazole O 3-hydroxy-5- CO 2~ LVA~r 6- CO H 1 3 ~
... `hy4h_.,~1 hydroybenzene ~ i--
NH NH CO Ihiazole O 3,5-dihyroxy CO 2- I,oAf 6- CO H 1 3 ~ CJ~
benzene hydroxy benzene w
21 NH NH CO isoxazole O 3,5-d' . IhuAy CO 2~ boAy 6- CO H 1 3 ~ i~
phenyi hydroxy benzene l~
22 NH NH CO pyrazole O 3,5~dihydroxy CO 2- bvAy 6- CO H 1 3
phenyi hydroxy benzene
23 NH NH CO i~ r~l',' 'e O 3,5-dihydroxy CO 2~- LUA~6- CO H 1 3
benzene hydroxy benzene
24 NH NH CO benzene O 3,5~" . hvAy CO 2~ bOAY 6- CO H 1 3 phenyi hydroxy phenyl
NH NH CO methyl benzene O 3-hydroxy-5- CO 2,6-dimethyl CO H 1 3 ~?
,.. _lh,1~ h_.. ~1 phenyi r~
26 NH NH CO dimethyl benzene O 3,5~ ./ CO 2,6~di,.. _1hvAy CO H 1 3 0
phenyl phenyi i~-
27 NH NH CO trimethyi benzene O 3,5-dihydroxy CO 2~arboxy-6- CO H 1 3
benzene hydroxy benzene
~.:~ o
Cmpd. A dl ¦ ~ Z ¦ D E F ~I ¦ X K m ¦ n ~ æ
28 NH NH CO Mr~.. h~1 0 3-hydroxy-5- CO 2~arboxy~ CO H 1 3
hyl~,h~" /l hydroxy benzene
29 NH NH CO ethyl benzene 0 3,5-dihydroxy CO 2-carboxy~ CO H 1 3
benzene hydroxy phenyl
NH NH CO tetraethyl benzene 0 3-hydroxy CO 2~ OAY 6- CO H 1 3
phenyl hydroxy benzene
31 NH NH CO propyl benzene 0 3,5-dihydroxy CO 2,6-dihydroxy CO H 1 3
phenyl phenyl
32 NH NH CO tetra propyl 0 3,5-dihydroxy CO 2-carboxy~ CO H 1 3
benzene benzene methyl phenyl
33 NH NH CO butyl benzene 0 3-hydroxy-5- CO 2-carboxy-6- CO H 1 3
methoxy phenyl hydroxy benzene
34 NH NH CO tetrabutyl benzene 0 3,5-dihydroxy CO 2~ I,oAt 6- CO H 1 3
benzene hydroxy benzene
NH NH CO pentyl benzene 0 3,5-dihydroxy CO 2-carboxy-6- CO H 1 3
benzene hydroxy benzene
36 NH NH CO t~ benzene 0 3,5-dihydroxy CO 2,6t'. hùAy CO H 1 3
phenyl phenyl
37 NH NH CO methoxy benzene 0 3,5-dihydroxy CO 2,6-dimethyl CO H 1 3
benzene phenyl
38 NH NH CO di~ hUAy benzene 0 3,5~i,.. 4hUAt CO 2~ LOAY 6- CO H 1 3
phenyl methylphenyl
39 NH NH CO td~ thoAy benzene 0 3,5-dlhydroxy CO 2~ I~OAY 6- CO H 1 3
benzene methoxy phenyl u
NH NH CO tetra methyl 0 3-hydroxy-5- CO 2-carboxy~- CO H 1 3 ~,
benzene methoxy phenyl hydroxy phenyl x~
41 NH NH CO ethoxy benzene 0 3,5-dihydroxy CO 2; t~oAy 6- CO H 1 3
benzene hydroxy phenyl
Cmpd ¦ A E1¦ d l Z ¦ D E F G X K m n
42 NH NH CO dlethoAy benzene 0 3-hydroAy-S- CO 2-- boA~ 6- CO H 1 3
methyl phenyi methoxy phenyi
43 NH NH CO nitro benzene 0 3,5-dihydroAy CO 2,6-dimethyl CO H 1 3
benzene phenyl
44 NH NH CO dinitro benzene 0 31 ~h y 5- CO 2~ bUAY 6- CO H 1 3
methyi phenyi hydroxy benzene
NH NH CO halo benzene 0 3,5-dihydroAy CO 2~arboAy4- CO H 1 3
benzene hydroxy phenyi
46 NH NH CO dihalo benzene 0 3-hydroxy CO 2-carboxy4- CO H 1 3
phenyl hydroxy phenyi
47 NH NH CO trihalo benzene 0 3,5-dihydroxy CO 2-carboxy4- CO H 1 3
phenyl hydroxy phenyi I 1
4~ NH NH CO tetrahalo benzene 0 3,5-dihydroxy CO 2~ bUA~ 6- CO H 1 3
benzene methoxy phenyl --
49 NH NH CO benzene_ bUA~1 0 3,5-~L. huAy CO 2-carboxy4- CO H 1 3 1
acid phenyi methoxy phenyi
NH NH CO benzene 0 3,5-dihydroxy CO 2~arhoAy4- CO H 1 3
d;~IIIUAYI;~ acid benzene methyi phenyi
51 NH NH CO b~" .id~ amido 0 3 m 'h /5- CO 2-hydroxy4- CO H 1 3
benzene methyi phenyl methyi phenyi
52 NH NH CO benzene dbmide 0 3-hydroxy CO 2,6-dimethyi CO H 1 3
phenyi phenyi
53 NH NH CO phenol 0 3,5-dihydroxy CO 2,6-d;,,,~lhuAy CO H 1 3 c
benzene phenyi u
54 NH NH CO dihydroxy benzene 0 3,5-dihydroAy CO 2,6~ihydroxy CO H 1 3
phenyi phenyl
NH NH CO trihydroxy benzene 0 3,5-dihydroxy CO 2-carboxy4- CO H 1 3
benzene methoxy phenyi
Cmpd. A dl E2 ¦ Z D ¦ E F G X K m n l~
56 NH NH CO pe... 'hJd~vA~ O 3hJd~uA~t5- CO 2~ LUA~6- CO H 1 3
benzene methyi phenyi methoxy phenyi
57 NH NH CO triethoAy benzene 0 3,5-dlhydroxy CO 2,6-dimethyi CO H 1 3
benzene phenyi
58 NH NH CO tetra ethoxy O 3-hydroxy CO 2,6-dihydroxy CO H 1 3
benzene phenyi phenyi
S9 NH NH CO propoxy benzene 0 3,5-dihydroxy CO 2 carboxy~ CO H 1 3
benzene hydroxy phenyi
NH NH CO ~ lU~UAy benzene 0 3n.- .y5- CO 2~arboAy~ CO H 1 3
methyi phenyi hydroxy phenyi
61 NH NH CO t~i~,u~oAybenzene O 3,5-dihydroxy CO 2-t~rboxy-6- CO H 1 3
benzene methyi phenyi
62 NH NH CO tetra propoAy O 3,5-dihydroxy CO 2,6-" hùA~t CO H 1 3
benzene phenyi phenyi tJ
63 NH NH CO amidine 0 3,5-dihydroAy CO 2 carboxy~ CO H 1 3
phenyi hydroxy phenyi
64 NH NH CO dbmino benzene 0 3,5-dihydroxy CO 2-1,/.' UAY 6- CO H 1 3
benzene methyi phenyi
NH NH CO methoxy pyridine 0 3,5-dihydroxy CO 2-o bVA'Y 6- CO H 1 3
benzene methyi phenyi
66 NH NH CO di,.. _lhuAy pyndine 0 3-hydroxy-5- CO 2-n jJUXY 6- CO H 1 3
methoxy phenyi methoxy phenyi
67 NH NH CO hydroxy pyridine 0 3,5-dihydroxy CO 2,6-dihydroxy CO H 1 3 c
benzene phenyi v
68 NH NH CO dihydroxypyridine O 3,5-dihydroxy CO 2,6t-i.. ,~ll.GAy CO H 1 3
benzene phenyi
69 NH NH CO ethoAy pyrrole 0 3,5-dihydroxy CO 2-hydroxy-6- CO H 1 3
phenyi methyi phenyi
¦Cmpd. A El ¦ E2 D E F G X ¦ K m n
NH NH CO dihydroxy pyrrole 0 3,5-dihydroAy CO 2-carboAy4- CO H 1 3
benzene hydroAy phenyl
71 NH NH CO dim h~JAy Indole 0 3-hydroAy-S- CO 2~arboAy4- CO H 1 3
methoxy phenyl hydroxy phenyl
72 NH NH CO hydroAy purine 0 3,5-dihydroAy CO 2-. bUA~ 6- CO H 1 3
benzene me~hyl phenyl
73 NH NH CO ~ hUAy ~uran 0 3: ~h J5- CO 2~ bUAy 6- CO H 1 3
methyl phenyl me~hoAy phenyl
74 NH NH CO hydro)yll,k~h~.,e 0 3,5-dihydroAy CO 2,6-dime~hyl CO H 1 3
benzene phenyl
NH NH CO melhoAy p~ e 0 3,5-dihydroAy CO 2.6-dihydroxy CO H 1 3
benzene phenyl
76 NH NH CO 'i~.. lhùAy O 3-hydroAy CO 2,6-~- hùAy CO H 1 3 ~ r~
~".iJ~;"e phenyl phe~nyl ~1
77 NH NH CO hydroxy~,.;'".e O 3,5-dihydroAy CO 2-carboAy4- CO H 1 3 1 -~
phenyl hydroAy phenyl
78 NH NH CO diamido pf~ . e O 3,5-dihydroAy CO 2-carboxy4- CO H 1 3 J~
benzene hydroxy phenyl
79 NH NH CO amido pyrazine 0 3,5-'.lhuA1 CO 2~ bùAt 6- CO H 1 3
phenyl methoAy phenyl
NH NH CO dielho)ty pyrazine 0 3 5-dihydroAy CO 2-carboAy4- CO H 1 3
benzene hydroxy phenyl
81 NH NH CO p-hydroAy phenyl O pyndine CO 2-~rboAy4- CO H 1 3 c
melhyl phenyl v
82 NH NH CO p-methoAy phenyl O pyrrole CO 2-hydroAy4- CO H 1 3
me~hyl phenyl
83 NH NH CO p-chloro phenyl O oxazole CO 2,6-~" huAy CO H 1 3
phenyl
Cmpd. A E~ Ez Z D E F G X K m n ~,
84 NH NH CO~ hUA~ phenyi O indole CO 2,6-dihydroxy CO H 1 3
phenyi
NH NH CO .. hJd~UA~ phenyi O purine CO 2- buAf 6- CO H 1 3
hydroxy phenyi
86 NH NH COm-chloro phenyi O furan CO 2-. buA~6- CO H 1 3
hydroxy phenyi
87 NH NH CO o-methoxy phenyi O Ihlu~ .. e CO 2-hydroxy4- CO H 1 3
methyi phenyi
88 NH NH CO o-hydroxy phenyi- O ~,.; ' ,e CO 2,6-dimethyi CO H 1 3
phenyi
89 NH NH CO o-chloro phenyi O ~".i ' .e CO 2,6-. . hùAy CO H 1 3
phenyi
NH NH CO p-hydroxy phenyi O pyrazine CO 2-carboxy4- CO H 1 3 o
methoxy phenyi I
91 NH NH CO m-methoAy phenyi O imidazole CO phenyi CO H 1 3
92 NH NH CO m-hydroxy phenyi O Ihiazole CO 2-carboxy4- CO H 1 3
methyi phenyi
93 NH NH CO p-methoAy phenyi O Isoxazole CO 2-carboxy4- CO H 1 3
hydroxy phenyi
94 NH NH CO o-methoAy phenyi O pyrazole CO 2-carboxy4- CO H 1 3
hydroxy phenyi
NH NH CO m-hydroA~ phenyi O Is~tl,' 'e CO 2-carboxy4- CO H 1 3
hydroxy phenyi u
96 NH NH CO o-methoAy phenyi O benzene CO 2-carboxy4- CO H 1 3 o
methyi benzene
97 NH NH CO o-chloro phenyi O methyibenzene CO 2-carboxy4- CO H 1 3
methyi benzene
CmDd. A E1 E2 Z D E F ~ X ¦ K m n
98 NH NH C0 p-hydroxy phenyl O dimethyl CO 2~ bUA'y6- CO H 1 3
benzene hydroxy phenyl
99 NH NH C0 p-chloro phenyt O trlmethyl CO 2-carboxy-6- CO H 1 3
benzene me~hyl phenyl
100 NH NH C0 m-hydroxy phenyl 0 t- ... lh~ CO 2-l,JJ~uA~6- CO H 1 3
methyl phenyl
101 NH NH C0 ... m lhuAy phenyl O ethylbenzene CO 2,6-Ji~ hUAy CO H 1 3
phenyl
102 NH NH C0 p-hydroAy phenyl O tetraethyl CO 2-carboxy-6- CO H 1 3
benzene methoxy phenyl
103 NH NH C0 p-methoAy phenyl O propylbenzene CO 2.6-dihydroxy CO H 1 3
benzene phenyl
104 NH NH C0 m-hydroxy phenyl O tetrapropyl CO 2-hJJ~UAy6- CO H 1 3 0 ~-~
benzene methyl phenyl ~ ~n
105 NH NH C0 o-hydroxy phenyl O butyl benzene CO 2,6-dihydroxy CO H 1 3
phenyl
106 NH NH C0 m-hydroxy phenyl O tetrabutyl CO 2-carboxy-6- CO H 1 3 l~a
benzene me~hyl benzene
107 NH NH C0 m-hydroxy phenyl O pentytbenzene CO 2-carboAy-6- CO H 1 3
methyl benzene
108 NH NH C0 o-hydroxy phenyl O t , ,t~ CO 2~rboxy-6- CO H 1 3
benzene hydroxy phenyl
109 NH NH C0 o-chloro phenyl O methoxy CO 2-carboAy-6- CO H 1 3 c
benzene methoxy phenyl ~O
110 NH NH C0 p-methoxy phenyl O :' huAy CO 2-hydroxy-6- CO H 1 3 jR
benzene methyl phenyl . ~
Cmpd. ¦ A E~ E~ D E F G X K m n ¦ ~ ~ g
111 NH NH CO p-chloro phenyi O t~lr.. hoAy CO 2,6-dimethyi CO H 1 3
benzene phenyi
112 NH NH CO p-hydroxy phenyi O tetramethyi CO 2,6-dihydroxy CO H 1 3
benzene phenyi
113 NH NH CO p-rnethoxy phenyi O ethoxybenzene CO 2-carboxy4- CO H 1 3
methyi benzene
114 NH NH CO p-hydroxy phenyi O diethoxy CO 2-carboxy4- CO H 1 3
benzene hydroxy phenyi
115 NH NH CO m-hydroxy phenyi O nitrobenzene CO 2-carboxy4- CO H 1 3
methoxy phenyl
116 NH NH CO p-chloro phenyi O dinitrobenzene CO 2,6-dimethyi CO H 1 3
phenyi
117 NH NH CO o-methoxy phenyi O halobenzene CO 2-hydroxy4- CO H 1 3 1.
methyi phenyi O
118 NH NH CO p-methoxy phenyi O dihalobenzene CO 2-carboxy4- CO H 1 3
methyi phenyi
119 NH NH CO p-hydroxy phenyi O trihalobenzene CO 2-hydroxy4- CO H 1 3
methyi phenyi
120 NH NH CO o-chloro phenyi O tetrahalo CO 2-carboxy4- CO H 1 3
benzene methoxy phenyi
121 NH NH CO m-hydroxy phenyi O carboxyacid CO 2-carboxy4- CO H 1 3
benzene hydroxy phenyi
122 NH NH CO m-chloro phenyi O ' bOAj ld CO 2-carboxy4- CO H 1 3 c
benzene ethyi benzene v
123 NH NH CO p-hydroxy phenyi O amido benzene CO 2,6-di".~t'loiy CO H 1 3 ,~
phenyi 00
124 NH NH CO o-methoxy phenyi O diamido CO 2-c~rboxy4- CO H 1 3
benzene hydroxy phenyi
-
Cmpd. A¦ B1 ¦ B2 ¦ Z D E F G ¦ X K m n ¦ o
125 NH NH CO p-hydroxy phenyi O phenol CO 2-carboxy4- CO H 1 3
hydroxy phenyl
126 NH NH CO 1.. hùAy phenyl O dihydroxy CO 2-; LUAY6- CO H 1 3
benzene hydroAvy phenyi
127 NH NH CO p-methoAy phenyi O trihydroAy CO 2-carboxy4- CO H 1 3
benzene propoxy benzene
128 NH NH CO 1.. 3~ UA~ phenyl O l~,t~ hJJ~UAy CO 2carboxy4- CO H 1 3
benzene hydroxy phenyi
129 NH NH CO p-chloro phenyi O triethoxy CO 2,6-dimethyi CO H 1 3
benzene phenyi
130 NH NH CO p-methoAy phenyi O tetraethoxy CO 2-carboxy4- CO H 1 3
benzene methoxy phenyi I
131 NH NH CO p-hydroxy phenyi O pentoAy CO 2,6-dihydroxy CO H 1 3 1~ C r~
benzene phenyl ~ _~
132 NH NH CO p-hydroxy phenyi O dipentoxy CO 2,6~ . t~10AY CO H 1 3
benzene phenyl
133 NH NH CO m-methoAy phenyl O tripentoAvy CO 2- buAy6- CO H 1 3
benzene ethyi benzene
134 NH NH CO o-hydroxy phenyi O I ~r~ Y CO 2-carboxy4- CO H 1 3
benzene . hydroxy phenyi
135 NH NH CO o-hydroxy phenyi O aniline CO 2-carboxy4- CO H 1 3
hydroxy phenyi
136 NH NH CO o-chloro phenyi O dbmino CO 2,6-~ thoAy CO H 1 3 C
benzene phenyi ~,
137 NH NH CO p-methoAy phenyi O methoxy CO 2-carboxy4- CO H 1 3
pyridine propyi benzene A
WO 94/20062
PCT/US94/02283
-- 112 --
2157~
E
_
y
I
O O O O O O O O O O O O O
N E N N N E N ~ N N N E N N ~D N _ N E N
O O O O O O O O O O O O O
~ C ~ t ~ 2 E
C~
O O O O O O O O O O O O O
~ ~ C G C ~ lii c ~ CL ~ t~. G
G G ~ O O O
m O O O O O O O O O O O O O
m
Z Z Z Z Z Z Z Z Z Z Z Z Z
I
Z Z Z Z Z Z Z Z Z Z Z Z Z
c
E 0 0 -- N ~ ~ It~ O
WO 94/20062 215 7 412 PCT/US94/02283
-- 113 --
e~ e~ e~ e~ e!~ e~ e!~ e~ e~ eq e!~ e~ e~
E _ _ _ _ _ _ _ _ _ _ _ _ _
Y
- O O O O O O O O O O O O O
~ '~
e~ e~
'. '. eD~U ~U 'U
N . N . G C~ O .E ~ 2 : L
O O O O O O O O O O O O O
-
2 2 3~ 3O~ ~ 3O~
LU ,. eD ~ g~ U g, ~ U ~ l U ~ I
e~ Ce!~ _ e!~ _ e!~ _er~ _ e!~ . eq . e~ _ e!~ . eq . e!~ _ eq .
O O OO O O O O OO O O O
~U ~ eD eu eu eeu ~ eu e ,eu eu eu
O OG CL ~ O C~
m O OO O O O O OO O O O O
~ o
m
Z Z Zz z z z z zZ Z z z
I
Z Z Zz z z z Z Zz Z z z
c u~ eD r~ e~ u~~OD ~ ~ eD
C~ ~ _ __ _ _ _ _ __ _ _ _ _
Cmpd. ¦ A B1 B2 Z D E F G X K m n ¦ O
166 NH NH CO p4hloro phenyl 0 3,5-dihydroxy CO thbzole CO H 1 3
benzene
167 NH NH CO m-methoAy phenyl 0 3,5-dihydroxy CO Isoxazole CO H 1 3
benzene
168 NH NH CO p-chloro phenyl 0 3,5-dihydroxy CO pyrazole CO H 1 3
benzene
169 NH NH CO p-hydroxy phenyl O benzene isutl,' '~ CO H 1 3
170 NH NH CO p-methoAy phenyl 0 3,5-dihydroxy CO benzene CO H 1 3
benzene
171 NH NH CO o-hydroxy phenyi 0 3,5-dihydroxy CO methyibenzene CO H 1 3
benzene
172 NH NH CO o-chloro phenyl 0 3,5-dihydroxy CO dimethytbenzene CO H 1 3 ~'
benzene ,~
173 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO trimethylbenzene CO H 1 3
benzene
174 NH NH CO p-me~hoxy phenyi 0 3,5-dihydroxy CO t~ tl,~l CO H 1 3
benzene
175 NH NH CO ,.. n lhoAy phenyl 0 3,5~ihydroxy CO ethylbenzene CO H 1 3
benzene
176 NH NH CO p-hydroxy phenyl 0 3,5-dihydro~ CO tetraethylbenzene CO H 1 3
benzene
177 NH NH CO m-hydroxy phenyt 0 3,5-dihydroxy CO propyibenzene CO H 1 3 c
benzene benzene u
173 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO tetrapropyi CO H 1 3
benzene benzene
179 NH NH CO p-chloro phenyl 0 3,5-dihydroxy CO butylbenzene CO H 1 3
WO 94/20062 215 7 412 PCT/US94/02283
-- 115 --
_
Y
- O O O O O O O O O O O O O
~ "
~ n ~ ~. .. ~
~ , . C . ~ . ~ _ o
o o o o o o o o o o o o o
J~ ' ; ~ ~L ~ o ~ O ~ o2 2
o o o o o o o o o o o o o
,c C c~ C~ C C~ c 1~ ~ Q, G 'c~ ~
c~ ~ G ~ c~ ~. G ~ G
,o ,o ,o
C- ~ O O G CL G i o
8 8 8 8 8 8 8 8 8 8 8 8 8
m
I
E 0 0 '`~ D ~ 0 ~ o,
Cmpd. A ¦ B~ B2 Z D E F G X K m n ~ 2
193 NH NH CO ... h~J~UAy phenyi 0 3,5-dihydroAy CO trihalobenzene CO H 1 3
benzene
194 NH NH CO m-chloro phenyi 0 3,5-dihydroAy CO tetrahalobenzene CO H 1 3
benzene
195 NH NH CO p-hydroxy phenyi 0 3,5"hyJ,JA~ CO carboxyadd CO H 1 3
benzene benzene
196 NH NH CO p-chloro phenyi 0 3,5-dihydroAy CO " ~ ~,f ld CO H 1 3
benzene benzene
197 NH NH CO p-methoAy phenyi 0 3,5-dihydroxy CO amido benzene CO H 1 3
benzene
198 NH NH CO m-hydroA~y phenyi 0 3,5-dihydroAy CO diamido benzene CO H 1 3
benzene
199 NH NH CO m-chloro phenyi 0 3,5-dihydroxy CO 3,5-dihydroxy CO H 1 3 1'
benzene benzene a~
200 NH NH CO o-methoAy phenyi 0 3,5-dihydroA~y CO trihydroxybenzene CO H 1 3
benzene
202 NH NH CO o-methoAy phenyi 0 3,5-dihydroA~y CO t `r~hJJ~uAy CO H 1 3
benzene benzene
202 NH NH CO p-chloro phenyi 0 3,5-dihydroAy CO p~.~' hJJ~uA~ CO H 1 3
benzene benzene
203 NH NH CO p-chloro phenyi 0 3,5-dihydroAy CO triethoAybenzene CO H 1 3
benzene
204 NH NH CO o-hydroAy phenyi 0 3,5-dihydroxy CO tetraethoAy CO H 1 3 c
benzene benzene r~
205 NH NH CO p-hydroAy phenyi 0 3,5-dihydroAy CO pentoxybenzene CO H 1 3
benzene ~,
206 NH NH CO p-hydroxy phenyi 0 3,5-dihydroxy CO dipentoxybenzene CO H 1 3
benzene
Cmpd. A B~ ¦ J2 ¦ ¦ E F G X K m n
207 NH NH CO p-methoAy phenyl 0 3,5~ihydroxy CO tripen~oxybenzene CO H 1 3
benzene
208 NH NH CO m-hydroxy phenyl 0 3,5-dihydroxy CO l~t,., , y CO H 1 3
benzene benzene
209 NH NH CO o-methoAy phenyi 0 3,5-dihydroxy CO amino benzene CO H 1 3
benzene
210 NH NH CO o-chloro phenyi 0 3,5-dihydroxy CO diamino benzene CO H 1 3
benzene
211 NH NH CO p-methoxy phenyi 0 3,5-dihydroxy CO methoxypyridine CO H 1 3
benzene
212 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO Ji",_lhuA~pyridine CO H 1 3
benzene I ~V
213 NH NH CO p-methoxy phenyi 0 3,5-dihydroxy CO hydroAypyridine CO H 1 3
benzene
214 NH NH CO p-methoxy phenyl 0 3,5-dihydroxy CO dihydroxypyridine CO H 1 3 1 ~_~
benzene l~
215 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO ethoAypyrrole CO H 1 3
benzene
216 NH NH CO ... hJJ~uAy phenyi 0 3,5-dihydroxy CO dihydroxypyrrole CO H 1 3
benzene
217 NH NH CO p-chloro phenyi 0 3,5-dihydroxy CO ' . `i~.. A~Hndole CO H 1 3
benzene
218 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO hydroAypurine CO H 1 3 c
benzene u
219 NH NH CO p-methoxy phenyi 0 3,5-dihydroxy CO d~.,._lhuAyfuran CO H 1 3
benzene
220 NH NH CO p-hydroxy phenyl 0 3,5-dihydroxy CO hydroxyll, ~ .,t CO H 1 3
benzene
~ o
Cmpd. A ~1¦ i32 Z D E F G X K m n Wg
221 NH NH C0 m-chloro phenyi O benzene CO methoAyl".; ' ,e CO H 1 3
222 NH NH C0 p-hydroAy phenyi 0 3,5-dihydroAy CO 'i.. IhuAy CO H 1 3
benzene IJ j . ;~;. ,e
223 NH NH C0 p-chloro phenyl O benzene CO hydroAyp,.; " ,e CO H 1 3
224 NH NH C0 m-chloro phenyi 0 3,5-dihydroAy CO diamido py.; .;di,.e CO H 1 3
benzene
225 NH NH C0 o-hydroAy phenyi 0 3,5-dihydroxy CO amido pyrazine CO H 1 3
benzene
226 NH NH C0 m-hydroAy phenyl 0 3,5-dihydroA~y CO diethoAypyrazine CO H 1 3
benzene
227 NH NH C0 quinoline 0 3,5-dihydroAy CO phenyi CO H 1 3 1--
benzene
22a NH NH C0 methoxy 0 3,5-dihydroxy C0 phenyi CO H 1 3
quinoline benzene
229 NH NH C0 ~'. hùAy 0 3,5-dihydroAy C0 phen~ CO H 1 3
quinoline benzene
230 NH NH C0 I,i hUA~ O 3,5-dihydroAy CO phenyl CO H 1 3
quinoline benzene
231 NH NH C0 hydroAy 0 3,5-dihydroxy C0 phenyi CO H 1 3
quinoline benzene
232 NH NH C0 dihydroAy 0 3,5-dihydroAy C0 phenyl CO H 1 3 c
quinoline benzene u
233 NH NH C0 ethoAy 0 3,5-dihydroxy C0 phenyl CO H 1 3 ~;
quinoline benzene
234 NH NH C0 amino 0 3,5-dihydroAy CO phen~ CO H 1 3
quinoline benzene
Cmpd. A 31 B2 7 D E ¦ F G ¦ X ¦ K m n
235 NH NH CO dbmido 0 3,5-dihydroxy CO phenyi CO H 1 3
qulnoline benzene
236 NH NH CO trihalo 0 3,5-dihydroAy CO phenyi CO H 1 3
quinoline benzene
237 NH NH CO quinolinecarboA~yic O 3,5-dihydroAy CO phenyi CO H 1 3
acid benzene
238 NH NH CO ~,ln l.e O 3,5-dihydroAy CO phenyi CO H 1 3
benzene
239 NH NH CO methoAy O 3,5-dihydroAy CO phenyi CO H 1 3
~il ' , " ,e benzene
240 NH NH CO . ~lh~AY O 3,5-dihydroAy CO phenyi CO H 1 3
'il' . ' ,e benzene
241 NH NH CO l~i\, Ihv~ O 3,5-dihydroAy CO phenyi CO H 1 3 ~ r~
il' ,- ',e benzene ~D
242 NH NH CO hydroxy O 3.5-dihydroAy CO phenyi CO H 1 3 1 _~
, ', ' ,e benzene ~p.
243 NH NH CO t~;hy.' VAY O 3,5-dihydroxy CO phenyi CO H 1 3
~,-ln !ine benzene
244 NH NH CO t hv~y O 3,5-dihydroAy CO phenyi CO H 1 3
e benzene
245 NH NH CO diamino 0 3,5-dihydroxy CO phenyi CO H 1 3
il~ln ' ,e benzene
246 NH NH CO triamido 0 3,5-dihydroAy CO phenyi CO H 1 3
q~lr ' ,e benzene ~
247 NH NH CO tetrahalo 0 3,5-dihydroAy CO phenyi CO H 1 3 O
, ', 'i. ~e benzene
248 NH NH CO 'i"~ ~'h,e O 3,5-dihydroxy CO phenyi CO H 1 3
Ji~_. i VAyi- acid benzene
WO 94/20062 PCT/US94/02283
21S7~ 120-
E _ _ _ _ _ _ _ _ _ _ _ _ _ _
O O O O O O O O O O O O O O
C C C C ~ &
G 1~ C~ ~ G C~ C~ ~ C~ 1~ D ~ C~ C~
LL
8 8 8 8 8 8 8 8 8 8 8 8 8 8
ID
o ~ ~ O ~
O O O O O O O O O O O O O O
C ~ ~ C C ~D ~ G C~ ~ ~ G
: : : . : :
G G t~ o o ; G ~ C t~
m O O O O O O O O O O O O O O
m
Z Z Z Z Z Z Z Z Z Z Z Z Z Z
ZZZZZZZZZZZZZZ
-
E ~ O -- N 0 ~ U) ~ ~-- 0 CO O _ N
~le N N N N N N N N N N N N N N
WO 94/20062 215 7 ~ 12 PCT/US94/02283
-- 121 --
E _ _ _ _ _ _ _ _ _ _ _ _ _
Y
x 8 ~, ~, ~, ~, 8 ~, ~, ~ ~
G G C~ Cl. ~ G ~ CL ' ' C
O O O O O O O O O O O O O
o ' o ! ' 7` J ~ ~ _ r
O O O o o o o o o o o o o
-- C~' c ~ S c
Q ~ G ~ G ~ ~, ~ ~,
o ' o 1' o
C C o
Q. O O 11 t~ G O O O
N
m O O O O O O O O O O O O O
m
Z Z Z Z Z Z Z Z Z Z Z Z Z
-
T T I I I T I I ~
Z Z Z Z Z Z Z Z Z Z Z Z Z
~
C.) N N N N N NN N N ~N` N C~l N
WO 94/20062 PCTtUS94/02283
21~7412 - 122 -
E
Y
O O O O O O O O O O O O O O
2 ~ o ~ o : : o ~ .E
8 8 8 ~, 8 8 8 8 8 8 8 8 8 8
2 ,~ D ~ ~ C ~ S
O O O O O O O O O O O O O O
G 0- G ~ ~ CL , ~ ~ ' ~ c G
c c~ G ~ c o o o
`.
m O O O O O O O O O O O O O O
m
ZzzzzzzzzzzzzZ
ZZZZZZZZZZZZZZ
E ~D ~ 0 0 c~ _ N C" ~ U~ ~ ~ 0 0
C~ ~1. N N N N N0 N0 N0 N N0 N0 N0 N N N0
WO 94120062 21~ 7 412 PCTIUS94/02283
-- 123 --
E _ _ _ _ _ _ __ _ _ _ _
Y
8 8 8 8 8 8 8 8 8 8 8 8 8
~ ~ ,,. . ~
_ 8 8 8 o 8 8 8 8 8 8 8 8 8
o o o o o o o o o o o o o
G ,c ~ ~ ~ G G ,c
. . O : '
G ~ . C G O
m O O O O O O O O O O O O O
m I I I I I I I I I I
Z Z Z O O O ~ Z c~ ~o Z Z Z Z Z
~ ' N N N N 0 N 0 r-- 0 0 0-- N
cn
~`~
Cmpd. A B 1 B 2 Z D E F G X K m n
f
303 NH O CO m-hydroxy phenyi O 3,Wihydroxy CO phenyi CO H 1 3
benzene
304 NH N-ethyi CO o-chloro phenyi O 3,5-dihydroAy CO phenyi CO H 1 3
benzene
305 NH N- CO ... mlhuAy phenyi O 3,5-dihydroxy CO phenyi CO H 1 3
methyi benzene
306 N-anisly N- CO p-hydroxy phenyi O 3,5-dihydroAy CO ;l~ln 'ine CO H 1 3
phenyi benzene " bGAJr': acid
307 NH NH CS p-chloro phenyi O 3,5 :"hJ ' UAy CO phenyi CO H 1 3
benzene
308 NH NH CO p-hydroxy phenyi CH2 3,5-dihydroAy CO phenyi CO H 1 3
benzene
309 NH NH CO p-hydroxy phenyi N-ethyi 3,5-dihydroAy CO phenyi CO H 1 3 N
310 NH NH CO p-methoxy phenyi N- 3,5-dihydroAy CO phenyi CO H 1 3
phenyi benzene
311 NH NH CO o-methoxy phenyi N- 3,5-dihydroxy CO phenyi CO H 1 3
propyi benzene
312 NH NH CO m-chloro phenyi O 3,5-dihydroxy CS phenyi CO H 1 3
benzene
313 NH NH CO m-chloro phenyi O 3,5-dihydroxy CH(OC phenyi CO H 1 3
benzene H ~ ~,
314 NH NH CO o-chloro phenyi O 3,5-dihydroAy CH(O- phenyi CO H 1 3
benzene phenyi) u
315 NH NH CO o-hydroxy phenyi O benzene CH2 phenyi CO H 1 3 O
316 NH NH CO m-chloro phenyi O 3,WihydroAy O phenyi CO H 1 3
benzene
WO 94/20062 215 7 ~ 1~ PCT/US94/02283
-- 125 --
r.~ -- r,~l ~ . _ _ N N r.~ r,~ _ _ N
-- _ _ _ N r~ ~ N e~ N
Y I I I I I T
8 8 8 8 8 8 8 8 8 8 8 8 ~ 8 ~
~ ;~,
f ~ f f f f f ~ ~D f f ~~D
~ 8 8 8 8 8 8 8 8 8 8 ~ 8 ~ 8
~i b ~ C C ~ D
~ . ~ . r, ~, r,.~, r, ~ rq _ ~ _ ~ . ~ . _ . C~ . .
O O O O O O O O O O O O O O O
c c ~ ~ ~ ~ ~ _ ~
~1 ~ ~ ~, ~ ~ ~. ~ ~, ~ rD
G G C~ i O C~ G 2 . : : .
_ 8 8 8 8 8 8 8 8 8 8 8 o 8 ~
m I I I I T
Z Z Z z z z z z z z a~ Z o. Z Z Z
E ~ N N CNU N N N N N N N c~ C~
WO 94/20062 PCT/US94/02283
2157 ~ 126 -
N N _N _ 0 0 0 0 _ N 0
E __ _ _ _ _ _ _ _ _ _ _ N
Y II I I I I T I I I I T
Otn Ou~ O a~ O u~ Z, ,~ ~ O O
Il . D
!I~
u
~ . :, o
.~ c
., ~
fB o z 8 t~ 8 z E 8 z c~ ~, z 8
E . c
OOOOOOOOOOzOOO
N ,
~ c ~ :
E cL '~ . . .
m u~ O tn ~ ~ ~
m OO z ,~ ~ , ,~ . ~ I I
~ TT T T I C u~ ~ e ,-- T
E NC~ ~ u~ ` 0 O) O _ N 0 ~ It7
WO 94/20062 21~ ~ 112 PCT/US94/02283
-- 127 --
--C~J N C'~ N ~ r~ _
Y
O O O O O o~ O a~ Ou~ O c/~ O a~
D ~ D 'D _
E _ 2 ~
~ . . . ~
0~ O~n
O O O O U~ I _ I ~ O O O
O O U~ C O ~
UJ ' ' ~ _ ~. 'D ~ D ~ ~ D ~ _ O . O ` O
o : o .~: o ~' . ~ ~ E
, " C ~
O O O O O O O O O O O O O O
'D
'D ~D _ . .
Nc ID
N ~ ~ ~ ~D
~; ~ : n : ~ ~ O
E ~ c : . E ~ -
m O o a~ ~
m ~ ~ ii; I ~ ~ I
C~
E ~ u~ N t'~
WO 94t20062 PCTtUS94/02283
-- 128 --
2157ll~2
N ~t _ _ _ N N c~ _ _ N
E _ _ N ~ ~ N C'~ N
Y
O a~ O a~ Z~,~U O O o o cn O O O
'U
'D 'D_ ~
~ o . O !- .
.~ o
~ .~ ~ . .
O ~n O O o o o oc~ o o o 'o
'U ~ .~ IU ~ U ~ C ~ ~U
o _
C ~
OOOOZOOOOOOOO
~U ~ IU C ~J C lU C C S C ~U ~1~
c .~ r~ ..
CL C~ O I:L O 1~ G o Q
,.,
m O 0~ o 0 0 0 0 O u~ O O O O
m t~ O a~ z '' z E ' ~ ' ~
~,C E ~ Q ~
Za~Z~ZZZZOt~ZZZZ
c-- Co _ N 7 ~ In tD
WO 94/20062 215 7 ~12 PCT/US94102283
-- 129 --
N ~ N ~ _ _
E _ _ _ _ _ _ _ _ _ _ _ N C'~
I T I I I I I T T T I I T
X N
O O O tn O o~ O a~ O o~ o 0, z
C~ ~ ,'D ~ o ~ ; ~ ; ; ; ,
O O O O O cn O O a~ O
O Z G ~1)
~ 2 ~ 2 ~ o '
G ' Q ~ Q ~ ~ b . ~ . ~ . ~ . ~ . ~ ~ c -
o o o o o o o o o o o o o
G Q Q ~Q 'G C ~ D C ~ ~D
N ~ ~ Q ~ ~ ~, ,~
: . - . . : . '
--~ _ _ _ _
G O Q O O O G Q C
N
m O O O o O~n O O ~ O O O O
- N ~ ~ ~
I I I I IC~ O O Z 'Q Z E IZ Z ~CQ Z Q
N C,'7 111E ' Q ~ I I I I z
E ~ o _ N C~
Cmpd. A ¦ 8 1 ¦ d 2 D E F (1 X K m n W
386 CH2 NH CS o-methoxy phenyi O methoxy CS amino benzene CS H 4
e
387 0 NH CO o-hydroxy phenyi O h 'hJAY CO diamino benzene CH(CH3 H 2 2
~".; ' ' .e CH3~
388 S NH CO ... n, Iho,~y phenyi O hydroxy N- methoxypyridine CS H 3 2
.. IJi"e methyi
389 N-SO z NH CO p-chloro phenyi NH dbmido CO d,,,,~lhùxypyridine C=N- H 2 3
phenyi 1,,.; .. Ji"e phenyi
390 N-methy NH CO p-chloro phenyi O amido pyrazine N- hydroxypyridine CS H 1 3
pyridine
391 N-butyi NH CO p-hydroxy phenyi NH diethoxy CO dihydroxypyridine CO H 1 3
pyrazine
392 N-phenyi NH CO p-chloro phenyi NH phenyi N- phenyi CO H 1 3 1'
purine o
393 S NH CO p-hydroxy phenyi NH phenyi CO phenyi CO H
394 N-SO z NH CS hydroxytl,', h~ne NH dihydroxy CH2 phenyi CO H
phenyi benzene
395 N-methyi NH CO methoxy~,.'' '.e CH2 dihydroxy CH2 phenyi CO H 1 4
benzene
396 N-butyl NH CS ~" . 'huAy O dihydroxy O phenyi CO H 2
~,.;JaL;"e benzene
397 N-phenyi CH2 CO hydroxypf~ ' " ,e O dihydroxy O phenyi CO H 3 1 ~
benzene 6
398 N-pyridy O CS diamido ~".; ,;di"e N- dihydroxy S pheny CS H 4 1
phenyi benzene
399 0 0 CO amido pyrazine N- dihydroxy CH2 phenyi CO H 2 2
methyi benzene
WO 94/20062 21~ 7 ~12 PCT/US94/02283
._
-- 131 --
N ~ ~r _ _ N _ N N _ N _ C'7
E ,., N
Y
8 8 8 ~ ~ 8 ~ 8
' . ,
X~ ", X~ o I ~ ~, ;
D D ~D
_ _ -- -- C
8 8 8 8 t~ O G ~ 8 8 8 8 ~, o
x x r O ~ r
~D ~ ~ -- S _ ~
Z D , '~ I I I I I N ~ _
'` 2 ~ ' ' ~ ~5, 'D o ~D D
~ . ~ . ~ _ ~ . Cl. ~ ~ G o
m O O o m O o m O cn O m O o~
m , _ ' E I , rD .
~7 D E ,0 D E
c~ z z z z O o u~ Z ~ z z Z z cL
Eo _ N ~ ~ cn O-- N
~,~ ~ ~ Sj! ~ ~ ~ ~o ~ ~o ~
WO 94120062
PCT/US94/02283
21S7~12 - 132 -
~ ~ _ N ~t . _ _ N N ~ ~
E
N ~ ~ N C'~ N
o~ O ~ O cn O a~ O cn O O O c~ O
:
~L
` - E ~E ~ c G
1~ ~C
~ c~ , cn 8 ~ o z ~ c~ ~ c~ z E
`
E " o ^ o , " E
, c . ~, I I I I T
!J . n
J ~ ~
a, ~ ; ~ .E
L . . . _ . . . . _ .
m o ~ O m O m O m O O O O O O
m , .~ , ti; I , .~ , ,, I
T T T z ~ ~ o ~ z ~ z z z C
E' ~ ~ m ~ ~ 0 ~N N N ~ ~tN N
wo g4/20062 2 1 ~ 7 Ll 1 2 PCT/US94/02283
-
-- 133 --
N ~ N ~ _ _ _ N
E _ _ _ _ _ _ _ _ _ _ N C'~ ~t N
Y
~, 8 ~, 8 8 ~, 8 ~, 8 o 8 ~, 8 c~
8 z ~ 8 z ~ 8 o o o o 0 o 8 8 8
' ' ~ ' ' E
Z ~ Z E z Z C z ~ I I I I
J
~ 1~ ~
Q . ~, ~ ~ , ' ; ; ; ~ _
~ O ' ~ c e
N
m 0 0 0 0 0 0 0 0 0 0 0 0 0 0
m T ~ O O Z ~ ' E I Z G Z ~ I
0 ,~ E o G ~
Z ~ Z Z Z Z O ~ Z Z Z Z (~ O 0
E r~ CD ~ ~ ~ N ~ ~ 10
WO 94/20062
PCTIUS94/On83
2157~ 134-
N rr~ r.~ _ _ N _ _ N _ r.~7 rr,
E ,~, N
Y I T I T I I I T T T T T
X I ~ --I U~ Z, ~,, o o o o o o
~D ~D
~ ~ G O G ~ C~ CL Q ~ C C~ Q
$ o c~ 8 ~ 8 u) 8 ~ o z ~ 8
J ,J . . ". ! rD !
o D D C r~ ' C
, r , . , D I , . ~ ~ I I I z
E '~
o
O ~ ' U '
8 8 8 8 8 8 c~ 8 8 8 8 8
m I ~ r z s . D D Z S ' ~
<7 ~ E ~ ~ 1~ C 1 T T T T
E
Cmpd. A ¦ E1 ~2 Z D E F G ¦ X K m n F~
454 N-SO 2- NH CO p_., hy ' UAt NH benzene CS dihydroxy CS H 1 3
phenyi benzene benzene
455 N-methyi NH CO triethoxy benzene NH phenyi N- dihydroxy CO H 1 3
methyl benzene
456 N-but~1 NH CO tetraethoxy O phenyi N- dihydroxy CO H
benzene methyi benzene
457 N-phenyl NH CO pentoxy benzene O phenyl CO benzene CO H 1 2
458 N-pyridyi NH CS dipentoxy benzene O phenyi N-ethyi dihydroxy CO H 1 4
benzene
459 NH NH CO tripentoxy benzene O phenyi CO dihydroxy CS H 2
benzene
460 S NH CO l-,t~ O phenyi N- dihydroxy CO H 3 1 1' ~3
benzene purine benzene Ul t~
461 N-SO 2 NH CS aniline O phenyi CO dihydroxy CO H 4 1 1 ~
phenyi benzene ~"
462 N-methyi NH CO diamino benzene O phenyi CH2 dihydroxy CS H 2 2
benzene
463 N-buty NH CO methoxy pyridine O phenyi CH2 dihydroAy CO H 3 2
464 N-phenyi CH2 CO 'i,. `h ./ pyridine O phenyi O dihydroxy CO H 2 3
benzene
465 N-pyridyi O CO hydroxy pyridine O phenyi O dihydroxy CO H 1 3
benzene v
466 0 0 CS dihydroxy pyridine O phenyi S dihydroxy CO H 1 1 0
benzene
467 CH2 N- CO ethoxy pyrrole O phenyi CH2 dihydroxy CS H 1 3
phenyi benzene
WO 94/20062
2 15 7 4 1 2 - 13 6 - PCT/US94/02283
r~ _ N ~ _ _ _ N N e~
---- -- -- N ~ ~t N'~ N _ ~ _
Y
O O O O O O O a~ O O a~ O O
D
o ~ O
O O O U7 I c I Ç~ O O O O ~1~ 0
~D )
tD ~D
D e~~D 'C~ ~D
u~ ! . ~ I ~ I : ~D 'C I,, I 'D ~CD ~. 'D
C ~C
O O O o o o o O O O O z ~ z E
'D 'D
:
D
N ,~ ~ . _ ~'D ~
'' O D , ~,
E ~
_ ` ` : . ` ~ E
-
m O O O O O o cn O ~ O C~ O O
Z E Z Z ~ Z ~ Z Z Z Z Z O O O Z c
, C E D ~ ~ I
c
E ~ 0 -- N ~ ~ O
WO 94/20062 215 ~ 412 PCT/US94/02283
-- 137 --
N _ N N _ N _ ~ C'~ _ N ~
E _ _ _ _ _ _ _ _ _ _ _ _ _ _
Y
X Z ~ Z C
t~ 8 ~ 8 t~ 8 ~ 8 ~ O ~ c~
ID ID : I ! ! I '
rL
o
E :
~ 8 ~ O Z G 8 ~, 8 z E 8 z 8 z E 8
~D I CD C CD CJ ~D >'
Z z c, z c, z z z z z ~ z O z c, z E z
CD C ,~D ~D ~D ~D C CL~C C 'CL CL ~D
N.. ~ ~ ~, ~ CL ~ ~ CL ~ CL o~ ~D
O : : , ~
G CL CL O O CL CL CL o o E~,
m O O O 0~ o m O m O m O m O o~
~ C
m ~ ~E Z c, ~ z z z
T T 1 T <~ A 4 4 T
CD ~ 0 00 _ N
WO 94/20062
2 1 5 ~ ~ 1 2 - 13 8 - PCT/US94/02283
-- -- -- C~J N C'~
E N C') ~ N ~ N
T I T
C~ O O O O O
D
'` O ~ 'I~
E e
0 ~ ~-
1-- N N N
O O U~ (.)
~u2 ' 2 ' 2 '
~ ~ a
Z c~ O Z ~ z E ~ Z z
O I . O x, x, ~' O
-- -- '~ ~ 'D . 2 _
N
8 o 8 8 8 8
m N
Z C Z Z Z Z O
E 5~ g~ '~ ~ ~ o
WO 94/20062 21-5 7 412 PCT/US94/02283
-- 139 --
y
T
ooooooooooooo
N . N . N ~ N . N N N Q N . N . N . N N . N Q
O O O O O O O O O O O O O
U~ C ~ C "~ S ~ C ~ C ~
~ Q r~ Q c~) Q c~ . ~ Q C~ Q ~ Q ~ Q ~ Q ~ Q ~ G ~ Q ~ Q
O O O O O O O O O O O O O
~ ' ~ ' ' ' c '
Q 'Q Q Q Q Q Q Q
m O O O O O O O O O O O O O
m
ZZZZZZZZZZZZZ
", ." ~
I
,c I I I I I I C) c
Q
o oN ~ o U~ ~o ~ ~ a~ --o -- --N Cl~
c~ o
Cmpd. A B 1 B 2 Z D E F G X K M N ~ o
514 NH NH CO p-hydro y phenyl O 3,5-d hOAY CO 2,641hYdrAY CO H 1 3
phenyi phenyl
515 NH NH CO p-hydroxy phenyl NH 3,5-dihydroxy CO 2-- LOAY6- CO H 1 3
phenyi hydroxy phenyi
516 NH NH CO p-hydroxy phenyl NH 3,5- ' hoAy CO 2- LOAY 6- CO H 1 3
phenyi hydroxy phenyl
517 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2-carboxy~ CO H 1 3
phenyi hydroxy phenyi
518 NH NH CO p-hydroxy phenyl O 3,5 ' hoAy CO 2,6-d' lhoA~y CO H 1 3
phenyi phenyi
519 NH NH CO p-hydroAy phenyl O 3,5-dihydroxy CO 2,6~' lhoA) CO H 1 3
phenyi phenyl
520 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2~arLoAy CO H 1 3 1'
phenyi cyclohe: ,e O
521 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2,6-dihydroxy CO H 1 3
phenyl phenyi
522 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 1-hJdl~.A~r 2- CO H 1 3
phenyi naphthyi
523 NCH2 NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2-carboxy-6- CO H 1 3
phenyi phenyi hydroxy phenyi
524 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2,6-dichloro phenyl CO H 1 3
phenyl
525 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2-carboxy CO H 1 3
phenyl c)_luh- ,e u
526 NH NH SO2 p-methyl phenyl O 3,5-dihydroxy CO 2-carboxy~ CO H 1 3
phenyi hydroxy phenyi j~,
527 NH NH CO p-hydroxy phenyl O 3,5-dihydroxy CO 2 I 'hOA1 6- CO H 1 3
phenyl hydroxy phenyi
WO 94/20062 215 7 412 PCT/US94/02283
-- 141 --
y
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l O O O O O O O O O O O O O
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WO 94/20062 PCT/US94/02283
215~12 - 142 -
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WO 94/20062 215 7 412 PCT/U594/02283
. _
-- 143 --
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PCT/US94/02283
-- 144 --
Z
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WO 94/20062 215 7 4 1 2 PCT/US94/02283
-- 145 --
_____________
T I T I I T I I I T I T
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WO 94120062 PCT/US94/02283
2157~ 146-
Y
C) ~ I I I I I I I I I T
8 8 8 8 8 8 8 8 8 8 8 8 8
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8 8 8 8 8 8 8 8 8 8 8 8
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WO 94/20062 215 7 412 PCT/US94/02283
-- 147 --
y
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WO 94/2006t PCT/US94102283
21 j~3 i~ - 148 - --
Z _ _ _ _ _ _ _ _ _ ~ _
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WO 94/20062 2 1 5 ~ ~ 1 2
PCT/US94/02283
-- 149 --
~ _ _ _ _ _ _ _ _ _ _ _ _
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2 ~ ~2 PCT/US94/02283
-- 150 --
_ _ _ _ _ _ _ _ _ _ _
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WO 94t20062 215 7 412 PCT/US94/02283
.
-- 151 --
y
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O O O O O O O O O O O O
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WO 94t20062 PCTtUS94/02283
~157412 - 152 -
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WO 94/20062 2 i ~ ~ 412 ~CT/US94/02283
-- 153 --
Z _ ~ 7 N
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WO 94/20062 PCT/US94/02t83
21S7~ 154- `
Z N N N ~ C~
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WO 94120062 215 7 412 PCT/US94/02283
-
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y
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N J-- N . N N . N ~D N ~D N U~ N ~D N . N q~ N . N U~ N
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WO 94/20062 PCT/US94/02283
215741~ - 156-
Z _ _ _ ~ _ _ _ _ _ ~ ~ r~ ~
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WO s4noo62 21~ 7 41~ pcTluss4/o22s3
-- 157 --
Z__________~
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WO 94/20062 PCT/US94/02283
2157412 - 158 -
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WO 94/20062 215 7 412 PCT/US94/02283
-- lS9 --
Z _ _ _ .,., _ _ _
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WO94/2~K2 PCT~S94/0~83
- 160 -
21~ ~12 The following, nonlimiting Examples illustrate
preferred methods for preparing the compounds for use in the
method of the present invention and the data demonstrating PKC
inhibitory activity of the Compounds used in the present
invention.
EXAMPL~8
3-Acetyl~minohexahydro-l-phenylmethylazepin-2,~-dione
A solution of hexahydro-l-phenylazepin-2,3,4-trione-3-
oxime (1.23 g, 5mmol) in 4:1 acetic acid/acetic anhydride (20
ml) was treated with Raney nickel (Aldrich, one-half tsp) in a
Parr bottle and subjected to hydrogenation for eighteen hours
at 40-45 psi and room temperature. The mixture was carefully
evacuated of hydrogen and filtered through Celite~. The filter
pad was then washed with methanol (with care taken not to let
the filter pad become dry). The filtrate was concentrated in
vacuo and the residue was diluted with toluene and further
concentrated to remove most of the acetic acid. The residue
was chilled on an ice bath and treated with saturated sodium
bicarbonate carefully to avoid excessive bubbling. The cloudy
aqueous solution was extracted with methylene chloride (3 x 50
ml), and the combined organic solution was dried (Na2SO4) and
concentrated in vacuo. The residue was flash chromatographed
on silica gel (eluted with 19:1 methylene chloride/methanol) to
afford 3-acetylaminohexahydro-1-phenylmethylazepin-2,4-dione
(1.11 g, 81~) as a white solid.
syn-3-Aminohexahy~ro-~-hy~roxy-l-phenylmethylazepin-2-one
A solution of 3-acetylaminohexahydro-1-
phenylmethylazepin-2,4-dione (0.82 g, 3.0 mmol) in absolute
ethanol (lS ml) was treated with sodium borohydride (0.23 g, 6
mmol) and stirred for thirty minutes. The solution was then
treated with water (5 ml) and concentrated in vacuo, and taken
up in 2:1 ethanol/water (7.5 ml). Concentrated hydrochloric
acid (2.5 ml) was added, and the mixture was refluxed for two
hours and partially concentrated, then diluted with water (25
ml). The aqueous acidic mixture was extracted with ether (25
2157412
W094l20062 PCT~S94/02283
- 161 -
ml), and the aqueous solution was basified with 30% sodium
hydroxide and extracted with methylene chloride (3 x 40 ml).
The combined methylene chloride extracts were washed with water
(25 ml), dried (Na2S04), and concentrated in vacuo, to a yellow
solid, which was recrystallized from ethyl acetate to afford
syn-3-aminohPYAhydro-4-hydroxy-1-phenylmethylazepin-2-one (0.42
- g, 60%) as a white solid.
syn-3-Aminohexahydro-~,-hydroxy-l-phQnylmethylazepine
A cooled (S-C) solution of lithium aluminum
hydride/tetrahydrofuran (Aldrich, 1Ø N, 5.1 ml) under
nitrogen was treated with syn-3-aminohexahydro-4-hydroxy-1-
phenlmethylazepin-2-one (0.40 g, 1.7 mmol) in portions so that
the pot temperature did not exceed 15-C. The mixture was
refluxed for 6.5 hours, cooled on an ice bath, and carefully
treated with water (0.21 ml), 15% sodium hydroxide (0.21 ml),
and water (0.63 ml). The suspension was allowed to stir for
five days, during which time the product partially decomposed
(optimal time is 2-5 hours). The suspension was filtered, and
the filtrate was concentrated in vacuo and chromatographed on
silica gel (eluted with 90:8:2 methylene
chloride/methanol/triethylamine). The appropriate fractions
were concentrated in vacuo to afford syn-3-aminohexahydro-4-
hydroxy-1-phenyl-methlazepine (0.22 g, 58%%) as a colorless
oil.
syn-~exahydro-~-hydroxy-3-(~-phenylmethoxy)benzoyl~mino-1-
phenylmethyl~zepine
A solution of 4-benzyloxybenzoic acid (0.183 g, 0.8
mmol) in anhydrous tetrahydrofuran (2 ml) and N, N-
dimethylformamide (0.5 ml) was treated with N,N'-
carbonyldiimidazole ((0.15 g, 0.9 mmol) and stirred at roomtemperature for l.Sh. The solution was treated with syn-3-
aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.20 g, 0.9
mmol) in anhydrous tetrahydrofuran (1 ml). The mixture was
stirred for eighteen hours, and then concentrated in vacuo.
the residue was taken up in lN sodium carbonate (20 ml), and
W094l2~62 PCT~S94/02283
21~7~12 - 162 -
the aqueous mixture was extracted with toluene (2 X 25 ml)
containing a small amount of 2-propanol. The combined organic
extracts were dried (Na2SO4) and the concentrated residue was
flash chromatographed on silica gel (eluted with 3:1 ethyl
acetate/hexane) to afford syn-hexahydro-4-hydroxy-3-(4-
penylmethoxy)-benzoylamino-l-phenylmethylazepine (0.17 g, 50%)
as a viscous oil.
3,5-Bis~phenylmethoxy)-~-~2-phenylmethoxybenzoyl benzoic ~ci~
ester with syn-Hexahy~ro-~-hydroxy-3-~4-phenylmethoxy benzoyl-
~mino-l-phenylmethylazepine
A solution of 3,5-bis(phenylmethoxy)-4-(2-
phenylmethoxybenzoyl)benzoic acid (0.245 g, 0.45 mmol) in
anhydrous methylene chloride (1.5 ml) was treated with N,N-
dimethylformamide (two drops), then with 2 N oxalyl
chloride/methylene chloride (Aldrich, 0.30 ml, 0.60 mmol), and
stirred for one hour under nitrogen. The solution was
concentrated in vacuo and placed under high vacuum for one
hour. syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoyl-
amino-l-phenylmethylazepine (0.170 g, 0.40 mmol) was dissolved
in anhydrous methylene chloride (3 ml), treated with 4-
dimethyl-aminopyridine (0.01 g) and triethylamine (0.12 ml, 1.2
mmol), and cooled in an ice bath under nitrogen. The acid
chloride was removed from high vacuum and dissolved in
anhydrous methylene chloride (2 ml), and was then added to the
cooled solution. The mixture was allowed to warm to room
temperature, stirred for one hour, and was partially
concentrated in vacuo. The residual solution was deposited on
a silica gel column and eluted (first with 2:1 hexane/methylene
chloride, then with 1:1 hexane/methylene chloride) to afford
(after concentration of the appropriate fractions) 3,5-
bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)- benzoic acid
ester with syn-hexahydro-4-hydroxy-3-(4-
phenylmethoxy)benzoylamino-l-phenylmethylazepine (0.29 g 77~)
as a white foam.
N O 94/20062 21~ 7 412 PCTrUS94tO2283
- 163 -
syn- (~- (3,5-Dihydroxy-~-(2-h~.osybenzoyl)benzoyloxy))hexhy~ro
-3-(~-hydroxybenzoylamino)azepine
A cloudy suspension of 3,5-bis(phenylmethoxy)-4-(2-
phenylmethoxybenzoyl-benzoic acid ester with syn-hexahydro-4-
hydroxy-3-(4-phenylmethoxy)benzoylamino-l-phenylmethylazepine
(0.29 g, 0.30 mmol) in reagent ethanol (25 ml) was treated with
- Pd(OH)2/C (Aldrich, Pearlman's catalyst, 0.20 g, 20%) in a Parr
bottle, then subjected to hydrogenation for twenty four hours
at 50-52 psi. The hydrogen was evacuated and the solution was
carefully filtered through Celite~ under nitrogen, and the
filter pad was washed with methanol (not to dryness). The
filtrate was concentrated in ~acuo to crude material, which was
flash chromatographed on a short column of silica gel (eluted
with 1:1 CHClJEtOH) to afford a product (0.13 g) as a pale
yellow foam. This was triturated from ether/ acetonitrile to
afford syn-(4-(3,5-dihydroxy-4-(2-hydroxybenzoyl-benzoyl-
oxy))hexahydro-3-(4-hydroxybenzoylamino) azepine (0.195 g, 62%)
as a pale yellow powder (dihydrate): mp 177-179-C; Rf (1:1
CHCl3/EtOH on silica) 0.45; IR (KBr) 1623 cm , lH NMR (d6-DMSO)
~ 8.20 (d, lH, J = 8Hz), 7.10 (s, 2H), 7.02 (d, lH, J - 9 Hz),
6.91 (t, lH, J -= 8Hz), 6.78 (d, 2H, J = 9Hz), 5.39 (br d, lH,
J = 7Hz), 4.48 (m, lH), 3.00-3.20 (m, 4H), 2.05-2.20 (M, lH),
1.70-2.00 (m, 3H). Anal. Calcd. for C27H26N2O8-2H20: C, 59.77; N,
5.57; H, 5.16. Found: C, 59.83; H, 5.39; N, 5.46.
anti-4-~3,5-Dihydroxy-~-~2-phenylcarbonyl-benzoyloxy-3-(~-
hydroxybenzamido)azepine
To a solution of tran-4-(3,5-dibenzyloxy-4-phenyl
carbonyl-benzoyloxy-3-(4-benzyloxybenzamido)-N-benzylazepine
(40 mg, 0.042 mmol) was added Pd(OH)2 (Pearlman's catalyst) 20
mg, 50% on weight basis) and introduced H2 gas at atmosphere
pressure.
Trans-l-(~-hydroxybenzamido)-2-~4-benzyl-3,5-
dihydroxybenzoyloxy) cycloheptane
The catalyst Pd(OH)2 on carbon (20%, moist, 20 mg) was
added to a solution of trans-1-(4-benzoyloxybenzamido)-2-)4-
W094l2~62 PCT~S94/02283
21S7~1Z - 164 -
benzoyl-3,5-dibenzyloxybenzoyloxy)cycloheptane (215 mg, 0.28
mmol) in methanol (8.4 ml). The mixture was stirred vigorously
at room temperature under 1 atm H2 contained in a balloon for
sixteen hours. The solid catalyst was removed by flash
chromatography (sioz~ 2:2:1/diethyl ether:hexane:methylene
chloride) to give a white powder (112 mg, 84%): mp 224-226-C;
lH NMR (CD30D) ~ 7.52-7.56 (m, 2H), 7.02-722 (m, 5H), 6.94 (s,
2H), 6.71-6.75 (m, 2H), 5.10 (tm, J = 9.1 Hz, lH), 4.35 (tm, J-
9.3 Hz, lH`), 3.93 (s, 2H), 1.56-2.02 (m, lOH) IR (KBr) cm
3389, 1687, 1626. Anal. calcd. for C28H29O6N: C, 70.72: H, 6.15:
N, 2.95. Found C, 70.39; H, 6.37; N, 2.67.
Trans~ -hydroxybenzamido)-2-[~-(2-hydroxybenzoyl)-3,5-
dihy~roxybenzoyloxy] cycloheptane
The catalyst Pd(OH)2 on carbon (20%, moist, 9 mg) was
added to a solution of trans-1-(4-benzyloxybenzamido)-2-t4-(2-
benzyloxybenzoyl)-3,5-dibenzyloxybenzoyloxy~ cycloheptane (112
mg, 0.13 mmol) in methanol (3.9 ml) and ethyl acetate (1.3 ml).
The mixture was stirred vigorously at room temperature under 1
atm H2 contained in a balloon for seventeen hours. The solid
catalyst was removed by filtration through Florisil. The
filtrate was evaporated and purified by flash chromatography
(SiO2 2:2:1/ethyl acetate:hexane:methylene chloride) to give a
pale yellow powder (40 mg, 61~): mp 234-236-C; lH NMR (CD30D)
~ 7.56-7.59 (m, 2H), 7.47 (t, J-7.1 Hz, lH), 7.23 (d, J=8.0 Hz,
lH), 7.00 (s, 2H), 6.96 (d, J=8.2 Hz, lH), 6.74-6.78 (m, 2H),
5.15 (tm, J=9.3 Hz, lH), 4.40 (tm, J=9.3 Hz, lH), 1.58-2.05 (m,
lOH) IR (KBr) cm 3392, 1700, 1678, 1626. Anal. calcd. for
C28H29O6N: C, 66.53; H, 5.38 N, 2.77. Found: C, 66.37; H,
5.56; N, 2.47.
wos4/20062 _ 165215 7 4 12 PCT~S94/02283
(+)-Tr~ns-4-[~-(2-Ca.LGa~ 6-hy~roxybenzoyl)-3,5- ~ihy~roxy-
benzoyloxy]-3-~-hydroxybenzami~o)-1-~methylsulfonyl)
pyrrolidine ~COMPO~ND 501)
HO N31 H2' 10h Pd/C HO~ ~N~
~j CH30 H , ~ ~OBn
N 2. THF, 2 N KOH N
- S 02CH3ClOC~OBn S 02CH3
C 02Bn
NE~, l o OBn
DMAP, C3~COCI
OBn
HO~ N~3~ EP~dOOHHT2FA BnO~O NJ~3
h OH 0~ h OBn
H SO2CH3 BnO2C~b 8BBn N
S01 600
~+)-Tr~ns-~-~ydroxy-3-~-benzylGxyLr~z~mido)-l-~methylsulfonyl)
-pyrrol~dine
To 10% palladium on carbon (41 mg) wetted with
methanol (1.0 ml) was added the azide (412 mg, 2.00 mmol) in
methanol (9.0 ml). The flask was evacuated and filled with H2
twice then allowed to stir under H2 (1 atm) for 4 h. The
mixture was filtered through Celite and washed through with
methanol (100 ml). The methanol was evaporated, providing an
off-white solid which was used without characterization.
To a O C solution of the above amino alcohol in THF
(6.0 ml) was added 2 N KOH (1.0 ml). The ice bath was removed,
and the acid chloride (approx. 0.75 eq) added portionwise over
2.5 h, until the starting material was gone as evidenced by
thin layer chromatography. The mixture was diluted with CH2Cl2
(30 ml) and poured into H2O (60 ml). Methanol (20 ml) was
W094/2~62 PCT~S94/02283
21S7~2 - 166 -
added to disperse the emulsion, the layers were separa~ed, and
the aqueous layer extracted with CH2C12 (4 x 50 ml). The
organic lyaers were combined, dried (MgSO4) filtered and
evaporated to provide the title compound as a white solid (593
mg, 76% over two steps).lH NMR (CD30D) ~ 7.60 (d, J = 8.7 Hz,
2H), 7.25-7.10 (m, 5H), 6.85 (d, J = 9.0 Hz, 2H), 4.94 (s, 2H),
4.18-4.12 (m, 2H), 3.58-3.42 (m, 4H), 2.71 (s, 3H).
(~)-Tr~ns-~-[~-~6-benzyloxy-2-(benzyloxycarbonyl)benzoyl]-3,5-
~ibenzyloxybenzoyloxy)-3-(~-~enzyloxybenzami~o)-1-~methyl
sulfonyl)-pyrrolidi~e ~COMPOUND 600)
To a 0-C solution of the product of the previous step
(202 mg, 0.518 mmol), diisopropylethylamine (98 ~1, 1.2 eq.,
0.565 mmol) and 4-dimethylaminopyridine (58 mg, 1.0 eq., 0.471
mmol) in CH2Cl2 (8.0 ml) under N2 was added a solution of the
acid chloride (0.471 mmol) in CH2C12 (4.0 ml). The reaction was
allowed to warm to room temperature with stirring over 16 h.
The cloudy reaction mixture was diluted with CH2Cl2 (50 ml) and
washed with satd. NaHCO3 (30 ml) then brine (30 ml). The
aqueous layers were extracted with CH2C12 (50 ml) each. The
organics were combined, dried (MgSO4), filtered and evaporated
to a light yellow oil. Flash column chromatography (1:1
hexane:ethyl acetate) provided the title product as an off
white foam (371 mg, 75%): mp 73-79-C; IR (KBr) 3367, 1723,
1660, 1230, 1112, 745, 697 cm-l; lH NMR (CDCl3) ~ 7.80 (d, J =
8.7 Hz, 2H), 7.40-7.10 (m, 20 H), 7.10-7.00 (m, 8H), 6.98 (s,
2H), 6.97 (d, J = 8.3 Hz, 2H), 5.48 (m, lH), 5.15 (s, 2H), 5.09
(s, 2H), 4.79 (s, 4H), 4.74 ( m, lH), 4.72 (s, 2 H), 3.99 (dd,
J = 12.3, 5.5 Hz, lH), 3.78 (dd, J = 11.0, 6.0 Hz, lH), 3.58 (
dd, J = 11.0, 3.3 Hz, lH), 3.51 (dd, J = 12.3, 2.1 Hz, lH) 2.78
(s, 3H); LRMS (M + H) 1051 (65), 797 (21), 661 (100), 571
(38); HRMS calcd for C62H55N2Ol2S (M + H) 1051.3476, found
1051.3433; Anal. Calcd. for C62H5~N2Ol2S-H20: C, 69.65; H, 5.28;
N 2.62; S, 3.00; found C, 69.79; H, 5.30; N, 2.59: S, 2.82.
Wo 94/20062 215 7 41~ PCT/US94102283
_
-- 167 --
~ Tran~-~-[~-~2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxy
benzoyloxy]-3-(4-hydroxybenzamido)-1-Imethylsulfonyl)
pyrrolidine (COMPOUND 501)
To Compound 600 (323 mg, 0.307 mmol) and Pd(OH)2 (50
mg of a 20% by weight powder) under N2 were added ethanol (28
ml) then trifluoroacetic acid (28 ~1). The flask was evacuated
and filled with H2 three times, then stirred under H2 27 h. The
mixture was filtered through Celite, washed with methanol (40
ml) and the filtrate evaporated to a yellow glass. A DMF
solution of the reaction product was purified by reverse phase
HPLC (21 x 250 mm C18 column) to provide Compound 501 (137 mg,
74%) as a yellow powder after lyophilization: mp 167-170,
179-187C (dec); IR (KBr) 3394, 1708, 1607, 1235, 762 cm ; H
NMR (CD30D) ~ 7.53 (d, J = 8.7 Hz, 2H), 7.29 (d, J = 7.6 Hz,
lH), 7.07 (dd, J = 7.9, 8.1 Hz, 1 H), 6.82 (d, J = 8.2 Hz, lH),
6.73 (s, 2H), 6.62 (d, J = 8.7 Hz, 2H), 5.29 (dt, J = 5.5, 3.1
Hz, lH), 4.48 (dt, J = 6.2, 4.2 Hz, lH), 3.73 (dd, J = 12.0,
5.5 Hz, lH), 3.67 (dd, J = 10.7, 6.9 Hz, lH), 3.35 (dd, J =
12.0, 2.8 Hz, lH), 3.28 (dd, J = 10.6, 4.4 Hz, lH), 2.75 (s,
3H); LRMS (M + H) 601 (100), 301 (54), 283 (56); HRMS calcd
for C27H25N2O12S (M + H) 601.1128, found 601.1831; Anal. Calcd.
for C2,H24N2O12S-2.5 H20: C, 50.23; H, 4.53; N, 4.34; S, 4.97;
found C, 50.22; H, 4.36; N, 4.37; S, 4.77.
W094/20062
- 168 - PCT~S94/02283
21~, -Tr~ns-~-t~-~2-Hy~roxycarbonyl-C-hy~roxybenzoyl)-3~5-
~ihy~roxybenzoyloxy]-3-(~-hy~roxybenz~mi~o)-1-tphenylsulfonyl)
pyrrolidine ~CONPOUND 502)
HO~N~ Na~CO~ ~ ~lOBn
H TFA S 02Ph
f ~2Bn
CH2C12 ~COCI
OBn
~0 ~N~ EtdOOH,H)2HF BnO~o N~
OH 0~ ~j OBn
H2cb~ooH SO2Ph 2 ~ OBn S02Ph
502 601
tl)-Tr~ns-3-(~-benzylG~ z~mi~o)-~-hy~roxy-l-(phenylsulfonyl)
-pyrrolidine
To a slurry of starting hydroxyamide (150 mg, 0.352
mmol) in H2O (8.8 ml) and CH2C12 (8.8 ml) were added anhydrous
Na2CO3 (112 mg, 3.0 eq, 1.06 mmol) then benzenesulfonyl chloride
(58 ~1, 0.458 mmol, 1.3 eq), and the mixture stirred at room
temperature 15 h. The solution was then diluted with CH2C12 (20
ml) and poured into H20 (20 ml) and methanol (4 ml). The
layers were separated and the aqueous layer extracted with
CH2Cl2 (3 x 30 ml). The organics were combined, dried (MgSO~),
filtered and evaporated to a white powder (159 mg, quant
yield): H NMR (CD30D) ~ 7.62 (d, J = 7.7 Hz, 2H), 7.43 (d, J
= 8.9 Hz, 2H), 7.35-7.30 (m, 3H), 7.25-7.10 (m, 5H), 6.80 (d,
J = 8.8 Hz, 2H), 4.95 (s, 2H), 4.06-4.00 (m, lH), 3.95-3.90 (m,
215~412
W094/20062 PCT~S94/02283
- 169 -
lH), 3.50-3.35 (m, 2H), 3.15 (dd, J = 10.6, 3.9 Hz, lH), 2.99
(dd, J = 10.8, 3.2 Hz, lH).
(+)-Tr~ns-4-[~-(6-benzyloxy-2-(benzyloxyc~rbonyl)benzoyl]-3,5-
dibenzyloxybenzoyloxy)-3-(~-benzyloxybenz~mi~o)-1- (phenyl
sulfonyl)-pyrrolidine (COMPOUND 601)
To a solution of the prior product (159 mg, 0.352
mmol) in CH2Cl2 (6.0 ml) were added 4-dimethylaminopyridine (43
mg, 0.352~mol, 1.0 eq), diisopropylethylamine (74 ~1, 0.42
mmol, 1.2 eq) then a solution of acid chloride (0.383 mmol, 1.1
eq) in CH2Cl2 (3.0 ml). The mixture was stirred at room
temperature under N2 14 h. The reaction mixture was then
diluted with CH2C12 (30 ml), and washed with 10% NaHCO3 (50 ml)
then brine (50 ml). The aqueous layers were combined and
extracted with CH2C12 (2 x 50 ml). The organics were combined,
dried (MgSO~), filtered and evaporated. Flash column
chromatoghraphy of the residue (2:1 hexane:ethyl acetate) on
silica gel provided the title compound (183 mg, 47%): 1H NMR
(CDCl3) ~ 7.77 (d, J = 6.7 Hz, 2H), 7.70 (d, J = 8.8 Hz, 2H),
7.47-7.16 (m, 14H), 7.15-7.05 (m, 6H), 7.04-6.94 (m, 3H),
6.90-6.83 (m, 3H), 6.42 (d, J = 6.7 Hz, lH), 5.30 (dt, J = 5.1,
2.6 Hz, lH), 5.18 (s, 2H), 5.13 (s, 2H), 4.78 (m, 2H), 4.76 (s,
2H), 4.72 (s, 2H), 4.64-4.60 (m, lH), 3.88 (dd, J = 12.6, 5.4
Hz, lH), 3.74-3.65 (m, lH), 3.60-3.38 (m, 2H).
(+)-Trans-~-t4-(2-Hydroxyc~rbonyl-6-hydrOXybenZOYll-3,5-
dih~k6~benzoyloxy]-3-(~-hydroxybenz~mido)-l-(phenylsulfonyl)
pyrrolidine (COMPOUND 502)
To a solution of Compound 601 (183 mg, 0.164 mmol) in
THF (7.4 ml) and ethanol (7.4 ml) was added Pd(OH)2 (92 mg, of
a 20% by weight powder). The flask was evacuated and filled
with H2 twice, then stirred under H2 (1 atm) for 20 h. The
suspension was filtered through Celite, washed through with
methanol (50 ml), and evaporated to a yellow oil. Purification
by HPLC (21 x 250 mm Cl8 column) provided Compound 502 (75 mg,
69%) as a fluffy yellow powder after lyophilization: mp
185-208-C; IR (KBr) 3402, 1709, 1636, 1608, 1232 cm-l; lH NMR
WOg4/2~62 PCT~S94/02283
(CD30D) ~ 7.53 (d, J = 8.2 Hz 2H), 7.49 (d, J = 8.7 Hz, 2H),
7.32 (d, J = 7.3 Hz, lH), 7.22-7.11 (m, 3H), 7.09 (dd, J = 8.1,
7.9 Hz, lH), 6.84 (d, J = 7.2 Hz, lH), 6.61 (d, J = 8.7 Hz,
2H), 6.35 (s, 2H), 4.99 (app t, J = 2.2 Hz, lH), 4.92 (dd, J =
5.6, 2.8 Hz, lH), 3.62 (dd, J = 13.0, 4.3 Hz, lH), 3.50 (dd, J
= 10.8, 6.0 Hz, lH), 3.39 (dd, J = 10.7, 2.4 Hz, lH), 3.32 (bd,
J = 13.1 Hz, lH); HRMS (M + H) calcd 663.1285, found
663.1302; Anal. Calcd. for C32H26N2Ol2S-l H2O: C, 56.47: H, 4.15;
N, 4.12; S, 4.71; found: C, 56.56; H, 4.17; N, 4.09; S, 4.58.
~09412~62 2 1 5 7 4 1 ~ PCT~S94/02283
Trans-4-[~-(2-Ethoxycarbonyl-6-hydroxybenzoyl)-3,5-
~ihydroxybenzoyloxy]-3-~-hy~roxybenz~mido)-~-~methylsulfonyl)
pyrrolidine ~COMPOUND 503)
(1)-Tr~s-4-[4-(2-Ethoxyc~ nyl-6-hy~roxybenzoyl)-3-etho~cy-S-
hy~roxybenzoyloxy~-3-(4-hydroxybenz~mido)-1-~methylsulfonyl)
pyrrolidine ~COMPOUND 50~)
~h ~
E102C~oo H N
H~ohN~3~ Etl, Na2C03 503
H02C~OHsO2CH3 acetone, DMF
501 X~ h ~
EtO2C~gH N
504
To a solution of Compound 501 (70 mg, 0.12 mmol) in
acetone (5.6 ml) under N2 were added Na2CO3 (anhyd, 25 mg, 0.23
mmol, 2.0 eq) then iodoethane (47 ~1, 0.58 mmol, 5.0 eq).
After stirring at room temperature 3h, a solid began to form.
After Sh, more iodoethane (0.47 ml, S0 eq) was added. After
22h, more iodoethane (0.47 ml, 50 eq) was added, and after
another 24 h DMF (1.0 ml) was added to force the precipitate to
dissolve. The clear yellow solution was stirred 20 h more,
evaporated to approx. 2 ml, then partitioned between H20 (30
ml) and CH2Cl2 (30 ml). The layers were separated, and the
aqueous layer extracted with CH2Cl2 (5 x 20 ml). The organics
were combined, dried (MgSO4), filtered and evaporated. The
yellow residue was purified by reverse phase HPLC (21 x 250 mm
Cle column) to provide Compound 503 (27 mg, 37%) as a yellow
WO 94/20062 PCT/US94/02283
2157~12 - 172 -
power after lyophilization, as well as Compound 504 (11 mg,
14%) as a light yellow powder after lyophilization.
Data for Compound 503 are mp 146-162-C; IR (KBr)
3404, 3364, 2361, 1714, 1637, 1576, 1300, 1231 cm; H NMR
(CD30D) ~ 8.33 (d, J = 6.6 Hz, lH), 7.53 (d, J = 8.7 Hz, 2H),
7.27 (d, J = 7.7 Hz, lH), 7.08 (dd, J = 8.1, 7.8 Hz, lH), 6.83
(d, J = 8.2 Hz, lH), 6.74 (s, 2H), 6.62 (d, J = 8.7 Hz, 2H),
5.29 (dd, J = 5.9, 3.2 Hz, lH), 4.53-4.47 (m, lH), 3.93 (q, J
= 7.1 Hz, 2H), 3.73 (dd, J = 12.1, 5.6 Hz, lH), 3.68 (dd, J =
10.3, 6.5 Hz, lH), 3.35 (dd, J = 12.0, 2.9 Hz, lH), 3.28 (dd,
J = 10.7, 4.5 Hz, lH), 2.76 (s, 3H), 0.92 (t, J = 7.1 Hz, 3H);
LRMS (M + H) 629 (100), 555 (10), 203 (41); HRMS (M + H)
calcd 629.1441, found 629.1476; Anal. Calcd. for
C29H28N2Ol2S-1.25 H2O: C, 53.49; H, 4.72; N, 4.30; S, 4.92; found:
C, 53.66; H, 4.61; N, 4.29; S, 4.87.
Data for Compound 504 are mp 129-134, 140-148 C (dec);
IR (KBr) 3404, 2362, 1715, 1633, 1573, 1300, 1229 cm 1; lH NMR
(CD30D) ô 7.53 (d, J = 8.7 Hz, 2H), 7.29 (d, J = 7.7 Hz, lH),
7.12 (dd, J = 8.1, 7.9 Hz, lH), 7.00 (d, J = 1.5 Hz, lH), 6.84
(d, J = 8.8 Hz, lH), 6.76 (d, J = 1.5 Hz, lH), 6.63 (d, J = 8.8
Hz, 2H), 5.30 (dd, J = 5.4, 3.1 Hz, lH), 4.57-4.50 (m, lH),
3.92 (q, J = 7.1 Hz, 2H), 3.76-3.68 (m, 2H), 3.56 (q, J = 6.9
Hz, 2H), 3.39 (dd, J = 11.9, 3.1 Hz, lH), 3.28 (dd, J = 10.7,
4.6 Hz, lH), 2.78 (s, 3H), 0.91 (t, J = 7.1 Hz, 3H), 0.50 (t,
J = 6.9 Hz, 3H); HRMS (M + H) calcd 657.1754, found 657.1619;
Anal. Calcd. for C31H32N2Ol2S-H2O: C, 55.19; H, 5.08; N, 4.15; S,
4.75; found: C, 55.18; H, 4.92; N, 4.10; S, 4.66.
`W094/20062 21~ 7 ~1~ PCT~S94/0~83
- 173 -
~yn~ 3,5-Dihydroxy-~-~2-hyd o~yLa~zoyl)benzoyloxy)hex~hydro-
3-~-hydroxybenzoyl~mino)azepine ~COMPOUND 505)
3-Acetyl~minohexahydro-1-phenylmethylazepin-2,~-dione
A solution of hexahydro-l-phenylazepin-2,3,4-trione-3-
oxime (1.23 g, 5 mmol) in 4:1 acetic acid/acetic anhydride (20
ml) was treated with Raney nickel (Aldrich, one-half tsp) in a
Parr bottle and subjected to hydrogenation over 18 h at 40-45
psi and room temperature. The mixture was carefully evacuated
of hydrogen and filtered through Celite. The filter pad was
then washed with methanol (with care taken not to let the
filter pad become dry). The filtrate was concentrated in vacuo
and the residue diluted with toluene and further concentrated
to remove most of the acetic acid. The residue was chilled on
an ice bath and treated with saturated sodium bicarbonate
carefully to avoid excessive bubbling. The cloudy aqueous
solution was extracted with methylene chloride (3x50 ml, and
the combined organic solution dried (Na2S04) and concentrated
in vacuo. The residue was flash chromatographed on silica gel
(eluted with l9:1 methylene chloride/methanol) to afford 3-
acetylaminoheYA~ydro-l-phenylmethylazepin-2,4-dione (1.11 g,
81%) as a white solid.
syn-3-Aminohexahydro-~-hydroxy-l-phenylmethylazepin-2-one
A solution of 3-acetylaminohexahydro-1-
phenylmethylazepin-2,4-dione (0.82 g, 3.0 mmol) in absolute
ethanol (15 ml) was treated with sodium borohydride (0.23 g, 6
mmol) and stirred for 30 min, then treated with water (5 ml)
and concentrated in vacuo. The aqueous residue was extracted
with methylene chloride (3x25 ml) and the combined organic
extracts were dried (Na2SO~), concentrated in vacuo, and taken
up in 2:1 ethanol/water (7.5 ml). Concentrated hydrochloric
acid (2.5 ml) was added, and the mixture was refluxed for 2h
and partially concentrated, then diluted with water (25 ml).
The aqueous acidic mixture was extracted with ether (25 ml),
and the aqueous solution basified with 30% sodium hydroxide and
extracted with methylene chloride (3x40 ml). The combined
W094/2~62 PCT~S94/02283
~157412 - 174 -
methylene chloride extracts were washed with water (25ml),
dried (Na2S0~), and concentrated in vacuo to a yellow solid,
which was recrystallized from ethyl acetate to afford syn-3-
aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.42 g,
60%) as a white solid.
syn-3-Aminohexahydro-4-hydroxy-1-phenylmethylazepine
A cooled (5-C) solution of lithium aluminum
hydride/tetrahydrofuran (Aldrich, 1.0 N, 5.1 ml) under nitrogen
was treated with syn-3-aminohexahydro-4-hydroxy-1-
phenylmethylazepin-2-one (0.40 g, 1.7 mmol) in portions so that
the pot temperature did not exceed 15-C. The mixture was
refluxed for 6.5h, cooled on an ice bath, and carefully treated
with water (0.21 ml), 15% sodium hydroxide (0.21 ml), and water
(0.63 ml). The suspension was allowed to stir for 5 days,
during which time the product partially decomposed (optimal
time is 2 - 5 hours). The suspension was filtered, and the
filtrate was concentrated in vacuo and chromotographed on
silica gel (eluted with 90:8:2 methylene
chloride/methanol/triethylamine). The appropriate fractions
were concentrated in vacuo to afford syn-3-aminohexahydro-4-
hydroxy-1-phenylmethylazepine (0.22 g, 58%) as a colorless oil.
syn-Hexahydro-~-hydroxy-3-~-phenylmethoxy)benzoylamino-1-
phenylmethylazepine
A solution of 4-benzyloxybenzoic acid (0.183 g, 0.8
mmol) in anhydrous tetrahydrofuran (2 ml) and N,N-
dimethylformamide (0-5 ml) was treated with N~Nl-
carbonyldiimidazole (0.15 g, 0.9 mmol) and stirred at room
temperature for 1.5h. The solution was treated with syn-3-
aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.20 g, 0.9
mmol) in anhydrous tetrahydrofuran (1 ml), and the mixture was
stirred for 18 h, then concentrated in vacuo. The residue was
taken up in lN sodium carbonate (20 ml), and the aqueous
mixture was extracted with toluene (2x25 ml) containing a
little 2-propanol. The combined organic extracts were dried
(Na2S0~) and the concentrated residue was flash chromatographed
WO 94/20062 215 7 412 PCT/US94102283
-- 175 --
on silica gel (eluted with 3:1 ethyl acetate/hexane) to afford
syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-
phenylmethylazepine (0.17 g, 50%) as a viscous oil.
3,5-B~ 8 (phenylmethoxy)-~.-(2-phenylmethoxybenzoyl)benzoic acid
ester with syn-}lexahy~ro-~-hy~roxy-3-(~-phenylmethoxy)benzoyl
~no-l-phenylmethylazepine
A solution of 3,5-bis(phenylmethoxy)-4-(2-
phenylmethoxybenzoyl)benzoic acid (0.245 g, 0.45 mmol) in
anhydrous methylene chloride (1.5 ml) was treated with N,N-
dimethylformamide (2 drops), then with 2.0 N oxalyl
chloride/methylene chloride (Aldrich, 0.30 ml, 0.60 mmol), and
stirred for one hour under nitrogen. The solution was
concentrated in vacuo and placed under high vacuum for one
hour. syn-Hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-
1-phenylmethylazepine (0.170 g, 0.40 mmol) was dissolved in
anhydrous methylene chloride (3 ml), treated with 4-
dimethylaminopyridine (0.001 g) and triethylamine (0.12 ml, 1.2
mmol), and cooled on an ice bath under nitrogen. The acid
chloride was removed from high vacuum and dissolved in
anhydrous methylene chloride (2 ml), then added to the cooled
solution, and the mixture was allowed to warm to room
temperature, stirred for one hour, and partially concentrated
in vacuo. The residual solution was deposited on a silica gel
column and eluted (first with 2:1 hexane/methylene chloride,
then with 1:1 hexane/methylene chloride) to afford (after
concentration of the appropriate fractions) 3,5-
bis(phenylmethoxy)-4-(2-phenylmethoxybenzoyl)benzoic acid ester
with syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-
phenylmethylazepine (0.29 g, 77%) as a white foam.
Qyn-(~.-(3,5-Dih~0~c",~ 4-(2-hy~.c,J,~e..zoyl)benzoyloxy))hexahydro
-3-(~-hydroxybenzoyl~ino)~zepine (COMPO~ND 505)
- A cloudy suspension of 3,5-bis(phenylmethoxy)-4-(2-
phenylmethoxybenzoyl)benzoic acid ester with syn-hexahydro-4-
hydroxy-3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine
(0.29 g, 0.30 mmol) in reagent ethanol (25 ml) was treated with
WO 94/20062 PCT/US94/02283
2 La ~ ~12 -176 - ~-
20% Pd(OH)2/C (Aldrich, Pearlman's catalyst, 0.20 g) in a Parr
bottle, then subjected to hydrogenation for 24h at 50 - 52 psi.
The hydrogen was evacuated and the solution was carefully
filtered through Celite under nitrogen, and the filter pad was
washed with methanol (not to dryness). The filtrate was
concentrated in vacuo to crude material, which was flash
chromatographed on a short column of silica gel (eluted with
1:1 CHCl3/EtOH) to afford 0.13 g of product as a pale yellow
foam. This was triturated from ether/acetonitrile to afford
syn- ( 4-(3,5-dihydroxy-4-(2-hydroxybenzoyl) benzoyloxy))
hexahydro-3-(4-hydroxybenzoylamino)azepine (0.95 g, 62%) as a
pale yellow powder (dihydrate); mp 177-179-C. R (1 1
CHCl3/EtOH on silica) 0.45; IR (KBr): 1623 cm 1; ~H NMR (d6-DMSO)
8.20 (d, lH, J = 8 Hz), 7.65 (d, 2H, J = 9 Hz), 7.57 (dt, lH,
J = 8, 1.5 Hz), 7.29 (dd, lH, J = 8, 1.5 Hz), 7.10 (s, 2H),
7.02 (d, lH, J = 8 Hz), 6.91 (t, lH, J = 8 Hz), 6.78 (d, 2H, J
= 9 Hz), 5.39 (br d, lH, J = 7 Hz), 4.48 (m, lH), 3.00 - 3.20
(m, 4H), 2.05 - 2.20 (m, lH), 1.70 - 2.00 (m, 3H).Anal.
Calcd. for C27H26N20~-2H2O: C, 59.77; H, 5.57; N, 5.16. Found:
C, 59.83; H, 5.39; N, 5.46.
W094l20062 21 S 7 412 PCT~S94tO~83
- 177 -
Anti-4-t3,5-Dihy~roxy-~-(2-hy~roxyphenylc~rbonyl)]benzoyloxy-
3-(~-hydroxybenz~mido)azepine ~COMPOUND 506)
To a solution of anti-3-(4-benzyloxybenzamido)-4-
[3,5-dibenzyloxy-4-(2-benzyloxyphenylcarbonyl)~benzoyloxy-N-
benzylazepine (150 mg, 0.156 mmol) in EtOAc/EtOH (6 ml, 1:1)
was added Pd(OH)2 (Pearlman's catalyst) (90 mg, 60% on weight
basis), and then H2 at atmospheric pressure. After stirring
vigorously for 24h at room temperature, the reaction mixture
was filtered through a pad of celite. The filtrate was
concentrated and purified on flash column (silica gel: 50 ml;
eluted with 20% ethanol in methylene chloride). CompoUnd 506
was obtained as yellow powder (30 mg, 38%): mp 174 - 176-C; lH
NMR (DMSO) ~ 7.65 (d, J = 8.64, 2H, ArH), 7.54 (td, lH, ArH),
7.26 (dd, J = 1.6, 7.9 Hz, lH, ArH), 6.70 (d, J = 6.48, lH,
ArH), 6.98 (s, 2H, ArH), 6.87 (td, lH, ArH), 6.77 (d, J = 8.67
Hz, 2H, ArH), 5.18 (m, lH, 4CH), 4.19 (m, lH, 3 CH), 2.94 -
2.88 (dd, lH, CH2N), 2.83 - 2.73 (m, 3H, CH2N), 1.91 (m, 2H, 6
CH2), 1.74 and 1.64 (m and m, 2H, 5 CH2); IR (KBr) cm1 3394,
1704, 1623, 1609, and 1504. Anal. calcd. for C2~H26N2O8-1 1/4
H2O: C, 61.30; H, 5.43; N, 5.29. Found: C, 61.33; H, 5.29;
N, 4.96.
W094l2~62 PCT~S94/0~83
21~7~12 - 178 -
Tran~ -benzyloxybenz~mido)-2-t~-(2-benzyloxycarbonylbenzo
yl)-3,5-dibenzyloxybenzoyloxyl)cyclohept~ne (COMPOUND 507)
An oven-dried 25ml 3-neck round bottom flask, under
N2~ was charged with trans-2-(4-benzyloxybenzamido)
cycloheptanol (204 mg, 0.6 mmol), 4-(2-benzyloxycarbonyl
benzoyl)-3,5-dibenzyloxybenzoic acid (376 mg, 0.66 mmol), 1,3-
dicyclohexylcarbodiimide (136 mg, 0.66 mmol), 4-dimethyl-
aminopyridine (74 mg, 0.66 mmol), and dry methylene chloride
(4.5 ml). The resultant suspension was stirred at room
temperature for 17 hours, diluted with methylene chloride
(6ml), washed with water (5 ml x 3), and dried MgSO4. The
solvent was evaporated and the residue was chromatographed
(SiO2, 1:1:2/ diethyl ether:methylene chloride:hexane) to give
a white powder (429 mg, 80%): mp 153-154-C; ~H NMR (CDCl3)
6.89 - 7.68 (m, 30H), 6.33 (d, J = 8.6 Hz, lH), 5.16 (s, 2H),
5.13 (tm, J = 9.4 Hz, lH), 5.04 (s, 2H), 4.95 (ABq, J = 14.2,
12.2 Hz, 4H), 4.45 (tm, J = 9.3 Hz, lH), 1.55 - 2.08 (m, 10H)
IR KBr 3466, 3367, 1735, 1717, 1679 cm 1. Anal. calcd for
CS7HS1O~ C, 76.58; H, 5.75; N, 1.57. Found: C, 76.61; H,
5.82; N, 1.35.
21~7~1~
WOg4/20062 - 179 - PCT~S94/02283
Tr~ns~ -(2-Carboxy-6-hy~roxybenzoyl)-3,5-dihydroxy
benzoyloxy)-3-(~-hy~roxybenz~mido)~zepineTr~fluoroaceticAci~
8alt (COMPO~ND 508) Trifluoroacetic Aci~ 8alt of Balanol
BnO2C~
o~
C02~n BnO~_~ OBn
B~ 1~ (COCI)2 ~ OBn
r¦ T ~ 2) NEt3, DMAP, r
HO2C~OBnOBn ~' N~ OBn ~ H~ OBn
SPC-104034 B B
HO~
O ~
H2 ~ o =( OH OH
Pd(OH)2 \ OH
HO ~ ~ CF3CO2H
o O HN o
(~NH 508
Trans-N-Benzyl~ -(6-benzyloxy-2-(benzyloxyc~rh~nyl)benzoyl)-
3,5-~ih6nzyloxybenzoyloxy)-3-(4-benzyloxybenzamido)azepinQ
(COMPOUND 602)
A solution of 84 mg (0.12 mmol) of 4-(6-benzyloxy-
2-(benzyloxycarbonyl)benzoyl)-3,5-dibenzyloxybenzoic acid in 2
ml of methylene chloride containing a trace (approximately 0.5
~L) of dimethylformamide was cooled to 0-C. Oxalyl chloride
(11.9 ~L, 0.136 mmol) was added, and the mixture was stirred
under a nitrogen atmosphere for 1.5 h. An additional 11.9 ~L
of oxalyl chloride was added, and the mixture was stirred for
an additional 1.5 h. The reaction mixture was evaporated, and
the residue was evaporated twice from 15 ml of methylene
chloride. The residue was dissolved in 2 ml of methylene
chloride, and was added to a solution of 59.1 mg (0.137 mmol)
of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine,
19.0 ~1 (0.136 mmol) of triethylamine, and 3 mg of DMAP in 1.5
W094l2~62 PCT~S94/02283
2157~12 - 180 _
ml of methylene chloride at 0CC. The mixture was stirred at
room temperature under a nitrogen atmosphere for 22 h, after
which it was diluted with 30 ml of methylene chloride, washed
with saturated sodium bicarbonate and brine, dried over
magnesium sulfate, and evaporated to give 139 mg of the crude
product. Chromatography on silica gel eluting with 6/4 hexane
- ethyl acetate gave 89.4 mg (66%) of Compound 602 as a yellow
oil, which was used directly in the next step.
Tr~ns-4-(4-(2-CarhoYy-6-hy~roxybenzoyl)-3,5-di~y ~x~oenzoyloxy)
-3-(4-hydroxy~6n~Amido)azepine Trifluoroacetic Acid Salt
(COMPO~ND 508)
A solution of 65 mg (0.060 mmol) of Trans-N-benzyl-4-
(4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-
dibenzyloxybenzoyloxy)-3-(4-benzyloxybenzamido)azepinein30ml
of 1/2/2 methanol/ethanol/methylene chloride was treated with
17 ~1 of trifluoroacetic acid and evaporated. The residue was
dissolved in 12 ml of 3/1 ethanol/methanol, 15.6 mg of moist
10% palladium hydroxide on carbon was added, and the mixture
was ch~ken on a Parr apparatus under 50 psi of hydrogen for 5
h. The mixture was filtered, evaporated, and the residue was
chromatographed on a 21 x 250 mm C18 column (solvent A: 95:5
water/acetonitrile + 0.1% TFA; solvent B: 100% acetonitrile;
gradient: 0-100% B over 60 min, flow: 15 ml/min). The pure
fraction was evaporated and then lyophilized from water to give
11.2 mg (33%) of the title compound as a yellow fluffy solid.
IR (KBr) : 1701, 1674, 1636, 1608, 1425, 1234, 1200 cm-l;
FABMS: m/z 573 (M + Na), 551 (M+H); HRMS: calcd for C28H27N2Olo
551.1665, found 551.1697.
WO 94/20062 - 181 21~ 7 ~12 PCT/US94/02283
ent-B~l~nol (coMpo~ D 509 )
E3n,N~'NH $~CF3 (N~ oCH33 ~%&F3
~ Cl Bn NH + Bn NH
~OB CH2CI2 ~ d~
OBn OBn
1) SilicaChi~""atography
3% NEt3, 30% EtOAc
`~ in hexanes
2) KOH, MeOH
~-OH ~OH
B,N ~ B~N ~
od~ o5~
OBn OBn
HO2C~
~OH ~
Bn,N~NH ~HO~_ oOH
o ~ ~d 509
~ 0~
OBn ~~~. N~ OH
HN
~8)-Mosher's a¢id chloride
Note that the chirality label is changed going from
acid to acid chloride. (R)- (+)--Methoxy--trifluoromethyl
acetic acid (5.1 g, 21.8 mmol) was slurried in hexane (3 ml).
DMF (2 drops) was added followed by a 2M methylene chloride
solution of oxalyl chloride (33 ml, 66 mmol). The solution was
refluxed for 3 h, cooled to rt, concentrated and distilled by
W094/20062 PCT~S94/0~83
21S7~12 - 182 -
kugelrohr (T = 45C at 0.1 mm Hg) to give the product as a
clear oil.
Trans-N-Benzyl-3-~-benzyloxybenz~mido)-~ R)-2-methoxy-2-
trifluoromethylacetoxy)hex~methyleneimine
Trans-N-Benzyl-3-(4-benzyloxybenzamido)-4-hydroxy-
hexamethyleneimine (1.2 g, 2.79 mmol), DMAP (4 mg, 0.3 mmol)
and triethylamine (2.25 g, 3.1 ml, 22.3 mmol) were dissolved in
methylene chloride (10 ml) and treated with (S)-Mosher's acid
chloride (1.8 g, 1.3 ml, 7 mmol). When TLC indicated complete
reaction, the mixture was concentrated and flashed (7 x 15 cm,
3% triethylamine in 4/1 : ethyl acetate/hexanes). The products
were separated into clean upper, mixed and clean lower
fractions. All three fractions were each again chromatographed
on a Dynamax-60 silica column (41.4 mm ID X 30 cm length)
using a linear gradient from 20% to 60% B (A = hexanes, B = 10%
triethyl amine in ethyl acetate) over 60 m at 25 ml/min. The
clean upper HPLC fractions from the upper and mixed runs were
combined (490 mg) for hydrolysis lH-NMR (300 MHz, CDCl3)
1.6-1.8 (2H, m), 1.94 (2H, m), 2.48 (lH, m), 2.77 (lH, m),
2.9-3.0 (2H, m), 3.50 (lH, d, J = 13 Hz), 3.54 (3H, s), 3.72
(lH, d, J = 13 Hz), 4.1-4.2 (lH, m), 5.14 (2H, s), 5.28 (lH,
m), 6.84-7.65 (14H, m).
The clean lower HPLC fractions from the lower run were
combined (260 mg) for hydrolysis. H-NMR (300 MHz, CDCl3)
1.64-1.8 (2H, m), 1.8-1.94 (2H, m), 2.53 (lH, m), 2.77 (lH, m),
2.9-3.0 (2H, m), 3.50 (lH, d, J = 13 Hz), 3.52 (3H, s), 3.72
(lH, d, J = 13 Hz), 4.1 (lH, m), 5.13 (2H, s), 5.28 (lH, m),
6.84-7.54 (14H, m).
The upper ester fraction (490 mg, 0.76 mmol) was
dissolved in methanol (5 ml) and treated with 85% potassium
hydroxide (97 mg, 1.52 mmol) dissoved in methanol (5 ml) and
stirred for 16 h. The mixture was treated with water (15 ml)
and extracted with methylene chloride (2 x 25 ml). The organic
layer was concentrated and chromatographed (2.5 x 10 cm, ethyl
acetate) to give the chiral alcohol (266 mg) as an oil.
W094/2~62 2 1 5 7 4 1 2 PCT~S94tO~
- 183 -
The lower ester fraction (260 mg, 0.40 mmol) was
dissolved in methanol (5 ml) and treated with 85% potassium
hydroxide (53 mg, 0.8 mmol) dissoved in methanol (5 ml) and
stirred for 48 h. The mixture was treated with water (15 ml)
and extracted with methylene chloride (2 x 25 ml). The organic
layer was concentrated and chromatographed (2.5 x 10 cm, ethyl
acetate) to give the chiral alcohol (154 mg) as an oil.
Balanol benzophenone (255 mg, 375 ~mol) was dissolved
in methylene chloride (3 ml) and treated with DMF (3 drops)
followed by a 2M methylene chloride solution of oxalyl chloride
(244 ~L, 62 mg, 488 ~mol). After stirring for 1 h, the mixture
was concentrated and put under vacuum. The residue was
dissolved in methylene chloride (5 ml) and added to chiral
amidoalcohol (150 mg, 375 ~mol), DMAP (5 mg), triethylamine
(157 ~L, 114 mg, 1.13 mmol) in methylene chloride (5 ml).
After stirring for 16 h, the mixture was chromatographed
directly (silica gel, 2.5 x 10 cm, 2/3 : ethyl acetate
/hexanes) to give the ester (150 mg)as a glass. The glass was
dissolved in 1/1 ethanol/methanol (10 ml), treated with
trifluoroacetic acid (100 ~L) and Pearlman's catalyst
(palladium hydroxide, 15 mg) and stirred under a hydrogen
atmosphere (balloon) for 16 h. The catalyst was filtered off
and the mixture concentrated. The residue was chromatographed
on a Dynamax-60 C18 column (21 X 250 mm) using a linear
gradient from 100% A (0.1% TFA and 5~ acetonitrile in water) to
100% B (pure acetonitrile) over 60 m at 15 ml/min. The clean
product, which eluted in 24 m., was concentrated to remove
acetonitrile and freeze-dried to give a light yellow powder (45
mg, 22%), Compound 509, identical to Balanol by NMR, IR, CHN
and analytical HPLC. Different rotation [~]D25 = +97.8 (c =
0.319 in CH30H).
W094/20062
PCT~S94/02283
~57~ 184 -
~-)-Trans~ -(2-carboxy-6-hy~roxybenzoyl)-3,5-dihy~roxy
benzoyloxy)-3-(~-hy~roxybenz~mi~o)azepineTrifluoroaceticAci~
8alt, (-)-Balanol (COMr~uNv 510)
8nO
) (COCI)2 ~ ~ OBn
ll l 2) IPr2NEt, DMAP, I
HO2C ~OBnOBn CH ~ OBn ~ H~ OBn
Ho2~3
HO~ OH
H2 ~ OH
pd(OH)2 o ~
~NS' HO~ OH
H
510
Trans-N-Benzyl-~(~-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)
-3,5-~ibenzyloxybenzoyloxy)-3-(~-benzyloxybenz~mido)azepine
A solution of 356 mg (0.583 mmol) of 4-(6-benzyloxy-2-
(benzyloxycarbonyl)benzoyl)-3,5-dibenzyloxybenzoic acid in 10
ml of methylene chloride contAjning a trace (approximately 1
~L) of dimethylformamide was cooled to O-C. A 2.0 M solution
of oxalyl chloride (0.35 ml, 0.70 mmol) was added, and the
mixture was stirred under a nitrogen atmosphere for 2 h. An
additional 0.35 ml of oxalyl chloride was added, and the
mixture was stirred for an additional 1 h. The reaction
mixture was evaporated, and the residue was evaporated twice
from 20 ml of methylene chloride. The residue was dissolved in
5 ml of methylene chloride, and was added to a solution of 251
mg (0.583 mmol) of trans-N-benzyl-3-(4-benzyloxybenzamido)
-4-hydroxyazepine, 122 ~L (0.700 mmol) of diisopropyl-
~094/2~62 215 7 412 PCT~S94/02283
- 185 -
ethylamine, and 4.1 mg of DMAP in 9 ml of methylene chloride at
0-C. The mixture was stirred at room temperature under a
nitrogen atmosphere for 16 h, after which it was diluted with
75 ml of methylene chloride, washed with saturated sodium
bicarbonate and brine, dried over magnesium sulfate, and
evaporated to give 690 mg of the crude product. Chromatography
on silica gel eluting with 1/1 hexane - ethyl acetate gave 381
mg (60%) of the title compound as a yellow oil. IR (KBr) :
1719, 1655, 1605, 1581, 1456, 1321, 1248, 1111, 744, 697 cm-l.
Anal. Calcd for C~oH62N2Olo-l~5 H2O: C, 75.18; H, 5.86; N, 2.51.
Found: C, 75.16; H, 5.88; N, 2.74.
~-)-Trans~ 2-Carboxy-6-hydroxybenzoyl)-3,5-dihy~roxy
benzoylo~y)-3-(~-hydroxybenz~mi~o)azepine tr~fluoroaceticacid
~lt (COHPOUND 510)
A solution of 363 mg (0.333 mmol) of trans-N-benzyl-4-
(4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)-3,5-
~ihe~yloxybenzoyloxy)-3-(4-benzyloxybenzamido)azepinein30ml
of ethanol was treated with 31 ~L of trifluoroacetic acid,
cooled to 0-C, and 53.3 mg of moist 10% palladium hydroxide on
carbon was added. The mixture was stirred under an atmosphere
of H2 for 22 h. The mixture was filtered, evaporated, and the
residue was chromatographed on a 41 x 250 mm C18 column
(solvent A; 95:5 water/acetonitrile +0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-50% B over 60 min, flow: 25 ml/min).
The pure fractions were pooled and evaporated and then
lyophilized from water to give 75.7 mg (32%) of Compound 510 as
a yellow fluffy solid, together with an additional 69 mg (29%)
of material which was 96% pure by NMR. mp>200 C; [~]25D = -
104- (c = 0.111, methanol); IR (KBr) : 1679, 1607, 1509, 1426,
1369, 1241, 1202, 763 cm . Anal. Calcd for C28H26N2O1o 3 H2O -
TFA: C, 50.14; H, 4.63, N, 3.90. Found: C, 50.11; H, 4.40; N,
4.01.
W094/2~62 PCT~S94/02283
~157~12 - 186 -
t+) -anti-3- ~ 4-hydroxybenzamido) -~- [ 3, 5-dihy~roxy-~- ~ 2 -
hydroxynaphthyl) carbonyl]benzoyloxyazepine trifluoroacetic acid
salt ~COMPOIJND 511)
To (+)anti-3-(4-benzyloxybenzamido)-4-[3,5-dibenzyloxy
-4-(2-benzyloxynaphthyl)carbonyl] benzoyloxy-N-benzylazepine
(338 mg, 0.366 mmol) dissolved in absolute ethanol (18 ml)
under an atmosphere of nitrogen was added trifluoracetic acid
(30 ~1, 0.386 mmol) followed by Pearlman's catalyst (135mg, 40
% by wt). An atmosphere of hydrogen was introduced and the
mixture was allowed to stir for 48 h. The catalyst was removed
by filtration and the volatiles removed under reduced pressure.
The product was chromatographed on a Dynamax-60 C18 column
(41.4 mm ID X 30 cm length) using a linear gradient from 100%
A (0.1% TFA and 5% acetonitrile in water) to 50% B (pure
acetonitrile) over 60 m at 25 ml/min. The product elutes in 50
minutes. Removal of the volatiles provided Compound 511 as a
yellow solid (99 mg, 38%), mp 165-168-C. IR KBr (disc) cm
3397, 3274, 3121, 2874, 1796, 1776, 1680, 1633, 1606, 1544,
1510, 1461, 1426, 1369, 1344, 1202, 1142, 1109, 1054, 986, 910,
827, 802, 762, 723, 671. Anal. Calcd for C3lH28N20, 2CF3C02H:
C, 54.69; H, 3.93: N, 3.64. Found: C, 54.46; H, 4.06; N,
3.65.
W094l20062 215 7 412 PCT~S94/02283
- 187 -
~rans-4-(~-~2-Ca Loh~ 6-hy~roxybenzoyl)-3,5-dihy~roxybenzoylo
xy)-3-(~-hydroxybenzam~do)-1-(phenylaminocarbonyl)azepine
(COMPOUND 512)
HO~ HO~
(~. N~_ OH ~~, NH~_ OH
O NH
512
A solution of 25.2 mg (0.035 mmol) of (-)-trans-4-(4-
(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-
(4-hydroxybenzamido)azepine trifluoroacetic acid salt in 0.30
ml of dry pyridine was treated with 6.5 ~L (7.1 mg, 0.060 mmol)
of phenylisocyanate. The mixture was stirred for 3 h at room
temperature, after which the reaction was quenched by the
addition of 0.5 ml of methanol. The mixture was evaporated to
a residue which was chromatographed on a 21 x 250 mm C18 column
(solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-100% B over 60 min, flow: 15 ml/min).
The pure fractions were pooled and evaporated and then
lyophilized from water to give 23.8 mg (95%) of Compound 512 as
W094/20062 - 188 - PCT~S94/022~
a yellow fluffy solid, mp >200 C. IR (KBr) : 1705, 1635,
1545, 1506, 1363, 1240, 760 cm . Anal. Calcd for C35H3lN3O11 -
2.5 H2O: C, 58.82 H, 5.08; N, 5.88. Found: C, 59.04; H, 4.94;
N, 5.77.
WOg4/20062 21~ 7 412 PCT~S94tO2283
- 189 -
(+)-~nti-3-~-hy~roxybenzami~o)-~-t3,5-~ihy~roxy-~-
(2,3,5,6-tetramethylphenyl)carbonyl]benzoyloxyazep~ne
trifluoro~cetic ~cid salt (CONPOUND 513)
To(+)-trans-3-(4-benzyloxybenzamido)-4-[3,5-~ihen7yl-
oxy-4-(2,3,5,6-tetramethylphenyl)carbonyl]benzoyloxy-N-benzyl-
azepine (330 mg, 0.364 mmol) dissolved in 1:1 absolute
ethanol:ethyl acetate (100 ml) under an atmosphere of nitrogen
was added trifluoracetic acid (42 ~1, 0.546 mmol) followed by
Pearlman's catalyst (66 mg, 20 % by wt). An atmosphere of
hydrogen was introduced and the mixture was allowed to stir for
24 h. The catalyst was removed by filtration and the volatiles
were removed under reduced pressure. The product was
chromatographed on a Dynamax~-60 C18 column (41.4 mm ID X 30 cm
length) using a linear gradient from 100% A (0.1% TFA and 5%
acetonitrile in water) to 50% B (pure acetonitrile) over 60 m
at 25 ml/min. The product elutes in 58 minutes. Removal of the
volatiles under reduced pressure provided Compound 513 as a
yellow solid (185 mg, 75%), mp 157-160-C. IR KBr (disc) cm
3084, 1717, 1682, 1637, 1609, 1559, 1542, 1509, 1474, 1457,
1425, 1375, 1339, 1270, 1234, 1199, 1140, 1108, 1076, 1007,
938, 848, 813, 767, 723, 669. Anal. Calcd for C3lH28N207 -
CF3COzH H20: C, 58.40; H, 5.50; N, 4.13. Found: C, 58.26;
H, 5.28; N, 4.03.
wos4l2~K2 PCT~S94/02283
7 157 412 - lgo -
tl)-Anti-4-t3~5-~imethoxy-4-~2~6-~ihydroxyphenylcarbonyl)]-3
(4-hydro~ybenz~mi~o)perhydroazepine (COMPOUND 514)
H3CO~¢~OCH3 BuU & o~CH~`C02+ o~ OCH~ ~+ Ht~OOH
CO2+
&OBn O~CH CO~H ~ (~N~ o ~d OBn
OH OCH3 O 603
H2 ~ C~O
OH OCH3 ~- N~ OH
H
514
The above compounds were synthesized using reactions
indicated. Compound 514 was isolated as a yellow powder (37
mg, 5.2% overall yield): mp 165-175 C; NMR (D6 DMSO); IR (KBr)
cm : 3431 (OH); 1710 (ester); 1625 (ketone). Anal. calc. for
C2~H3oN2~-l-5 H2O: C, 60.31; H, 5.76; N, 4.85. Found: C, 60.53;
H, 5.85; N, 4.91. Intermediate Compound 603 was also isolated.
_ W094/2~62 21~ 7 412 PCT~S9410~
-- 191 --
syn-4- ( ~- ( 2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyl-
amino)hexahy~ro-3-(4-hydroxybenzoylamino)azepine, trifluoro-
acetic aci~ salt (COMPOUND 516)
Hexahydro-3-(~-phenylmethoxy)benzoyl~mino-1-phenylmethyl-~zepin
~-one
~ A 25 ml 3-neck round bottom flask under nitrogen was
charged with 2.0 N oxalyl chloride/methylene chloride (Aldrich,
1.125 ml, 2.25 mmol), diluted with anhydrous methylene chloride
(2 ml), cooled (-65-C), and treated dropwise with anhydrous
dimethylsulfoxide (0.35 g, 4.5 mmol) in anhydrous methylene
chloride (1.2 ml) at a rate to keep the pot temperature below
-60-C. The mixture was stirred at -65 + 5C for 30 min, then
treated dropwise with a solution of syn-hexahydro-4-hydroxy-3-
(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.645 g,
1.5 mmol) in anhydrous methylene chloride (1.5 ml) at a rate to
keep the pot temperature below -55-C. The mixture was stirred
at 55+5-C for 2 h, then treated dropwise with triethylamine
(1.5 ml), warmed to room temperature over one hour, and diluted
with methylene chloride (10 ml). The organic solution was
washed with water (10 ml), saturated aqueous sodium bicarbonate
(10 ml), dried (Na2SO4), and concentrated in vacuo. The residue
was chromatographed on silica gel (eluted with 5% acetone/
methylene chloride) to afford hexahydro-3-(4-phenyl-methoxy)
benzoylamino-l-phenylmethylazepin-4-one (0.53 g, 82%) as a
viscous colorless oil.
~exahydro-3-(~-phenylmethoxy)benzoylamino-1-phenylmethyl-
azepin-~-one, oxime
A solution of hexahydro-3-(4-phenylmethoxy)-
benzoylamino-l-phenylmethyl-azepin-4-one (0.87 g, 2.03 mmol) in
ethanol (12 ml) was treated with hydroxylamine hydrochloride
(0.19 g, 2.73 mmol), followed by 25% methanolic sodium
methoxide (Aldrich, 0.20 g, 0.93 mmol), and was heated to 50C
for one hour. The mixture was cooled to room temperature and
treated with additional 25~ methanolic sodium methoxide (0.42
g, 1.94 mmol), then concentrated in vacuo to afford hexahydro-
WOg4/2~62 PCT~S94/022~
21S7~12 - 192 -
3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepin-4-one
oxime (0.89 g, 99%) as a colorless foam.
syn-4-(3,5-Bis~phenylmethoxy)-4-~2-ca~hophenylmethoxy-6-phenyl-
methoxybenzoyl)benzoylaminohexahydro-3-~4-phenylmethoxy)-
benzoyl~mino-1-phenylmethylazepine ~COMPOUND 615)
A solution of hexahydro-3-(4-phenylmethoxy)-benzoyl
amino-l-phenylmethyl-azepin-4-one oxime (0.40 g, 0.90 mmol) in
reagent methanol (25 ml) in a Parr bottle was treated with
Raney Nickel (Aldrich, quarter tsp.), then subjected to
hydrogenation at 49-50 psi for six hours. The solution was
carefully evacuated of hydrogen, filtered through celite, and
the filtrate was concentrated in vacuo to afford 4-
aminohexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenyl
methylazepine, 1:1 mixture of isomers, which was kept-under
nitrogen. Meanwhile, 2'-carbobenzyloxy-2,6,6'-tribenzyloxy
benzophenone-4-carboxylic acid (SPC-104034, b.37 g, 0.S5 mmol)
was placed in a round-bottom flask and repeatedly covered with
toluene and concentrated in vacuo to remove all water and other
persistent solvents. Finally, the residue was dissolved in
anhydrous methylene chloride (2 ml) under nitrogen, treated
with dimethylformamide (3 drops), then with 2.0 N oxalyl
chloride/methylene chloride (0.4 ml, 0.8 mmol), and stirred at
room temperature for one hour. The solution was concentrated
in vacuo, placed under high vacuum for one hour, then dissolved
in methylene chloride (3 ml) and added to the 4-aminohexahydro-
3-(4-phenylmethoxy)-benzoylamino-1-phenylmethylazepine prepared
above. Sodium hydroxide (1.0 N, 1.5 ml) was added, and the
mixture was stirred for one hour and separated. The aqueous
layer was extracted with methylene chloride (2x10 ml), and the
combined organic layer and extracts were washed with saturated
sodium chloride (10 ml), dried (Na2SO~), and concentrated in
vacuo. The residue was chromatographed (flash) on silica gel
(eluted successively with 3% acetone/methylene chloride, 5%
acetone/methylene chloride, and 8% acetone/methylene chloride)
to afford, initially, syn-4- ( 3,s-bis(phenylmethoxy)-4- ( 2-
carbophenylmethoxy-6-phenylmethoxybenzoyl)-benzoylamino
_ V094/2~ 2 1 ~ 7 4 1 2 PCT~S94/0~
- 193 -
hexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenylmethyl-
azepine (0.26 g, 43%), then anti-4-(3,5-bis(phenylmethoxy)-4-
(2-carbophenyl-methoxy-6-phenylmethoxybenzoyl)benzoylamino
hexahydro-3-(4-phenyl-methoxy)benzoylamino-1-phenylmethyl
azepine (0.21 g, 35%) as colorless foams. The combined yield
was 0.47 g (78%).
syn~ - (2-Carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyl-
~m~no)hexahydro-3-(~-hy~roxybenzoyl~m~no)azepine, trifluoro-
acetic acid salt (CO~PO~ND 516)
A solution of anti-4-(3,5-bis(phenylmethoxy)-4-(2-
carbophenylmethoxy-6-phenylmethoxybenzoyl)benzoylaminohexahydro
-3-(4-phenylmethoxy)-benzoylamino-1-phenylmethylazepine (0.20
g, 0.183 mmQl) in reagent ethanol (9 ml) and ethyl acetate (1
ml) in a 2-neck 25-ml round bottom flask under nitrogen was
treated with Pearlman's catalyst (20% Pd(OH)2/C, 50 mg) and
trifluoroacetic acid (42 mg, 0.37 mmol). The flask was fitted
with a balloon and a balloon valve, purged with hydrogen, and
placed under positive hydrogen pressure for 18 h, then
evacuated of hydrogen and purged for several minutes with
nitrogen. The solution was carefully filtered through celite
(wash filter pad with ethanol) and the filtrate was
concentrated in vacuo to a yellow foam. This was dissolved in
methanol (20 ml), diluted with deionized water (60 ml), and
concentrated in vacuo to remove the methanol. Freeze drying
(withoutchromatography)affordedsyn-4-(4-(2-carboxy-6-hydroxy
benzoyl)-3,5-dihydroxybenzoylamino)hexahydro-3-(4-hydroxy
benzoylamino)azepine (94 mg, 68%) as a voluminous yellow solid;
mp >300-C(dec). Rf (4% acetic acid/ethanol) 0.50; IR (KBr)
1683, 1636, 1604 cm1; 1H NMR (d6-DMS0) ~ 11.66 (s, 2H), 10.07
(s, lH), 9.86 (s, lH), 8.60 - 9.00 (br s, 2H), 8.33 (d, lH, J
= 8 Hz), 8.00 (d, lH, J = 7 Hz), 7.70 (d, 2H, J = 9 Hz), 7.37
(d, lH, J = 8 Hz), 7.27 (t, lH, J = 8 Hz), 7.06 (d, lH, J = 8
Hz), 6.82 (d, 2H, J = 9 Hz), 6.68 (s, 2H), 4.55 (m, lH), 4.40
(m, lH), 3.10 - 3.50 (m, 4H), 1.70 - 2.10 (m, 4H). Anal.
Calcd. for C2~H27N30~-l.5(C2H02F3)-2.0(H20): C, 49.21; H, 4.33; N,
5.55. Found: C, 48.82; H, 4.59; N, 5.79.
WO 94/20062
2 1 5 ~ 4 1 ~ - 194 - PCT/US94/02283
syn- ~ - t ~ - ( 2 -hydroxycarbonyl- 6-hy~roxybenzoyl ) - 3, 5-0 ihydroxy
benzoyloxy] -3- (~-hy~roxybenzamido) perhydroazepine
trifluoroacetic aci~ salt (CONPO~rND 517 )
BnO2C O OBn BnO2C O OBn
OH OBn O~ COCI ~ H OBn
~,H~ Et3N, DMAP. CH2C12 (~Nb~
Bn Bn
1. Pd(OH)2-C H ~ ~OH 610
EtOH. EtOAc OH OH ~, O
NH CF3CO2H
517
COMPO~lND 610
To a chilled solution (0-5 C) of 4-(6-benzyloxy-2-
(benzyloxycarbonyl)benzoyl)-3,5-dibenzyloxybenzoic acid (0.52
mmol, 350 mg) in 2 ml of methylene chloride under nitrogen
atmosphere was added oxalyl chloride (0.75 mmol, 95 mg) in one
portion. N,N-Dimethylformamide (1 drop) was added and the
light brown solution was stirred at 0 C for 1.5 h. The
solvent and excess oxalyl chloride were then removed in vacuo,
the resulting brown oil was redissolved in methylene chloride
(2 ml) and added to a cooled solution (0-5 C) of syn-N-
benzyloxycarbonyl -3 - ( 4 -hydroxybenzamido ) -4 -
hydroxyperhydroazepine (0.62 mmol, 226 mg), triethylamine
(1.29 mmol, 130 mg) and 4-dimethylaminopyridine (approximately
10 mg) in 2 ml methylene chloride (nitrogen atmosphere). The
reaction mixture was allowed to warm to room temperature and
~0 94l20062 2 1 !~ 7 ~ 1 2 PCT/US94/02283
-- 195 --
stirred overnight (approximately 18 h) under a nitrogen
atmosphere, after which it was diluted with 50 ml of methylene
chloride, washed with saturated sodium bicarbonate solution (lo
ml), water (10 ml), and brine (10 ml), dried over anhydrous
sodium sulfate, filtered, and concentrated under vacuum to give
440 mg of the crude product. Chromatography on silica gel
eluting with 4:1-hexane:ethyl acetate gave 399 mg (71%) of the
coupled product as a light yellow solid, which was used
directly in the next step.
To a solution of syn-N-benzyloxycarbonyl-4-[4-(2-
benzyloxycarbonyl-6-benzyloxybenzoyl) -3, 5-dibenzyloxy
benzoyloxy]-3-(4-benzyloxybenzamido)perhydroazepine (0.16 mmol,
180 mg) in ethyl acetate (5 ml) and absolute ethanol (10 ml)
was added moist 20% palladium hydroxide on carbon (20% w/w, 36
mg). The reaction flask was fitted with a hydrogen balloon and
the grey suspension was stirred at room temperature for 18
hours. The reaction flas~c was then purged with nitrogen gas
and the solution diluted with chloroform (50 ml), filtered over
celite, treated with 1 ml of trifluoroacetic acid, and
concentrated in vacuo to give 87 mg of the crude product. The
material was purified by HPLC chromatography with a 21 x 250 mm
C18 column (solvent A: 95:5 water/acetonitrile + 0.1% TFA;
solvent B: 100% acetonitrile; gradient 0-50% B over 60
minutes, flow: 15 ml/min). The purified fractions were
concentrated and lyophilized from water to give 67 mg (63%) of
the title compound as a yellow fluffy solid. IR (KBr): 1704,
1688, 1677, 1632, 1608, 1427, 1235, 1201 cm 1; EA (calculated
for C28H26N2Ol0-1.2 C2HF3O2-2.0 H2O): C, 50.48; H, 4.35 N, 3.87.
Found: C, 50.51; H, 4.46; N, 3.88.
W094/2 62 - 196 - PCT~S94/0~
I+)-anti-3-t4-hydroxybenzamido)-4-t3,5-dimethoxy-~-(2,6-
dimethoxy)benzoyl]benzoyloxylperhydroazepine trifluoroAcetic
aci~ salt (COMPOUND 518)
To (+)-anti-3-(4-benzyloxybenzamido)-4-t3,5-dimethoxy-
4-(2,6-dimethoxy)benzoyl]benzoyl~oxy-N-benzylperhydroazepine
(221 mg, 0.291 mmol) dissolved in ethyl acetate (50 ml) under
an atmosphere of nitrogen was added trifluoracetic acid (35 ~L,
0.437 mmol) followed by Pearlman's catalyst (44 mg, 20 % by wt
on carbon). An atmosphere of hydrogen was introduced and the
mixture allowed to stir for 48 h. The catalyst was removed by
filtration and the volatiles were removed under reduced
pressure. The product was chromatographed on a Dynamax~-60 C18
column (41.4 mm ID X 30 cm length) using a linear gradient from
100% A (0.1% TFA and 5% acetonitrile in water) to 100% B (pure
acetonitrile) over 60 m at 25 ml/min. The product elutes in 58
minutes. Removal of the volatiles under reduced pressure
provided Compound 518 as a white solid (62 mg, 26%), mp
134-137-C. IR KBr (disc) cml 3430, 3105, 3016, 2948, 2843,
1772, 1677, 1592, 1544, 1508, 1474, 1436, 1411, 1331, 1235,
1203, 1182, 1127, 1111, 950, 917, 848, 798, 766, 720, 620, 602.
Anal. Calcd for C31H34N2Og 2CF3CO2H: C, 52.11; H, 4.50; N,
3.47. Found: C, 52.30; H, 4.50; N, 3.47.
(+)-~nti-3-(~-benzyloxybenzami~o)-~-t3,5-dimethoxy-~-(2,6-
dimethoxy)benzoyl]benzoyloxy-N-benzylperhydroazepine
~COMr~uN~ 60~)
To a solution of 3,5-dimethoxy-4-(2,6-
dimethoxybenzoyl)benzoic acid (221 mg, 0.639 mmol) in anhydrous
dichloromethane (10 ml) under an atmosphere of nitrogen at 0-C
was added oxalyl chloride (436 ~L, 2 M in dichloromethane,
0.872 mmol) dropwise over 5 minutes followed by anhydrous
dimethylformamide (3 drops). The ice bath was removed and the
suspension rapidly turned into a clear yellow solution. The
reaction mixture was allowed to stir for 0.5 h at room
temperature. The volatiles were removed under reduced pressure
and the remaining solid was dried under vacuum for 2.5 h.
_ ~094/2~K2 2 ~ 5 7 412 pcT~ss4lo22~
- 197 -
To a solution of (+)-anti-3-(4-benzyloxybenzamido)-4-
hydroxy-N-benzylperhydroazepine (250 mg, 0.581 mmol),
triethylamine (267 ~L, 1.92 mmol), and dimethylaminopyridine
(7.1 mg, 0.0581 mmol) in anhydrous dichloromethane (10 ml)
under an atmosphere of nitrogen at 0C was added a solution of
the above generated acid chloride in anhydrous dichloromethane
(10 ml) dropwise over 0.5 h. After allowing to stir while
warming to room temperature overnight the reaction mixture was
diluted with dichloromethane (200 ml) and washed with water (75
ml). The dichloromethane layer was dried over magnesium
sulfate, filtered, and the volatiles were removed under reduced
pressure to give a crude white solid. The solid was purified
using flash column chromatography (silica gel, 9 : 1
dichloromethane and ethyl acetate) to provide Compound 604 as
a white solid (266 mg, 60%), mp 82-85-C. IR KBr (disc) cm
3481, 3029, 2939, 2837, 1711, 1676, 1644, 1591, 1530, 1498,
1474, 1407, 1327, 1248, 1177, 1111, 1024, 995, 913, 845, 795,
744, 700, 605. Anal. Calcd for C45H46N20~: C, 71.22; H, 6.11;
N, 3.69. Found: C, 71.08; H, 6.15; N, 3.64.
3,5-dimethoxy-4-(2,6-dimQthoxy)bQnzoylbenzoiG acid
To t-butyl-3,5-dimethoxy-4-(2,6-dimethoxybenzoyl)
benzoate (2.26 g, 5.62 mmol) under an atmosphere of nitrogen at
O C was added formic acid (25 ml) dropwise. The ice bath was
removed and the reaction mixture was allowed to stir at room
temperature for 8 h. The volatiles were removed under reduced
pressure. Recrystallization of the crude solid with 1:1 ethyl
acetate:hexanes provided the title compound as a white solid
(1.69 g , 87%), mp 209-212-C. H NMR (DMSO-d6) ~ 7.32 (t, 1 H,
J = 8 Hz), 7.16 (s, 2 H), 6.64 (d, 2 H, J = 8 Hz), 3.67 (s, 6
H), 3.61 (s, 6 H); IR KBr (disc) cm1 3432, 3199, 3103, 3016,
2936, 2838, 1731, 1653, 1589, 1475, 1429, 1409, 1313, 1258,
1222, 1185, 1128, 1113, 1028, 944, 897, 869, 801, 773, 713,
690, 617. Anal. Calcd for C1~Hl~O7: C, 62.42; H, 5.24. Found:
C, 62.43; H, 5.30.
t-butyl-3,5-dimethoxy-4-(2,6-dimQthoxybenzoyl)benzoatQ
W094/2~62 PCT~S94/0~
2157 ~ 198 -
To a solution of t-butyl-3,5-dimethoxy-4-[(2,6-
dimethoxyphenyl)hydroxymethyl]benzoate (3.10 g, 7.70 mmol) in
acetone (50 ml) at O C was added Jones reagent dropwise until
the reaction mixture retained the orange color of the Jones
reagent. The reaction mixture was diluted with dichloromethane
(500 ml) and washed with water (150 ml). The dichloromethane
layer was dried over anhydrous magnesium sulfate, filtered and
the volatiles were removed under reduced pressure. The crude
residue was purified using flash column chromatography (silica
gel, 1 : 8 ethyl acetate / hexane) to provide the title
compound as white solid (2.36 g, 88%), mp 54-57-C. lH NMR
(DMSO-d6) ~ 7.32 (t, 1 H, J = 8.5 Hz), 7.11 (s, 2 H), 6.63 (d,
2 H, J = 8.5 Hz), 3.67 (s, 6 H), 3.61 (s, 6 H) 1.55 (s, 9 H);
IR KBr (disc) cm1 2974, 2839, 1711, 1685, 1590, 1473, 1434,
1406, 1369, 1330, 1292, 1253, 1163, 1126, 1111, 1032, 957, 915,
849, 796, 765, 705, 669, 610. Anal. Calcd for C22H26O7: C,
65.66; H, 6.51. Found: C, 65.67; H, 6.49.
t-butyl-3,5-dimethoxy-~-~(2,6-dimethoxyphenyl)hydroxy-
methyl]benzoate
Toasolutionoft-butyl-3,5-dimethoxy-4-bromobenzoate
(3.50 g, 11.0 mmol) in anhydrous tetrahydrofuran (60 ml) under
an atmosphere of nitrogen with an internal temperature of -78-C
(ether/dry ice) was added n-butyllithium (7.58 ml, 1.6 M in
hexanes, 12.1 mmol) dropwise at a rate which did not allow the
internal temperature to rise above -65-C. To the reaction
mixture was added a solution of 2,6-dimethoxybenzaldeyde (1.83
g, 12.1 mmol) in anhydrous tetrahydrofuran (20 ml) dropwise at
a rate which did not allow the internal temperature to rise
above -65-C. The reaction mixture was allowed to stir while
warming to room temperature over 2 h. The reaction mixture was
quenched with solid ammonium chloride and the volatiles were
removed under reduced pressure. The crude residue was diluted
with ethyl acetate (500 ml) and washed with water (200 ml).
The ethyl acetate layer was dried over anhydrous magnesium
sulfate, filtered, and the volatiles were removed under reduced
pressure. The crude residue was purified by recrystallization
W094/2~62 215 ~ 412 PCT~S94/0~
-- 199 --
from 1 : 1 ethyl acetate : hexane which provided a white solid
of the title compound (3.2 g, 72%), mp 131-133-C. H NMR
(DMSO-d6) ~ 7.14 (t, 1 H, J = 8.5 Hz), 7.08 (s, 2 H), 6.58 (d,
2 H, J = 8.5 Hz), 6.43 (d, 1 H, J = 10.5 Hz), 5.37 (d, 1 H, J
= 10.5 Hz) 3.76 (s, 6 H), 3.70 (s, 6 H) 1.63 (s, 9 H);IR KBr
(disc) cml 3541, 3445, 2978, 2843, 1701, 1651, 1588, 1542,
1477, 1455, 1417, 1366, 1328, 1241, 1161, 1119, 1030, 958, 847,
809, 770, 667. Anal. Calcd for Cz2H2~O7: C, 65.33; H, 6.98.
Found: C, 65.19; H, 6.99.
t-butyl-~-bromo-3,5-~imethoxybenzoate
To a solution of 4-bromo-3,5-dimethoxybenzoic acid
(9.20 g, 35.2 mmol) in anhydrous dimethylformamide (200 ml)
under an atmosphere of nitrogen was added N,N-carbonyl
diimidazole (6.29 mg, 38.8 mmol). After the reaction mixture
was allowed to stir for 1 h at room temperature, DBU (5.80 ml,
38.8 mmol), and t-butanol (9.97 ml, 106 mmol) were added. The
reaction mixture was heated at 80-C for 2 h. The reaction
mixture was quenched by the slow addition of water (300 ml).
The solid which formed was collected by suction filtration and
washed with water (3 X 30 ml) which provided the title compound
as a white solid (5.8 g, 52%), mp 119-121-C. lH NMR (DMSO-d6)
~ 7.17 (s, 2 H), 3.89 (s, 6 H), 1.57 (s, 9 H); IR KBr (disc)
cm-l 2979, 2936, 2836, 1708, 1590, 1456, 1408, 1366, 1337,
1258, 1231, 1174, 1124, 1033, 960, 8S7, 798, 761, 643. Anal.
Calcd for C13Hl7BrO4: C, 49.23; H, 5.40. Found: C, 49.05; H,
S.36.
~-bromo-3,5-dimethoxybenzoic acid
To methyl 4-bromo-3,5-dimethoxybenzoate (10 g, 36.4
mmol, Pharmatech International) was added sodium hydroxide (75
ml, 10 N) and methanol (50 ml). The reaction mixture was
heated at 70-C for l.S h. The reaction mixture was cooled to
Q-C and slowly acidified using HCl (6N). The solid was
collected by suction filtration to provide a white solid of the
title compound (9.31 g, 98%), mp 22S-228-C. 1H NMR (DMSO-d6)
~ 13.3 (br s, 1 H), 7.23 (s, 2 H), 3.90 (s, 6 H); IR KBr
~ 5 7 4 1~ PCT~S94/022~ _
- 200 -
(disc) cml 3400, 3064, 2969, 2838, 2745, 1689, 1586, 1461,
1407, 1329, 1276, 1230~, 1189, 1127, 1039, 935, 857, 764, 730,
667, 639. Anal. Calcd for CgHgBr04: C, 41.41; H, 3.47. Found:
C, 41.44; H, 3.40.
_ W094/20~2 2 1 ~ 7 412 PCT~S94/02283
~3R,~R)-[~-(2,6-Dimethoxybenzoyl)-3,s-~ihy~roxybenzoyloxy~-3-
(~-hy~roxybenz~mido)perhy~ro~zepine trifluoro~cetic ~cid ~lt
(COMPOUND 519)
Br
BnO~OBn1)gULi,THF,-72C OMe O ~
~Me ~CO2t-Bu
CO2t-Bu2) ~ OMe OBn
OMe
Jones
Reagent
OMeo OMe o
~COOH , HCOOH ~ CO2t-Bu
OMe OBn OMe OBn
1) (cOc1)2
CH2CI2/DMF
,OH
~) (;;~Nb~OBn
~ Ph PH320~7A
Et3N, DMAP
CH2CI2
0~ 0~
O BnO 1)TFA
~S--HN ,~OBn 2) H2 ~tNb~O H
~NJ O PdoH)2 / C ~NJ O
- Fh TFA
605 519
WO94t2~62 215 7 ~12 PCT~S94/0~
- 202 -
l,l-Dimethylethyl 3,5-dibenzyloxy-~-[~2,6-dimethoxyphenyl)-
hydro~ymethyl) benzoate
2.5 ml (7.02 mmole) of 2.5 M n-butyllithium in hexane
was added dropwise to a cold solution (-72-C) of t-butyl-4-
bromo-3,5-dibenzyloxybenzoate (2.69 g, 5.47 mmole) in 40 ml of
anhydrous tetrahydrofuran (THF) under nitrogen. The
temperature of the solution was maintained below -70-C
throughout the addition and the solution was stirred in the
cold for ten minutes. A solution of 1 g (6.02 mmole) of 2,6-
dimethoxybenzaldehyde in 15 ml of THF was added dropwise
maintaining the temperature below -65-C throughout the
addition. The solution was allowed to warm to room temperature
as it stirred for two hours. The solution was partitioned
between 20 ml of 1 N hydrocholoric acid and 50 ml of ethyl
acetate while stirring fifteen minutes. The organic layer was
separated, washed with water, saturated brine and dried over
magnesium sulfate. The solvent was removed in vacuo. The
residue was recrystallized from ethyl acetate-hexane to give
0.83 g (26%) of the title compound as white crystals, mp
152-155-C. Anal. Calcd for C34H36O7: C, 73.36; H, 6.51. Found:
C, 73.04; H, 6.54.
l,1-Dimethylethyl 4-(2,6-dimethoxybenzoyl)-3,5-dibenzoyl
oxybenzoate
To a cold solution (O-C) of 0.7 g (1.25 mmole) of 1,1-
dimethylethyl 3,5-dibenzyloxy-4-[(2,6-dimethoxyphenyl)-
hydroxymethyl) benzoatein 15 ml of acetone was added 4 ml of
Jones reagent. The solution was stirred in the cold for two
hours. To the solution was added 10 ml of isopropyl alcohol to
destroy the Jones reagent. The reaction mixture was filtered
through celite and washed through with acetone. The filtrate
was concentrated in vacuo. The residue was recrystallized from
ethanol-water to yield 0.47 g (68%) of the title compound as
tan crystals. Anal. Calcd for C34H3407: C, 73.63: H, 6.18.
Found: C, 73.15; H, 6.38.
WO94/2~K2 21~ 7 412 PCT~S94/022~
- 203 -
~-(Z,6-Dimethoxybenzoyl)-3,5-~ibenzyloxybenzoic acid
A solution of 0.46 g (0.83 mmole) of l,l-dimethylethyl
4-(2,6-dimethoxybenzoyl)-3,5-dibenzyloxybenzoate in 10 ml of
formic acid was stirred at room temperature for two hours.
After stirring for one hour a precipitate formed. The reaction
mixture was poured over ice water, and the resultant
precipitate was collected and dried to yield 0.36 g (87%) of a
tan solid.
Trans-N-Benzyl-4-(~-(2,6-dimethoxybenzoyl)-3,5-~;~^n~yloxy)-3-
(4-benzyloxybenz~mido)~zepine (COMPOUND 605)
A solution of 0.36 g (0.72 mmole) of 4-(2,6-
dimethoxybenzoyl)-3,5-dibenzyloxybenzoic acid in 10 ml of
methylene chloride containing a trace (approximately 1 ~L) of
dimethylformamide was cooled to 0-C. A 2.0 M solution of
oxalyl chloride in methylene chloride (0.41 ml, 0.82 mmole) was
added, and the mixture was stirred under a nitrogen atmosphere
for ninty minutes. The reaction mixture was evaporated and the
residue was evaporated twice from 15 ml of methylene chloride.
The residue was dissolved in 8 ml of methylene chloride and was
added to a solution of 0.35 g (0.82 mmole) of trans-N-benzyl-3-
(4-benzyloxybenzamido)-4-hydroxyazepine, 0.06 ml (0.82 mmole)
triethylamine, and 4.0 mg of DMAP in 10 ml of methylene
chloride. The solution was stirred at room temperature under
nitrogen for sixteen hours. The solution was diluted with 30
ml of methylene chloride and washed with saturated sodium
bicarbonate, saturated brine, dried over magnesium sulfate, and
the solvent was removed in vacuo. The residue was
chromatographed on silica gel eluting with hexane-ethyl acetate
(70:30). Yield 0.33 g (43%) of a glassy oil which solidified
on standing.
(3R,~R)-t~-(2,6-Dimethoxybenzoyl)-3,5-dihy~roxybenzoyloxy]-3-
(~-hydroxybenzamido)perhydroazQpine trifluoroacetic acid salt
(COMPO~ND 519)
A solution of 0.33 g (0.36 mmole) of trans-N-benzyl-4-
(4-(2,6-dimethoxybenzoyl)-3,5-dibenzoyloxy)-3-(4-benzyloxy
W094/20062 - 204 - PCT~S94/02283
benzamido)azepine in 20 ml ethanol-methylene chloride (1:1) was
treated with 0.055 ml (0.720 mmole) of trifluoroacetic acid.
The solution was stirred for five minutes. The solvent was
evaporated. The ethanol-methylene chloride solvent was added
twice more and evacuated in order to remove excess
trifluoroacetic acid. The residue was taken up in 15 ml of
ethanol, cooled to O-C, and 0.5 g of moist 10% palladium
hydroxide on carbon was added. The mixture was then stirred
under an atmosphere of hydrogen for six hours at room
temperature. The mixture was filtered, evaporated, and the
residue was chromatographed on a 41 X 250 mm C 18 column
(solvent A: 95 : 5 water / acetonitrile + 0.1% TFA; solvent B:
100% acetonitrile; gradient : 0 - 50% B over 60 min., flow: 25
ml / min.). The pure fractions were pooled and evaporated to
yield 0.158 g (60%) of Compound 519, a yellow powder, mp
189-193 C. IR (KBr); 1678, 1605, 1508, 1427, 1370, 1250, 1200,
765 cml. Anal. Calcd for C29H30N2O~ 2H2O B 1-.3 TFA: C, 51.65;
H, 4.84: N, 3.81. Found: C, 51.27; H, 4.68; N, 3.62.
WO 94/20062 215 7 412 PCT/US94/02283
-- 205 --
Trans-4- ~4- (2-cis-carboxycyclohexylcarbonyl) -3, 5-
~ihydroxybenzoyloxy) -3 - ( 4 -hy~roxybenzamido ) ~zepine
trifluoroacetic ~cid salt (COMPO~rND 519)
HOOC O OBn
BnO~,OBn l)BULi. THF, -72Qc [~C02t Bu
CO2t-Bu 2) ~O
BnBr
O K2C3
DMF
BnOOC O OBn BnOOC O OBn
[J~ , HCOOH ~¢,~
BnO CO2H BnOCO2t-Bu
1) (cOc1)2
CH2C12 / DMF
~O H
2) ( )~...N~OBn
N O
~Ph
~OBn 1)TFA 0~
(~... N~OBnPd(OH)2 / C (~S~N~O H
- ~ Fh TFA
606 520
1, 1-Dimethylethyl 3, 5-dibenzyloxy-4- (2 -Ci9-
carboxycyclohexylcarbonyl ) benzoate
2 . 8 ml (7.03 mmole) of 2.5 N n-butyllithium in hexane
was added dropwise to a cold solution (-72 C) of 3.0 g (6.39
mmole) of 1,1-dimethylethyl 4-bromo-3,5-dibenzyloxybenzoate in
W094/2~62
Z 15 ~ 412 PCT~S94/0~
- 206 -
40 ml of anhydrous tetrahydrofuran (THF) under nitrogen. The
solution was stirred in the cold for ten minutes. A solution
of 1.2 g (7.03 mmole) of cis-1,2-cyclohexanedicarboxylic
anhydride in 10 ml of THF was added dropwise maintaining the
temperature below -70 C. The solution was stirred in the cold
for two hours. The solution was poured into 150 ml of
saturated ammonium chloride and 350 ml of ether. The reaction
mixture was stirred for thirty minutes. The organic layer was
separated, washed with 0.1 N hydrochloric acid, saturated brine
and dried over magnesium sulfate. The solvent was evaporated
to yield 3.4 g (98 % ) of a clear oil.
1,1-Dimethylethyl ~ 2-cis-benzyloxycyclohexylcarbonyl))-
3,5-dibenzyloxybenzoate
To a solution of 3.40 g (6.20 mmole) of 1,1-
d im e t h y l e t h y l 3, 5 - d i b e n z y l o xy -4 - (2 - c i s -
carboxycyclohexylcarbonyl)benzoate in 20 ml of dry DMF was
added 0.43 g (3.10 mmole) of potassium carbonate and 0.33 ml
(3.10 mmole) of benzyl bromide. The solution was stirred at
room temperature under nitrogen for eight hours. Starting
material was still present in the reaction. Therefore, an
additional 0.16 ml (1.5 mmole) of benzyl bromide was added and
stirring was continued for sixteen hours. The solution was
poured over 100 ml of ice water, extracted twice with 150 ml
portions of ether. The ether solution was washed with water,
saturated brine and dried over magnesium sulfate. The solvent
was evaporated and the residue was chromatographed on silica
gel eluting with 5 % - 10 % ethyl acetate - hexane to yield 1.0
g (25 %) of a clear oil.
4-(4-~2-cis-benzyloxycyclohQxylc~rbonyl)-3,5-~ih~nzyloxybenzoic
~ci~
A solution of 0.45 g (0.71 mmole) of 1,1-dimethylethyl
4-(4-(2-cis-benzyloxycyclohexylcarbonyl)-3,5-
dibenzyloxybenzoate in 10 ml of formic acid was stirred at room
temperature for four hours. The solution was poured over ice
water, the resultant precipitate was collected and dried to
yield 0.34 g (82.8%) of a white solid. Anal. Calcd for C36H34O7
: C, 74.72 ; H, 5.92. Found : C, 74.46 ; H,5.99.
wo g4t20062 2 1 ~ ~ 4 1 2
PCT~S94/0
- 207 -
Trans-N-benzyl-4-(4-~2-cis-bQnzyloxycyclobexylcarbonyl)-3-~4-
benzyloxybenz~mido)azepine ~COMPOUND 606)
A solution of 0.40 g (0.69 mmole) of 4-(4-(2-cis-
benzyloxycyclohexylcarbonyl)-3,5-dibenzyloxybenzoic acid in 8
ml of methylene chloride containing a trace (amount
approximately 1 ~L) of dimethylformamide was cooled to o oc.
A 2.0 M solution of oxalyl chloride in methylene chloride (0.59
ml, 1.18 mmole) was added and the solution was stirred under
nitrogen for 2.5 hours. The reaction mixture was evaporated,
and the residue was evaporated twice from 15 ml of methylene
chloride. The residue was dissolved in 8 ml of methylene
chloride and added to a solution of 0.34 g (0.78 mmole) of
trans-N-benzyl-3-(4-benzyloxybenzamido)-4-hydroxyazepine,0.08
ml (0.78 mmole) of triethylamine and 4.0 mg of DMAP in 10 ml of
methylene chloride. The solution was stirred at room
temperature under nitrogen for sixteen hours. The solution was
diluted with 30 ml of methylene chloride and washed with
saturated sodium bicarbonate, saturated brine and dried over
magnesium sulfate. The solvent was evaporated and the residue
was chromatographed on silica gel eluting with ethyl
acetate-hexane (1:4). Yield of 0.5 g (73%) of a clear oil.
Trans-4-~ 2-cis-carboxycyclohexylcarbonyl)-3,5-
dihydroxybenzyloxy)-3-(~-hy~roxybenzamido) a zepine
trifluoroacetic acid salt (CO~POUND 520)
A solution of 0.38 g (0.38 mmole) of trans-N-benzyl-4-
(4-(2-cis-benzyloxycyclohexylcarbonyl))-3-(4-
benzyloxybenzamido)azepine in a 25 ml mixture of methanol,
ethanol and methylene chloride (1 : 2 : 2 ) was treated with
0.06 ml (0.77 mmole) of trifluoroacetic acid for five minutes.
The solvent was evaporated, and the methanol, ethanol,
methylene chloride solvent was added twice more and evaporated.
The residue was taken up in 15 ml of ethanol, cooled to 0 C
and 0.05 g of palladium hydroxide on carbon (20% by wt.) was
added. The mixture was then stirred under an atmosphere of
hydrogen for six hours at room temperature. The mixture was
filtered, evaporated and the residue was chromatographed on a
WO 94/20062 PCTtUS94tO2283
2157 ~ 208 -
41 X 250 mm C18 column (solvent A: 95: 5 water / acetonitrile
~ 0.1% TFA; solvent B : 100% acetonitrile ; gradient O - 50% B
over 60 min., flow 25 ml / min.). The pure fractions were
pooled to yield 57 mg (21%) of a yellow powder, mp 122 - 127
C. IR (KBr): 1676, 1607, 1508, 1427, 1365, 1276, 1203, 757
cm . Anal. Calcd for C2ôH32N209 2H20 1.2 TFA : C, 51.18 ; H,
5.25 ; N, 3.92. Found : C, 51.46 ; H, 5.37 ; 3.92.
_ N094/20062 2 15 7 412 PCT~S94/0~
~ Anti-~-t3~5-~ihy~roxy-~-(2~6-~ihy~.ohyLo~zoyl)]hexahy~ro-3
(~-hydroxybenzoyl~mine)~zepine (COMPOUND 521)
OBn O OBn
BnO ~ OBn
OBn OBn
OBn O OBn OBn O OBn
HCOOH ~J~ (I)BnBr ~
OH OBn C02H (2)KOH OBn OBn CO2H
OH O OH
(I)Courlin~ ~
(2)H~ ~ ~
OH OH O CO ~ OH
,~
) 52
N~ H
tert-butyl 4-(2,6-dibenzyloxybenzoyl)-3,5-
dibenzyloxybenzoate (360 mg, 0.51 mmol) was placed in formic
acid(10 ml). The resulting suspension was stirred for 20 min
and intermittantly heated with a heat gun. The reaction was
poured over water (300 ml) and stirred. The solids that
precipitated were then filtered. Next, the solids were
dissolved in ethyl acetate, and dried over sodium sulfate. The
sodium sulfate was filtered off, and the filtrate was
concentrated in vacuo and recrystalized in hexane: ethyl
acetate to yield a light yellow solid (Acid 175 mg). This
~ ~ 7 ~1 ~ PCT~S94/0~
- 210 -
material was dissolved in dimethylformamide (2 ml). Potassium
carbonate (2.2 eq.) was then added. Next, benzyl bromide (5
eq) was added. The reaction was stirred at R.T. for 2 h. The
reaction was taken up in ethyl acetate and lN HCl, and placed
in a separatory funnel. The organic layer was isolated, dried
over sodium sulfate, concentrated in vacuo, and flash
chromatographed eluting with hexane:ethyl acetate / 90:10. The
major product was isolated as a white foam. This material was
then dissolved in a solution of methanol:isopropyl
alcohol:water / 45:45:10 with 5% potassium hydroxide. The
solution was stirred for 4 h at R.T. The organic solvents were
then removed in vacuo, and to the remaining aqueous solution
was acidified with lN HCl. Ethyl acetate (100 ml) was then
added. The biphasic solution was then placed in a separatory
funnel and the organic phase was isolated, dried over sodium
sulfate, filtered, and concentrated in vacuo. The residue was
recrystalized in hexane:ethyl acetate to yield a light yellow
solid (92 mg). This benzophenone acid was then converted to
Compound 521. Compound 521 was prepared through coupling and
reduction in the same was as for Compound 547. A light yellow
solid was obtained through reverse phase C-18 HPLC and
subsequent lyophilization (7.5 mg). The structure was
identified as Compound 521 through 1H NMR.
~094/2~K2 2 1 S 7 ~ 1 2
- 211 - PCT~S94/022~
~ anti-2-(~-~yd~hyL~hzamido)-3-t3,s-dihy~roxy-~-(2-hy~roxy-
6-c~rboxyphenylcarbonyl)benzoyloxy]-N-benzylcaprolactam
(COMPOUND 523)
OBn 1. (COCI)2 cat. DMF, ~OBn
B~OOC~L OBn HO N~ 2. Et3N, DMAP, BnO~ OBn
=~ OB~ C~'F CH2CI2 ~O N
BnO~ N~_ S1% ~XF
o~OH ~-anti ~ ~-anti ~~ h
612
HOOC ¢~L OH
O I~OH OH H2, Pd(OH)2
HO ~ H _~ 95% THF-TFA
CCF
~-anU N~
523
COMPOUND 612
To a solution of 4-(2-benzyloxy-6-benzyloxycarbonyl)-
3,5-dibenzyloxybenzoic acid (304.7 mg, 0.449 mmol) in CH2Cl2
(3ml) was added 2 drops of DMF and oxalyl chloride (2.0 M
solution in CH2C12, 561 ~L, 1.123 mmol) at room temperature.
The mixture was kept stirring at room temperature for 1 hr.
Solvents were removed and the acid chloride residue was taken
into CH2C12 (5ml) after drying under vaccuum for 1 hr.
A solution of the lactam alcohol (200 mg, 0.449 mmol),
Et3N (227.19 mg, 312.9 ~L, 2.245 mmol) and DMAP (10.97 mg,
0.089 mmol) in CH2C12 (5ml) was treated with the acid chloride-
CH2Cl2 solution made above at 5-C. The reaction mixture was
allowed to stir at room temperature for 3 hr and then
chromatographed on silica gel eluting with 5:3 to 1:1
hexane:EtOAc. The product was obtained as white solid (250 mg,
51%).
W24f~67 4 - 212 - PCT~S94/02283
COMPO~ND 523
Compound 612 (225 mg, 0.204 mmol) was dissolved in THF
(20 ml) and treated with a few drops of TFA and Pd(OH)2 (70 mg,
30% by weight on carbon). The mixture was subject to
hydrogenolysis with a H2 balloon for 5 hr. THF was removed in
vacuo and the residue taken into MeOH. The MeOH solution was
concentrated after filtering through a pad of celite to give
Compound 523 as a yellow solid (126 mg, 95%). m.p. 174-176
(dec)-C; lH NMR (CD30D) ~ 7.90 (d, J = 8.4Hz, lH, NH), 7.42
(d, J = 8.7Hz, 2H, ArH), 7.26 (d, J = 7.7Hz, lH, ArH),
7.14-7.12 (s, br, 5H, ArH), 7.04 (t, J = 8.1Hz and J = 7.7Hz,
lH, ArH), 6.78 (d, J = 8.1Hz, lH, ArH), 6.65 (s, 2H, ArH), 6.57
(d, J = 8.6Hz, 2H, ArH), 5.22 (m, lH, H-3). 4.92 (m, lH, H-2),
4.52 (d, J = 14.5Hz, lH, NCHPh), 4.42 (d, J = 14.5Hz, NCHPh),
3.59 (t, br, lH, H-6), 3.22 (dd, br, lH, H-6), 1.87 (m, 2H,
H-5), 1.62 (m, lH, H-4), and 1.32 (m, lH, H-4); IR (KBr) cm1
3398, 1717, 1708, 1635, 1606, and 1543. Anal. Calcd. for
C2SH30NZOll-l-25H2O C, 61.67; H, 4.88; N, 4.12. Found: C, 61.78;
H, 5.02; N, 3.92. LRFAB (M + 1) : 655.
_ W 0 94/20062 2 132 1 5 7 ~ 1 2
- - PCTAUS94/02283
~- R~-4- t~2,6-Dichlorohen~oyl)-3,5- ~ihy ~. GxyL e~zoyloxy]-3- R~- t~-
hy~roxybenzamido)perhydro~zepine trifluoro~cetic ~cid
tCOM PO~nND 52 ~)
Br
BnO~_OBn1)BuLi,THF,-72c Cl O HO~
- ~J l~ C H O ~CO2t-Bu
- CO2t-Bu 2) r~ ~ Cl OBn
~CI
Jones
Reagent
Cl O OBn Cl O OBn
~COOH , HCOOH ~CO2t-Bu
Cl OBn Cl OBn
1)(C O C1)2
CH2CI2/DMF
,OH
2) (~HN ~OBn
~ Ph PH320~7A
Et3N, DMAP
CH2C12
0~ 0~
O BnO 1)TFA
~HN ~ OBn ' ~H ~OH
- ~NJ O Pd(OH)2 t C ~ J o
Ph TFA
608 524
W094/2~62 21~ 7 4 ~ ~ PCT~S94/02283
- 214 -
1,1-Dimethylethyl-3,5-~; hen ~yloxy-~-(2,6-~ichloromethylphenyl-
hydroxy)benzoate
2.80 ml (7.03 mmole) of 2.5 M n-butyllithium was added
dropwise to a cold solution (-72- C) of 3.00 g (6.39 mmole) of
1,1-dimethylethyl-4-bromo-3,5-dibenzyloxybenzoate in 40 ml of
anhydrous tetrahydrofuran (THF) under nitrogen. The
temperature of the solution was maintained below -70 C
throughout the addition and the solution was stirred in the
cold for ten minutes. A solution of 1.20 g (7.03 mmole) of
2,6-dichlorobenzaldehyde in 10 ml of THF was added dropwise to
the solution maintaining the temperature below -65 c
throughout the addition. The solution was allowed to warm to
room temperature as it stirred for two hours. The solution was
partitioned between 20 ml of lN hydrochloric acid and 50 ml of
ethyl acetate while stirring for five minutes. The organic
layer was separated, washed with water, saturated brine and
dried over magnesium sulfate. The solvent was evaporated and
the residue was recrystallized from ethanol to yield 1.90 g
(55%) of white crystals. Anal. Calcd for C30H30Cl205: C, 67.97;
H, 5.35. Found: C, 67.99: H, 5.20
1,1-Dimethylethyl-~-(2,6-dichlorobenzoyl)-3,5-
dibenzyloxybenzoate
To a cold solution (0- C) of 1,1-dimethylethyl-3,5-
dibenzyloxy-4-(2,6-dichlorophenylmethylhydroxy)benzoate (1.00
g, 1.77 mmole) in 20 ml of acetone was added 5 ml of Jones
reagent. As the Jones reagent was being added the solution
became very thick. Therefore, an additional 20 ml of acetone
was added. The reaction mixture was allowed to stir at room
temperature for five hours. Isopropyl alcohol was added to the
solution to destroy the Jones reagent. The reaction mixt~re
was filtered through celite and washed with acetone. The
solvent was removed in vacuo. The residue was taken up in 200
ml of ether. The ether solution was washed with saturated
sodium bicarbonate, saturated brine and dried over magnesium
sulfate. ~he solvent was evaporated to yield 0.79 g (79%) of
a white solid.
2157412
~094/20062 PCT~S94/02283
- 215 -
4-(2,6-DichlorobQnzoyl)-3,5-~ihQnPyloxybenzoic acid
A solution of 0.95 g of the prime compound (1.70
mmole) in 20 ml of formic acid was stirred at room temperature
for three hours. The solution was poured over ice water,
collected and dried. Recrystallized from toluene to yield 0.75
g (87%) of yellow crystals.
Trans-N-Benzyl-4-t~-~2,6-~ichlorobenzoyl)-3,5-dibenzyloxy-
benzoyloYy]-3-(~-benzyloxybenz~mido)perhydroazepine
(COMPOUND 608)
A solution of 0.40 g (0.79 mmole) of 4-(2,6-
dichlorobenzoyl)-3,5-dibenzyloxybenzoic acid in 20 ml of
methylene chloride containing a trace (approximately 1 ~L) of
dimethylformamide was cooled to 0-C. A 2.0 M solution of
oxalyl chloride (0.45 ml, 0.89 mmole) was added and the
solution was stirred under a nitrogen atmosphere at room
temperature for two hours. The solvent was removed in vacuo.
The residue was taken up twice in 20 ml portions of methylene
chloride and the solvent was removed in vacuo. The residue was
dissolved in 8 ml of methylene chloride and added dropwise to
a solution of 0.38 g (0.89 mmole) of trans-N-benzyl-3-(4-
benzyloxybenzamido)-4-hydroxyperhydroazepine, 0.06 ml (0.89
mmole) of triethylamine and 4.00 mg of DMAP in 10 ml of
methylene chloride. The solution was stirred at room
temperature under nitrogen for twenty-four hours. The solution
was diluted with 30 ml of methylene chloride and washed with
saturated sodium bicarbonate, saturated brine and dried over
magnesium sulfate. The solvent was removed in vacuo The
residue was chromatographed on silica gel eluting with hexane
- ethyl acetate (70 : 30). Yield 0.30 g (41%) of a clear oil.
4-R*-4-[(2,6-dichlorohenPoyl)-3,5-~ihydl GAYb~ZOY1OAY] - 3 - R* - ( ~ -
hy~roxybenz~mido)perhy~roazepine trifluoroacetic aci~
(COMPOUND 524)
A solution of 3.00 g (0.33 mmole) of trans-N-benzyl-4-
[4-(2,6-dichlorobenzoyl)-3,5-dibenzyloxybenzoyloxy)-3-(4-
benzyloxybenzamido)perhydroazepine in 15 ml of ethanol was
W094/20062 ~15 7 41~ PCT~S94/02283
- 216 -
treated with 0.05 ml (0.65 mmole) of trifluoroacetic acid. The
solution was stirred at room temperature for five minutes. The
solvent was removed in vacuo. The residue was treated twice
with 15 ml portions of ethanol and evaporated to remove the
excess trifluoroacetic acid. The residue was taken up in 15 ml
of ethanol, cooled to 0- C, and 0.05 g of moist 10% palladium
hydroxide on carbon was added. The mixture was then stirred
under an atmosphere of hydrogen for sixteen hours at room
temperature. The mixture was filtered, evaporated and the
residue was chromatographed on a 41 X 250 mm C 18 column
(solvent A: 9S : 5 water/acetonitrile + 0.1% TFA; solvent B:
100% acetonitrile;gradient 0-50% B over 60 min., flow 25 ml /
min.). The pure fractions were pooled to yield 22.7 mg (10%)
of a yellow powder, of Compound 524, mp 179 - 183-C. IR (KBr):
3425, 2875, 1677, 1607, 1508, 1429, 1370, 1239, 1200, 777 cm~l;
Anal Calcd for C27H24C12N2O7-H20-1.4 C2HF302: C, 48.56; H, 3.74; N,
3.80. Found: C, 48.57; H, 4.02; N, 3.46.
W094/20062 ~5 7 4 12 PCT~S94102283
Tr~ns-~-R~-t~-(2-tr~ns-c~rboxycyclohexylc~rbonyl~-3,5-
~ihydroxybQnzyloxy]-3-R~ -hy~roxybenz~mi~o)perhydro~zepine
trifluoro~cetic ~cid s~lt (COMPOUND 525)
- HOOC O OBn
BnO~3,0Bn 1)BuLi. THF, -72Qc (~CO2t-Bu
CO2t-Bu 2) C~
BnBr
O K2C3
DMF
BnOOC O OBn BnOOC O OBn
[~ , HCOOH ~
BnO CO2H BnO CO2t-Bu
1) (COCI)2
CH2CI2/ DMF
OH
N~OBn
o
~Ph
~OEn 1)TFA , O~
~$ HN~OBnPd(OH)2 / C'~S H~_o H
L HTFA
Ph 609
525
2157~12
W094/2~62 PCT~S94/Ot283 -
- 218 -
~ Dimethylethyl-3,5-~iben~yloxy-4-~2-tr~ns-c~ y~lohe
carbonyl)benzoate
2.8 ml (7.03 mmole) of 2.5 M n-butyllithium was added
dropwise to a cold solution (-72 C) of 3.0 g (6.39 mmole) of
1,1-dimethylethyl-4-bromo-3,5-dibenzyloxybenzoate in 40 ml of
anhydrous tetrahydrofuran (THF) under nitrogen. The solution
was stirred in the cold for ten minutes. A solution of 1.2 g
(7.03 mmole) of trans-1,2-cyclohexanedicarboxylic anhydride in
10 ml of THF was added dropwise maintaining the temperature
below -70 C. The solution was stirred in the cold for two
hours. The solution was poured into lSO ml of saturated
ammonium chloride and 350 ml of ether. The reaction mixture
was stirred for thirty minutes. The organic layer was
separated, washed with 0.1 N hydrochloric acid, saturated brine
and dried over magnesium sulfate. The solvent was evaporated
to yield 3.4 g (98 % ) of a clear oil.
1,1-Dimethylethyl-~ -(2-trans-benzyloxycarbonyl-
cyclohexylcarbonyl))-3,5-dibenzyloxybenzoate
To a solution of 3.40 g (6.20 mmole) of 1,1-
dimethylethyl-3,5-dibenzyloxy-4-(2-trans-carboxycyclohexyl-
carbonyl)benzoate in 20 ml of dry DMF was added 0.90 g (6.20
mmole) of potassium carbonate and 0.70 ml (6.20 mmole) of
benzyl bromide. The solution was stirred at room temperature
under nitrogen for two hours. The solution was poured over 100
ml of ice water, extracted twice with 150 ml portions of ether.
The ether solution was washed with water, saturated brine and
dried over magnesium sulfate. The solvent was evaporated and
the residue was chromatographed on silica gel eluting with 5 %
- 10 % ethyl acetate - hexane to yield 1.0 g (25 % ) of a clear
oil.
4-~ 2-trans-benzyloxycarbonylcyclohexylcarbonyl)-3,5-
dibenzyloxybenzoic acid
A solution of 1.10 g (1.17 mmole) of 1,1-
dimethylethyl-4-(4-(2-trans-benzyloxycarbonylcyclohexyl-
carbonyl)-3,5-dibenzyloxybenzoate in 10 ml of formic acid was
21~74~2
W094/2~62 PCT~S94/0~283
- 219 -
stirred at room temperature for four hours. The solution was
poured over ice water, collected and dried to yield 0.78 g
(82.8%) of a white solid. Anal. Calcd for C36H34O7 : C, 73.63 ;
H, 6.18. Found : C, 73.75 ; H,6.08.
Tr~ns-N-benzyl-~ -(2-tr~ns-benzyloxyc~rbonylcyclohexyl-
c~rbonyl)-3-~-benzyloxybenz~mi~o)perhydroazepine
(CONPO~ND 609)
A solution of 0.40 g (0.69 mmole) of 4-(4-(2-trans-
benzyloxycarbonylcyclohexylcarbonyl)-3,5-dibenzyloxybenzoic
acid in 8 ml of methylene chloride containing a trace (amount
approximately 1 ~L) of dimethylformamide was cooled to 0-C. A
2.0 M solution of oxalyl chloride (0.59 ml, 1.18 mmole) was
added and the solution was stirred under nitrogen for 2.5
hours. The reaction mixture was evaporated, and the residue
was evaporated twice from 15 ml of methylene chloride. The
residue was dissolved in 8 ml of methylene chloride and added
to a solution of 0.34 g (0.78 mmole) of trans-N-benzyl-3-(4-
benzyloxybenzamido)-4-hydroxyazepine, 0.08 ml (0.78 mmole) of
triethylamine and 4.0 mg of DMAP in 10 ml of methylene
chloride. The solution was stirred at room temperature under
nitrogen for sixteen hours. The solution was diluted with 30
ml of methylene chloride and washed with saturated sodium
bicarbonate, saturated brine and dried over magnesium sulfate.
The solvent was evaporated and the residue was chromatographed
on silica gel eluting with ethyl acetate - hexane (1 : 4 ).
Yield of 0.39 g (57%) of a clear oil.
Tr~ns-~-R~ 2-trans-¢arboxycyclohexylc~rbonyl)-3,5-
~ihydroxybenzyloxy)-3-R~-(4-hy~roxybenz~mido)perhydroazepine
trifluoroacetic aci~ salt ~COMPOUND 525)
A solution of 0.39 g (0.39 mmole) of trans-N-benzyl-4-
(4-(2-trans-benzyloxycarbonylcyclohexylcarbonyl))-3-(4-
benzyloxy-benzamido)perhydroazepine in a 20 ml solution of
methanol, ethanol and methylene chloride (1:2:2) was treated
with 0.06 ml (0.77 mmole) of trifluoroacetic acid for five
minutes. The solvent was evaporated, and the methanol,
W094/20062 ~lS 7 ~ ~ ~ PCT~S94/02283
- 220 -
ethanol, methylene chloride solvent was added twice more and
evaporated. The residue was taken up in 15 ml of ethanol,
cooled to 0C and 0.05 g of palladium hydroxide on carbon was
added. The mixture was then stirred under an atmosphere on
hydrogen for six hours at room temperature. The mixture was
filtered, evaporated and the residue was chromatographed on a
41 X 250 mm C18 column (solvent A: 95 : 5 water / acetonitrile
+ 0.1% TFA; solvent B : 100% acetonitrile ; gradient 0 - 50% B
over 60 min., flow 25 ml / min.). The pure fractions were
pooled to yield 57 mg (21%) of Compound 525 as a yellow powder,
mp 199 - 204 C. IR (KBr) : 1676, 1607, 1508, 1427, 1365,
1276, 1203, 757 cml. Anal. Calcd for C28H3zN20~ H20 1.2 TFA
: C, 52.50 ; H, 5.10 ; N, 4.02. Found : C, 52.83 ; H, 5.45 :
3.97.
NO94/2~62 215 7 41 2 PCT~S94/02283
- 221 -
Trans-~-t4-(2-carboxy-6-hydroxybenzoyl)-3-benzoyloxy-5-
hydroxybenzoyloxy]-3-(~-hy~roxybenzamido)perhydroazepine
Trifluoroacetic Aci~ Ralt (COMPOUND 528)
HO~
(~. N~ p~ in~ (~ N~ OH
H H
528
A solution of 9.1 mg (0.013 mmol) of (-)-trans-4-(4-
(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-
hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in
0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016
mmol) of benzoyl chloride. The mixture was stirred for 24 h at
room temperature, after which an additional 6.4 ~L of benzoyl
chloride was added. After an additional 24 h, 4 ml of benzoyl
chloride was added, and the mixture was allowed to stand for a
final period of 24 h, after which the reaction was quenched by
the addition of 2 ml of methanol. The mixture was evaporated
to a residue which was chromatographed on a 21 x 250 mm C18
column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent
B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15
ml/min). The pure fractions were pooled and evaporated and
then lyophilized from water to give 4.8 mg of Compound 528 as
a white fluffy solid. FABMS: m/z 655 (M + H).
W094/2~62 21 S ~ ~1 2 PCT~S94/02283
- 222 -
N-Benzoyl-tr~ns-~-[~-(2-carboxy-6-hy~roxybenzoyl)-3,5-
dihydroxybenzoyloxy]-3-(~-hydroxybenzamido)perhydroazepine
~COMr~uNv 529)
~ H~ py~ n~ (~. N~
o~3
529
A solution of 9.1 mg (0.013 mmol) of (-)-trans-4-(4-
(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-
hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in
0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016
mmol) of benzoyl chloride. The mixture was stirred for 24 h at
room temperature, after which an additional 6.4 ~L of benzoyl
chloride was added. After an additional 24 h, 4 ~L of benzoyl
chloride was added, and the mixture was allowed to stand for a
final period of 24 h, after which the reaction was quenched by
the addition of 2 ml of methanol. The mixture was evaporated
to a residue which was chromatographed on a 21 x 250 mm C18
column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent
B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15
ml/min). The pure fractions were pooled and evaporated and
then lyophilized from water to give 4.6 mg of Compound 529 as
a white fluffy solid. FABMS: m/z 677 (M + Na), 655 (M + H).
2157412
W094/20062 PCT~S94/02283
- 223 -
- N-Benzoyl-trans-~ 2-~ArhQxy-6-hydlG~yLehzoyl)-3-benzoyloxy-
5-hydroxybenzoyloxy]-3-(4-hydroxybenzAmido)perhydroazepine
~COMPOUND 530)
(~, N~ OH I~cOC~
o~0
530
A solution of 9.1 mg (0.013 mmol) of- (-)-trans-4-(4-
(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-
hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in
0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016
mmol) of benzoyl chloride. The mixture was stirred for 24 h at
room temperature, after which an additional 6.4 ~L of benzoyl
chloride was added. After an additional 24 h, 4 ~L of benzoyl
chloride was added, and the mixture was allowed to stand for a
final period of 24 h, after which the reaction was quenched by
the addition of 2 ml of methanol. The mixture was evaporated
to a residue which was chromatographed on a 21 x 250 mm C18
column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent
B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15
ml/min). The pure fractions were pooled and evaporated and
then lyophilized from water to give 5.3 mg of Compound 530 as
a white fluffy solid. FABMS: m/z 781 (M + Na), 759 (M + H).
21~7~1~
W094/2~62 - 224 - PCT~S94/02283
N-Benzoyl-tr~ns-~-t~-(2-~ 6-hy~ L~zoyl)-3-benzoyloxy-
5-hydroxybenzoyloxy]-3-(~-benzoyloxybenz~mi~o)perhy~roazepine
~COMPOUND 531)
HO~
H~_~ OH PhCOCI ~ H~ O
H O~
531
A solution of 9.1 mg (0.013 mmol) of (-)-trans-4-(4-
(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-
hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in
0.15 ml of dry pyridine was treated with 1.8 ~L (2.2 mg, 0.016
mmol) of benzoyl chloride. The mixture was stirred for 24 h at
room temperature, after which an additional 6.4 ~L of benzoyl
chloride was added. After an additional 24 h, 4 ~L of benzoyl
chloride was added, and the mixture was allowed to stand for a
final period of 24 h, after which the reaction was quenched by
the addition of 2 ml of methanol. The mixture was evaporated
to a residue which was chromatographed on a 21 x 250 mm C18
column (solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent
B: 100% acetonitrile; gradient: 0-50% B over 60 min, flow: 15
ml/min). The pure fractions were pooled and evaporated and
then lyophilized from water to give 2.9 mg of Compound 531 as
a white fluffy solid. FABMS: m/z 885 (M + Na), 863 (M + H).
W094/2~62 21~ 7 ~12 PCT~S94/02283
- 225 -
Tr~ns-N-benzyl-~-t~-(3-hydroxyphthalido)-3~5-dihydroxybenzoyl-
oXy]-3-(~-hyd.Gh~Le~z~;do)perhydroazepine Trifluoroacetic Acid
8alt Hy~rate (COMPOUND 535)
~ OH PhCHO ~
(~, N~ NaCNBH3 ~. N~ OH
H M~OH N
510 535
A solution of 15.6 mg (0.022 mmol) of (-)-trans-4-(4-
(2-carboxy-6-hydroxybenzoyl)-3,5-dihydroxybenzoyloxy)-3-(4-
hydroxybenzamido)perhydroazepine trifluoroacetic acid salt in
0.30 ml of methanol was treated with 4.4 ~L (4.6 mg, 0.043
mmol) of benzaldehyde and 21.7 ~1 (0.022 mmol) of a 1 M
solution af sodium cyanoborohydride in tetrahydrofuran. The
mixture was stirred for 24 h at room temperature, after which
the reaction was quenched by the addition of 0.30 ml of TFA.
The mixture was stirred for 3 h and then evaporated to a
residue which was chromatographed on a 21 x 250 mm C18 column
(solvent A: 95:5 water/acetonitrile ~ 0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min).
The pure fractions were pooled and evaporated and then
W094/20062 2 ~ 5 7 ~12 PCT~S94/02283
- 226 -
lyophilized from water to give 10.1 mg of the title compound as
a white fluffy solid. FABMS: m/z 625 (M + H). Anal. Calcd
for C35H32N2O~ 3 H2O TFA: C, 56.06; H, 4.96; N, 3.53. Found:
C, 55.76; H, 4.87; N, 3.82.
21~74 12
NO 94/20062 PCT/US94/02283
-- 227 --
~+) -anti-3- (~.-c~rboxybenz~mi~o) -~- t3, s-~ihy~roxy-~- ~2, 6-
~ihy~roxy) phenylc~rbonyl] benzoyloxyhex~hy~ro~zepine
trifluoro~cetic ~ci~ s~lt (COMPO~D 536)
OBn ~
, Br~BnO~L OBn
~ DIBAL-H ~qBnOJ~OBu t I OBn
BnO~fSL BenzeneBnO~ OBn n-BuU HO~
CN 43%CHO ~ BnO~OBu-
48% O
based on 26 % of
aldehyde recovery
quan. Jones
1. (COC1)2 cat. DMF, ~,
CH2C12 BnOJ~ OBn 1~1
2. Et3N, DMAP, ~ BnO~ ~ OBn
CH2C12 O ~ OBn KOH-EtOH OBn
~COOBn BnO~ 60% ~1
HO N ~1 ~ o~OH based on 28 % of BnO ~ OBu t
61% ( ~ ester recovery
~ NBn ~-anti
BnO ~L OBn ~ OH
~ COOBn H2, Pd(OH)2 s~ N ~ COOH
BnO ~ H ~ lF-TFA HO ~ X~ ~~
~X j2% ~-anti ~,NH CF~OH
~-anV ~_NBn 617 536
The 2,6-dibenzyloxybenzonitrile (4.0 g, 12.68 mmol)
was dissolved in benzene (30 ml) and cooled to 5-C. The
DIBAL-H (1.0 M in Hexane, 15.2 ml, 15.2 mmol) was then added
and the reaction was allowed to warm up to room temperature and
stirred for 4 days. H20 (2 ml) was slowly added- to the
reaction, followed by 2N HCl until pH was 3Ø Solids
precipitated were filtered off and washed with H20 and EtOAc.
WOg4/2~62 215 7 41~ PCT~S94/02283
- 228 -
The organic layer was washed with brine, dried over Na2SO4, and
concentrated. The residue was flash chromatographed on silica
gel eluting with 6:1 Hexane:EtOAc to yield yellow oil (1.69 g,
43~), which was taken to the next, coupling reaction.
To a solution of t-butyl ester bromide (1.9 g, 4.08
mmol) in THF (40 ml), precooled to -75-C, was added n-BuLi
(2.5M, 1.79 ml, 4.49 mmol). The resulting purple solution was
allowed to stir at -75C for 30 min, and a solution of aldehyde
(1.3 g, 4.08 mmol) in THF (20 ml) was added in a period of 10
min. The mixture was allowed to warm up to -65-C in 20 min.
The color of reaction changed from purple to yellow. The
reaction mixture was poured into 0.5N HCl (50 ml) after warming
up to 0-C, extracted with EtOAc (100 ml), and washed with
brine. The crude material after concentration was purified on
a silica gel column eluting with 8:1 Hexane:EtOAc to recover
starting material aldehyde (0.5 g, 26~) and yield pure coupling
product (1.0 g, 48%, based on 26% of recovered aldehyde).
The coupling product (1.0 g, 1.41 mmol) was dissolved
in acetone (40 ml) and treated with Jones's reagent (ca. 2ml)
at 5-C until the color of the reaction remained essentially the
same color as the Jone's reagent. The reaction was then
stirred at room temperature for lhr. Acetone was removed in
vacuo and residue was taken into EtOAc, washed with brine,
dried over Na2SO4, and concentrated. The pure product was
obtained as bright yellow foam from a short silica gel column
eluting with 5:1/Hexane:EtOAc (997 mg, 100%).
The t-butyl ester of benzophenone (565mg, 0.779 mmol)
was suspended in EtOH-H20 (9:1, 10 ml) and treated with KOH
(2N, 8 ml). The resulting cloudy mixture was warmed to 70-C
for 4 hr. Ethanol was removed in vacuo. The aqueous residue
was diluted with EtOAc and water, and acidified by lN HCl. The
organic layer was washed with brine, dried over Na2SO4, and
concentrated. Flash chromatography of the crude with 5:1 to
1:1 /Hexane:EtOAc afforded recovered starting material ester
(160 mg, 28%) and yielded the corresponding acid as a yellow
solid (220 mg, 60%, based on 28~ of recovered ester).
W094/2~62 215 7 ~12 PCT~S94/02283
- 229 -
COMPOUND 617
To a solution of benzophenone acid (200 mg, 0.307
mmol) in CH2Cl2 (3ml) was added cat. DMF and oxalyl chloride
(2.0 M solution in CH2Cl2, 384 ~L, 0.768 mmol) at room
temperature The mixture was stirred at room temperature for 1
hr. Solvents were removed and the acid chloride residue was
taken into CH2Cl2 (5ml) after drying over vacuum for lhr.
A solution of azepine alcohol (SPC 104101, 140.8 mg,
0.307 mmol), Et3N (31.1 mg, 43 ~L, 0.307 mmol) and DMAP (7.5
mg, 0.06 mmol) in CH2C12 (5ml) was treated with the freshly made
acid chloride-CH2Cl2 solution at 5-C. The reaction mixture was
allowed to stir at r. t. for 3 hr and then chromatographed on
silic gel eluting with 3:2 / hexane:EtOAc. The product was
obtained as pale yellow foam-like solid (205 mg, 61%).
COMPOUND 536
The prior product (200 mg, 0.183 mmol) was dissolved
in THF (20ml) and treated with few drops of TFA and 10% Pd(OH)2
(120mg, 62 mol %). The mixture was subjected to hydrogenolysis
at 50 psi for 30 hr. THF was removed in vacuo and the residue
taken into MeOH. The MeOH solution was concentrated after
filtering through a pad of celite and chromatographed on 41 x
300 mm C18 column (solvent A: 95:5 water/acetonitrile + 0.1%
TFA: solvent B: 100% acetonitrile; gradient: 0-100% B over 60
min, flow: 25 ml/min). The pure fractions were evaporated to
give two yellow solids. The minor product (15.0 mg) remained
identified. The major product was found to be Compound 536
(63.5 mg, 52%). m.p. 176-178 (dec)-C; lH nmr (CD30D) ~ 8.06
(d, J = 8.4Hz, 2H, ArH), 7.80 (d, J = 8.5Hz, 2H, ArH) 7.18
(t, J = 8.3 and 8.3 Hz, lH, ArH), 6.94 (s, 2H, ArH), 6.26 (d,
J = 8.3Hz, 2H, ArH), 5.42 (m, lH, C~-H), 4.54 (m, lH, C3-H),
3.50 (d, br, 2H, C~-H or C2-H), 2.30 and 2.09 (m and m, lH and
3H, C5-H and C6-H); IR (KBr) cml 3392, 1707, 1676, 1626, and
1593. Anal. Calc. for C28H26N2Ol0-3.0H20-1.0TFA: C, 50.14; H,
4.63; N, 3.90. Found: C, 50.26; H, 4.33; N, 4.21. LRFAB (M +
1) : 551.
W094t20062 215 ~ 412 PCT~S94/02283
- 230 -
anti-~- t ~ - ~ 2 -carboxy- 6 -hydroxybenzoyl ) - 3, 5-
dihy~roxybenzoyloxy]-bexahydro-3- ~indole-5-c~rh~y~mido) azepine,
trifluoroacetic acid salt ~COMPO~ND 538)
anti-hexahy~ro-~.-hydroxy-3- ~in~ole-5-carboxamido)-1-
phenylmethylazepine
A cooled (5-C) solution of lithium aluminum
hydride/tetrahydrofuran (Aldrich, l.ON, 6 ml, 6 mmol) under
nitrogen was treated with anti-3-aminohexahydro-4-hydroxy-1-
phenylmethylazepin-2-one (0.47g, 2.0 mmol) at a rate to keep
the pot temperature below lO-C. The mixture was stirred for
16h at room temperature, for 2h at reflux, then cooled on an
ice bath. The reaction mixture was treated sequentially
dropwise with water (0.23 ml), 15% sodium hydroxide (0.23 ml),
and water (0.70 ml), and filtered. The filter cake was rinsed
with tetrahydrofuran and the filtrate was concentrated in vacuo
to afford the crude perhydroazepine. Meanwhile, a solution of
indole-5-carboxylic acid (0.45 g, 2.8 mmol) in anhydrous
tetrahydrofuran (8 ml) under nitrogen was treated with 1,1'-
carbonyldiimidazole (0.46 g, 2.8 mmol); some bubbling ensued,
which subsided after a few minutes. The solution was stirred
for 1.5 h, then combined with the crude perhydroazepine. The
mixture was stirred at room temperature for 40 h and
concentrated in vacuo. The residue was dissolved in methanol
(8 ml) and treated with potassium hydroxide (1.0 g) in water
(2.0 ml). The solution was stirred at room temperature for 2h,
partially concentrated in vacuo, and diluted with water (10
ml). The aqueous suspension was extracted with methylene
chloride (3 x 25 ml) containing some 2-propanol, and the
combined organic extracts were dried (Na2SO4), concentrated in
vacuo, and chromatographed on silica gel (eluted with ethyl
acetate) to afford anti-hexahydro-4-hydroxy-3-(indole-5-
carboxamido)-l-phenylmethylazepine (0.47 g, 65%) as a white
solid.
~0 94/20062 215 7 41~ PCT/US94/02283
-- 231 --
anti-~. - t 4 - ~ 2 -carbophenylmethoxy- 6 -phenylmethoxybenzoyl ) - 3,5 -
bis- (phenylmethoxy) benzoyloxyl hex~hydro-3- ( indole-5-
carbo~c~ido) -1-phenylmethylazepine (COMPO~ID 619)
A solution of 4- (2-carbophenylmethoxy-6-
phenylmethoxybenzoyl ) -3,5-bis- (phenylmethoxy) benzoic acid
(0.24g, 0.35 mmol) in anhydrous methylene chloride (1.2 ml) was
treated with N,N-dimethylformamide (2 drops), then with 2.0 N
oxalyl chloride/methylene chloride (0.25 ml , 0.50 mmol), and
stirred for one hour under nitrogen. The solution was
concentrated in vacuo, placed under high vacuum for one hour,
and dissolved in anhydrous methylene chloride (1.5 ml ) . Anti-
hexahydro-4-hydroxy-3- ( indole-5-carboxamido) -1-
phenylmethylazepine (0.145g, 0.40 mmol) was suspended in
anhydrous methylene chloride (1.0 ml), then treated with
4-dimethylaminopyridine ( lOmg), triethylamine (0.10 ml , 0.72
mmol ), and the acid chloride solution prepared above . The
mixture was stirred under nitrogen for 17h and concentrated in
vacuo . S il ica gel chromatography ( eluted with 3 %
acetone/methylene chloride, then 5% acetone/methylene chloride)
af forded anti -4 - t 4 - ( 2 -carbophenylmethoxy-6-
phenylmethoxybenzoyl ) -3,5-bis (phenylmethoxy ) benz oyloxy ] hexa-
hydro-3-(indole-5-carboxamido)-1-phenylmethylazepine (0.18g,
50%) as a colorless foam.
anti-~- t ~- ~ 2-carboxy- 6-hydroxybenzoyl ) -3, 5-
dih~ ~,o..~ Lehzoyloxy] -hexaLy~.o 3- ~ indole-5-c~ rhQY ~mido ) azepine,
trifluoroacetic acid salt ~COMPO~JND 538)
A solution/suspension of anti-4 - [4 - (2-
carbophenylmethoxy-6-phenylmethoxybenzoyl) -3, 5-
bis (phenylmethoxy) benzoyloxy ] hexahydro-3 - ( indole-
5-carboxamido) -1-phenylmethylazepine (0.13g, 0.127 mmol) in
reagent ethanol (5ml) in a 25 ml 2-neck flask under nitrogen
was treated with trifluoroacetic acid (30 mg, 0.26 mmol) and
with ethyl acetate (0.20 ml, for solubility), then with 20%
Pd(OH)JC (Pearlman's catalyst, 50mg). The flask was fitted
with a baloon valve connected to a baloon containing hydrogen,
W O 94/20062 215 7 ~1 2 PCTrJS94/02283
-- 232 --
then purged with hydrogen and placed under positive hydrogen
pressure for 20h. The flask was carefully purged with nitrogen
and the solution filtered through celite (wash filter cake with
ethanol ), then the f iltrate was concentrated in vacuo . The
residue was taken up in methanol ( 15 ml ) and trifluoroacetic
acid (0. 5 ml), gravity filtered, and the filtrate was diluted
with de-ionized water (75 ml). The mixture was partially
concentrated ~ n vacuo, and the aqueous solution was
freeze-dried for 18h. Collection of the yellow solid Compound
538, anti-4 - [ 4- ( 2-carboxy-6-hydroxybenzoyl ) -3, 5-
dihydroxybenzoyloxy ~ hexahydro-3 - ( indole-5-carboxamido) -azepine,
trifluoroacetic acid salt (71mg, 68%); mp 170-180-C. IR (KBr)
1680 , 1635 , 1607 cm; mass spectrum (FAB): m/z 574 ; H N~
(d6-DMS0) ~ 11.67 (s, 2H), 11.37 (s, lH), 9.87 (s, lH), 9.00 -
9.20 (m, 2H), 8.64 (d, lH, J = 8 Hz), 8.06 (s, lH), 7.56 (d,
lH, J = 8 Hz), 7.43 (s, lH), 7.30 - 7.45 (m, 2H), 7.27 (t, lH,
J = 8 Hz), 7.05 (d, lH, J = 8 Hz), 6.80 (s, 2H), 6.52 (br s,
lH), 5 . 30 (m, lH), 4 . 55 (m, lH), 3 . 25 - 3 . 50 (m, 2H), 3 . 10 -
3 . 25 (m, 2H), 2 . 05 - 2 . 20 (m, lH), 1 . 80 - 2 . 05 (m, 3H) . Anal .
Calcd. for C30Hz7N3Og- 1. 9 (C2HO2F3) 2 . 0 (H2O): C, 49 .14; H, 4 . 01; N,
5.09. Found: C, 49.10; H, 4.38; N, 4.88.
NO94/2~62 215 7 412 PCT~S94/02283
- 233 -
anti-~- t ~- ~ 2-~ ~ hY~h ~ 6-hydroxybenzoyl)-3,5-~ihydroxybenzoyl-N-
methylamino]hexahy~ro-3-(4-hy~roxybenzoyl~mino)azepine~
trifluoroacetic acid salt ~CONPOUND 539)
Ant$-hexahy~ro-3-(4-phenylmethoxy)benzoyl~mino-1-phenylmethyl-
~-trlfluoroacetyl~minoazepine
~~ A solution of hexahydro-3-(4-phenylmethoxy)-
benzoylamino-l-phenylmethyl-azepin-4-one (0.87 g, 2.03 mmol) in
ethanol (12 ml) was treated with hydroxylamine hydrochloride
(0.19 g, 2.73 mmol), followed by 25% methanolic sodium
methoxide (Aldrich, 0.20 g, 0.93 mmol), and was heated to 50-C
for one hour. The mixture was cooled to room temperature and
treated with additional 25% methanolic sodium methoxide (0.42
g, 1.94 mmol), then concentrated in vacuo to afford hexahydro-
3-(4-phenylmethoxy)benzoylamino-1-phenylmethylazepin-4-one
oxime (0.89 g, 99%) as a colorless foam. A solution of this
oxime (1.065 g, 2.4 mmol) in reagent ethanol (45 ml) in a Parr
bottle was treated with Raney Nickel (Aldrich, one tsp.) and
subjected to hydrogenation in a Parr apparatus at 47 - 51 psi
for 5h. The bottle was carefully evacuated of hydrogen and the
contents filtered through celite (washed with ethanol under
nitrogen). The filtrate was gently concentrated in vacuo at
45-C, diluted with toluene, and further concentrated at -45-C
to remove the remaining ethanol. Meanwhile, a solution of
trifluoroacetic acid (0.33 g, 2.9 mmol) in anhydrous
tetrahydrofuran (6 ml) under nitrogen was treated with
l,l'-carbonyl diimidazole (0.50 g, 3.1 mmol). Some bubbling
ensued, and the mixture was stirred for two hours, cooled on an
ice bath, and combined under nitrogen with the residual amine
prepared above (an additional 2 ml of tetrahydrofuran was used
~ to rinse the CDI adduct into the reaction vessel). The mixture was stirred at room temperature for 18h and concentrated in
vacuo, then the residue was chromatographed on silica gel
(eluted with 3% acetone/methylene chloride, then with 8%
acetone/methylene chloride) to afford, initially, syn-
hexahydro-3-(4-phenylmethoxy)benzoylamino-1-phenylmethyl-4-
trifluoroacetylaminoazepine (0.27 g) followed by anti-
WO 94/20062 21 S 7 ~ :12 PCT/US94/02283
- -- 234 --
hexahydro-3-(4-pllenylmethoxy)benzoylamino-1-phenylmethyl-4-
trifluoroacetylaminoazepine (0.40 g). The total yield of
trifluoroacetamides was 0.67 g (53%); the anti-isomer could be
recrystallized from acetonitrile.
~ti-hexahydro-4-~methylamino)-3-(4-phenylmethoxy)benzoyl~ino-
l-phenylmethylazep~ne
An ice-cooled (5-C) solution of anti-hexahydro-3-(4-
phenylmethoxy) benzoyl-amino-1-phenylmethyl-4-
trifluoroacetylaminoazepine (0.20 g, 0.38 mmol) in anhydrous
N,N-dimethylformamide (2 ml) under nitrogen was treated
dropwise with 1.0 N potassium t-butoxide/tetrahydrofuran
(Aldrich, 0.40 ml, 0.40 mmol), then stirred for 20 min at room
temperature and recooled (5-C). Dimethyl sulfate (38 ~L, 0.40
mmol) was added via syringe, and stirring was continued at 5 C
for 3h. The solution was added to a rapidly stirred mixture of
methylene chloride (10 ml) and saturated sodium bicarbonate (5
ml) and the organic layer was separated. The aqueous layer was
extracted with methylene chloride (10 ml) and the combined
organic solution was dried (Na2S04) and concentrated in vacuo.
The residue was chromatographed on silica gel (eluted
successively with 2%, 3%, and 4% acetone/methylene chloride) to
afford the methylated intermediate (0.15 g) as a viscous
colorless oil. This was dissolved in reagent methanol (1.5
ml), treated with a solution of potassium hydroxide (0.25 g,
4.5 mmol) in water (0.25 ml), and stirred at room temperature~
for 2.5h. The solution was partially concentrated to remove
most of the methanol, diluted with water (5 ml), and extracted
with methylene chloride (2x15 ml). The combined organic
extracts were dried (Na2S04) and concentrated in vacuo to afford
anti-hexahydro-4-(methylamino)-3-(4-phenylmethoxy)benzoylamino-
l-phenylmethylazepine (0.105 g, 62%) as a colorless oil.
WO 94/20062 215 7 112 PCT/US94/02283
-- 235 --
anti-~- t3,5-Bi~ (phenylmethoxy)-~-~2-carbophQnylmethoxy-6-
phenylmethoxybenzoyl)benzoyl-N-methyl~mino]hexahydro-3-(~-
phenylmethoxy)benzoyl~mino-l-phenylmethyl~zepine lC-I.r~uNv
620)
A solution of 3,5-bis (phenylmethoxy) -4- (2-
carbophenylmethoxy-6-phenylmethoxybenzoyl)benzoic acid (0.204
g, 0.30 mmol ) in anhydrous methylene chloride (1.0 ml ) was
treated with N,N-dimethylformamide (2 drops), then with 2.0 N
oxalyl chloride/methylene chloride (Aldrich, 0.22 ml, 0.44
mmol ) and stirred for one hour under nitrogen . The solution
was concentrated in vacuo, placed under high vacuum for 45 min,
then dissolved in methylene chloride (2 ml ) and combined with
anti-hexahydro-4- (methylamino) -3- (4-phenylmethoxy) benzoylamino-
l-phenylmethylazepine (0.10 g, 0.225 mmol). The mixture was
treated with 1.0 N sodium hydroxide (1.0 ml) and stirred for
two hours, then diluted with methylene chloride (15 ml ) and
water (5 ml). The organic layer was separated and the aqueous
solution was extracted with methylene chloride (2x15 ml). The
combined organic solution was dried (Na2S0~) and concentrated
in vacuo. The residue was chromatographed on silica gel
(eluted successively with 5%, then 10% acetone/methylene
chloride, then with 3% methanol/methylene chloride) to afford
anti-4- [3,5-bis (phenylmethoxy) -4- (2 -carbophenylmethoxy-6-
phenylmethoxybenz oyl ) benz oy 1 -N -methy 1 amino ] hexahydro- 3 - (4 -
phenylmethoxy)benzoylamino-l-phenylmethyl-azepine (0.22g, 88%)
as a viscous colorless oil.
anti-4-t4-~2-c~ 6-hy~.oa~L3nzoyl)-3,5-dihy~roxybenzoyl-N-
methyl~mino]hexahydro-3-(~-hydroxybenzoyl~mino)~zepine,
trifluoroacetic ~ci~ s~lt (CONPO~ND 539)
A solution/suspension of anti-4- [3, 5-
bis (phenylmethoxy ) -4 - ( 2 -carbophenylmethoxy-6-
phenylmethoxybenzoyl ) benzoyl-N-methylamino ] hexahydro-3 - (4 -
phenylmethoxy) benzoylamino-l-phenylmethylazepine (0.22 g, 0.20
mmol ) in reagent ethanol (9 ml ) in a 25 ml 2 -neck round bottom
flask was treated with trifluoroacetic acid (50 mg, 0.43 mmol),
then with ethyl acetate (1 ml, for solubility) . Pearlman ' s
WO 94l20062 21~ 7 412 PCT/US94/02283
-- 236 --
catalyst (20% Pd(OH)2/C, so mg) was added, then the flask was
quickly purged with nitrogen and fitted with a baloon valve and
baloon containing hydrogen. The mixture was purged with
hydrogen and kept under positive hydrogen pressure for 20h.
The flask was carefully evacuated of hydrogen and purged for
several minutes with nitrogen. The solution was filtered
through celite (wash filter cake with ethanol) and the filtrate
was concentrated in vacuo and dissolved in
N,N-dimethylformamide (1 ml). The solution was loaded onto a
41x250 mm C18 HPLC column and eluted as follows: A-0.1%
TFA/95:5 water:acetonitrile, B-acetonitrile, 100% A to 50:50
A:B over 60 min collected at 25 ml/min. The appropriate
fractions were combined and partially concentrated in vacuo,
then freeze-dried overnight to afford anti-4-[4-(2-carboxy-6-
hydroxybenzoyl)-3,5-dihydroxybenzoyl-N-methylamino]-hexahydro-
3-(4-hydroxybenzoylamino)azepine, trifluoroacetic acid salt
(101 mg, 68%) as a voluminous pale yellow solid; mp 285-295-C
(dec). Rf(6:1:1 n-BuOH/AcOH/H20) 0.45; IR (KBr): 1682, 1633,
1620 cm; H NMR (d6-DMSO) ~ 11.68 (br s, 2H), 10.07 (br s,
lH), 9.92 + 9.851(s, lH), 8.95 + 8.751 (br s, 2H), 8.28 +
8.181(d, lH, J = 9 Hz), 7.67 + 7.581(d, 2H, J = 9 Hz), 7.20 -
7.40 (m, 2H), 7.00 - 7.10 (m, lH), 6.78 - 6.85 (m, 2H), 6.00
(s, 2H), 4.40 - 4.80 (m, 2H), 3.00 - 3.40 (m, 4H), 2.85 +
2.771(s, 3H), 1.80 - 2.10 (m, 4H); mass spectrum(FAB): m/z 564.
Anal. Calcd. for C29H29N3Og-1~1(C2HO2F3)-3~0(H2O) C, 50.43; H,
4.90; N, 5.66. Found: C, 50.32; H, 4.74; N, 5.74.
21~7~12
~094/2~62
- 237 - PCT~S94/02283
~-R~-~- [ ( ( (3-hy~roxyc~rbonyl) -2-pyri~inyl) c~rbonyl) -3, 5-
Oihy~roxybenzoyloxy] -3-R~ -hy~.o,.yL3nzami~0) perhy~roazepine
trifluoroacetic ~ci~ s~lt (COIIrOu~v S~l)
BnOOC O OBn BnO
1) (COC1)2 ,~\ ~
BnO CO2HCH2CI2 / DMF . o~oBnN
2) (~---N~OBn ~ NHb~OBn
Ph 622
1 ) TFA
2) H2
Pd(OH)2 / C
EtOH
0~
(~,...N~O H
H
TFA
541
Trans-N-benzyl-~- t ~ ( (3-benzyloxyc~rbonyl) -2-
pyridinyl ) c~rbonyl ) -3, 5-dibenzyloxybenzoyloxy] -3- ( ~-
benzyloxybenzamido) perhydro~zepine (coMpo~rv 622 )
Carbonyldiimidizole (0.11 g, 0.65 mmole) was added to
a solution of 4-[((3-benzyloxycarbonyl)-2-pyridinyl)carbonyl]-
3,5-dibenzyloxybenzoic acid ( 0.25 g, 0.44 mmole) in 5 ml of
215 7 '1~ ~ PCT/US94102283
-- 238 --
methylene chloride and the solution was stirred at room
temperature for sixty minutes under nitrogen. The solution was
added to a solution of 0.19 g (0.44 mmole) of trans-N-benzyl-3-
(4-benzyloxybenzamido)-4-hydroxyperhydroazepine, 0.12 ml
triethylamine, and 5 mg DMAP in 8 ml of methylene chloride.
The solution was stirred at room temperature for twenty hours.
The solution was diluted with 30 ml of methylene chloride,
washed with water, saturated brine and dried over magnesium
sulfate. The solvent was removed in vacuo. The residue was
chromatographed on silica gel eluting with a gradient of 5% -
10% - 20% ethyl acetate - hexane to yield 70 mg of a clear oil.
~.-R~-~- [~3-hy~roxycarbonyl)-2-pyridinyl)c~rbonyl)-3,5-
dihydro~cybenzoylo~cy]-3-R~-~4-hy~roxybenzamido)perhydro~zepine
trifluoro~cetic acid
A solution of 0.070 g (0.071 mmole) of trans-N-benzyl-
4-~(((3-benzyloxycarbonyl) -2-pyridinyl) carbonyl) -3, 5-
dibenzyloxybenzoyloxy]-3-(4-benzyloxybenzamido)perhydroazepine
in 8 ml of ethanol/methylene chloride (1:1) was treated with 10
L (0.142 mmole) of trifluoroacetic acid. The solution was
stirred at room temperature for fifteen minutes. The solvent
was evaporated and the ethanol/methylene chloride solvent was
added twice more and evaporated in order to remove the excess
trifluoroacetic acid. The residue was taken up in 10 ml of
absolute ethanol and cooled to 0 C under nitrogen, and 0.030
g (0.025 mmole) of palladium hydroxide on carbon was added.
The reaction mixture was stirred under an atmosphere of
hydrogen for twenty-four hours. The reaction mixture was
filtered, evaporated and the residue was chromatographed on a
21 X 250 mm C18 column (solvent A: 95: 5 water/acetonitrile +
0.1% TFA; solvent B: 100% acetonitrile; gradient: 0 - 50% B
over 60 min., flow 15 ml/min). The pure fractions were pooled
and evaporated to yield 0.010 g of a yellow powder, mp 198-205
C. Anal. Calcd for C27H25N30~-2H20-1.8TFA: C, 47.32; H, 4.00; N,
5.41. Found: C, 47.27; H, 3.86, N, 5.47.
W094/2~62 215 7 4 1 ~ PCT~S94/02283
- 239 -
~+)-Trans-3-~-Hydroxy)benzamido-~ 3,5-~ihydroxy)-~-
phenylcarbonyl]benzoyloxyperhydroazepine trifluoroacetic acid
salt ~COMPOUND 544)
OBn
H~ O OOn O OH
Bn (~)-trans ~X~ ~F OH
1. (COC1)2 ,_~ N~f H2/Pd(OH)2
2. Et3N,DMAP ~ ~ (--~-` O
O OBn. Bn ~-~rans H CF3COOH
COOH 627
COMPOIJN`D 627
To a mixture of N-benzylated azepine intermediate (0.3
g, 0.697 mmol), Et3N (352.6 mg, 487 ~1, 3.485 mmol), and DMAP
(17.0 mg, 0.139 mmol) in CH2Cl2 (5 ml) was added a freshly made
solution of benzophenone acid chloride (corresponding
benzophenone acid: 336 mg, 0.766 mmol; oxalyl chloride: 2.0 M
solution in CH2C12, 0.697 ml, 1.394 mmol and cat. DMF) in CH2Cl2
(5 ml) at room temperature. The reaction mixture was stirred
at room temperature overnight. Flash chromatography of the
reaction mixture on silica gel using 3:2/Hexane:EtOAc as an
eluent gave white solid product (469 mg, 71%).
CO~POUND 5~
The preceeding compound (300 mg, 0.315 mmol) in
EtOAc:MeOH (1:1, 25 ml) was treated with CF3COOH (37.76 mg,
W O 94t20062 2 ~ 5 7 ~ 12 PCTrJS94/02283
-- 240 --
25.5 ~Ll, 0.33 mmol) and 20% Pd(OH)2 on activated carbon (150
mg, 50% on weight basis). The mixture was subjected to
hydrogenolysis at 45 psi for 15 hr. The crude product after
filtration and concentration was taken into DMF (0.5 ml) and
purified on Cl8-HPLC column eluting with 596-50% acetonitrile in
H2O containing 0.1% CF3COOH. The title compound was obtained
as white powder (118 mg, 62%). m.p. 204-206 (dec)-C; 1HNMR
(DMSO-d6) ~ 8.42 (d, lH, NHCO), 7.58 (d, J = 7.5 Hz, 2H, ArH),
7.28-7.15 (m, 4H, ArH), 6.82 and 6.80 (s and s, 2H, ArH), 6.72
(d, J = 8.1 Hz, 2H, ArH), 5.18 (s, br, lH, CH--4), 4.45 (s, br,
lH, CH-3), 3.30 and 3.10 (s and s, br, 4H, CH2N-2,7), 2.10-1.70
(m, 4H, CH2-5,6); IR (KBr) cm 1 3429, 1717, 1703, 1637, 1608,
and 1509. Anal. Calcd. for C27H26N2O7 - 1.5 H2O - 1.0 TFA: C,
55.15; H, 4.79; N, 4.44. Found: C, 54.97; H, 5.08; N, 4.06.
_ WO94/2~K2 21~ 7 412 PCT~S94/02283
- 241 -
Trans-2- ~-Benzoyl-3, 5-dihy~roxybenzoyloxy) -1-
hydroxybenzamido) cyclohexane ~COMPO~IND 545)
Trans-2-(4-benzoyl-3,5-dibenzyloxybenzoyloxy)-1-(4-
benzyloxybenzamido)-cyclohexane (172 mg, 231 ~mol) was
dissolved in ethyl acetate (5 ml) and added to a stirring
mixture of Pearlman's catalyst (Pd(OH) 2, 17 mg) in ethyl
acetate (3 ml). The flask was evacuated under house vacuum and
filled with H2(g) via balloon. After 3 h, additional
Pearlman's catalyst (17 mg in 2 ml of EtOH) was added and left
to stir overnight. TLC showed complete reaction (Starting
material Rf=0.90, product Rf=0.41 in 50% EtOAc/hexanes.) The
reaction mixture was filtered through Celite and concentrated
to give the product as a yellow glass. The glass was
triturated with water to give a pale yellow powder (122 mg,
86%): lH-NMR (DMSO, 300 MHz) ~ 1.27-1.57 (4H, m), 1.63-1.77
(2H, s), 1.84-1.89 (lH, m), 2.05-2.17 (lH, m), 4.05-4.18 (lH,
m), 4.91-5.01 (lH, m), 6.75 (2H, d, J = 8 Hz), 6.97 (2H, s),
7.24-7.36 (2H, m), 7.48 (3H, t), 7.69 (2H, d, J = 8 Hz), 8.10
(lh, d, J = 9 Hz), 9.85 (lH, s), 9.95 (2H, s): 13C NMR (DMSO,
300 MHz) ~ 24.14, 24.52, 24.57, 31.20, 31.79, 51.73, 75.70,
107.68, 108.19, 114.98, 125.96, 126.29, 126.35, 128.35, 129.11,
129.15, 129.45, 132.08, 133.80, 137.24, 155.75, 160.29, 165.47,
166.13, 194.87. IR (KBr) cml 3360, 3271, 2943, 2858, 2360,
1708, 1659, 1610, 1541, 1507, 1450, 1424, 1369, 1347, 1278,
1240, 1177, 1106, 1047, 1011, 847, 771, 595. Anal. Calcd. for
C27H2sNO7 - H20: C, 65.71; H, 5.51; N, 2.84. Found: C, 66.16: H,
5.49; N, 2.76.
W O 94/20062 215 7 ~12 PCTrUS94/02283
- 242 -
Anti-3-(~-benzyloxybenzami~o)-~-[3,5-dibenzyloxy-~-~3,~-
dibenzyloxyphenylcarbonyl)benzoyloxy]-N-benzylperhy~roazepine
~CONPOUND 546)
3,5-Dibenzyloxy-4-[(3,4-dibenzyloxy)benzoyl]benzoic
acid (500 mg, 0.768 mmol) was dissolved in anhydrous
dichloromethane (6 ml). Anhydrous dimethylformamide (0.10 ml)
was then added to the solution followed by oxalyl chloride (2
N in dichloromethane, 0.50 ml, 0.99 mmol). This solution was
stirred for 1 h and then concentrated in vacuo. The resulting
yellow oil was placed under high vacuum for a period of 1 h to
make sure all of the excess oxalyl chloride was removed. The
residue was dissolved in anhydrous dichloromethane (S ml), and
a solution of dimethylaminopyridine (67 mg, 0.5S0 mmol),
triethylamine, and anti-N-benzyl-3-(4-benzyloxybenzamido)-
4-hydroxyazepine (SPC-103853, 215 mg, 0.500 mmol) in anhydrous
dichloromethane (5 ml) was added under nitrogen. The reaction
was stirred at room temperature for 1.5 h. At this point
dichloromethane (100 ml) and sodium hydroxide (0.5 N in water,
30 ml) were added to the reaction. The aqueous and organic
layers were separated and the organic phase was washed with
brine (100 ml). The organic phase was isolated and dried with
magnesium sulfate. The magnesium sulfate was filtered off, and
the solution was concentrated in vacuo to yield a yellow solid.
The yellow solid was flash chromatographed on silica gel
eluting with hexanes:ethyl acetate/9:1, 4:1, and 1:1 to yield
a white solid of the title compound (400 mg, 75%): mp 65-C: lH
NMR (CDCl3) ~ 1.80 (m, 2H, CH2), 2.04 (m, 2H, CH2), 2.73 (m, 2H,
NCH2), 3.01 (m, 2H, NCH2), 3.59 (d, J = 12.5 Hz, lH, NCH2Ph),
3.80 (d, J = 12.9 Hz, lH, NCH2Ph), 4.32 (m, lH, CH), 5.03 (s,
4H, CH2Ph), 5.12 (s, 2H, CH2Ph), 5.13 (s, 2H, CH2Ph), 5.22 (m,
lH, CH), 5.24 (s, 2H, CH2Ph), 6.74 (d, J = 7.8 Hz, lH, NH),
6.86-7.56 (m, 39H, ArH); IR (KBr) cm 1580 (COO-), 1654 (CO).
Analysis calculated for C69H62N2O~: C, 77.95; H, 5.88; N, 2.63.
Found: C, 77.70; H, 5.99: N, 2.60.
W094/20062 21~ 7 412 PCT~S94/02283
- 243 -
Anti-~-[3~s-dihydroxy-~-~3~-dihydroxyphenylc~rbonyl)]
benzoyloxy-3-l~-hy~roxybenz~mido)perhydroazepine
(COMPOUND 547)
Anti-3-(4-benzyloxybenzamido)-4-[3,5-benzyloxy-4-(3,4-
dibenzyloxyphenylcarbonyl)benzoyloxy]-N-benzylazepine (240mg,
0.23 mmol) and acetic acid were dissolved in methanol:ethyl
acetate/2:1 in a 500 ml Parr bottle. Next, 5~ palladium on
activated carbon (45 mg) was added under nitrogen. The
reaction mixture was placed on a Parr hydrogenator for 3 h.
The mixture was then filtered over celite, and the filtrate was
concentrated in vacuo to yield a yellow solid. The solid was
flash chromatographed on silica gel eluting with
dichloromethane:methanol/8:2 to yield a yellow solid of the
title compound (57 mg, 48%): mp 176-C; H NMR (D6 DMSO) ~ 1.65
(m, lH, CH2), 1.78 (m, lH, CH2), 1.90 (2, 2H, CH2), 2.85 (m, 4H,
N(CH2)2), 4.20 (m, lH, CH), 5.18 (m, lH, CH), 6.70-7.70 (m, 9H,
ArH), 8.16 (d, J = 8.4 Hz, lH, NH); IR (KBr) cm1 1607 (CO),
1704 (COO-), 3431 (OH). Anal. calcd. for C27H26N2O8 H20: C,
59.50; H, 5.27; N, 5.14. Found: C, 59.54; H, 5.33; N, 4.93.
~1 L ~'7
W094/2~62 ~lJ J ~ PCT~S94102283
- 244 -
Trans-3-t3,5-Dihy~roxy-~-(2-hydroxy)phenylc~hQnyl]benzami~o-4-
~-hydroxy)benzoyloxyperhy~ro~zepine ICOMr~u~v 548)
To a solution of trans-N-benzyl-3-[3,5-dibenzyloxy-4-
(2-benzyloxy)phenylcarbonyl]benzamido-4-(4-benzyloxy)-
benzoyloxyazepine (65 mg, 0.068 mmol) in EtOAc/MeOH (5/15 ml)
was introduced Pd(OH)z on carbon (35 mg of 20% Pearlman's
catalyst). The reaction mixture was subjected to
hydrogenolysis at 45 psi for 24 h at room temperature. The
catalyst was filtered off through a pad of celite and washed
with MeOH. The combined filtrate was concentrated to dryness
and purified by flash column chromatography (SiO2: 60 ml,
eluted with 10% to 20% MeOH in CH2C12). The title compound was
obtained as a yellow solid (17 mg, 50%): mp 172-175-C, 1H NMR
(CD30D) ~: 7.65 (d, J = 8.7 Hz, 2H, ArH), 7.26 (td, lH, ArH),
7.10 (dd, lH, ArH), 6.74 (dd, lH, ArH), 6.60 (td, lH, ArH),
6.59 (d, J = 8.7 Hz, 2H, ArH), 6.50 (s, 2H, ArH), 5.03 (m, lH,
CH-4), 1.19 (m, lH, CH-3), 2.95-2.70 (m, 4H, CH2N-2,7),
1.91-1.60 (m, 4H, CH2-5,6); IR (KBr) cm1 3434, 1700, 1623,
1610, 1542; low resolution FAB: (M + 1) 507.
_ W094/2~62 215 7 412 PCT~S94/02283
- 245 -
Tr~ns-3-t3,5-~ihy~roxy-4-~2-hy~roxyphenylra-honyl)]benz~mido-~-
(4-hydroxy)benzoyloxy-N-isopropylperhydroazepine ~COMPOUND 549)
To a solution of trans-N-benzyl-3-[3,5-dibenzyloxy-4-
(2-benzyloxy)phenylcarbonyl]benzamido-4-(4-benzyloxy)-
benzoyloxyperhydroazepine (190 mg, 0.199 mmol) in EtOAc-MeOH
(1:1, 30 ml) was introduced Pearlman's catalyst 20% Pd(OH)2 (70
mg). The mixture was subjected to hydrogenolysis at 50 psi for
17 hours. The catalyst was carefully filtered off through a
pad of celite and washed with MeOH. The filtrate was
concentrated and chromatographed on silica gel, eluting with 5%
MeOH in CH2C12. The title compound was obtained (500 mg, 50%);
the Parr bottle was contaminated with acetone and acid. All
spectra ( H, C, APT, DEPT, MS) and CHN analysis fully support
the structure. lH NMR (DMSO-d6) ~ 11.95 (s, brs, lH, OH), 10.25
(s, lH, OH), 10.00 (s, 2H, 20H), 8.20 (d, lH, NH), 7.78 (d, J
= 8.67 Hz, 2H, ArH), 753 (td, lH, ArH), 7.26 (dd, lH, ArH),
6.99 (d, lH, ArH), 6.87 (t, lH, ArH), 6.82 (d, J = 8.73 Hz,
ArH), 6.74 (s, 2H, ArH), 5.01 (s, br, lH, CH-4), 4.18 (s, br,
lH, CH-3), 2.90 (s, br, lH, CH(CH3)3), 2.68 (s, br, 4H,
CH2N-2,7), 1.90 (s, br, 3H, CH2-6, CH-5), 1.63 (s, br, lH,
CH-5), 1.00 (dd, 6H, 2CH3); 13C NMR (DMSO-d6 + D2O) ~ 55.02
(CH-(CH3)3), 18.66 and 18.40 (CH3)2CH); high resolution FAB M +
1: 549.2224; calculated for C30H32N2Oe: 549.2228. Anal. calcd.
for C30H32N2O~ 1.25 H2O: C, 63.09; H, 6.09; N, 4.90. Found: C,
63.06; H, 5.95; N, 4.66.
WOg4/2~62 21~ 7 4 ~ 2 PCT~S94/02283 --
- 246 -
anti-4- ~3,5-Dihy~roxy-~- ~2-hy~roxybenzoyl)-
benzoyl~mino)hexahy~ro-3-(4-hy~roxybenzoylamino)azepine,
complex with water:acetonitrile ~1:1.7:0.3) (COMPOUND 550)
~8YNTg~8I8 OF COMPOUND 550)
8yn-3-Aminohexahy~ro-~-hydroxy-l-phenylmethylazepi~-2-one
A solution of 3-acetylaminohexahydro-1-phenylmethyl-
azepin-2,4-dione (0.82 g, 3.0 mmol) in absolute ethanol (15 ml)
was treated with sodium borohydride (0.23 g, 6 mmol) and
stirred for 30 min, then treated with water (5 ml) and
concentrated in vacuo. The aqueous residue was extracted with
methylene chloride (3 x 25 ml) and the combined organic
extracts were dried (Na2SO~), concentrated in vacuo, and taken
up in 2:1 ethanol/water (7.5 ml). Concentrated hydrochloric
acid (2.5 ml) was added. The mixture was refluxed for 2 h and
partially concentrated, then diluted with water (25 ml). The
aqueous acidic mixture was extracted with ether (25 ml). The
aqueous solution was basified with 30% sodium hydroxide and
extracted with methylene chloride (3 x 40 ml). The combined
methylene chloride extracts were washed with water (25 ml),
dried (Na2SO~), and concentrated in vacuo to a yellow solid,
which was recrystallized from ethyl acetate to afford syn-3-
aminohexahydro-4-hydroxy-1-phenylmethylazepin-2-one (0.42 g,
60%) as a white solid.
8yn-3-Aminohexahydro-~-hydroxy-1-phenylmethylazepine
A cooled (5-C) solution of lithium aluminum
hydride/tetrahydrofuran (Aldrich, 1.0 N, 5.1 ml) under nitrogen
was treated with syn-3-aminohexahydro-4-hydroxy-1-
phenylmethylazepin-2-one (0.40 g, 1.7 mmol) in portions so that
the pot temperature did not exceed 15-C. The mixture was
refluxed for 6.5 h, cooled on an ice bath, and carefully
treated with water (0.21 ml), 15~ sodium hydroxide (0.21 ml),
and water (0.63 ml). The suspension was allowed to stir for 5
days (optimal time is 2-5 hours). The suspension was filtered,
and the filtrate was concentrated in vacuo and chromatographed
_ W094/2~62 215 7 412 PCT~S94/02283
- 247 -
on silica gel (eluted with 90:8:2 methylene chloride/methanol/
triethylamine). The appropriate fractions were concentrated in
vacuo to afford syn-3-aminohexahydro-4-hydroxy-1-
phenylmethylazepine (0.22 g, 58%) as a colorless oil.
8yn-~exahydro-~-hy~roxy-3-(4-phenylmethoxy)benzoyl~mino-1-
phenylmethyl~zepine
A solution of 4-benzyloxybenzoic acid (0.183 g, 0.8
mmol) in anhydrous tetrahydrofuran (2 ml) and N,N-
dimethylformamide (0.5 ml) was treated with N,N'-
carbonyldiimidazole (O.lS g, 0.9 mmol) and stirred at room
temperature for 1.5h. The solution was treated with syn-3-
aminohexahydro-4-hydroxy-1-phenylmethylazepine (0.20 g, 0.9
mmol) in anhydrous tetrahydrofuran (1 ml). The mixture was
stirred for 18 h, then concentrated in vacuo. The residue was
taken up in lN sodium carbonate (20 ml), and the aqueous
mixture was extracted with toluene (2x25 ml) cont~ining a
little 2-propanol. The combined organic extracts were dried
(Na2S0~) and the concentrated residue was flash chromatographed
on silica gel (eluted with 3:1 ethyl acetate/hexane) to afford
syn-hexahydro-4-hydroxy-3-(4-phenylmethoxy)benzoylamino-1-
phenylmethylazepine (0.17 g, 50%) as a viscous oil.
syn-~-Aminohex~hydro-3-(~-phenylmethoxy)benzoyl~mino-1-
phenylmethylazepine
A solution of syn-hexahydro-4-hydroxy-3-(4-
phenylmethoxy)-benzoylamino-l-phenylmethylazepine(0.645g,1.5
mmol) in anhydrous tetrahydrofuran (3 ml) under nitrogen was
cooled (ice bath, 5-C) and treated with triphenylphosphine
(0.495 g, 1.8 mmol) and diethylazodicarboxylate (0.313 g, 1.8
mmol). The mixture was then treated dropwise over 15 min with
a solution of diphenylphosphoryl azide (0.495 g, 1.8 mmol) in
anhydrous tetrahydrofuran (3 ml). The reaction was stirred at
room temperature for 18 h, then concentrated in vacuo,
dissolved in a little methylene chloride, and passed through a
short column of silica gel (eluted with 10% acetone/methylene
chloride). The early fractions containing chromophoric
W094/2~62 215 7 ~1~ PCT~S94/02283
- 248 -
material were concentrated in vacuo and dissolved in
ethanol/acetic acid/water (6:1:1, 12 ml), then treated with
zinc powder (0.50 g, 7.5 mmol). After 30 min, the mixture was
filtered and the filtrate was concentrated in vacuo. The
residue was taken up in 1.0 N sodium hydroxide (40 ml), and the
aqueous mixture was extracted with methylene chloride (3 x 35
ml). The combined extracts were dried (Na2SO4) and concentrated
in vacuo to afford crude syn-4-aminohexahydro-3-(4-
phenylmethoxy)benzoylamino-l-phenylmethylazepine (0.76 g).
syn-4-~3,5-Bis~phenylmethoxy)-~-~2-phenylmethoxybenzoyl))-
benzoyl~minohexahydro-3-~-phenylmethoxy)benzoyl~mino-1-
phenylmethylazepine ~COMPOUND 628)
A solution of 2,2',6-tribenzyloxybenzophenone-4-
carboxylic acid (0.545 g, 1.0 mmol) in anhydrous methylene
chloride (3 ml) and N,N-dimethylformamide (0.1 ml) under
nitrogen was treated with 2.0 N oxalyl chloride/methylene
chloride (0.7 ml, 1.4 mmol) and stirred for one hour. The
solution was concentrated in vacuo to a yellow solid, which was
kept under high vaccuum for one hour. A solution of crude syn-
4-aminohexahydro-3-(4-phenylmethoxy)-benzoylamino-1-
phenylmethylazepine (0.76 g, from above) in methylene chloride
(5 ml) was added to the acid chloride above, followed by 1.0 N
sodium hydroxide (4 ml). The mixture was stirred for 1.5 h,
diluted with methylene chloride (15 ml) and separated. The
aqueous layer was extracted with methylene chloride (15 ml) and
the combined organic solution was washed with saturated aqueous
sodium chloride, dried (Na2SO~), and concentrated in vacuo. The
residue was chromatographed on silica gel (eluted initially
with 2.5% acetone/methylene chloride, then with 7.5%
acetone/methylene chloride) to afford syn-4-(3,5-
bis(phenylmethoxy)-4-(2-phenylmethoxy-benzoyl))-
benzoylaminohexahydro-3-(4-phenylmethoxy)-benzoylamino-1-
phenylmethylazepine (0.52 g, 54%) as an opaque gum.
_ WO 94/20062 21 S 412 PCT/US94/02283
-- 249 --
syn~ 3, 5-Dihydroxy-~- ~ 2-hydroxybenzoyl ) -
benzoylamino)hexahy~ro-3-14-hydroxybenzoylamino)azepine,
complox with water:acetonitril~ ~1:1.7:0.3) (COMPO~ID 550)
A solution of syn-4- (3,5-bis (phenylmethoxy) -4- (2-
phenylmethoxybenzoyl ) ) benzoylaminohexahydro-3 - (4 -
phenylmethoxy) benzoylamino-l-phenylmethylazepine (0.39 g, 0.41
mmol) in ethanol/ethyl acetate (1: 1, 30 ml) was placed in a
Parr bottle and treated (under nitrogen) with Pearlman ' s
catalyst (Aldrich, 150 mg), then subjected to hydrogenation in
a Parr apparatus for 18 h at 48-50 psi. The reaction mixture
was carefully purged of hydrogen and the solution was filtered
through celite (care taken not to let filter cake dry). The
filtrate was concentrated in vacuo to afford crude material,
which was chromatographed on silica gel (eluted with 2 :1
methylene chloride/isopropanol) to give syn-4- (3,5-dihydroxy-4-
( 2 -hydroxybenzoyl ) benzoylamino ) hexahydro-3 - ( 4 -
hydroxybenzoylamino)azepine, complex with water:acetonitrile
(1:1.7:0.3) (0.055 g, 27%) as a yellow solid, which was
triturated with acetonitrile to afford a yellow powder: mp 210
- 215-C; Rf (2: 1 methylene chloride/isopropanol on silica)
0.50; IR (KBr) 1624 cm 1; lH NMR (d6-DMSO) ~ 8.33 (d, lH, J = 7
Hz), 7.97 (d, lH, J = 7 Hz), 7.73 (d, 2H, J = 9 Hz), 7.55 (m,
lH), 7.30 (m, lH), 7.00 (m, lH), 6.89 (m, lH), 6.84 (s, 2H),
6.82 (d, 2H, J = 9 Hz), 4.25 - 4.40 (m, 2H), 3.00 - 3.10 (m,
2H), 2.80 - 3.00 (m, 2H), 1.85 - 1.95 (m, lH), 1.65 - 1.80 (m,
3H). Anal. calcd. for C2~H27N307 1.7H20 0.3(CH3CN): C, 60.44;
H, 5.75; N, 8.43. Found: C, 60.34; H, 5.56; N, 8.42.
W094/20062 215 7 41~ PCT~S94/02283
- 250 -
Hexahydro-3-(~-phenylmethoxy)benzoylamino-1-phenylmethyl-
azepin-~-one (COMPOUND 551)
A 25 ml 3-neck round bottom flask under nitrogen was
charged with 2.0 N oxalyl chloride/methylene chloride (Aldrich,
1.125 ml, 2.25 mmol), diluted with anhydrous methylene chloride
(2 ml), cooled (-65-C), and treated dropwise with anhydrous
dimethylsulfoxide (0.35 g, 4.5 mmol) in anhydrous methylene
chloride (1.2 ml) at a rate to keep the pot temperature below
-60-C. The mixture was stirred at -65 + 5C for 30 min, then
treated dropwise with a solution of syn-hexahydro-4-hydroxy-3-
(4-phenylmethoxy)benzoylamino-1-phenylmethylazepine (0.645 g,
1.5 mmol) in anhydrous methylene chloride (1.5 ml) at a rate to
keep the pot temperature below -55-C. The mixture was stirred
at 55 + 5-C for 2 h, then treated dropwise with triethylamine
(1.5 ml), warmed to room temperature over 1 h, and diluted with
methylene chloride (10 ml). The organic solution was washed
with water (10 ml), saturated aqueous sodium bicarbonate (10
ml), dried (Na2S0~), and concentrated in vacuo. The residue was
chromatographed on silica gel (eluted with 5% acetone/methylene
chloride) to afford hexahydro-3-(4-phenylmethoxy)benzoylamino-
l-phenylmethylazepin-4-one (0.53 g, 82%) as a viscous colorless
oil.
~-Aminohexahydro-3-(~-phenylmethoxy)benzoylamino-1-phenyl-
methylazepine, 1:1 mixture of syn and anti i~omers
A solution of hexahydro-3-(4-phenylmethoxy)-
benzoylamino-l-phenylmethylazepin-4-one (0.37 g, 0.86 mmol) in
ethanol (5 ml) was treated with hydroxylamine hydrochloride (80
mg, 1.15 mmol) and 25% methanolic sodium methoxide (Aldrich, 70
mg, 0.32 mmol) and heated to 50-C for 1 h. The mixture was
cooled to room temperature, treated with additional 25% sodium
methoxide (0.18 g, 0.83 mmol), then stirred for 10 min and
partially concentrated in vacuo. The residue was taken up in
0.5 N sodium hydroxide (5 ml) and extracted with methylene
chloride (3 x 15 ml). The combined organic extracts were
washed with saturated aqueous sodium chloride, dried (Na2SO4),
and concentrated in vacuo. This residue was dissolved in 95%
W094/20062 2 1 ~ 7 4 ~ ~ PCT/US94/022~
- 251 -
ethanol (20 ml), treated with Raney Nickel (Aldrich, 1/4
teaspoon) in a Parr bottle, and subjected to hydrogenation at
30 psi for 2.25 h (more time needed). The solution was
carefully evacuated of hydrogen, filtered through celite (do
not allow to dry), and the filtrate was concentrated in vacuo
and chromatographed on silica gel (eluted first with 15%
methanol/methylene chloride, then with 25%, and finally with
33%) to afford 4-aminohexahydro-3-(4-phenylmethoxy)benzoyl-
amino-l-phenylmethylazepine, 1:1 mixture of syn and anti
isomers (0.15 g, 40%) as a viscous colorless oil which was
stored under nitrogen.
~nt~-4-~3,5-B$~phenylmethoxy)~ 2-phenylmethoxybenzoyl))-
benzoyl~minohexahydro-3-(~-phenylmQthoxy)benzoyl~mino-l-
phenylmethylazepine ~COMPO~ND 629)
A solution of 2,6,2'-tribenzyloxybenzophenone-4-
carboxylic acid (0.245 g, 0.45 mmol) in anhydrous methylene
chloride (1.5 ml) under nitrogen was treated with
N,N-dimethylformamide (3 drops), then with 2.0 N oxalyl
chloride/methylene chloride (0.30 ml, 0.60 mmol). The vigorous
bubbling soon subsided and the mixture was stirred for 1 h,
concentrated in vacuo, and kept under high vacuum for 1 h to
ensure removal of excess oxalyl chloride. The acid chloride
was dissolved in methylene chloride (2.5 ml) and treated with
4-aminohexahydro-3-(4-phenylmethoxy)benzoylamino-1-
phenylmethylazepine, 1:1 mixture of syn and anti isomers (0.14
g, 0.325 mmol) followed by 1.0 N sodium hydroxide (1.5 ml).
The biphasic mixture was rapidly stirred for 2 h and separated.
The aqueous layer was extracted with methylene chloride (2 x 7
ml), and the combined organic extracts and organic layer were
washed with saturated aqueous sodium chloride (10 ml), dried
(Na2SO~), and concentrated in vacuo. Flash chromatography on
silica gel (eluted first with 3% acetone/methylene chloride,
then with 7%, and finally with 10%) afforded anti-4-(3,5-
bis(phenylmethoxy)-4-(2-phenylmethoxy-benzoyl))-
benzoylaminohexahydro-3-(4-phenylmethoxy)benzoylamino-1-
WO 94/20062 215 7 il ~ 2 PCT/US94/02283
-- 252 --
phenylmethylazepine (0.13 g, 42%) as a colorless foam, followedby the syn isomer (0.13 g, 42%) .
~nti-4-~3,5-Dihydroxy-~-~2-hydroxybenzoyl)benzoyl~mino)hexa-
hydro-3-~-hydroxybenzoylamino)azepine, complex with
water:2-prop~nol ~1:1.5:0.~) ~COMPOUND 551)
A solution of syn-4- (3,5-bis (phenylmethoxy) -4- (2-
phenylmethoxybenzoyl) ) benzoylaminohexahydro-3- (4-
phenylmethoxy) benzoylamino-l-phenylmethylazepine (0.13 g, 0.136
mmol) in ethanol/ethyl acetate (1: 1, 25 ml) was placed in a
Parr bottle and treated (under nitrogen) with Pearlman ' s
catalyst (Aldrich, 120 mg), then subjected to hydrogenation in
a Parr apparatus for 18 h at 50 psi. The reaction mixture was
carefully purged of hydrogen and the solution was filtered
through celite (care taken not to let filter cake dry). The
filtrate was concentrated in vacuo to afford crude material,
which was chromatographed on silica gel (eluted with 1: 1
methanol/methylene chloride). This concentrated product was
dissolved in 2-propanol, filtered to remove inorganic material,
re-concentrated in vacuo, and triturated from ether to afford
anti-4- ( 3, 5-dihy~roxy-4- ( 2 -hydroxybenzoyl ) -
benzoylamino) hexahydro-3- (4-hydroxybenzoylamino) azepine complex
with water:2-propanol (1:1.5:0.4) (56 mg, 75%) as a yellow
powder: mp 207 - 210 C; Rf (2: 1 methylene chloride/2-propanol
on silica) 0.20: IR (KBr) cm 1 1639, 1607; H NMR (d6-DMSO)
8.32 (d, lH, J = 7 Hz), 7.96 (d, lH, J = 7 Hz), 7.66 (d, 2H, J
= 9 Hz), 7.52 (m, lH), 7.27 (m, lH), 6.97 (m, lH), 6.86 (m,
lH), 6.77 (d, 2H, J = 9 Hz), 6.72 (s, 2H), 4.00 - 4.15 (m, 2H),
2.60 - 3.00 (m, 4H), 1.50 - 1.90 (m, 4H) . Anal . calcd. for
C2~Hz~N307 1-5 H20 0.4 (C3H80): C, 60.85; H, 6.01; N, 7.55.
Found: C, 60.68; H, 5.68; N, 7.22.
~15~ 2
W094/20062
- PCT~S94/0~R~
- 253 -
~ Anti-3-~ minobenzam~do)-~-[3,5-dihy~roYy-~-(2-
hy~roxyphenylcarbonyl)]benzyloxyperhy~roazepine (COMPOUND552)
OE~ 8110 OH HO
OH H ` O~ ~ o
(';;)-- ,C, ~ N2 ~ C~ NOz -- (;~-- ~ C~ NH2
630 H 552
3,5-dibenzyloxy-4-~(2-benzyloxybenzoyl]benzoic acid
(218 mg, 0.40 mmol) was dissolved in anhydrous dichloromethane
(2.0 ml), and dimethylformamide (0.05 ml). The reaction was
placed under nitrogen, and oxalyl chloride (0.3 ml, 2M in
dichloromethane) was slowly added via a syringe. The reaction
was stirred at room temperature for 1 h and then concentrated
in vacuo and placed under high vacuum for 1 h. The residue was
redissolved in anhydrous dichloromethane (2 ml) and stirred
under nitrogen. Next, a solution of (+)-anti-3-(4-
nitrobenzamido)-4-hydroxybenzylperhydroazepine (120 mg, 0.32
mmol) and dimethylaminopyridine (44 mg,0.36 mmol) in
triethylamine (0.9 ml) and anhydrous dichloromethane (3 ml)
were added. The reaction was then stirred for 1 h at room
temperature. At this point, dichloromethane (50 ml) and 0.5 N
sodium hydroxide (20 ml) were added, and the reaction was
transferred to a separatory funnel. The aqueous layer was
removed, and brine was added. The organic layer was then
isolated, dried over sodium sulfate, filtered, and concentrated
in vacuo to yield a yellow oil. The oil was flash
chromatographed eluting with hexane:ethyl acetate/4:1. Pure
fractions of the major product were concentrated in vacuo to
yield 180 mg of white solid (COMPOUND 636). The solid was then
dissolved in methanol:ethyl acetate/2:1 (30 ml) and acetic acid
~094/20062 2 1 ~ 7 4 1 2 PCT~S94/02283
- 254 -
(0.2 ml) in a 500 ml Parr bottle along with 5% palladium on
activated carbon (70 mg). The Parr bottle was then placed on
the Parr hydrogenator for 24 h. The reaction mixture was
filtered to remove the catalyst, and then concentrated in vacuo
to yield a yellow solid. The solid was flash chromatographed
eluting with dichoromethane:methanol/9:1 to yield a yellow
solid of the title compound (60 mg, 61%): mp 175 - 185-C; NMR
(CD30D) consistant with structure; IR (RBr) cm1 3389 (OH); 2361
(alkyl); 1704 (ester); 1625 (ketone). Anal. calcd. for
C27H27N3O7 1.0 H2O 1.0 acetic acid: C, 59.69; H, 5.70; N,
7.20. Found: C, 59.94; H, 5.70; N, 7.31.
~09~/20062 21~7412
_ PCT~S94/0
- 255 -
~ + ) -Anti-3- ( ~- f luoro~^n~mi~o)-4-t3,5-dihydroxy-~-(2-hy~roxy-
phenylc~rbonyl)]benzyloxyperhydroazepine (COMPOUND 53)
OBn i3nO OH HO
0,.~~ o ,~_,0,~
o~ F _ (;;)-- C~ F (;;~_ ~H OH F
631 553
3,5-Dibenzyloxy-4-t(2-benzyloxybenzoyl]benzoic acid
(545 mg, 1.0 mmol) was dissolved in anhydrous dichloromethane
(5.0 ml), and dimethylformamide (0.05 ml). The reaction was
placed under nitrogen, and oxalyl chloride (1.0 ml, 2 M in
dichloromethane) was slowly added via a syringe. The reaction
was stirred at room temperature for 2 h and then concentrated
in vacuo and placed under high vacuum for 1.5 h. The residue
was redissolved in anhydrous dichloromethane (3 ml) and stirred
under nitrogen. Next, a solution of (+)-anti-3-(4-
fluorobenzamido)-4-hydroxy-N-benzylperhydroazepine (300 mg,
0.93 mmol) and dimethylaminopyridine (125 mg, 1.02 mmol) in
triethylamine (0.9 ml) and anhydrous dichloromethane (2 ml)
were added. The reaction was then stirred for 20 min at room
temperature. At this point, dichloromethane (50 ml) and 0.5 N
sodium hydroxide (30 ml) were added, and the reaction was
transfered to a separatory funnel. The aqueous layer was
removed, and brine was added. The organic layer was then
isolated, dried over sodium sulfate, filtered, and concentrated
in vacuo to yield a light brown foam. The foam was flash
chromatographed eluting with hexane:ethyl acetate/2:1. Pure
fractions of the major product were concentrated in vacuo to
yield 440 mg of white solid (COMPOUND 631). 260 mg of the
W094/20062 2 1 5 7 4 ~ ~
PCT~S94/02283
- 256 -
solid was dissolved in methanol:ethyl acetate/5:1 (30 ml) and
acetic acid (0.2 ml) in a 500 ml Parr bottle along with 5%
palladium on activated carbon (100 mg). The Parr bottle was
then placed on the Parr hydrogenator for 4.5 h. The reaction
mixture was filtered to remove the catalyst, and then
concentrated in vacuo to yield a yellow solid. The solid was
flash chromatographed eluting with dichoromethane:methanol/8:2
to yield a yellow solid of the title compound (110 mg, 72%):
mp 189-C; IR (KBr) cm1 3349 (OH); 2362 (alkyl); 1704 (ester);
1626 (ketone). Anal. calcd. for C27H25N2O7F 1.0 H2O 1.0
acetic acid: C, 59.38; H, 5.33; N, 4.78. Found: C, 59.09; H,
5.00; N, 4.71.
W094/20062 2 1 5 7 ~ 1 2
- PCT~S94/022
- 257 -
Tr~ns-3-~4-hy~,ox~z~m~do)-~-t~-(2-hy~oh~ 6-hy~ c~rhonyl-
benzoyl)-3,5-~ihy~roxybenzoyloxyl]perhy~rothiepine (COMPOUND
55~)
N~NHa 13~OH m~ DMF O~OTBDMS )O~H~ ~OH
HO~OH Ms Cl. EI~N ~AsO~OM~ L~MS EI~N TBDIA5~N,
H NH~ 1) ~~ CO~ OBn ~
I.JAIH~THF ~ CDI, THF H ~ COCI
H~ ~N~
\_~S 2)lNN OH H~O ~S El~N DM~P,CH~
OBn OE~n H~ Pd(OHb~ ~(OH OH
l~nO~_ H ~ ~ OH THF HO ~_
~N~ O ~ O
S
632 554
A mixture of 1,3-cyclohexadienemonoepoxide (9.61 g,
100 mmol, for preparation see: J. K. Crandall et àl., J. Org.
Chem., 1968, 33, 423), NaN3 (26 g, 400 mmol), NH4Cl (10.7 g, 200
W094/20062 215 ~ PCT~S94/02283
- 258 -
mmol), MeOH (300 ml), and H2O (50 ml) was stirred at 65 C for
16h. The resultant slurry was evaporated to remove all the
volatile materials and the residue was treated with 1 N NaOH
(100 ml), and extracted with CH2C12 (3 x 60 ml). The combined
CH2C12 extracts were washed with H20 (2 x 100 ml), dried
(MgSO4), evaporated, and chromatographed on a SiO2 column
(Et2O:hexane = 1:4) to give a colorless oil (7.308 g, 53%).
A mixture of the product of the previous reaction
(7.23 g, 52 mmol), TBDMS-Cl (8.227 g, 55 mmol), and imidazole
(5.308 g, 78 mmol) in N,N-dimethylformamide (50 ml) was stirred
at room temperature for 16h. Et2O (250 ml) was added and the
resultant mixture was washed with H2O (5 x 100 ml), dried
(MgSO~), and evaporated to give a colorless oil (12.76 g, 97%)
which was used in the next step without further purification.
A solution of oil (7.63 g, 30 mmol) in MeOH (150 ml)
and CH2C12 (30 ml) was cooled to -78-C, stirred, and treated
with a mixture of air and 03 until a blue color persisted (ca.
1.5h). Excess O3 was removed by bubbling N2 through the
solution, and then NaBH~ (4.54 g, 120 mmol) was added. After
being stirred at -78-C for 30 min. followed by 30 min. at room
temperature the resultant mixture was poured into 1 N HCl (100
ml) and then evaporated to remove the volatile components. The
aqueous residue was extracted with Et2O (4 x 50 ml), and the
combined ether extracts were washed with HzO (100 ml), brine
(100 ml), dried (MgSO~), evaporated, and crystallized from
Et2O:hexane to give a white solid (6.772 g, 78%).
Methanesulfonyl chloride (2.67 ml, 34 mmol) was added
dropwise to a stirred solution of the borohydride reduction
product (4.01 g, 14 mmol) and Et3N (S.76 ml, 41 mmol) in CH2Clz
(40 ml) at 5-C. After 30 min. the cooling bath was removed and
stirring was continued for 16h. The resultant mixture was
diluted with CH2C12 (20 ml), washed with 1 N HCl (3 x 30 ml),
H2O (2 x 30 ml), brine (2 x 30 ml), dried (MgSO4), and
evaporated. The residue was chromatographed (SiO2, Et2O:hexane
= 1:1 followed by Et2O:hexane:CH2Cl2 = 1:1:0.25) to give a pale
yellow oil (5.885 g, 96%).
UO9~/20062 2 1 5 7 ~ 1 2 PCT~S94/02283
- 259 -
A mixture of the product of the previous reaction
(2.24 g, 5 mmol), lithium sulfide (253 mg, 5.5 mmol), and Et3N
(2.09 ml, 15 mmol) in EtOH (100 ml) was stirred under reflux
for 16h. The resultant mixture was evaporated and the residue
was chromatographed (SiO2, Et2O:hexane = 1:30) to give a
colorless oil (515 mg, 36%).
A solution of LiAlH4 in Et2O (1 M, 3.58 ml) was added
to the colorless oil from the reaction above (515 mg, 1.79
mmol) in Et2O (9 ml) and the mixture was stirred at room
temperature for 2h. 5N aq NaOH (5 ml) was added cautiously and
stirring was continued at room temperature for 16h. The phases
were separated and the aqueous phase was saturated with NaCl
and extracted with CH2C12 (3 x 10 ml). The combined organic
layers were dried (MgSO4), and evaporated to give a colorless
oil which solidified on standing (250 mg, 95%). This material
was used in the next step without further purification.
A mixture of 4-benzyloxybenzoic acid (427 mg, 1.87
mmol) and l,l'-carbonyldiimidazole (302 mg, 1.87 mmol) in THF
(5 ml) was stirred at room temperature for 2h and then the
product of the previous reaction (250 mg, 1.70 mmol) in THF (5
ml) was added. The mixture was stirred at room temperature for
36h, evaporated, dissolved in CH2C12 (30 ml), washed with H2O,
and dried (MgSO~). The resultant solution was diluted with
hexane (10 ml) and rotaevaporated at 0-C to ca. 10 ml. The
white precipitate was collected, dried under vacuum, and
dissolved in a mixture of MeOH (10 ml) and THF (8 ml). 1 N aq.
NaOH (1 ml) was added and the mixture was stirred at room
temperature for 4h. The volatile components were evaporated
and the residue was dissolved in CH2C12 (25 ml), washed with H2O
(3 x 5 ml), dried (MgSO4), and filtered. The filtrate was
diluted with hexane (lO ml) and rotaevaporated at 0 C to ca. 5
ml. The precipitate was collected, washed with hexane, and
dried under vacuum to give a white powder (382 mg, 67%).
To a solution of 4-(2-benzyloxy-6-benzyloxycarbonyl-
benzoyl)-3,5-dibenzyloxybenzoic acid (305 mg, 0.45 mmol) and a
drop of N,N-dimethylformamide in CH2C12 (1.3 ml) at 5C was
w094~20062215 7 ~12 PCT/US94/02283
- 260 -
added a solution of oxalyl chloride in CH2Cl2 (2 M, 0.25 ml).
The resultant mixture was stirred at room temperature for 2h
and then evaporated. The residue was dried under vacuum for 2h,
dissolved in CH2C12 (0.8 ml), and added to a mixture of the
above intermediate (143 mg, 0.4 mmol), Et3N (61 mg, 0.6 mmol),
and 4-N,N- dimethylaminopyridine (5 mg, 0.04 mmol) in CH2Cl2
(1.2 ml). The resultant solution was stirred at room
temperature for 17h, diluted with CH2C12 (10 ml), washed with
H2O (3 x 5 ml), dried (MgSO~), and evaporated. The residue was
chromatographed (SiO2, Et2O:hexane:CH2Cl2 = 1:1:0.5) to give a
colorless oil (347 mg, 76%).
This intermediate (105 mg, 0.1 mmol), Pd(OH)2 on
carbon (20 wt%, contains > 50% moist, 281 mg, 0.2 mmol), THF (1
ml), and MeOH (1 ml) was stirred and treated with 1 atm H2 at
room temperature for 40h. The resultant mixture was filtered
through a Florisil pad and the Florisil pad was washed with
MeOH (15 ml). The combined filtrate and wash were evaporated
to give a yellow solid (25 mg, 35%) (COMPOUND 554). IR (KBr,
cm ): 1706, 1689, 1633.
2157412
0 94/20062 PCT/US94/02283
-- 261 --
1, 1-Dioxo-trans-3- ~ ~-hydroxybenz~mi~o ) -~- t ~.- ( 2-hy~roxy-6-
hydroxycarbonylbenzoyl ) -3, 5-~lihy~roxybenzoyloxyl ] -
perhydrothiepine ~COMPO~JND 555)
~o ~o
CH,CO,H ~_
~ ~CH,CO,H. ct'.,a, 11l1
632 633
~ OH OH
H,. Pd~OH), C
C~so
o
555
Figure AA
Peroxyacetic acid (32 wt% in acetic acid, 37 mg, 0.155
mmol) was added to a solution of Compound 632 (75 mg, 0. 074
mmol, see Compound 554 for preparation) in CH2Cl2 (0.7 ml). The
resultant mixture was stirred at room temperature for lh,
diluted with CH2C12 (10 ml), and washed with sat. aq. K2CO3 ( 3
x 5 ml). The organic layer was dried (MgSO4) and evaporated to
give a white solid which was recrystallized from hot EtOAc (10
ml, contains ca. 2 ml of THF and 3 ml of hexane) to give a
white powder (61 mg, 79~) (COMPOUND 633).
21~7412
~094/20062 PCT~S94/02283
- 262 -
A mixture of Compound 633 (61 mg, 0.058 mmol), Pd(OH)2
on carbon (20 wt%, contains > 50% moist, 8 mg, 0.0058 mmol),
THF (1 ml), and MeOH (1 ml) was stirred and treated with 1 atm
H2 at room temperature for 3 hr. The resultant mixture was
filtered through Florisil and the filtrate was evaporated to
give the title compound as a yellow solid (33 mg, 95~). IR
(KBr, cm ): 1718, 1686, 1635.
_ WO9~/20062 21~7~
- 263 - PCT/US94/02283
Tr~ns-4-(4-(2-c~rboxy-6-hydroxybenzoyl)benzoyloxy)-3-(4-
hy~roxybenz~mi~o)perhydro~zepine Trifluoro~cetic Aci~ ~lt
Hy~r~te ~COMPOUND 556)
OBn ( CO2H
~ t~ BuU ~ PDC, DMF
MeO~OMeO~SJ oan MeO~I OBn
CO2Bn
BnB- ~ KOH, DMF
KDCMO~3 MeO ~ OBn
o
CO2Bn BnO~
)~\ 1) (coa)2 ~, OBn
"~/ 2) IPr2NEI, DMAP, ~/
HO2C OBn OH O ~~
(~ . N ~ OBn ~ H~ OBn
Bn Bn
6~
HoO
H2 ~ OH
Pd(OHh
. N~ OH
H
556
W094/20062 215 7 ~12 PCT~S94/02283
- 264 -
Methyl-4-(6-Benzyloxy-2-hy~roxymethylbenzoyl)benzoate
To a solution of 1.07 g (5.00 mmol) of 3-
benzyloxybenzyl alcohol in 15 ml of toluene at -5CC under an
atmosphere of nitrogen was added 5.8 ml (12.2 mmol) of a 2.1
M solution of butyllithium in hexanes over 15 min. The
solution was stirred at -5C for 6 h, after which it was cooled
to -78-C, and a solution of 1.00 g (5.03 mmol) of 4-
(methoxycarbonyl)benzoyl chloride in 5 ml of tetrahydrofuran
was added. The mixture was stirred for 1 h, after which it was
poured onto 200 ml of ether and 100 ml of saturated aqueous
ammonium chloride, and this mixture was stirred for 10 min.
The layers were separated, and the organic phase was washed
with saturated aqueous sodium bicarbonate and brine, dried over
magnesium sulfate, and evaporated to give the crude product.
Flash chromatography on silica gel eluting with 3/1 ethyl
acetate -hexane afforded 0.68 g (36~) of the title compound as
a white solid, which was carried on to the next step.
Methyl-~-~6-Benzyloxy-2-ca,~6~benzoyl)benzoate
To a solution of 0.63 g (1.7 mmol) of methyl 4-(6-
benzyloxy-2-hydroxymethylbenzoyl)benzoate in 20 ml of
dimethylformamide was added 4.41 g (11.7 mmol) of pyridinium
dichromate. The solution was stirred at room temperature under
a nitrogen atmosphere for 4 days, after which it was poured
onto 300 ml of ether and washed with 200 ml of water, 150 ml of
2 M HCl, and 150 ml of brine, and dried over magnesium sulfate.
Evaporation of the solvent afforded 0.47 g (72%) of the crude
product. This material was sufficiently pure for further use
and was carried directly to the next step.
~ethyl-4-~6-Benzyloxy-2-~benzyloxyc~rbonyl)benzoyl)benzoate
To a c ~ution of 0.47 g (1.2 mmol) of methyl 4-(6-
benzyloxy-2-carboxybenzoyl)benzoate in 20 ml of dry
dimethylformamide was added 501 mg (3.62 mmol) of potassium
carbonate and 0.158 ml (227 mg, 1.32 mmol) of benzyl bromide.
The solution was stirred at room temperature under a nitrogen
atmosphere for 18 h. The mixture was then poured onto 300 ml
WO9-1/20062 215 7 ~12 PCT~S94/02~83
- 265 -
of ether and washed with two 200 ml portions of water and then
with 150 ml of brine, and dried over magnesium sulfate.
Evaporation of the solvent afforded 0.57 g of the crude
product, which was chromatographed on silica gel, eluting with
3/1 hexane-ethyl acetate to give 0.32 g (54%) of the title
compound as a colorless oil. This material was used directly
in the next step.
~-(6-Benzyloxy-2-(benzyloxycarbonyl)benzoyl)benzoic Acid
A solution of 0.301 g (0.614 mmol) of methyl 4-(6-
benzyloxy-2-(benzyloxycarbonyl)benzoyl)benzoate in 7 ml of DMF
was treated with 0.337 ml of a 2 M aqueous solution of
potassium hydroxide under an atmosphere of nitrogen for 20 h.
The mixture was then poured onto 100 ml of ethyl acetate and
washed with 60 ml each of 0.2N HCl, water, and brine. The
organic extracts were dried over magnesium sulfate and
evaporated to give 0.32 g of the crude product, which was
chromatographed on a 41 x 250 mm C18 column (solvent A: 95:5
water/acetonitrile + 0.1% TFA solvent B: 100% acetonitrile:
gradient: 0-100% B over 60 min, flow: 25 ml/min). The pure
fractions were pooled and evaporated and then lyophilized from
water to give 70 mg of the title compound as a white solid,
which was carried on as is to the next step.
Tr~ns-N-Benzyl~ -(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)
benzoyloxy)-3-(~-benzyloxybenz~mido)perhydroazepine
A solution of 68 mg (0.143 mmol) of 4-(6-benzyloxy-2-
(benzyloxycarbonyl)benzoyl)benzoic acid in 5 ml of methylene
chloride containing a trace (approximately 1 ~L) of
dimethylformamide was cooled to O-C. A 2.0 M solution of
oxalyl chloride (0.11 ml, 0.22 mmol) was added, and the mixture
was stirred under a nitrogen atmosphere for 2 h. An additional
0.17 ml of oxalyl chloride was added, and the mixture was
stirred for an additional 2 h. The reaction mixture was
evaporated, and the residue was evaporated twice from 10 ml of
methylene chloride. The residue was dissolved in 3 ml of
methylene chloride, and was added to a solution of 69.6 mg
21~1 2
WO 94/20062 PCT/US94/02283 --
-- 266 --
(0.162 mmol) of trans-N-benzyl-3-(4-benzyloxybenzamido)-4-
hydroxyazepine, 29.8 ,uL (0.17 mmol) of diisopropylethylamine,
and 5.4 mg of DMAP in 5 ml of methylene chloride at O-C. The
mixture was stirred at room temperature under a nitrogen
atmosphere for 17 h, after which it was diluted with 30 ml of
methylene chloride, washed with saturated sodium bicarbonate
and brine, dried over magnesium sulfate, and evaporated to give
146 mg of the crude product. Chromatography on silica gel
eluting with 1/1 hexane-ethyl acetate gave 46 mg (37%) of the
title compound as a yellow oil, which was taken directly to the
next step.
Trans~ (2-Carboxy-6-hy~roxybenzoyl)benzoyloxy)-3~
hydro~cybenzami~o)perhydroazepine Trifluoroacetic Acid Salt
Hydrate
A solution of 46 mg (0.052 mmol) of trans-N-benzyl-4-
(4-(6-benzyloxy-2-(benzyloxycarbonyl)benzoyl)benzoyloxy)-3-(4-
benzyloxybenzamido)azepine in 10 ml of ethanol was treated with
8.1 ~L of trifluoroacetic acid, cooled to O-C, and 22 mg of
moist 10% palladium hydroxide on carbon was added. The
mixture was then stirred under an atmosphere of hydrogen for 19
h at room temperature. The mixture was filtered, evaporated,
and the residue was chromatographed on a 21 x 250 mm C18 column
(solvent A: 95:5 water/acetonitrile + 0.1% TFA; solvent B: 100%
acetonitrile; gradient: 0-50% B over 60 min, flow: 15 ml/min).
The pure fractions were pooled and evaporated and then
lyophilized from water to give 9.8 mg (28%) of the title
compound as a white fluffy solid. FABMS: m/z 519 (M + H).
Anal. Calc. for C28H26N208 2.5 H20 TFA; C, 53.18 H, 4.76; N,
4.13. Found: C, 53.43: H, 4.64; N, 4.30
Wo 94/20062 215 7 412 PCTtUS94tO2283
Trans 4 - t 4 - ~ 2 -Hy~roxycarbonyl - 6-hy~roxybenzoyl ) - 3, 5-
dih~ oa~enzoyloxy~-3- ~4-hydlohyLe"z~ o) -1- ~phenyl~ulfonyl) -
pyrrolidine ~COMPO~ND S62 )
CHzCIz H20 ~, OBn
H-~FA S 02Ph
C02Bn
Pr~El, I OBn
DMAP,~.
OBn
Hæ~O~N~ EIOH. ~F ~ h OBn
b~gH S02Ph BnO2Cb_OBn N
562 639
Figure AC
+ ) -Trans-3- ~ ~-benzyloxybenzamido) -~-hydroxy-l-
~phenylsul f onyl ) pyrrolidine
To a slurry of (+)-Trans-3-(4-benzyloxybenzamido)-4-
hydroxypyrrolidine TFA (150 mg, 0.352 mmol) in H20 (8.8 ml)
and CH2Cl2 (8.8 ml) were added anhydrous Na2C03 (112 mg, 3.0 eq,
1.06 mmol) then benzene sulfonyl chloride (58 ~1, 0.458 mmol,
1.3 eq), and the mixture stirred at room temperature 15 h. The
solution was then diluted with CH2Cl2 (20 ml) and poured into
H20 (20 ml) and methanol (4 ml). The layers were separated and
the aqueous layer extracted with CH2C12 (3 x 30 ml). The
organics were combined, dried (MgS0~), filtered and evaporated
to a white powder (159 mg, quant yield): 1H NMR (CD30D) ~ 7.62
- (d, J = 7.7 Hz, 2H), 7.43 (d, J = 8.9 Hz, 2H), 7.35-7.30 (m,
2~57412
W094/20062 PCT~S94102283
- 268 -
3H), 7.25-7.10 (m, 5H), 6.80 (d, J = 8.8 Hz, 2H), 4.95 (s, 2H),
4.06-4.00 (m, lH), 3.95-3.90 (m, lH), 3.50-3.35 (m, 2H), 3.15
(dd, J = 10.6, 3.9 Hz, lH), 2.99 (dd, J = 10.8, 3.2 Hz, lH).
Trans-4-[4-~6-benzyloxy-2-~benzyloAyc~rbonyl)benzoyl]-3,5-
dibenzyloAybenzoyloAy)-3- ~-benzyloxybenzamid,o)-l-
(phenylsulfonyl)pyrrolidine (CONPO~nlD 639)
To a solution of the previous product (159 mg, 0.352
mmol) in CH2Cl2 (6.0 ml) were added 4-dimethylaminopyridine (43
mg, 0.352 mmol, l.o eq), diisopropylethylamine (74 ~1, 0.42
mmol, 1.2 eq) then a solution of acid chloride (0.383 mmol, 1.1
eq) in CH2Cl2 (3.0 ml). The mixture was stirred at room
temperature under N2 14 h. The reaction mixture was then
diluted with CH2C12 (30 ml), and washed with 10% NaHCO3 (50 ml)
then brine (50 ml). The aqueous layers were combined and
extracted with CH2C12 (2 x 50 ml). The organics were combined,
dried (MgSO~), filtered and evaporated. Flash column
chromatoghraphy of the residue (2:1 hexane:ethyl acetate) on
silica gel provided the title compound (183 mg, 47%): lH NMR
(CDCl3) ~ 7.77 (d, J = 6.7 Hz, 2H), 7.70 (d, J = 8.8 Hz, 2H),
7.47-7.16 (m, 14H), 7.15-7.05 (m, 6H), 7.04-6.94 (m, 3H),
6.90-6.83 (m, 3H), 6.42 (d, J = 6.7 Hz, lH), 5.30 (dt, J = 5.1,
2.6 Hz, lH), 5.18 (s, 2H), 5.13 (s, 2H), 4.78 (m, 2H), 4.76 (s,
2H), 4.72 (s, 2H), 4.64-4.60 (m, lH), 3.88 (dd, J = 12.6, 5.4
Hz, lH), 3.74-3.65 (m, lH), 3.60-3.38 (m, 2H).
Trans 4- t 4 - ~ 2 -Hydroaycarbonyl- 6-hydroAybenzoyl ) -3, 5-
diL~oa~anzoyloAy]-3- (~-h2~roA~enz~i~o) -1- ~phenylsulfonyl) -
pyrrolidine ~COMP0~1ND 562 )
To a solution of the previous product (183 mg, 0.164
mmol) in THF (7.4 ml) and ethanol (7.4 ml) was added Pd(OH)2
(92 mg. of a 20% by weight powder). The flask was evacuated
and filled with H2 twice, then stirred under H2 (1 atm) for 20
h. The suspension was filtered through Celite, washed through
with methanol (50 ml), and evaporated to a yellow oil.
Purification by HPLC (21 x 250 mm Cl8 column) provided the title
compound (75 mg, 69%) as a fluffy yellow powder after
~09412W62 21~ 7 412 PCT~S94/022~
- 269 -
lyophilization: mp 185-208C; IR (KBr) 3402, 1709, 1636, 1608,
1232 cm ; H NMR (CD30D) ~ 7.53 (d, J = 8.2 Hz, 2H), 7.49 (d,
J = 8.7 Hz, 2H), 7.32 (d, J = 7.3 Hz, lH), 7.22-7.11 (m, 3H),
7.09 (dd, J = 8.1, 7.9 Hz, lH), 6.84 (d, J = 7.2 Hz, lH), 6.61
(d, J = 8.7 Hz, 2H), 6.35 (s, 2H), 4.99 (app t, J = 2.2 Hz,
lH), 4.92 (dd, J = 5.6, 2.8 Hz, lH), 3.62 (dd, J = 13.0, 4.3
Hz, lH), 3.50 (dd, J = 10.8, 6.0 Hz, lH), 3.39 (dd, J = 10.7,
2.4 Hz, lH), 3.32 (bd, J = 13.1 Hz, lH); HRMS (M + H) calcd
663.6315, found 663.1302; Anal. Calcd- for C32H26N20l2S 1 H2O:
C, 56.47; H, 4.15; N, 4.12; S, 4.71; found: C, 56.56; H, 4.17;
N, 4.09; S, 4.58.
-094/20062 2 1 5 7 1 1 2 PCT~S94/0~
- 270 -
Tr~ns-1-(4-hy~-o~yLehz~mi~o)-2-l4-~2-hy~roxy-6-meth
benzoyl)-3,5-dihydroxybenzoyloxyl]cycloheptane ~COMPOUND 566)
HO OM
O O
H~ ~ H~ , K,CO,, HMPA ~ H~
b o ~ o
588 566
Figure AD
A mixture of Compound 588 (55 mg, 0.1 mmol),
iodomethane (0.05 ml. 0.8 mmol), and K2CO3 (28 mg, 0.2 mmol) in
HMPA (0.2 ml) was stirred at 40-C for 1.5h, and the reaction
was judged incomplete by TLC. Additional iodomethane (0.025
ml, 0.4 mmol) was added and stirring was continued for 2h. at
40 C. EtOAc (15 ml) was added and the resultant mixture was
washed with H2O (3 x 10 ml) and brine (10 ml), dried (MgSO4),
and evaporated. The residue was purified by preparative TLC
(SiO2, multi-elution with CH2C12:5% MeOH in EtOAc = 4:1) to give
a yellow solid (34 mg, 61%). IR (KBr, cm1): 1712, 1634, 1607.
FBMS: M/Z = 564 (M + 1).
W094/20062 21~ 7 ~ 12 PCT~S94/02283
- 271 -
Trans-~-t~-~2-Hydroxycarbonyl-6-hydroxybenzoyl)-3,5-
~ihydko~L~zoyloxy]-3-(4-hyd-~yL~z~mido)-l-(5-dimethyl~min
l-naphthalene sulfonyl)perhydroazepine (COMPOUND 569)
O HO~
~OH OH,CH3 ~OH OH
H~o~ H ~ Cl ~I TEA ~ HHo~ Q~
,~ O N~ +~ S~ MeOH ~ ~O~N~ CH3
CF,CO2H NH CF,C02H \~ N~ SJ3~ ~ C~
569 Oo
Racemic balanol (preparation described in Compound
508; 100 mg, 147 ~mol) was dissolved in methanol (1 ml) and
treated with triethylamine (204 ~1, 1.47 ~mol) and dansyl
chloride (39.5 mg, 146.5 ~mol) in methylene chloride (1 ml).
After stirring at room temperature for 3 h, the mixture was
concentrated under vacuum to a yellow film. The residue was
dissolved in DMF (2 ml) and chromatographed on a Dynamax-60 C18
column (41 mm ID x 25 cm length) using a linear gradient from
100% A (0.1% TFA and 5% acetonitrile in water) to 100% B (pure
acetonitrile) over 60 m at 25 ml/min. The clean product, which
eluted in 40 m, was freeze-dried to give a yellow powder (33
mg, 29%): m.p. 185-187-C dec; 1H-NMR (DMSO, 300 MHz)
1.73-2.12 (4H, m), 2.85 (6H, s), 4.30-4.41 (lH, m), 5.20 (lH,
pseudo t), 6.75-6.87 (4H, m), 7.07 (lH, d, J = 8 Hz), 7.30 (2H,
t), 7.39 (lH, d, J = 8 Hz), 7.62-7.70 (4H, m), 8.06 (lH, d, J
= 8 Hz), 8.24 (lH, d, J = 9 Hz), 8.50 (lH, d, J = 9 Hz), 9.91
(lH, 8), 11.68 (lH, s); IR (KBr): cm1 3399, 2361, 2340, 1702,
1677, 1635, 1607, 1542, 1507, 1462, 1425, 1371, 1319, 1283,
1237, 1200, 1139, 1103, 1064, 991, 920, 848, 793, 767, 723,
669, 581, 542, 532. Anal. Calcd. for C~oH37N3O12S 2H2O .92TFA
W094/20062 215 7 ~12 PCT~S94/022~
- 272 -
.15CH3CN: C, 54.37; H, 4.58; N, 4.73; S, 3.44. Found: C,
57.35; H, 4.51; N, 4.63; S, 3.17. LRMS (FAB) m/z 784.0 (783.2
calcd for C40H37N3O12S).
21~741~
wos~/20062 PCT~S94/02283
- 273 -
Tr~ns- ~ - t ~- ~ 2 -Carboxy-6-hydroxy~enzoyl ) -3, 5-
dihydroxybenzoyloxy] -3- ~ 4 -hydroxy~enz~mido ) -1- ~ 2 -
nitrobenzenesulfonyl)perhydroazepine, trifluoracetic acid salt
COMPOIIND 57 3 )
o HO~=
~OH OH ~OH OH
H~o~ H~ Cl~ ~ TEA , H
~ o s ~ MeOH ~ O H~
CF,CO2H NH
CF,CO2H S~
573 O O NO2
Racemic balanol (preparation described in Compound
508; 100 mg, 147 ~mol) was dissolved in methanol (1 ml) and
treated with triethylamine (204 ~1, 1.47 ~mol) and
2-nitrobenzene sulfonyl chloride (48.7 mg, 219.7 ~mol) in
methylene chloride (1 ml). After stirring at room temperature
for 3 h, the mixture was concentrated under vacuum to a yellow
film. The residue was dissolved in DMF (2 ml) and
chromatographed on a Dynamax-60 C18 column (41 mm ID x 25 cm
length) using a linear gradient from 100% A (0.1% TFA and 5%
acetonitrile in water) to 100% B (pure acetonitrile) over 60 m
at 25 ml/min. The clean product, which eluted in 33 m, was
freeze-dried to give a yellow powder (28 mg, 26%): m.p.
158-160-C dec; lH-NMR (DMSO, 300 MHz) ~ 1.81-1.97 (3H, m),
2.06-2.17 (lH, m), 4.43-4.57 (lH, m), 5.18-5.28 (lH, m), 6.75
- (2H, s), 7.04 (lH, d, J = 8 Hz), 7.22-7.31 (3H, m), 7.36 (lH,
d, J = 8 Hz), 8.05 (lH, t), 8.20 (lH, d, J = 8 Hz), 8.20 (lH,
d, J = 8 Hz), 8.85 (lH, d, J = 8 Hz), 9.87 (lH, s), 11.67 (lH,
s); IR (KBr): cm 3430, 3412, 1701, 1676, 1636, 1604, 1545,
W094/20062 PCT~S94/02283
21~7~1~
- 274 -
1496, 1425, 1370, 1290, 1237, 1201, 1148, 1104, 1063, 1016,
993, 921, 874, 856, 799, 761, 738, 724, 586. Anal. Calcd. for
C34H2~N30l4S 1-2 H20 1.1 TFA: C, 49.25; H, 3.71; N, 4.76; S,
3.63. Found: C, 49.22; H, 3.70; N, 4.72; S, 3.39. LRMS (FAB)
m/z 735.9 (735.68 calcd for C34H29N3014S).
w094/20062 213 ~ ~ ~ 2 PCT/US94/0~
- 275 -
Tran8-4- ~- (2-Carboxy-6-hy~roxybenzoyl)-3,5-
dihydroxybenzoyloxy]-3- (~-hydroxybenzamido) -1- ~
nitrobenzenesulfonyl)perhy~roazepine, trifluoracetic acid salt
COMPO~ND 57 ~ )
o HO~
~OH OH ~=~OH OH
~~ H~ Cl` ~ TEA ~ HHo
O .S ~ MeOH~ H~
~ ~ O O O ,~ j~ ,~,, NO2
CF~Co2H \~, NH CF~Co2H \ N` S
574 Oo
Figure AG
Racemic balanol (preparation described in Compound
508; 100 mg, 147 ~mol) was dissolved in methanol (1 ml) and
treated with triethyiamine (204 ~1, 1.47 ~mol) and
4-nitrobenzene sulfonyl chloride (48.7 mg, 219.7 ~mol) in
methylene chloride (1 ml). After stirring at room temperature
for 3 h, the mixture was concentrated under vacuum to a yellow
film. The residue was dissolved in DMF (2 ml) and
chromatographed on a Dynamax-60 C18 column (41 mm ID x 25 cm
length) using a linear gradient from 100% A (0.1% TFA and 5%
acetonitrile in water) to 100% B (pure acetonitrile) over 60 m
at 25 ml/min. The clean product, which eluted in 34 m, was
freeze-dried to give a yellow powder (12 mg, 11%): m.p.
186-188-C dec; lH-NMR (DMSO, 300 MHz) ~ 1.80-1.97 (3H, m),
2.07-2.18 (lH, m), 3.13 (2H, pseudo t), 4.42-4.53 (lH, m),
5.17-5.28 (lH, m), 6.75 (2H, d, J = 6 Hz), 7.03 (lH, d, J = 8
Hz), 7.19 (lH, d, J = 9 Hz), 7.26 (lH, t), 7.34 (lH, d, J = 8
Hz), 7.63 (lH, d, J = 8 Hz), 7.76 (lH, d, J = 9 Hz); 7.88 (lH,
d, J = 9 Hz), 8.15 (2H, d, J = 9 Hz), 8.43 (2H, d, J = 9 Hz),
W094/20062 ~15 7 41 ~ PCT~S94/022~
~ - 276 -
9.84 (lH, pseudo s, 11.71 (lH, pseudo s); IR (KBr): cm 3422,
3273, 3250, 3108, 3081, 2873, 2361, 2339, 1676, 1636, 1606,
1536, 1497, 1426, 1369, 1288, 1232, 1200, 1148, 1093, 1072,
1013, 960, 920, 874, 856, 761, 724, 681, 606, 568. Anal.
Calcd. for C34H29N3Ol4S 1 H2O 1.2 TFA .17 CH3CN: C, 49.17;
H, 3.67: N, 4.94; S, 3.57. Found: C, 49.16; H, 3.57; N, 4.64;
S, 3.36. LRMS (FAB) m/z 736.1 (735.68 calculated for
C34HZ9N3ol4s ) -
W094/20062 2 1 ~ 7 4 1 2
PCT~S94/02283
- 277 -
Tr~ns-3-(4-hy~roxybenz~mido)-~-t~-~2-hy~roxy-6-(2-methyl-
propyloxy)c~rbonylhen~oyl)-3,5-~ihy~roxybenzoyloxy]pyrroli~ine
trifluoro~cetic ~cid s~lt (COMPOUND 577)
BnO~n1, (COCI)2, DMF, CH2C12 BnO~O o o~n
OBn OFB~ HO", NJb~ ~, HJ~3
BOC BOC
DMAP, EtJN
H2, Pd(OH)~ / C
TFA
HO E~OH / EtOAc
~N)~ $N~ QH
BOC
646
TFA
~0
~ O OH
H~
TFA
577
21S741`~
WO 94/20062 PCT/US94/02283
~ ~ - 278 -
Trans-N-t-butoxycarbonyl-3-(~-benzyloxybenzam~ ~o)-~-t~-(2-
benzyloxy-6-benzyloxycarbonylbenzoyl)-3,5-dibenzyloxy]-
pyrrolidine
4-[4-(2-benzyloxy-6-benzyloxycarbonylbenzoyl)]-3,5-
dibenzyloxybenzoic acid (1.47 mmol, 996 mg) and 15 ml anhydrous
CH2Cl2 in a dry round-bottom flask were cooled in an ice/water
bath under N2. To this was added oxalyl chloride (2.87 mmol,
0.25 ml) and 5 drops of DMF. This was allowed to stir for 2
hours while the bath melted. TLC (2:1 hexanes:EtOAc) indicated
complete formation of the acid chloride. The solvent was
removed in vacuo.
In a 200 ml dry round-bottom flask was added trans-N-
t-butoxycarbonyl-3-(4-benzyloxybenzamido)-4-hydroxypyrrolidine
(1.26 mmol, 500 mg) in 12 ml anhydrous CH2C12 under N2. To this
was added triethylamine (3.6 mmol, 0.5 ml) and DMAP (150 mg).
A solution of the acid chloride generated above in 10 ml
anhydrous CH2Cl2 was added via cannula. This was allowed to
stir under N2 at room temperature overnight. The reaction
mixture was then diluted with CH2C12, washed with sat. NaHCO3,
brine, then dried over MgSO" and concentrated in vacuo. The
crude product was purified via flash column chromatography
using 5% acetone/CH2Cl2 as the eluent. Trans-N-t-
butoxycarbonyl-3-(4-benzyloxybenzamido)-4-[4-(2-benzyloxy-6-
benzyloxycarbonylbenzoyl) -3,5-dibenzyloxy benzoyloxy~
pyrrolidine (1.08 mmol, 1.15 g) was obtained in 8696 yield.
Trans-N-t-butoxyoarbonyl-3-(~,-hy~roxybenzamido)-~-[~-(2-
hy~roxy-6-¢~,bG.~benzoyl)-3,5-dihydroxybenzoyloxy]pyrrol~dine
To a 500 ml 3-neck round-bottom flask was added trans-
N-t-butoxycarbonyl-3-(4-benzyloxybenzamido)-4-[4-(2-benzyloxy-
6-benzyloxycarbonylbenzoyl) -3, 5-dibenzyloxy
benzoyloxy]pyrrolidine (1.02 mmol, 1.08 g) in 17 ml EtOAc and
70 ml ethanol under N2. To this was added trifluoroacetic acid
(2.55 mmol, 0.20 ml) and Pd(OH)2/C (730 mg) followed
immediately by introduction of H2 at 1 atmosphere. After a
reaction time of 3.5 hours, the reaction was flushed with N2
-~094/20062 21~ 7 ~ ~ 2 PCT/US94/02283
- 279 -
and filtered through Celite, rinsing with ethanol. Following
concentration in vacuo, crude product (644 mg) was obtained in
quantitative yield. A small portion was purified via HPLC (21
x 250 mm C18 column, gradient ~B = 0 to 50 over 60 min. where
A = 0.1~ TFA, 5% CH3CN in water and B = CH3CN, 15 ml/min. W =
254 nm) for characterization and the remainder was used crude
in subsequent reactions. m.p. 196C(dec). IR (KBr) 3375(br),
2978, 1704, 1660, 1637, 1607, 1506, 1426, 1368, 1231 cm . H
NMR CD30D, ~ 8.52 (d, lH), 7.72 (d, 2H), 7.49 (d, lH), 7.26 (t,
lH), 7.01 (d, lH), 6.91 (s, 2H), 6.80 (d, 2H), 5.40 (m, lH),
4.63 (m, lH), 3.87 (m, 2H), 3.50 (m, 2H), 1.47 (s, 9H). LRMS
(M + 1) cacld for C31H31N2O12 623.2, found 623.2. Anal. Calcd for
C3lH3lN2O12 1-5 H2O: C, 57.317; H, 5.120; N, 4.312. Found: C,
57.26; H, 5.18; N, 4.47.
Trsns-N-t-butoxyc~rbonyl-3-(~-hydroxybenz~mi~o)-~-t4-~2-
hydroxy-6-~2-methylpropyloxy)¢~rbonylbenzoyl)-3,5-
dihydroxybenzoyloxy]pyrrolid~ne (COMPOUND 6~6)
To a round-bottom flask was added trans-N-t-
butoxycarbonyl-3-(4-hydroxybenzamido)-4-[4-(2-hydroxy-6-
carboxylbenzoyl)-3,5-dihydroxybenzoyloxy]pyrrolidine (0.lS
mmol, 95 mg) in 5 ml DMF. To this was added NaHCO3 (0.23 mmol,
19 mg) and 1-iodo-2-methylpropane (0.75 mmol, 0.09 ml). This
was allowed to stir under N2 for 7 days with additional 1-iodo-
2-methylpropane being added each day. A total of 48 additional
equivalents (7.2 mmol, 0.83 ml) were added over the reaction
period. The reaction mixture was diluted with EtOAc and washed
with water 3 times. The aqueous layer was back extracted with
EtOAc and the organic layers combined and dried over MgSO4 then
concentrated in vacuo. The crude product was purified via HPLC
(21 x 250 mm C18 column, gradient %B = 25 to 100 over 60 min.
where A = 0.1% TFA, and 5% CH3CN in water, B = CH3CN, 15 ml/min.
W = 254 nm) to isolate trans-N-t-butoxycarbonyl-3-(4-
hydroxybenzamido)-4-[4-(2-hydroxy-6-(2-methylpropyloxy)-
carbonylbenzoyl)-3,5-dihydroxybenzoyloxy]pyrrolidine (46.5mg,
44% yield).
WO 9~/20062 215 7 4 ~ 2 PCT/US94/02283
280 --
Trans-3- ~4-hydroxybenzami~o) -4- t4- ~2-hydroxy-6- ~2-
methylpropyloxy)carbonylbenzoyl)-3,5-dihy~roxybenzoyloxy]-
pyrrolidine trifluoroacetic acid ~alt ~COMPOlrND 577)
Trans-N-t-butoxycarbonyl-3-(4-hydroxybenzamido)-4-[4-
(2-hydroxy-6- (methylpropyloxy) carbonylbenzoyl) -3, 5-
dihydroxybenzoyloxy]pyrrolidine (46.5 mg, 0.068 mmol) was
dissolved in 0.75 ml neat trifluoroacetic acid and allowed to
stir at room temperature under N2 for 45 minutes at which time
TLC (75% CH2C12, 24% MeOH, 1% (10% aq.) NH40H) indicated the
reaction was complete. This was diluted with toluene and
concentrated in vacuo to yield 42.7 mg (91% yield) of crude
product. Purification via HPLC (21 x 250 mm C18 column,
gradient %B = 0 to 100 over 60 min. where A = 0.1% TFA, 5%
CH3CN in water, B = CH3CN, 15 ml/min. W = 254 nm) yielded
trans-3- (4-hydroxybenzamido) -4- [4- (2-hydroxy-6- (2-
methylpropyloxy ) -carbony1benz oy1 ) - 3, 5 -
dihydroxybenzoyloxy~pyrrolidine trifluoroacetic acid salt
(Compound 577, 32 mg, 6896 yield) as a yellow solid. m.p.
150-156-C (dec.). IR (KBr) 3191 (br), 2968, 1677, 1607, 1508,
1426, 1373, 1293, 1201 cm-l. lH NMR, DMSO-d6, ~, 11.74 (s,
2H), 10.13 (s, lH), 10.03 (s, lH), 9.20 (br, NH), 8.51 (d, lH),
7.74 (d, 2H), 7.44 (d, lH), 7.35 (t, lH), 7.12 (d, lH), 6.95
(s, 2H), 6.84 (d, 2H), 5.50 (m, lH), 4.60 (m, lH), 3.88 (d,
2H), 3.72 (m, 2H), 3.52 (m, 2H), 1.80 (q, lH), 0.83 (d, 6H).
LRMS (M + 1) calcd for C30H3lN2Olo 579.20, found 579.1. Anal.
calcd for C30H30N2Olo C2HF3O2 1.3 H2O: C, 53-68; H~ 4-73; N~
3.91. Found: C, 53.78; H, 4.54; N, 3.99.
~o9~l2oo62 ~1574~2
- 281 - PCT/US94/02283
Tr~n8~ -hy~roxybenz~mi~o) -2- t~- (2-hy~roxy-6-
hy~roxyc~rbonylbenzoyl)-3,5-~ihy~roxybenzoyloxyl]cyclohept~ne
(COMPO~ND 588)
C~ N-~CO~ CH~ (9 I~OH ~O (~ N~ TBDI~tS Cl
OBn
OTB0MS H?~Pd-C OTBDMS . ' H ~)
N~ M OH (~ BnO~ CO~H, THf TBDMS~N~
CO~Bn
OBn OlloO Olln g~n O n
Bu,NF THF H~j o EI,N,DMAP,CH2CI, BnO O N~
~J ~ O
651
a~,H
H?~Pd(OH)~ C ~H ~ H
M~OH THF o ~
588
21~7~12
W094/20062 PCT/US94/02283 --
~ ; - 282 -
NaOAc (426 mg, 5.2 mmol) was dissolved in a solution
of peroxyacetic acid in acetic acid (32 wt. %, 21.9 ml, 104
mmol) and the resultant solution was added dropwise over 30
min. to a mixture of cycloheptene (10 g, 104 mmol) and Na2CO3
(44.1 g, 416 mmol) in CH2C12 (100 ml) at 5 C. The mixture was
allowed to stir at room temperature for 3h with occasional
cooling using a water bath. All solid material was removed by
filtration, and the filtrate was distilled at atmospheric
pressure using a vigreux column to remove most of the CH2Cl2,
giving a colorless liquid which was shown by Hl NMR to be a
mixture of the epoxide and CH2C12. The purity was estimated to
be ca. 73% in the epoxide (total 15.93 g, 100% crude yield).
This material was used without further purification in the next
step.
A mixture of the epoxide from the previous step (10
g, 65 mmol), NaN3 (27.5 g, 423 mmol), NH4Cl (10.44 g, 195 mmol),
MeOH (162 ml), and H2O (20 ml) was stirred at reflux for 24h.
The solid was removed by filtration and the filtrate was
evaporated. The residue was treated with 0.5 N aq. NaOH (50 ml)
and extracted with CH2C12 (3 x 30 ml). The combined CH2Cl2
extracts were washed with H2O (50 ml), brine (50 ml), dried
(MgSO4), and evaporated to give the azide (8.39 g, 83%).
The azide (7 g, 45 mmol) in N,N-dimethylformamide (10
ml) was added to a solution of t-butyldimethylsilylchloride
(6.8 g, 45 mmol) and imidazole (3.07 g, 45 mmol) in N,N-
dimethylformamide (35 ml). The resultant mixture was stirred at
room temperature for 16h, diluted with Et2O (70 ml), washed
with H2O (4 x 30 ml) and brine (50 ml), dried (MgSO4), and
evaporated to give a pale yellow oil (11.33 g, 93%).
A mixture of the product of the previous reaction (8.1
g, 29.95 mmol) and 5% Pd on carbon (1.59 g, 2.5 mol. %) in MeOH
was stirred vigorously under 1 atm H2 at room temperature for
20h. The catalyst was removed by filtration over Celite and the
filtrate was evaporated. The residue was purified by flash
chromatography (SiO2, Et2O: Hexane = 2:1) to give the amine as
a colorless oil (5.27 g, 72~).
~094/20062 213 7 ~1~ PCT/US94/02283
- 283 -
A mixture of 4-benzyloxybenzoic acid (3.287 gj 14.40
mmol) and 1,1'-carbonyldiimidazole (2.335 g, 14.40 mmol) in THF
was stirred at room temperature for 2h. To the resultant slurry
was added a solution of the amine product of the previous
reaction (3.07 g, 12.56 mmol) in THF and stirring was continued
at room temperature for 24h. The mixture was diluted with CH2Cl2
(40 ml), washed with H20 (3 x 15 ml) and brine (2 x 15 ml),
dried (MgS04), and evaporated. The residue was purified by
flash chromatography (SiO2, Et20 : hexane = 1:5) to give the
amide as a white powder (4.69 g, 82%).
Tetrabutylammonium fluoride in THF (lM, 10 ml, 10
mmol) was added to a solution of the amide product from the
previous reaction (3.8 g, 8.36 mmol) in THF and the resultant
yellow solution was stirred at room temperature for 2h. The
mixture was poured into CH2C12 (150 ml), washed with H20 (3 x 30
ml) and brine (2 x 30 ml), dried (MgS04), and concentrated to
about 30 ml. The precipitate was collected by filtration and
washed with CH2Cl2 (3 x 3 ml). The combined filtrate and washes
were concentrated to about 10 ml, and the resultant precipitate
was collected, washed with CH2C12 (2 x 2 ml), combined with the
first crop and dried under vacuum. This gave a white solid
(2.24 g, 79%). The mother liquors were combined and
chromatographed (SiO2, Et20 : hexane = 1:5 followed by CH2Cl2 to
give an additional 256 mg of the product, total yield: 89%.
To a stirred solution of 4-(2-benzyloxycarbonyl-6-
benzyloxy)benzoyl-3,5-dibenzyloxybenzoylchloride (350 mg, 0.52
mmol) in CH2Cl2 (1.5 ml) was added oxalyl chloride (0.067 ml,
0.77 mmol) and one drop of N,N-dimethylformamide. The
resultant solution was stirred at room temperature for 2h and
then evaporated to dryness. The residue was dissolved in CH2Cl2
(1 ml) and added to a mixture of the product of the preceding
reaction (177 mg, 0.52 mmol) and Et3N (105 mg, 1.04 mmol) in
CH2Cl2 (2 ml). The mixture was stirred at room temperature for
17h, diluted with CH2Cl2 (15 ml), washed with H20 (3 x 10 ml),
dried (MgS04), and evaporated. The residue was chromatographed
wos4/20062 215 7 ~12 PCT~S94/02283
- 284 -
(sio2~ Et2O: hexane = 1:1, followed by Et2O: hexane: CH2Cl2 =
1:1:1) to give a white solid (166 mg, 32%).
Pd(OH)2 on carbon (20 wt. % contains > 50% moisture;
22 mg, 0.016 mmol) and MeOH (1.6 ml) were added to a solution
of the product of the prec~ing reaction (160 mg, 0.16 mmol) in
THF (1.6 ml) and the resultant mixture was stirred under 1 atm
H2 at room temperature for 17h. The catalyst was removed by
filtration over Celite and the Celite pad was washed with MeOH
(15 ml). The filtrate and washes were combined and evaporated
to give a yellow solid (83 mg, 95%) (COMPOUND 588). m.p. 186-C
(dec). Anal. calcd for C29H2,NOlo-lH2O: C, 61.37; H, 5.15; N,
2.47. Found: C, 61.41; H, 5.29; N, 2.36.
2 1 ~ 741~
094/20062 - 285 - PCT/US94/02283
Trans-3- (~-hy~lohyLt:nz~lmido) -4- t4- ~2-hy~roxy-6-hy~ro2ycarbonyl
benzoyl) -3,5-~ihy~roxybenzoyloxyl]pyrroli0inium
trifluoroacetate (COMPO~ND 589) ~nd 1-}~exa~ecanoyl-trzms-3-
~hy~GhyL ~nzami~o) -~.- t 40 ~2-hy~roxy-6-hy~roxycarbonylbenzoyl ) -
3, 5-~ihy0roxybenzoyloxyl ] pyrrol i~ine ~ COMPOIJND S 9 0 )
co2~
e~~i C~ ~n ~H~en
EloN~ DMAP, CH,C~I
o~L OEln O 06n
652
CO~H
~ ~ c~(c~"coa
MeOH, THF O ~ O
CF,CO2H
589
C~2H
~,~OH D~ H
HO~ ~H~J
,~0,~ 0
0~ (CH~"C~
590
To a stirred solution of 4(2-benzyloxycarbonyl-6-
benzyloxy)benzyl-3,5-dibenzyloxybenzoylchloride (350 mg, 0.52
mmol) in CH2Cl2 (1.5 ml) was added oxalyl chloride (0.067 ml,
~og~/20062 215 7 412 PCT/US94/02283
- 286 -
0.77 mmol) and one drop of N,N-dimethylformamide. The
resultant solution was stirred at room temperature for 2h and
then evaporated to dryness. The residue was dissolved in CH2Cl2
tl.5 ml) and added to a-mixture of trans-3-(4-hydroxy-1-
benzyloxycarbonyl pyrrolidine (232 mg, 0.53 mmol; for
preparation see Compound 585), Et3N (0.14 ml, 1.04 mmol), and
4-(N,N-dimethylamino)pyridine (6 mg, 0.052 mmol) in CH2Cl2 (1.5
ml). The mixture was stirred at room temperature for 17h,
diluted with CH2Cl2 (15 ml), washed with H2O (3 x 10 ml), dried
(MgSO4), and evaporated. The residue was chromatographed (SiO2,
Et2O:hexane = 1:1, then EtzO:hexane:CH2Cl2 = 1:1:0.5) to give
the ester (230 mg, 40%) (COMPOUND 652).
To a solution of Compound 652 (200 mg, 0.18 mmol) in
THF (1.8 ml) were added MeOH (1.8 ml), Pd(OH)2 on carbon (20
wt.~, contains 2 50% moist; 25 mg, 0.018 mmol), and CF3CO2H (41
mg, 0.36 mmol). The mixture was stirred at room temperature
under 1 atm H2 for 17h. The volatile components were removed
by evaporation, and the residue was taken up in MeOH, filtered
through Celite, and evaporated to give a yellow solid (88 mg,
72~) (COMPOUND 589).
Palmitoyl chloride (10 mg, 0.038 mmol) was added to
a stirred solution of Compound 589 (24 mg, 0.038 mmol) in
pyridine (0.4 ml). The mixture was stirred at room temperature
for 16h and TLC showed that the reaction was incomplete. More
palmitoyl chloride (5 mg) was added and stirring was continued
for 16h. The reaction mixture was evaporated to remove
pyridine leaving a yellow syrup with some solid material which
was shown to contain starting material by lH NMR. This
material was dissolved in pyridine (0.4 ml) and treated with
palmitoyl chloride (15 mg). The mixture was stirred at room
temperature for 24h, evaporated, and chromatographed (SiO2,
EtOAc followed by S% HOAc in acetone) to give a yellow solid
(19 mg, 66%) which was shown by lH NMR to be the desired amide
(Compound 590) with some contamination. m.p. 175-178C (dec).
Anal. calcd for C42H52N2Oll-3H2O: C, 61.90; H, 7.17; N, 3.44.
Found: C, 61.85; H, 7.07; N, 3.62.
_~094/20062 - 287 - PCT~S94/02283
Tran~-3-~-hydL~L~z~mido)-4-[4-(2-hydroxy-n~phth-1-oyl)-3,5-
dihydroxybenzyoyloxyl]pyrrolidine trifluoroacetic ~ci~ s~lt
(COMPOUND 591)
H~;;5 E~,N. O~P, C~
0~ OE~n OJ` Oan
653
~H 01 1
U O~, THF
N CF~
591
To a mixture of 4-(2-benzyloxy-naphth-1-oyl)-3,5-
dibenzyloxylbenzoic acid (297 mg, 0.5 mmol) and 2 drops of N,N-
dimethylformamide in CH2C12 (1.5 ml) was added oxalyl chloride
(0.065 ml, 0.75 mmol) dropwise over 2 min. The resultant
mixture was stirred at room temperature for 2h and the volatile
components were evaporated. The remaining yellow solid was
dried under vacuum for lh, dissolved in CH2Cl2 (1.5 ml), and
added to a solution of trans-3-(4-hydroxybenzamido-4-hydroxy-1-
benzyloxycarbonyl pyrrolidine (168 mg, 0.38 mmol; for
preparation see Compound 585), Et3N (0.1 ml, 0.76 mmol), and 4-
(N,N-dimethylamino)pyridine (5 mg, 0.038 mmol) in CH2C12 at 5C.
wos4noo62 215 ~ 4 ~- ~ PCT/US94/022~
- 288 -
The mixture was stirred at room temperature for 16h, diluted
with CH2Cl2 (10 ml), washed with H2O (3 x 10 ml), dried (MgSO4),
and evaporated. The residue was chromatographed (SiO2,
Et2O:hexane = 1:1, followed by Et2O:hexane:CH2Cl2 = 1:1:0.5) to
give a white solid (336 mg, 66%) (COMPOUND 653).
Pd(OH)2 on carbon (20 wt.%, contains > 50% moist; 21
mg, 0.015 mmol), followed by CF3CO2H (34 mg, 0.3 mmol) and MeOH
(1.5 ml), was added to a solution of Compound 653 (155 mg, 0.15
mmol) in THF (1.5 ml). The resultant mixture was stirred at
room temperature under 1 atm H2 for 2Oh. The catalyst was
removed by filtration over Celite and the Celite pad was washed
with MeOH (20 ml). The combined filtrate and washes were
evaporated to give a yellow solid (85 mg, 88%) (COMPOUND 591).
~ ~O9~/20062 2157~12
- 289 - PCT/US94/02283
l-Isopropyl-tr~n~-3-(~-hydroxybenz~mi~o~ y~2~by ~ ~y~
hydroxyc~rbonylhe~7cyl)-3~5-dihy~roxybenzoyloxyl]pyrrolidinium
trifluoro~cet~te (COMPOUND 592)
OBn
O~n ,~ ~,O~;
~S ~, ~ o
~=/ NBH~. ~H)--\ O E~,~. DMAP, CHICI~
H~,
H CF,CO,H
~eO ~e o
~H~ H, Pd(OH~,-C ~o
c~ TF~ 0~. T\~F O ~ o
~ Cf~
6~ 592
W09~i20062 215 7 ~ PCT/US94/02283
- 290 -
To a solution of trans-3-(4-benzyloxybenzamido)-3-
hydroxy pyrrolidine (213 mg, 0.5 mmol) in acetic acid (2.5 ml)
was added NaBH4 (95 mg, 2.5 mmol) in small portions. After the
H2 evolution had ceased (ca. 10 min.) acetone (0.18 ml, 2.5
mmol) was added and the mixture was stirred at room temperature
for 16h. The reaction mixture was basified with 2N KOH and the
resultant cloudy mixture was extracted with CH2C12 (3 x 15 ml).
The combined CHzCl2 extracts were dried (MgSO4), and evaporated
to give a colorless oil (163 mg, 92%).
Oxalyl chloride in CH2C12 (2 M, 0.39 ml, 0.78 mmol)
was added dropwise to a solution of 4-(2-benzyloxy-6-
benzyloxycarbonylbenzoyl)-3,5-dibenzyloxybenzoic acid (353 mg,
0.52 mmol) and a drop of DMF in CH2C12 (2 ml) at 5 C. The
mixture was stirred at room temperature for 2h, then evaporated
to remove the solvent and excess oxalyl chloride. The residue
was dried in vacuo for lh, dissolved in CH2C12 (1 ml), and added
to a mixture of the product of the preceding reaction (142 mg,
0.4 mmol), Et3N (81 mg, 0.8 mmol), and DMAP (6 mg, 0.054 mmol)
in CH2Cl2 (2 ml) at 5-C. The mixture was stirred at room
temperature for 17h, diluted with CH2C12 (15 ml), washed with
H2O (3 x 10 ml), dried (MgSO4), and evaporated. The residue was
chromatographed (SiO2, Et2O:CH2Cl2:hexane = 1:1:1 followed by
Et2O:CH2Cl2:hexane = 2:2:1) to give a pale yellow oil (82 mg,
20%) (COMPOUND 654).
Pd(OH)2 on carbon(20 wt%, contains <50% moist, 11 mg,
0.008 mmol), trifluoroacetic acid (18 mg, 0.16 mmol), and MeOH
(1 ml) was added to a solution of Compound 654 (84 mg, 0.08
mmol) in EtOAc (1 ml) and the mixture was stirred under 1 atm
H2 contained in a balloon at room temperature for 16h. The
mixture was filtered through Celite and the Celite pad was
washed with MeOH (5 ml). The combined filtrates were
evaporated to give a yellow solid (32 mg, 59%) (COMPOUND 592).
IR (KBr, cm ):1705, 1676, 1636, 1607. FBMS: M/Z = 565 (M + 1).
2157~12
~O g~/20062 - 291 - PCT/US94/02283
Trans-~- t~- (2-carboxy-6-hydroxybenzoyl) -3,5-
~ihy~roxybenzoyloxy]-3- (4-hy~roxybenzami~o) -1-
[ (phenylamino) carbonyl] -pyrroli~ine (COMPO~JND 593)
NE~ OBn
H-TFA O~N
H
C 02Bn
iPr2NE~, ¦ OBn
DMAP, ~3 c OC 1
OBn
08n
H~OhN~ Pd(OHk ~ O OBn
H02C~ooH N 2 ~-08n I Fh
593 655
(~) -Trans-~-Hydroxy-3- (~-benzyloxybenz~mido)-l-
tphenylamino)carbonyl]-pyrrolidine
To a solution of (+) trans-4-hydroxy-3-(4-
benzyloxybenzamido pyrrolidine TFA (100 mg, 0.235 mmol) in
methanol (4.7 ml) were added triethyl amine (66 ~L, 0.470 mmol,
2.0 eq) then phenyl isocyanate (26 ~L, 0.235 mmol, 1.0 eq).
The cloudy mixture was stirred at room temperature under N2 45
min, then poured into H2O (30 ml) and extracted with CH2Cl2 (3
x 25 ml). The organic layers were combined, dried (MgSO4),
filtered and evaporated to a white solid (82 mg, 81%): 1H NMR
(300 MNz, CD30D ~ 7.60 (d, J = 8.8 Hz, 2H), 7.00-7.25 (m, llH),
6.85 (d, J = 8.9 Hz, 2H), 6.79 (d, J = 7.1 Hz, lH), 4.93 (s,
2H), 4.20-4.25 (m, lH), 4.16 (dd, J = 8.2, 3.1 Hz, lH), 3.73
W094/20062 21 S 7 ~12 ~ ~ PCT~S94/02283
- 292 -
(dd, J c 11, 6.4 Hz, lH), 3.57 (dd, J = 11.4, 5.1 Hz, lH), 3.12
(dd, J = 11.0, 3.5 ~z, lH), 3.26 (dd, J = 11.1, 2.8 Hz, lH).
( + ) -Tran~-~- t ~ - ~ 6-benzyloxy-2- ( benzyloxycarbonyl ) benzoyl ] - 3, 5-
~ibenzyloxybenzoyloxy) -3- (~.-benzyloxybenzamido) -1-
(phenylamino) c~rbonyl]-pyrrolidine (CON~uNv 655)
To a solution of the product of the preceding reaction
(80 mg, 0.19 mmol), 4-dimethylaminopyridine (26 mg, 0.21 mmol,
1.1 eq) and diisopropylethylamine (37 ~L, 0.21 mmol, 1.1 eq) in
CH2Cl2 (3.2 ml) was added a solution of acid chloride (0.21
mmol) in CH2C12 (1.6 ml). The mixture was stirred at room
temperature under N2 18 h, then poured into 5% HCl (30 ml) and
extracted with CH2Cl2 (3 x 25 ml). The organic layers were
combined, dried (MgSO4) filtered and evaporated. Flash column
chromatography of the golden residue (1.1 hexanes:ethyl
acetate) on silica gel provided the title product (110 mg, 53%)
as an off white foam: 1H NMR (300 MHz, CDCl3) ~ 7.89 (d, J = 8.8
Hz, 2H), 6.9-7.6 (m, 26H), 6.82 (d, J = 7.5 Hz, 2H), 5.4-5.5
(m, lH), 5.12 (s, 2H), 5.02 (s, 2H), 4.80 !m, lH), 4.76 (s,
4H), 4.68 (s, 2H), 4.0-4.05 (m, lH), 3.95-4.0 (m, lH), 3.65-3.7
(m, lH), 3.6-3.65 (m, lH).
Tr~ns- ~ - t 4 - ~ 2 -C~rboxy- 6-hydroxybenzoyl ) -3, 5-
dihydroxybenzoyloxy] -3- (4-hy~roxybenzami~o) -~-
~phenylamino) carbonyl]-pyrrolidine ~COMr~u~v 593)
To a round bottom flask containing Compound 655
(110 mg, 0.101 mmol) were added Pd(OH)2 (22 mg of a 20% powder)
then THF (2.2 ml) and ethanol (2.2 ml). The flask was
evacuated and filled with H2 twice, then allowed to stir under
H2 (1 atm) for 23 h. The suspension was filtered, washed
through with methanol (40 ml) and evaporated to a yellow solid.
Purification by reverse phase HPLC (618 column) provided the
title product as a yellow powder after lyophilization (24.9 mg,
39%): m.p. 183-208 (dec); IR (KBr) 3458, 3336, 1719, 1679,
1622, 1227, 760 cm ; lH NMR (300 MHz, CD30D) ~ 7.53 (d, J = 8.6
Hz, 2H), 7.28 (d, J = 7.7 Hz, lH), 7.20 (d, J = 7,7 Hz, 2H),
7.0-7.1 (m, 3H), 6.82 (d, J = 8.3 Hz, 2H), 6.73 (s, 2H), 6.62
215741~ ~
WO 94/20062 PCT/US94/02283
-- 293 --
(8.7, J = d Hz, 2H), 5.25-5.35 (m, lH), 4.5-4.6 (m, lH), 3.75-
3.9 (m, 2H), 3.4-3.5 (m, 2H): MS m/e calc'd for C33H2~N3Oll:
642.1732, found 642.1865; Analysis calc'd for C33H27N3O1l-0.5
TFA-1.25 H2O: C,56.63; H, 4.19; N, 5.83; found: C, 56.85; H,
~4.26; N, 5.96.
W094l20062 ~lS 7 4 1 2
- 294 - PCT~S94/02283
~)Trans-4t4-(2-Carboxy-6-hy~roxybenzoyl)-3,5-
dihydroxybenzoyloxy]-3-(4-hydroxybenzamido)-1-
t(methylamino)c~rbonyl]-pyrroli~ine (COMPOUND S9~)
HO~p~ , NE~3 ~ ~3`08n
H-lFA H
C02Bn
Pr2NEt, ¦ OB n
DMAP, ~'COCI
OBn
OBn
H 0~0 ~ EdlOHHkHF _~
~o h OH Oq~ ~j OBn
H02C~b~ o~N,CH3 BnO2C~_gBn N
656
(~)-Trans-4-Hydroxy-3-(~-benzyloxybenzamido)-1-
t(methylsmino)c~rbonyl]-pyrrolidine
To a solution of (+)-trans-4-hydroxy-3-(4-
benzyloxybenzamido)pyrrolidine TFA (100 mg, 0.235 mmol) in
methanol (4.7 ml) were added triethyl amine (66 ~1, 0.470 mmol,
2.0 eq) then methyl isocyanate (14 ~L, 0.235 mmol, 1.0 eq). The
cloudy mixture was stirred at room temperature under N2 1.5 h,
then more methyl isocyanate (14 ~L, 1.0 eq) was added. The
mixture was allowed to stir 10 min more, then poured into H20
(30 ml) and extracted with CH2C12 (3 x 25 ml). The organic
layers were combined, dried (MgS04) filtered and evaporated to
~ W094/20062 21~ 7 412 PCT/US94/02283
- 295 -
a white solid (83 mg, 95%): H NMR (300 MHz, CD30D) ~ 7.60 (d,
J = 8.8 Hz, 2H), 7.1-7.25 (m, 5M), 6.85 (d, J = 8.8 Hz, 2H),
4.66 (s, 2H), 4.15 (dd, J = 6.5, 3.4 Hz, lH), 4.10 (dd, J =
5.0, 3.4 Hz, lH), 3.57 (dd, J = 10.8, 6.5 Hz, lH), 3.43 (dd, J
= 11.1, 5.2 Hz, lH), 3.1-3.2 (m, 2H), 2.53 (s, 3H).
(+)-Trans-~-t~-~6-benzyloxy-2-~benzyloxycarbonyl)benzoyl]-3,5-
dibenzyloxybenzoyloxy) -3- ~-benzyloxybenzamido) -1-
[ ~methylamino) c~rhonyl~ -pyrrolidine ~COMPOUND CS6)
To a solution of the product of the prececeding
reaction (85 mg, 0.23 mmol), 4-dimethylaminopyridine (28 mg,
0.23 mmol, 1.1 eq) and diisopropylethylamine (40 ~L, 0.23 mmol,
1.1 eq) in CH2C12 (3.9 ml) was added a solution of acid chloride
(0.25 mmol) in CH2C12 (1.9 ml). The mixture was stirred at room
temperature under N2 16 h, then poured into 5% HCl (30 ml) and
extracted with CH2C12 (3 x 25 ml). The organic layers were
combined, dried (MgS04) filtered and evaporated. Flash column
chromatography of the golden residue (98:2 CH2Cl2:methanol) on
silica gel provided the title product (159 mg, 67%) as an off
white solid: lH NMR (300 MHz, CDCl3) ~ 7.84 (d, J = 8.7 Hz, 2H),
7.3-7.4 (m, 5H), 7.1-7.3 (m, 16H), 7.06-7.08 (m, 6H), 6.9-7.0
(m, 3H), 6.84 (d, J = 7.6 Hz, 2H), 5.49 (m, lH), 5.14 (s, 2H),
5.07 (s, 2H), 4.78 (s, 4H), 4.70 (m, lH), 4.70 (s, 2H), 4.40
(m, lH), 3.95-4.0 (m, 2H), 3.53-3.60 (m, 2H), 2.80 (d, J = 4.5
Hz, 3H).
~ + ) -Trans- ~ - t ~ - ~ 2 -Carboxy- 6 -hydroxybenzoyl ) - 3, 5 -
dihy~ro~cyenzoyloxy] -3- ~.-hy~roxybenzamido) -1-
~methyl~ino) carbonyl]-pyrrolidine ~COMPOIJND 594)
To a round bottom flask containing Compound 656
(159 mg, 0.154 mmol) were added Pd(OH)2 (40 mg of a 20% powder)
then THF (3.0 ml) and ethanol (3.0 ml). The flask was evacuated
and filled with H2 twice, then allowed to stir under H2 (1 atm)
for 21 h. The suspension was filtered, washed through with
methanol (50 ml) and evaporated to a yellow solid.
Purification by reverse phase HPLC (C18 column) provided the
title product as a yellow powder after lyophilization (68.7 mg,
~VO 94/20062 21~ 7 412 PCT/US94/02283
-- 296 --
77%): m.p. 178-198 (dec); IR (KBr) 3385, 1714, 1605, 1236, 763
cm; H NMR (300 MHz, CD30D) ~ 7.53 (d, J = 8.7 Hz, 2H), 7.30
(d, J = 7,7 Hz, lH), 7.06 (dd, J = 8.1, 7.9 Hz, lH), 6.80 (d,
J = 8.3 Hz, lH), 6.70 (s, 2H), 6.62 (d, J = 8.7 Hz, 2H), 5.2-
5.3 (m, lH), 4.4-4.5 (m, lH), 3.6-3.8(6 line mult, 2H), 3.2-3.4
(m, 2H), 2.53 (s, 3H); MS m/e calc'd for C28H25N3011: 580-1567~
found 580.1481; Analysis calc'd for C2~H25N3O11-0.3TFA 0.8H20:
C, 54.32; H, 3.96; N, 6.72; found: C, 54.61; H, 4.32; N, 6.68.
~O9~/20062 21 r 7 ~
- 297 - ~ PCT~S94/02283
~ tr~ns-3-~3,~-~ihy~roxybenz~mido)-~-t~-(2-c~rboxy-6-
hy~roxybenzoyl)-3,5-dihy~roxy]benzoyloxypyrroli~ine
trifluoro~cetic ~cid ~lt (COMPOUND 675)
BnO
-- ~ O Jonesox.
BnO ~ ~
~-J H 93%
OBn
H NH2 ~ OBn
- BnO H _
+ Bn ~ O NaOH,CH2C12 HO\ NL_
CB OH 99% N
Z B
z
BnO
O 1.(COCI)2cat.DMF,~ OBn OBn
OBnCH2CI2 ~ ~
2. Et3N, DMAP,BnLOnO ~ H ~ OBn
B~sOno~~OH CH2CI2.70%
O N
/ B
HO / 2. Pd(H)2
,~--o / EtOAc-EtOH, TFA
OH OH
H~ ~ H ~ OH
O ~ N ~
H CF3COOH
675
WO9~/20062 21~ 7 ~1 2 PCT/US94102t83
- 298 -
Jones reagent (6ml) was added to a solution of
3,4-dibenzyloxybenzaldehyde (Aldrich, 1.0 g, 3.14 mmol) in
acetone (20 ml) until the reaction remained the color of Jones
reagent. The excess of Jones reagent was destroyed by adding
i-PrOH and acetone was removed in vacuo. The slurry residue
was taken into EtOAc, washed with brine, dried with Na2SO4, and
concentrated to afford off-white solids (0.98 g, 93~).
To a suspension of 3,4-dibenzyloxybenzoic acid (0.467
mg, 1.40 mmol) in anhydrous CH2C12 (5 ml) was added cat. DMF and
oxalyl chloride (2.0 M solution in CH2C12, 1.92 ml, 3.85 mmol)
at room temperature The mixture was kept for stirring at room
temperature for 2 hr. Solvents were removed and the acid
chloride residue was taken into CH2C12 (5 ml) after drying over
the vacuum for lhr. To a biphasic reaction mixture of N-CBZ-3-
amino-4-hydroxypyrrolidine (0.74 M solution in CH2Cl2, 300 mg,
1.7 ml, 1.27 mmol) in CH2C12 (10 ml) and 1 N NaOH (13.0 ml) was
added a solution of 3-benzyloxybenzoic acid chloride in
anhydrous CH2C12 (5 ml). The resulting mixture was vigorously
stirred at room temperature for 3h. The reaction mixture was
diluted with EtOAc, washed with brine, and dried over Na2SO4.
The crude product after concentration was triturated in Et2O
and EtOAc to afford white solids (quantitative yield).
To a solution of benzophenone acid (265 mg, 0.39 mmol)
in CH2Cl2 (5 ml) was added cat. DMF and oxalyl chloride (2.0 M
solution in CH2C12, 0.488 ml, 0.976 mmol) at room temperature
The mixture was kept for stirring at room temperature for 2 hr.
Solvents were removed and the acid chloride residue was taken
into CH2Cl2 (5 ml) after drying over the vacuum for lhr.
A solution of amidoalcohol (215.5 mg, 0.39 mmol), Et3N
(197.3 mg, 272 ~L, 1.95 mmol) and DMAP (47.6 mg, 0.39 mmol) in
CH2Cl2 (5 ml) was treated with the freshly made acid
chloride-CH2Cl2 solution (5 ml) at 5C. The reaction mixture
was allowed to stir at room temperature for 3h and then
chromatographed on silica gel eluting with 2:3 to
l:l/EtOAc:Hexane. The product was obtained as fluffy white
solids (332 mg, 70%).
~O 94/20062 215 ~ PCT/US94/02283
-- 299 --
The product of the previous reaction (320 mg, 0.264
mmol) was dissolved in EtOAc-HOEt (1:1, 25 ml) and treated with
TFA (cat.) followed by 10% Pd(OH)2 (170 mg, 60 mol%). The
mixture was subject to hydrogenolysis at 50 psi for 20hr.
Solvents were concentrated after filtering through a pad of
celite, and the residue was dissolved in DMF (1.0 ml) and
loaded onto HPLC; conditions: A-0.1% TFA 5%CH3CN/H2O, B-100%
CH3CN, 0-50% B over 60 min, 25 ml/min, 41 x 300 mm C18 column.
Fractions (one/min) 37-40 were combined, partially
concentrated, and lyophilized to afford fluffy yellow solids
(113, 65%). (COMPOUND 675) m.p. 210-213 (dec)-C. IR (KBr)
cm 1 3391, 3246, 1717, 1676, 1636, and 1603. Anal. Calcd. for
C26H22N2O1l-2-0H2O-1.0C2HF3O2: C, 48.84; H, 3.95; N, 4.07. Found:
C, 48.75; H, 3.63; N, 4.07. LRFAB (M + 1): 579.
wosl/20062 21~ 7 ~ 12 PCT/US94/02283
- 300 -
(+)-Trans-2-t4-(6-hydroxy-2-(carboxyl)benzoyl)-3,5-
dihydroxybenzoyloxy]-1-(2-hydroxybenzamido)cyclopentane
~COMPOUND 681)
2 H2,PdonC HO
3.2NKOH,THF h
CIOC~3 ~ BnO
BnO
~0 OBn COCI
CH2CI2, DIPEA, DMAP
BnO
Ho2c~ N)~[3 H2 ~0b~1HlB~3
OH OBn
681 680
FIGURE AP
~2-Benzyloxybenzoyl)chloride
To a solution of 2-benzyloxybenzoic acid (684 mg, 3.00
mmol) in CH2Cl2 (15 ml) were added dimethylformamide (1 drop)
then oxalyl chloride (3.0 ml of a 2.0 M solution in CH2C12, 6.00
mmol, 2.0 eq). The solution was stirred at room temperature 1
h, then evaporated and the light yellow semi-solid was placed
on the vacuum pump before use.
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COMPREND PLUS D'UN TOME. - -
CECI EST LE TOME I DE
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