Note: Descriptions are shown in the official language in which they were submitted.
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TELOMERASE INHIBITORS AND METHODS OF THEIR USE
Field of the Invention
The present invention relates to thiazolidinone compounds that inhibit
telomerase
activity, to pharmaceutical compositions containing the compounds and to the
use of the
compounds and compositions, alone or in combination with other
pharmaceutically active
agents, in the treatment of telomerase-mediated conditions or diseases, such
as cancer.
Background of the Invention
Telomerase catalyzes the synthesis of telomeres. Telomeres are characteristic
tandem
repeats (TTAGGG in mammals) found at the ends of most eukaryotic chromosomes,
that may
be 15-25 kilobases long in human germline cells. With each. cell division,
about 60-100 bases
are lost from the ends of the chromosomes, and as the telomeres shorten, cells
eventually reach
crisis and apotosis is triggered. See Harley et al., (1991) Mutation Res. 256:
271-282}.
Telomerase acts to maintain the telomere length just above the crisis level,
and are thus
responsible for chromosome stability and are involved in the regulation of the
cell cycle.
Telomerase is a ribonucleoprotein reverse transcriptase that contains its own
RNA
template for the synthesis of telomeric DNA. See Blackburn, 1992, Annu. Rev.
Biochem.,
61:113-129. Telomerase is present in stem and germline cells of normal
tissues, and at much
higher levels in over 85% of tumors (Kim et al., 1994, Science, 266:2011-
2014). Thus, drugs
targeted towards telomerase potentially will have a high selectivity for tumor
over healthy
tissues. Consequently, telomerase inhibition has been proposed as a new
approach to cancer
therapy.
The inhibition of telomerase activity by antisense strategies directed towards
the
telomerase RNA component, for example peptide nucleic acids (Norton et al.,
(1996) Nature
Biotech. 14: 615-619) and phosphorothioate oligonucleotides has been reported.
Since
telomerase is a reverse transcriptase, the use of inhibitors of reverse
transcriptases, such as
AZT, and other nucleosides has also been reported. Telomerase inhibition by
cisplatin,
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possibly due to crosslinking of the telomeric repeat sequences, is also known
(Burger et al.,
(1997) Eur. J. Cancer 33: 638-644).
Thiazolidinediones comprise a group of structurally related antidiabetic
compounds that
increases the insulin sensitivity of target tissues (skeletal muscle, liver,
adipose) in insulin
resistant animals. In addition to these effects on hyperglycemia,
thiazolidinediones also reduce
lipid and insulin levels in animal models of NIDDM. Recently, the
thiazolidinedione
troglitazone was shown to have these same beneficial effects in human patients
suffering from
impaired glucose tolerance, a metabolic condition that precedes the
development of NIDDM, as
in patients suffering from NIDDM (Nolan et al., (1994) N. Eng. J. Med. 331,
1188-1193).
While their mechanism of action remains unclear, it is known that the
thiazolidinediones do not
cause increases in insulin secretion or in the number or affinity of insulin
receptor binding sites,
suggesting that thiazolidinediones amplify post-receptor events in the insulin
signaling (Colca,
J. R., and Morton, D. R. (1990) in New Antidiabetic Drugs (C. J. Bailey and P.
R. Flatt, eds.).
Smith-Gordon, New York, 255-261; Chang et al. (1983) Diabetes 32, 839-845).
Thiazolidinediones have been found to be efficacious inducers of
differentiation in
cultured pre-adipocyte cell lines (Hiragun et al. (1988) J. Cell Physiol. 134,
124-130; Sparks et
al. (1991) J. Cell. Physiol. 146, 101-109; Kleitzien et al. (1992) Mol.
Pharmacol. 41, 393-398).
Additionally, thiazolidinediones have been implicated in appetite regulation
disorders (see WO
94/25026 A1), and in increase of bone marrow fat content. In addition,
thiazolidinedione
compounds have been suggested for use in the treatment of psoriasis (IJ.S.
Patent No.
5,824,694) and climacteric symptoms and mesenchymal tumors (U.S. Patent No.
5,814,647).
Some of the compounds useful for practicing the method of the present
invention, and
methods of making some of these compounds are known. For example, some of
these
compounds are disclosed in WO 91/07107; WO 92/02520; WO 94/01433; WO 89/08651;
JP
Kokai 69383/92; EP 0 155845; EP 0 155848; EP 0 193256; EP 0 295828; and U.S.
Pat. Nos.
4,287,200; 4,340,605; 4,376,777; 4,438,141; 4,444,779; 4,461,902; 4,486,594;
4,572,912;
4,582,839; 4,687,777; 4,703,052; 4,725,610; 4,738,972; 4,775,687; 4,791,125;
4,812,570;
4,873,255; 4,897,393; 4,897,405; 4,918,091; 4,948,900; 5,002,953; 5,023,085;
5,053,420;
5,061,?17; 5,120,754; 5,132,317; 5,143,928; 5,194,443; 5,223,522; 5,232,925;
5,252,735;
5,260,445; 5,814,647; 5,824,694; and 5,874,454.
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The identification of compounds that inhibit telomerase activity provides
important
benefits to efforts at treating human disease. Compounds that inhibit
telomerase activity can be
used to treat telomerase-mediated disorders, such as cancer, since cancer
cells express
telomerase activity and normal human somatic cells do not possess telomerase
activity at
biologically relevant levels (i.e., at levels sufficient to maintain telomere
length over many cell
divisions). Unfortunately, few such compounds, especially compounds with high
potency or
activity and compounds that are orally bioavailable, have been identified and
characterized.
Hence, there remains a need for compounds that act as telomerase inhibitors
that have
relatively high potency or activity and that are orally bioavailable, and for
compositions and
methods for treating cancer and other diseases in which telomerase activity is
present
abnormally. The present invention meets these and other needs.
Summary of the Invention
The present invention provides methods, compounds and compositions that are
specific
and effective for treating telomerase-mediated disorders, such as malignant
conditions by
targeting cells having telomerase activity. The methods, compounds, and
compositions of the
invention can be applied to a wide variety of malignant cell types and avoid
the problems
inherent in current cancer treatment modalities which are non-specific and
excessively toxic.
In a first aspect, the present invention is based on the finding that certain
known
thiazolidinone compounds, as well as new thiazolidinone derivatives disclosed
herein, are
effective in the inhibition of telomerase enzyme activity, in vitro, ex vivo
and in vivo. Thus, in
certain aspects, the present invention provides methods of inhibiting
telomerase by contacting
telomerase with the compounds described herein. In particular embodiments, the
telomerase to
be inhibited is a mammalian telomerase, such as a human telomerase. A related
aspect of the
present invention is the discovery that thiazolidinone compounds inhibit the
proliferation of
cells that have telomerase activitiy, such as many cancer cells. Thus, this
aspect of the present
invention provides methods of inhibiting telomerase activity in a patient,
preferably a mammal,
suffering from a telomerase-mediated condition or disease, comprising
administering to the
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patient a therapeutically effective amount of a telomerase inhibiting
thiazolidinone compound,
or a pharmaceutically acceptable salt thereof.
In another aspect, the present invention provides methods, compounds and
compositions for inhibiting a telomerase enzyme, comprising contacting the
telomerase enzyme
with a composition or a compound of formula (I):
'~- ~2 R
H ~/> (I)
O . R~ L~~ 3
Ra
n
Or its pharmaceutically acceptable salts, wherein X is oxygen or sulfur, ~ is
a single or double
bond; A is aryl or heteroaryl; R, is hydrogen or lower alkyl; R2, R3 and R4
are independently
selected from the group consisting of hydrogen, halo, alkyl, aryl, hydroxyl,
alkoxyl, aryloxyl,
aralkoxyl, cyano, nitro, alkylcarbamido, arylcarbamido, dialkylcarbamido,
diarylcarbamido,
alkylarylcarbamido, alkylthiocarbamido, arylthiocarbamido,
dialkylthiocarbamido,
diarylthiocarbamido, alkylarylthiocarbamido, amino, alkylamino, arylamino,
dialkylamino,
diarylamino, arylalkylamino, aminocarbonyl, alkylaminocarbonyl,
arylaminocarbonyl,
dialkylaminocarbonyl, diarylaminocarbonyl, arylalkylaminocarbonyl,
alkylcarbonyloxy,
arylcarbonyloxy, carboxyl, alkoxycarbonyl, aryloxycarbonyl, sulfo,
alkylsulfonylamido,
arylsulfonamido, alkylsulfonyl, arylsulfonyl, alkylsulfinyl, arylsulfinyl and
heteroaryl; L is a
direct bond or a linking group having from 1 to 3 atoms independently selected
from
unsubstituted or substituted carbon, N, O or S; and n is 1 or 2.
In another aspect, the present invention provides methods, compounds and
compositions for inhibiting a telomerase enzyme, comprising contacting the
telomerase enzyme
with a compound, or its pharmaceutically acceptable salt, having the formula
(IV):
H ~ A~ (IV)
O 'Rs
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where X is O or S; ~ is a single or double bond; RS is H or lower alkyl; and
Ar is a substituted
or unsubstituted aryl, heteroaryl, aralkyl, heteroarylalkyl, arylalkeryl,
heteroarylalkeryl,
arylalkynyl or heteroarylalkynyl.
In another aspect, the present invention provides methods, compounds and
compositions for inhibiting a telomerase enzyme, comprising contacting the
telomerase enzyme
with a composition or a compound of formula (V):
O Rs L-A1
or its pharmaceutically acceptable salt, wherein X is O or S; R6 is H or lower
alkyl; W is
CH=CH, S, or -N=C-; R~ is OH, halogen, mercapto, vitro, cyano; lower
alkylthio, lower alkyl,
lower alkoxy, lower alkanoyloxy, NR,~R,2 (wherein R~1 and R,2 are
independently selected
from the group consisting of hydrogen, lower alkyl, lower alkanoyl, aryl,
heteroaryl,
heteroarylalkyl, or R" and R,Z form a substituted or unsubstituted
heterocyclic ring), CO2R13
(wherein R,3 is selected from the group consisting of hydrogen, lower alkyl,
aralkyl, and
heteroarylalkyl), CONR"R,Z, substituted or unsubstituted aryl, substituted or
unsubstituted
heteroaryl, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkyloxy, lower
alkanoyl, amyl, lower
alkenyl, arylthio, or lower alkynyl; and when W represents S, R' may also be
H; L is O, S, SO,
S02, OCHZ, SCHZ, SOCH2, SOZCHZ, or N(R,o)(CH2)m (wherein R,o is substituted or
unsubstituted aryl, heteroaryl, aralkyl, or heteroarylalkyl, and m is 0 or 1),
(CH2)N(R,o)(CHz)m,
or CR,3R,4 (wherein R,3 and R,4 are independently selected from the group
consisting of
hydrogen, hydroxy, aryl, and heteroaryl); and A~ is cycloarlkyl of formula
(A1):
-5-
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2
Z3 (A1 )
Z5 Z4
wherein Z, to ZS are independently selected from the group consisting of
hydrogen, lower
alkyl, lower alkenyl, lower alkanoyloxy, mercapto, alkylthio, NR"R,2, nitro,
cyano, COZR,3,
CONR"R,2, aryl, heteroaryl, aryloxy, heteroaryloxy, aralkyloxy,
heteroarylalkyloxy, halogen,
S and lower alkanoyl provided that when W is CH=CH, A may also by pyridyl.
The new compounds of the invention have many valuable uses as inhibitors of
deleterious telomerase activity, such as, for example, in the treatment of
cancer in mammals,
such as humans. The pharmaceutical compositions of this invention can be
employed in
treatment regimens in which cancer cells are killed, in vivo, or can be used
to kill cancer cells
ex vivo. Thus, this invention provides therapeutic compounds and compositions
for treating
cancer, and methods for treating cancer and other telomerase-mediated
conditions or diseases
in humans and other mammals (e.g., cows, horses, sheep, steer, pigs and
animals of veterinary
interest such as cats and dogs).
Detailed Description
I. Definitions
Unless otherwise defined below, the terms used herein have their normally
accepted
scientific meanings. Definition of standard chemistry terms may be found in
reference works,
including Carey and Sundberg (1992) "Advanced Organic Chemistry 3'°
Ed." Vols. A and B,
Plenum Press, New York.
The term "thiazolidinone" or "thiazolidinone derivative" as used herein refers
to
compounds of the general formula:
-6-
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Hey
O
wherein X is O or S. When X is O, the derivatives are thiazolidinedione
derivatives. When X
is S, the derivatives are the thiazolidinonethione derivatives also known as
rhodanines (see
Examples 25-28, below).
The term "alkyl" as used herein refers to a straight, branched, or cyclic
hydrocarbon
chain fragment or radical containing between about one and about twenty carbon
atoms, more
preferably between about one and about ten carbon atoms (e.g., methyl, ethyl,
n-propyl, iso-
propyl, cyclopropyl, n-butyl, iso-butyl, tert-butyl, cyclobutyl, adamantyl,
noradamantyl and the
like). Straight, branched, or cyclic hydrocarbon chains having eight or fewer
carbon atoms will
, also be referred to herein as "lower alkyl". The hydrocarbon chains may
further include one or
more degrees of unsaturation, i.e., one or more double or triple bonds (e.g.,
vinyl, propargyl,
allyl, 2-buten-1-yl, 2-cyclopenten-1-yl, 1,3-cyclohexadien-1-yl, 3-cyclohexen-
1-yl and the
like). Alkyl groups containing double bonds such as just described will also
be referred to
herein as "alkenes". Similarly, alkyl groups having triple bonds will also be
referred to herein
as "alkynes". However, as used in context with respect to cyclic alkyl groups,
the combinations
of double and/or triple bonds do not include those bonding arrangements that
render the cyclic
hydrocarbon chain aromatic.
In addition, the term "alkyl" as used herein further includes one or more
substitutions at
one or more carbon atoms of the hydrocarbon fragment or radical. Such
substitutions include,
but are not limited to: aryl; heterocycle; halogen (to form, e.g.,
trifluoromethyl, --CF3); nitro
(--N02); cyano (--CN); hydroxyl (also referred to herein as "hydroxy"),
alkoxyl (also referred
herein as alkoxy) or aryloxyl (also referred to herein as "aryloxy")(--OR);
thio or mercapto,
alkyl- or arylthio (--SR); amino, alkylamino, arylamino, dialkyl- or
diarylamino, or
arylalkylamino (--NRR'); aminocarbonyl, allcylaminocarbonyl,
arylaminocarbonyl,
dialkylaminocarbonyl, diarylaminocarbonyl or arylalkylaminocarbonyl (--
C(O)NRR');
carboxyl, or alkyl- or aryloxycarbonyl (--C(O)OR); carboxaldehyde, or aryl- or
alkylcarbonyl
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(--C(O)R); iminyl, aryl- or alkyliminyl (--C(=NR)R'); sulfo (--SOzOR); alkyl-
or arylsulfonyl
(--SOZR); ureido (--HNC(=O)NRR'); or thioureido (--HNC(=S)NRR'); where R and
R'
independently are hydrogen, aryl or alkyl as defined herein. Substituents
including heterocyclic
groups (i.e., heterocycle, heteroaryl, and heteroaralkyl) are defined by
analogy to the above-
described terms. For example, the term "heterocycleoxy" refers to the group --
OR, where R is
heterocycle as defined below.
The alkyl moiety of "lower alkanoyl", "lower alkoxy", "lower alkanoyloxy",
"lower
alkylthio", is the same as "alkyl" defined above.
The term "methylene" refers to the group --CHZ--.
The term "methine" refers to a methylene group for which one hydrogen atom has
been
replaced by a substituent as described above. The term "methine" can also
refer to a methylene
group for which one hydrogen atom is replaced by bond to form an sp2 -
hybridized carbon
center (i.e., >C=O).
The term "halo" or "halogen" as used herein refers to the substituents fluoro,
bromo,
chloro, and iodo.
The term "carbonyl" as used herein refers to the functional group --C(O)--.
However, it
will be appreciated that this group may be replaced with well-known groups
that have similar
electronic and/or steric character, such as thiocarbonyl (--C(S)--); sulfinyl
(--S(O)--); sulfonyl
(--SOz--), phosphonyl (--P01--), and methylidene (--C(=CH2)--). Other carbonyl
equivalents
will be familiar to those having skill in the medicinal and organic chemical
arts.
The term "aryl" as used herein refers to cyclic aromatic hydrocarbon chains
having
twenty or fewer carbon atoms, e.g., phenyl, naphthyl, biphenyl and
anthracenyl. One or more
carbon atoms of the aryl group may also be substituted with, e.g.: alkyl;
aryl; heterocycle;
formyl; halogen; vitro; cyano; hydroxyl, alkoxyl or aryloxyl; thio or
mercapto, alkyl-, or
arylthio; amino, alkylamino, arylamino, dialkyl-, diaryl-, or arylalkylamino;
aminocarbonyl,
alkylaminocarbonyl, arylaminocarbonyl, dialkylaminocarbonyl,
diarylaminocarbonyl or
arylalkylaminocarbonyl; carboxyl, or alkyl- or aryloxycarbonyl;
carboxaldehyde, or aryl- or
alkylcarbonyl; iminyl, or aryl- or alkyliminyl; sulfo; alkyl- or arylsulfonyl;
hydroximinyl, or
aryl- or alkoximinyl; ureido; or thioureido. In addition, two or more alkyl or
heteroalkyl
substituents of an aryl group may be combined to form fused aryl-alkyl or aryl-
heteroalkyl ring
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systems (e.g., tetrahydronaphthyl). Substituents including heterocyclic groups
(e.g.,
heterocycleoxy, heteroaryloxy, and heteroaralkylthio) are defined by analogy
to the above-
described terms.
The term "aralkyl" as used herein refers to an aryl group that is joined to a
parent
structure by an alkyl group as described above, e.g., benzyl, a-methylbenzyl,
phenethyl, and
the like. The aralkyl moiety of "aralkylsulfonyl" aralkyloxy is the same as
"aralkyl" defined
above.
The aryl moiety of "aroyl", "arylalkenyl", "arylalkenyl", "arylsulfonyl",
"arylthio",
"aryloxy", "arylalkenylsulfonyl", "arylalkynylsulfonyl" is the same as "aryl"
defined above.
The term "heterocycle" as used herein refers to a cyclic alkyl group or aryl
group as
defined above in which one or more carbon atoms have been replaced by a non-
carbon atom,
especially nitrogen, oxygen, or sulfur. Non-aromatic heterocycles will also be
referred to
herein as "cyclic heteroalkyl". Aromatic heterocycles are also referred to
herein as
"heteroaryl". For example, such groups include furyl, tetrahydrofuryl,
pyrrolyl, pyrrolidinyl,
thienyl, tetrahydrothienyl, oxazolyl, isoxazolyl, triazolyl, thiazolyl,
isothiazolyl, pyrazolyl,
pyrazolidinyl, oxadiazolyl, thiadiazolyl, imidazolyl, imidazolinyl, pyridyl,
pyridazinyl,
triazinyl, piperidinyl, morpholinyl, thiomorpholinyl, pyrazinyl, piperazinyl,
pyrimidinyl,
naphthyridinyl, benzofuranyl, benzothienyl, indolyl, indolinyl, indolizinyl,
indazolyl,
quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl,
quinazolinyl, quinoxalinyl,
pteridinyl, quinuclidinyl, carbazolyl, acridiniyl, phenazinyl, phenothiazinyl,
phenoxazinyl,
purinyl, benzimidazolyl, benzthiazolyl, and benzoxazolyl.
The heteroaryl moiety of "heteroarylalkyl", "heteroarylalkenyl",
"heteroarylalkynyl",
"heteroarylsulfonyl", "heteroarylalkylsulfonyl", "heteroarylalkenylsulfonyl",
"heteroarylalkynylsulfonyl", "heteroaryloxy", "heteroarylalkyloxy" is the same
as "heteroaryl"
defined above.
The above heterocyclic groups may further include one or more substituents at
one or
more carbon and/or non-carbon atoms of the heteroaryl group, e.g.: alkyl;
aryl; heterocycle;
halogen; nitro; cyano; hydroxyl, alkoxyl or aryloxyl; thio or mercapto, alkyl-
or arylthio;
amino, alkyl-, aryl-, dialkyl- diaryl-, or arylalkylamino; aminocarbonyl,
alkylaminocarbonyl,
arylaminocarbonyl, dialkylaminocarbonyl, diarylaminocarbonyl or
arylalkylaminocarbonyl;
_g_
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carboxyl, or alkyl- or aryloxycarbonyl; carboxaldehyde, or aryl- or
alkylcarbonyl; iminyl, or
aryl- or alkyliminyl; sulfo; alkyl- or arylsulfonyl; hydroximinyl, or aryl- or
alkoximinyl; ureido;
ar thioureido. In addition, two or more alkyl substituents may be combined to
form fused
heterocycle-alkyl or heterocycle-aryl ring systems. Substituents including
heterocyclic groups
(e.g., heterocycleoxy, heteroaryloxy, and heteroaralkylthio) are defined by
analogy to the
above-described terms.
The term "heterocyclealkyl" refers to a heterocycle group that is joined to a
parent
structure by one or more alkyl groups as described above, e.g., 2-
piperidylmethyl, and the like.
The term "heteroaralkyl" as used herein refers to a heteroaryl group that is
joined to a parent
structure by one or more alkyl groups as described above, e.g., 2-
thienylmethyl, and the like.
The compounds of the present invention may be used to inhibit or reduce
telomerase
enzyme activity and/or proliferation of cells having telomerase activity. In
these contexts,
inhibition and reduction of the enzyme or cell proliferation refers to a lower
level of the
measured activity relative to a control experiment in which the enzyme or
cells are not treated
with the test compound. In particular embodiments, the inhibition or reduction
in the measured
activity is at least a 10% reduction or inhibition. One of skill in the art
will appreciate that
reduction or inhibition of the measured activity of at least 20%, 50%, 75%,
90% or 100% may
be preferred for particular applications.
II. Telomerase Inhibitors
As noted above, the immortalization of cells involves inter alia the
activation of
telomerase. More specifically, the connection between telomerase activity and
the ability of
many tumor cell lines, including skin, connective tissue, adipose, breast,
lung, stomach,
pancreas, ovary, cervix, uterus, kidney, bladder, colon, prostate, central
nervous system (CNS),
retina and blood tumor cell lines, to remain immortal has been demonstrated by
analysis of
telomerase activity (Kim et al.). This analysis, supplemented by data that
indicates that the
shortening of telomere length can provide the signal for replicative
senescence in normal cells
(see WO 93/23572), demonstrates that inhibition of telomerase activity can be
an effective anti-
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cancer therapy. By "inhibition" is simply meant a reagent, drug or chemical
which is able to
decrease the activity of the telomerase enzyme in vitro or in vivo. Such
inhibitors can be
readily identified using standard screening protocols in which a cellular
extract or other
preparation having telomerase activity is placed in contact with a potential
inhibitor, and the
S level of telomerase activity measured in the presence or absence of the
inhibitor, or in the
presence of varying amounts of inhibitor. In this way, not only can useful
inhibitors be
identified, but the optimum level of such an inhibitor can be determined in
vitro for further
testing in vivo.
In a related aspect, the invention proves a method for inhibiting the ability
of a cell to
proliferate or replicate. In this method, one or more of the thiazolidinone
compounds of the
invention, that are capable of inhibiting telomerase enzyme activity, are
provided during cell
replication. As explained above, telomeres play a critical role in allowing
the end of the linear
chromosomal DNA to be replicated completely without the loss of terminal bases
at the 5'-end
of each strand. Immortal cells and rapidly proliferating cells use telomerase
to add telomeric
DNA repeats to chromosomal ends. Inhibition of telomerase will result in the
proliferating
cells not being able to add telomeres and they will eventually stop dividing.
As will be evident
to those of ordinary skill in the art, this method for inhibiting the ability
of a cell to proliferate
is useful for the treatment of a condition associated with an increased rate
of proliferation of a
cell, such as in cancer (telomerase-activity in malignant cells), and
hematopoiesis (telomerase
activity in hematopoietic stem cells), for example.
Thus, in one aspect, the present invention provides compositions and compounds
for the
prevention or treatment of many types of malignancies. In particular, the
compounds of the
present invention can provide a highly general method of treating many, if not
most,
malignancies, as demonstrated by the highly varied human tumor cell lines and
tumors having
telomerase activity. More importantly, the thiazolidinone compounds of the
present invention
can be effective in providing treatments that discriminate between malignant
and normal cells
to a high degree, avoiding many of the deleterious side-effects present with
most current
chemotherapeutic regimes which rely on agents that kill dividing cells
indiscriminately.
Representative known thiazolidinedione compounds include the glitazones, such
as, for
example, troglitazone (also known as CS-045 (Sankyo) and CI-991 (Park-Davis)),
pioglitazone
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(also known as AD-4833 and U-72107E), rosiglitazone (also known as BRL49653),
englitazone (also known as CP-68,722), and ciglitazone.
In another aspect, the present invention provides new compounds,
pharmaceutical
compositions and methods relating to the new compounds, or their
pharmaceutically acceptable
salts, for inhibiting a telomerase enzyme, comprising contacting the
telomerase enzyme with a
compound, or its pharmaceutically acceptable salt, having the formula (I):
R
.. L 3
R
1
Ra
n
wherein X is oxygen or sulfur, ~ is a single or double bond; A is aryl or
heteroaryl; R, is
hydrogen or lower alkyl; R2, R3 and R4 are independently selected from the
group consisting of
hydrogen, halo, alkyl, aryl, hydroxyl, alkoxyl, aryloxyl, aralkoxyl, cyano,
nitro,
alkylcarbamido, arylcarbamido, dialkylcarbamido, diarylcarbamido,
alkylarylcarbamido,
alkylthiocarbamido, arylthiocarbamido, dialkylthiocarbamido,
diarylthiocarbamido,
alkylarylthiocarbamido, amino, alkylamino, arylamino, dialkylamino,
diarylamino,
arylalkylamino, aminocarbonyl, alkylaminocarbonyl, arylaminocarbonyl,
dialkylaminocarbonyl, diarylaminocarbonyl, arylalkylaminocarbonyl,
alkylcarbonyloxy,
arylcarbonyloxy, carboxyl, alkoxycarbonyl, aryloxycarbonyl, sulfo,
alkylsulfonylamido,
arylsulfonamido, alkylsulfonyl, arylsulfonyl, alkylsulfinyl, arylsulfinyl and
heteroaryl; L is a
direct bond or a linking group having from 1 to 3 atoms independently selected
from
unsubstituted or substituted carbon, N, O or S; and n is 1 or 2.
In certain embodiments, the new compounds of the present invention have the
general
structure shown as formula II below:
- 12-
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/R3
H ~ \ /
O Ra
n
PCT/US00/18211
and their pharmaceutically acceptable salts, wherein X is O or S, and R2, R3,
R4, L, and n are as
defined above.
In the compounds of formula I, A may be aryl to form, for example, a phenyl
moiety.
S Alternatively, A may be heteroaryl, such as, for example, pyridine,
quinoline, isoquinoline,
thiophene, furan, imidazole, benzimidazole, pyrazole, and the like. In
presently preferred
embodiments, A is phenyl, as in formula (II). In other presently preferred
embodiments, when
n is l, R, can not be hydrogen. In yet other presently preferred embodiments,
at least one of RZ
and R3 is other than hydrogen. In presently particularly preferred
embodiments, at least one of
RZ and R3 is halo, and most preferably both RZ and R3 are halo to form a
dihalo-substituted
phenyl moiety.
As noted above, L may be a direct bond, or may be a 1 to 3 atom linking group
wherein
the atoms of the linking group independently selected from unsubstituted or
substituted carbon,
N, O or S. Representative linking groups useful in the compounds of the
invention include, for
example -O-, -S-, -NH-, -CHI-, -OCHZ-, -OC(O)-, -C02-, -NHC(O)-, -C(O)NH-, -
OC(O)CH2-,
-OC(O)NH-, and -NHC(O)NH-.
For purposes of illustration, representative compounds include, without
limitation, 5-(2-
(3,4-Dichlorophenyl)benzylidene)thiazolidine-2,4-dione, 5-(3-(3,4-
Dichlorophenyl)-
benzylidene)thiazolidine-2,4-dione, 5-(4-(3,4-
Dichlorobenzyloxy)benzylidene)thiazolidine-2,4-
dione, 5-(2-(3,4-Dichlorobenzyloxy)benzylidene)thiazolidine-2,4-dione, 5-(4-
(3,4-
Dichlorobenzamido)benzylidene)thiazolidine-2,4-dione, 5-(4-(N-3,4-
Dichlorophenyureido)-
benzylidene)thiazolidine-2,4-dione, 5-(2-(N-3,4-
Dichlorophenyureido)benzylidene)-
thiazolidine-2,4-dione, 5-(2-(N-3,4-
Dichlorophenylcarbamido)benzylidene)thiazolidine-2,4-
dione, 5-(3-(N-3,4-Dichlorophenylcarbamido)benzylidene)thiazolidine-2,4-dione,
5-(4-(N-3, 4-
Dichlorophenylcarbamido)benzylidene)thiazolidine-2,4-dione, S-(4-(N-3,4-
Dichlorophenylcarbamoyloxy)benzylidene)thiazolidine-2,4-dione, 5-(4-(3,4-
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Dichlorophenoxycarbonyl)benzylidene)thiazolidine-2,4-dione, S-(2-(3,4-
Dichlorophenoxycarbonyl)benzylidene)thiazolidine-2,4-dione, 5-(2-(3,4-
Dichlorophenylacetoxy)benzylidene)thiazolidine-2,4-dione, S-(3-(3,4-
Dichlorophenylacetoxy)-
benzylidene)thiazolidine-2,4-dione, 5-(4-(3,4-
Dichlorophenylacetoxy}benzylidene)thiazolidine-
2,4-dione, 5-(2-(3,4-Dichlorobenzoyloxy)benzylidene)thiazolidine-2,4-dione, 5-
(3-(3,4-
Dichlorobenezoyloxy)-benzylidene)thiazolidine-2,4-dione, 5-(4-(3,4-
Dichlorobenzoyloxy)-
benzylidene)thiazolidine-2,4-dione, 5-(3,4-Bis-(3,4-
dichlorobenzyloxy)benzylidine)-
thiazolidine-2,4-dione, 5-(2-(3,4-Dichlorophenoxy)benzylidine)thiazolidine-2,4-
dione, S-(4-
(3,4-Dichlorophenoxy)benzylidine)thiazolidine-2,4-dione, 5-(2,5-Bis-(3,4-
dichlorobenzyloxy)-
benzylidine)thiazolidine-2,4-dione, 5-(2,4-Bis-(3,4-
dichlorobenzyloxy)benzylidine)-
thiazolidine-2,4-dione, 5-(2-(3,4-Dichlorobenzylthio)-3H-pyrimidin-4-on-6-
ylmethylidene)-
rhodanine, 5-(2-(3,4-Dichlorobenzylthio)pyrimidin-4-ylmethylidene)rhodanine, 5-
(2-(3,4-
Dichlorobenzylthio)pyrimidin-4-ylmethylidene)rhodanine, 5-(3-Cyano-2-(3,4-
dichlorobenzylthio)pyridin-6-ylmethylidene)thiazolidine-2,4-dione and 5-(3-
(3,4-
Dichlorobenzyloxy}benzylidene)thiazolidine-2,4-dione.
In certain embodiments, the new compounds of the present invention have the
general
structure shown as formula III below:
z
(iii)
/R3
O R~ ~
Ra
and their pharmaceutically acceptable salts, wherein X is O or S, and R,, R2,
R3, R4, and L are
as defined above.
In another embodiment, the present invention provides methods, compounds and
compositions for inhibiting a telomerase enzyme, comprising contacting the
telomerase enzyme
with a compound, or its pharmaceutically acceptable salt, having the formula
(IV):
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H . Ar (IV)
O 'R5
where X is O or S; ~ is a single or double bond; RS is H or lower alkyl; and
Ar is a substituted
or unsubstituted aryl, heteroaryl, aralkyl, heteroarylalky, arylalkenyl,
heteroarylalkenyl,
arylalkynyl or heteroarylalkynyl.
Compounds of formula (IV) having the double bond can be obtained by reacting a
thiazolidine derivative with an aromatic carbonyl compound. The reaction can
be carried out
optionally in the presence of a base catalyst and optionally in a solvent. The
base catalyst,
usually present in about 0.1 to about 1 equivalent, may be piperidine,
piperidinium acetate,
diethylamine, pyridine, sodium acetate, potassium carbonate, sodium carbonate,
and the like.
IO The solvent may be an alcohol, such as methanol, ethanol, propanol, or the
like, an ether, such
as diethyl ether, tetrahydrofuran, dioxane, or the like, or a hydrocarbon,
such as benzene,
roluene, xylene, or the like, and mixtures thereof. The reaction is carned out
at a temperature
of about room temperature to about 200°C, preferably about 50-
100°C, and completes in about
one hour to about 50 hours. Compounds of formula (IV) wherein ~ is a single
bond can be
synthesized by reducing the double bond of the compound made above. Typically,
hydrogenation is carried out using a noble metal catalyst, such as palladium,
platinum,
rhodium, or the like, as is well known in the art.
In another aspect, the present invention provides methods, compounds and
compositions for inhibiting a telomerase enzyme, comprising contacting the
telomerase enzyme
with a compound, or its pharmaceutically acceptable salt, having the formula
(V):
W.~~ (v)
O R
s
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wherein X is O or S; R6 is H or lower alkyl; W is CH=CH, S, or N=C-; R~ is H,
OH, halogen,
mercapto, vitro, cyano, lower alkylthio, lower alkyl, lower alkoxy, lower
alkanoyloxy, NR"R12
(wherein R" and R12 are independently selected from the group consisting of
hydrogen, lower
alkyl, lower alkanoyl, aryl, heteroaryl, heteroarylalkyl, or R, ~ and R,2 form
a substituted or
unsubstituted heterocyclic ring), COZR13 (wherein R,3 is selected from the
group consisting of
hydrogen, lower alkyl, aralkyl, and heteroarylalkyl), CONRI ~R~2, substituted
or unsubstituted
aryl, substituted or unsubstituted heteroaryl, aryloxy, heteroaryloxy,
aralkyloxy,
heteroarylalkyloxy, lower alkanoyl, aroyl, lower alkenyl, arylthio, or lower
alkynyl; L is O, S,
SO, SO2, OCH2, SCHZ, SOCH2, S02CH2, or N(Rlo)(CHz)m (wherein R,o is
substituted or
unsubstituted aryl, heteroaryl, aralkyl, or heteroarylalkyl, and m is 0 or 1),
(CHZ)N(Rlo){CH2)m,
or CR,3R~4 (wherein R,3 and R,4 are independently selected from the group
consisting of
hydrogen, hydroxy, aryl, and heteroaryl); and A, is cycloarlkyl of formula
(A1):
1 2
Z3
Z5 Z4
wherein Z, to ZS are independently selected from the group consisting of
hydrogen, lower
alkyl, lower alkenyl, lower alkanoyloxy, mercapto, alkylthio, NR~,R,2, vitro,
cyano, C02R~3,
CONR11R,2, aryl, heteroaryl, aryloxy, heteroaryloxy, aralkyloxy,
heteroarylalkyloxy, halogen,
and lower alkanoyl provided that when W is CH=CH, A, may also by pyridyl.
Examples of the compound of the present invention represented by formulae I to
V are
shown in Tables 1 to 6 below, though compounds of the present invention are
not restricted
thereby.
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Table 1
R5
O I ~ R2
f.7"
. .V
Com ound X RS L Rz R3
No.
1 O 5-NOz 2-S H CH3
2 O 5-NHz 2-S H CH3
3 O 5-NHCOCH3 2-S H CH3
4 O 5-NOz 2-SO H CH3
O S-NOz 2-SOz H CH3
6 O H 2-S H Cl
7 O H 2-SO H C1
8 O H 2-SOz H Cl
9 O 3-NOz 4-S H CH3
O H 2-O H H
11 O H 3-O H H
12 O H 3-O H CH3
13 O H 3-O C1 CI
14 O H 4-O H H
O H 4-O H CH3
16 O 5-NOz 2-OCHz H H
17 O 5-NOz 2-OCHz Cl Cl
18 O 5-NOz 2-OCHz H CH3
19 O 4-NOz 3-OCHz H CH3
O 3-NOz 4-OCHz H CH3
21 O 2-NOz 5-OCHz H CH3
46 O H 4-N- 4-bromo hen 1 H Br
47 O 5-Ph 2-OCHz- H CH3
48 O 5- 2-thien 2-OCHz- H CH3
49 O 1 4-N- 4-h drox meth 1 hen H CH20H
H 1
50 O H 2-NCHz 4-bromo hen 1 H H
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Table 2
-,Rs
O Ri
R.,
' -J
Com ound No. X R, R6 L R3
22 O H H 2-OCHz CH3
23 O H 5-OCH3 2-OCH2 CH3
24 O H S-Cl 2-OCH2 CH3
25 O H 5-Br 2-OCH2 CH3
26 O H H 4-NC6H5 H
27 O H H 2- H
28 O H H 3- H
29 O H H 4- H
30 O H H 4-CHOH H
31 O H H 4-CO H
32 O H H 4-CHZ H
33 O H H 4-C OH C6H5 H
34 O H H 4-CHC6H5 H
35 O H H 4-CH2NC6H5 H
36 O H H 4-N CHZC6H5 H
CHZ
37 S H 5-N02 2-S C1
38 S H 5_NOZ 2-S CH3
39 O H S_NOZ 2-O CF3
40 O H 2-Br 5-OCHZ CH3
41 O H 2-OCHz -Tol S-OCH2 CH3
*
42 O CH3 S-Br 2-OCHZ CH3
*p-Tol = 4-methylphenyl
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Table 3
H
O
R3
Com ound No. R~ L R3
43 S-Br 2-S C1
44 S-C6Hs 2-S Cl
4S H 2-S CI
S1 S-Br 2-SO CI
S2 S-Br 2-SOz CI
S3 S-COzH 2-S Cl
S4 S-CONEtz 2-S Cl
SS S-CONHPh 2-S CI
S6 S-CO -meth I i erazine2-S CI
S~ S-CO-mo holine 2-S CI
S8 S-COZCH3 2-S Cl
Table 4
~ R~
H ~ ~
O ~ L-A
Com ound No. R~ L-A
S9 H 3-OPh
60 H 4-S- 4-chloro hen 1
61 4-S- 4-chloro S-S- 4-chloro hen 1
hen 1
62 H S-S- 4-chloro hen 1
63 4-Br S-S- 4-chloro hen 1
64 4-Br S-SO- 4-chloro hen 1
6S 4-CO- 4-chloro S-S- 4-chloro hen 1
hen I
66 4-CO- 4-chloro S-SOz- 4-chloro hen 1
hen 1
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Table 5
R6
H
~~L-A
Com and No. R6 L-A
67 5-NOZ 2-S 4-chloro hen 1
6g 5-NOZ 2-SO 4-chloro hen 1
69 5-NOZ 2-SOz 4-chloro hen 1
70 5-NOZ 2-S 3-chloro hen 1
71 5-N02 2-S 2-chloro hen 1
72 S-NOZ 2-S 3 4-dichloro hen 1
73 5-NOz 2-S 4-bromo hen 1
74 5-NOZ 2-S 4-methox hen I
75 5-NOZ 2-S 4-eth 1 hen I
76 5-NOZ 2-SCH2C6H5
77 5-N02 2-SOCHzC6H5
78 5-NOZ 2-S02CHZC6H5
79 5-NO2 2-SCHZ 4-chloro hen 1
3-NOZ 4-S 4-bromo hen 1
gl 3-N02 4-S 4-chloro hen 1
g2 3-NOZ 4-SO 4-meth 1 hen I
g3 3-NOZ 4-S02 4-meth 1 hen 1
g4 5-N02 2-S-c clohex I
gs 5-NOZ 2-SO-c clohex 1
g6 5-NOZ 2-SOZ-c clohex 1
87 5-Br 2-S 4-meth I hen 1
gg 3-Br 4-S 4-meth 1 hen 1
g9 5- 2- rid I 2-S 4-meth 1 hen 1
90 5- 2-fu 1 2-S 4-meth 1 hen 1
91 5- 2- 1 2-SO 4-meth 1 hen 1
92 5- 2-thien 1 2-S 4-meth 1 hen 1
93 5- 2-thien 1 2-SO 4-meth 1 hen I
94 5-CN 2-S 4-meth 1 hen 1
95 3-CN 4-S 4-meth 1 hen 1
96 S-CH20H 2-S 4-meth I hen 1
97 5-COCH3 2-S 4-meth I hen 1
9g 5-COCH3 2-SO 4-meth 1 hen I
99 6-CF3 2-S 4-meth 1 hen 1
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100 5-CF3 2-S 4-meth 1 hen 1
101 4-CF3 2-S 4-meth 1 hen 1
102 3-CF3 2-S 4-meth I hen 1
103 3-CF3 2-SO 4-meth I hen I
104 5-OCH3 2-S 4-meth 1 hen 1
105 4-OCH3 2-S 4-meth I hen 1
106 5-CI 2-S 4-meth 1 hen 1
107 5-CI 2-SO 4-meth I hen I
108 4-CI 2-S 4-meth 1 hen 1
109 3-CI ~ 4-S 4-meth 1 hen I
110 5-NOz 2-OC6Hs
111 5-NOZ 2-O 4-meth 1 hen 1
112 5-NOZ 2-O 4- 2',2'-dimeth 1-eth I
hen I
113 5-CF3 2-SO 4-meth 1 hen 1
114 5-CN 2-SO 4-meth 1 hen 1
115 5-NOz 2-S 4- trifluorometh 1 hen
I
116 5-N02 2-SO 4-methox hen 1
117 5-N02 2-SO 2-chloro hen 1
118 5-COZH 2-S 4-meth 1 hen 1
119 5-N02 2-S 2- rid I
120 5-NOZ 2-SO 4- rid 1
121 H 4-N C6Hs CH2C6Hs
122 5-NOZ 2-S 2-h drox eth I
123 5-N02 2-N ro 12
124 5-NOZ a
125 5-NOZ b
126 2-OCH3 4-OH
127 H 2-OCF3
129 S-N02 2-S 4-carbox 1 hen 1
130 5-NOZ c
131 5-N02 d
132 S-N02 2-S 4-meth lthio hen 1
133 5-N02 2-SO 4-eth I hen I
134 5-N02 2-SO 3-chloro hen 1
135 5-N02 2-SO 3 4-dichloro hen I
136 3-N02 4-S 4-bromo hen 1
137 3-OC6Hs 4-S 4-bromo hen I
138 3-OCH3 4-S 4-meth I hen 1
139 5-C02CH2C6Hs 2-S 4-meth I hen 1
140 3-CN 4-SO 4-meth I hen 1
141 3-Cl 4-SO 4-meth 1 hen I
142 5-CH OCH3 2 2-S 4-meth I hen 1
143 3-Br 4-SO 4-meth 1 hen 1
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144 5-CHO 2-S 4-meth 1 hen 1
145 5-CH=CHC02C CH3 2-S 4-meth I hen 1
3
146 5-CH=CHCOZH 2-S 4-meth 1 hen 1
147 3-CFA 4-S 4-meth 1 hen 1
148 3-CF3 4-SO 4-meth I hen 1
149 5-CON CZHS z 2-S 4-meth I hen I
150 5-CON C2H5 z 2-SO 4-meth 1 hen 1
151 a 2-S 4-meth 1 hen I
152 3-COCH3 _
2-S 3,4-dichloro hen 1
153 3-COCH3 2-SO 3 4-dichloro hen 1
154 5-NOz 2-S 2,3-dichloro hen 1
155 5-NOz 2-S 2,4-dichloro hen 1
156 5-NOz 2-SO 2,3-dichloro hen 1
157 5-NOz 2-SO 2,4-dichloro hen 1
158 5-NOz 2-SOz 2 3-dichloro hen 1
159 5-NOz 2-SOz 2 4-dichloro hen 1
160 5-NOz 2-S 4-h drox hen I
161 5-NOz 2-S 3 4-dimeth 1 hen I
162 5-NOz 2-SOz 3,4-dichloro hen 1
163 3-NOz 4-SO 4-chloro hen 1
164 3-NOz 4-S 4-eth 1 hen 1
165 3-NOz 4-SO 4-eth 1 hen 1
166 3-NOz 2-SOz 4-eth I hen I
167 3-NOz 4-S 3,4-dichloro hen I
168 3-N02 4-SO 3,4-dichloro hen 1
169 5-NOz 2-SO 2,3-dimeth I hen 1
170 5-NOz 2-SOz 2,3-dimeth 1 hen I
171 ~ 5-NOz
.
rn~ . ~ = 2' \ / ~N~c~~z d = 2 \
e= 5-co-n~ . f= \ ~ I
z_
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Table 6
Com ound No.
128
CI
III. Synthesis of Telomerase Inhibitors
The compounds of the present invention can be synthesized using techniques and
materials known to those of skill in the art, such as described, for example,
in March,
ADVANCED ORGANIC CHEMISTRY 4'" Ed., {Wiley 1992); Carey and Sundberg,
ADVANCED ORGANIC CHEMISTRY 3'd Ed., Vols. A and B (Plenum 1992), and Green and
Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 2"d Ed. (Whey 1991). Starting
materials for the compounds of the invention may be obtained using standard
techniques and
commercially available precursor materials, such as those available from
Aldrich Chemical Co.
(Milwaukee, Wis.), Sigma Chemical Co. (St. Louis, Mo.), Lancaster Synthesis
(Windham,
N.H.), Apin Chemicals, Ltd. (New Brunswick, N.J.), Ryan Scientific (Columbia,
S.C.),
Maybridge (Cornwall, England), Arcos (Pittsburgh, Pa.), and Trans World
Chemicals
(Rockville, Md.).
The procedures described herein for synthesizing the compounds of the
invention may
include one or more steps of protection and deprotection (e.g., the formation
and removal of
acetal groups). In addition, the synthetic procedures disclosed below can
include various
purifications, such as column chromatography, flash chromatography, thin-layer
chromatography (TLC), recrystallization, distillation, high-pressure liquid
chromatography
(HPLC) and the like. Also, various techniques well known in the chemical arts
for the
identification and quantification of chemical reaction products, such as
proton and carbon-13
nuclear magnetic resonance ('H and '3C NMR), infrared and ultraviolet
spectroscopy (IR and
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LJV), X-ray crystallography, elemental analysis (EA), HPLC and mass
spectroscopy (MS) can
be used as well. Methods of protection and deprotection, purification,
identification and
quantification are well known in the chemical arts.
Compounds of the class represented by formulas I, II and III can be
synthesized using
General Procedure 1 and General Procedure 2 described in detail in the
Examples below.
Detailed protocols from which the individual compounds described above can be
synthesized
are also provided in the Examples. The compounds of formula IV, where L is SO
or S02 can
be synthesised by oxidizing the corresponsing S compound in an inert solvent.
The inert
solvent may be dichloromethane, methanol, tetrahydrofuran, ether, hexane,
toluene,
cyclohexane, or the like, and mixtures thereof. The oxidizing agent may be m-
chloroperbenzoic acid, hydrogen peroxide, or the like. The reaction is carried
out at a
temperature in the range of about -78 °C to the boiling point of the
solvent, preferably from
about 0 °C to about 30 °C for about 0.5 to about 12 hours.
IV. Anti-Tumor Activity of the Telomerase Inhibitors of the Invention
The compounds of the present invention demonstrate inhibitory activity against
telomerase activity in vivo, as has been and can be demonstrated as described
below. The in
vitro activities of the compounds of the invention can also be demonstrated
using the methods
described herein. As used herein, the term "ex vivo " refers to tests
performed using living cells
in tissue culture.
One method used to identify compounds of the invention that inhibit telomerase
activity
involves placing cells, tissues, or preferably a cellular extract or other
preparation containing
telomerase in contact with several known concentrations of a test compound in
a buffer
compatible with telomerase activity. The level of telomerase activity for each
concentration of
test compound is measured and the ICsa (the concentration of the test compound
at which the
observed activity for a sample preparation was observed to fall one-half of
its original or a
control value) for the compound is determined using standard techniques. Other
methods for
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determining the inhibitory concentration of a compound of the invention
against telomerase can
be employed as will be apparent to those of skill in the art based on the
disclosure herein.
With the above-described methods, ICso values for several of the compounds of
the
present invention were determined, and found to be below 100 pM.
With respect to the treatment of malignant diseases using the compounds
described
herein, compounds of the present invention are expected to induce crisis in
telomerase-positive
cell lines. Treatment of telomerase-positive cell lines, such as HEK-293 and
HeLa cells, with a
compound of the invention is also expected to induce a reduction of telomere
length in the
treated cells.
Compounds of the invention are also expected to induce telomere reduction
during cell
division in human tumor cell lines, such as the ovarian tumor cell lines OVCAR-
5 and SK-OV-
3. Importantly, however, in normal human cells used as a control, such as BJ
cells of fibroblast
origin, the observed reduction in telomere length is expected to be no
different from cells
treated with a control substance, e.g., dimethyl sulfoxide (DMSO). The
compounds of the
invention also are expected to demonstrate no significant cytotoxic effects at
concentrations
below about 5 ~M in the normal cells.
In addition, the specificity of the compounds of the present invention for
telomerase can
be determined by comparing their activity (ICso) with respect to telomerase to
other enzymes
having similar nucleic acid binding or modifying activity similar to
telomerase in vitro. Such
enzymes include DNA Polymerase I, HeLa RNA Polymerase II, T3 RNA Polymerase,
MMLV
Reverse Transcriptase, Topoisomerase I, Topoisomerase II, Terminal Transferase
and Single-
Stranded DNA Binding Protein (SSB). Compounds having lower ICSO values for
telomerase as
compared to the ICSO values toward the other enzymes being screened are said
to possess
specificity for telomerase.
In vivo testing can also be performed using a mouse xenograft model, for
example, in
which OVCAR-5 tumor cells are grafted onto nude mice, in which mice treated
with a
compound of the invention are expected to have tumor masses that, on average,
may increase
for a period following the initial dosing, but will begin to shrink in mass
with continuing
treatment. In contrast, mice treated with a control (e.g., DMSO) are expected
to have tumor
masses that continue to increase.
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From the foregoing those skilled in the art will appreciate that the present
invention also
provides methods for selecting treatment regimens involving administration of
a compound of
the invention. For such purposes, it may be helpful to perform a terminal
restriction fragment
(TRF) analysis in which DNA from tumor cells is analyzed by digestion with
restriction
enzymes specific for sequences other than the telomeric (T2 AG3)N sequence.
Following
digestion of the DNA, gel electrophoresis is performed to separate the
restriction fragments
according to size. The separated fragments are then probed with nucleic acid
probes specific
for telomeric sequences to determine the lengths of the terminal fragments
containing the
telomere DNA of the cells in the sample. By measuring the length of telomeric
DNA, one can
estimate how long a telomerase inhibitor should be administered and whether
other methods of
therapy (e.g., surgery, chemotherapy and/or radiation) should also be
employed. In addition,
during treatment, one can test cells to determine whether a decrease in
telomere length over
progressive cell divisions is occurring to demonstrate treatment efficacy.
V. Telomerase Inhibiting Compositions and Methods for Treating Diseases
The present invention also provides pharmaceutical compositions for inhibiting
cell
proliferation of telomerase positive cells, and treating cancer and other
conditions in which
inhibition of telomerase is an effective therapy. These compositions include a
therapeutically
effective amount of a telomerase inhibiting compound of the invention in a
pharmaceutically
acceptable carrier or salt.
In one embodiment, the present invention provides methods, compounds and
compositions for inhibiting a telomerase enzyme, inhibiting proliferation of
telomerase postive
cells, and for treating cancer in a mammal. The compositions of the invention
include a
therapeutically effective amount of a compound of formulas I to V (or a
pharmaceutically
acceptable salt thereof) in a pharmaceutically acceptable carrier. The
compounds and
compositions of the present invention may also be used for the treatment of
other telomerase
mediated conditions or diseases, such as, for example, other
hyperproliferative or autoimmune
disorders such as psoriasis, rheumatoid arthritis, immune system disorders
requiring immune
system suppression, immune system reactions to poison ivy or poison oak, and
the like.
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In addition, it will be appreciated that therapeutic benefits for treatment of
cancer can be
realized by combining a telomerase inhibitor of the invention with other anti-
cancer agents,
including other inhibitors of telomerase such as described in U.S. Patent Nos.
5,656,638,
5,760,062, 5,767,278, 5,770,613 and 5,863,936. 'The choice of such
combinations will depend
on various factors including, but not limited to, the type of disease, the age
and general health
of the patient, the aggressiveness of disease progression, the TRF length and
telomerase
activity of the diseased cells to be treated and the ability of the patient to
tolerate the agents that
comprise the combination. For example, in cases where tumor progression has
reached an
advanced state, it may be advisable to combine a telomerase inhibiting
compound of the
invention with other agents and therapeutic regimens that are effective at
reducing tumor size
(e.g. radiation, surgery, chemotherapy and/or hormonal treatments). In
addition, in some cases
it may be advisable to combine a telomerase inhibiting agent of the invention
with one or more
agents that treat the side effects of a disease, e.g., an analgesic, or agents
effective to stimulate
the patient's own immune response (e.g., colony stimulating factor).
In one such method, a pharmaceutical formulation comprises a telomerase
inhibitor of
the invention with an anti-angiogenesis agent, such as fumagillin, fumagillin
derivatives, or
AGM-1470. The latter compound is available from Takeda Chemical Industries,
Ltd., while
the former compounds are described in Ingber, et al., 6 Dec. 1990, "Synthetic
analogues of
fumagillin that inhibit angiogenesis and suppress tumor growth", Nature
348:555-557. Other
combinations may include, but are not limited to, a telomerase inhibitor of
the invention in
addition to one or more antineoplastic agents or adjuncts (e.g., folinic acid
or mesna).
Antineoplastic agents suitable for combination with the compounds of the
present
invention include, but are not limited to, alkylating agents including alkyl
sulfonates such as
busulfan, improsulfan and piposulfan; aziridines, such as a benzodizepa,
carboquone,
meturedepa and uredepa; ethylenimines and methylmelamines such as altretamine,
triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide
and
trimethylolmelamine; nitrogen mustards such as chlorambucil, chlornaphazine,
cyclophosphamide, estramustine, iphosphamide, mechlorethamine, mechlorethamine
oxide
hydrochloride, melphalan, novembichine, phenesterine, prednimustine,
trofosfamide, and uracil
mustard; nitroso areas, such as carmustine, chlorozotocin, fotemustine,
lomustine, nimustine
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and ranimustine. Additional agents include dacarbazine, mannomustine,
mitobronitol,
mitolactol and pipobroman. Still other classes of relevant agents include
antibiotics, hormonal
antineoplastics and antimetabolites. Yet other combinations will be apparent
to those of skill in
the art.
Additional agents suitable for combination with the compounds of the present
invention
include protein synthesis inhibitors such as abrin, aurintricarboxylic acid,
chloramphenicol,
colicin E3, cycloheximide, diphtheria toxin, edeine A, emetine, erythromycin,
ethionine,
fluoride, S-fluorotryptophan, fusidic acid, guanylyl methylene diphosphonate
and guanylyl
imidodiphosphate, kanamycin, kasugamycin, kirromycin, and O-methyl threonine.
Additional
protein synthesis inhibitors include modeccin, neomycin, norvaline,
pactamycin,
paromomycine, puromycin, ricin, = sarcin, shiga toxin, showdomycin,
sparsomycin,
spectinomycin, streptomycin, tetracycline, thiostrepton and trimethoprim.
Inhibitors of DNA
synthesis, including alkylating agents such as dimethyl sulfate, mitomycin C,
nitrogen and
sulfur mustards, MNNG and NMS; intercalating agents such as acridine dyes,
actinomycins,
adriamycin, anthracenes, benzopyrene, ethidium bromide, propidium diiodide-
intertwining, and
agents such as distamycin and netropsin, can also be combined with compounds
of the present
invention in pharmaceutical compositions. DNA base analogs such as acyclovir,
adenine ~3-1-
D-arabinoside, amethopterin, aminopterin, 2-aminopurine, aphidicolin, 8-
azaguanine,
azaserine, 6-azauracil, 2'-azido-2'-deoxynucleosides, 5-bromodeoxycytidine,
cytosine ~3-1-D-
arabinoside, diazooxynorleucine, dideoxynucleosides, 5-fluorodeoxycytidine,
S-fluorodeoxyuridine, 5-fluorouracil, hydroxyurea and 6-mercaptopurine also
can be used in
combination therapies with the compounds of the invention. Topoisomerase
inhibitors, such as
coumermycin, nalidixic acid, novobiocin and oxolinic acid, inhibitors of cell
division,
including colcemide, colchicine, vinblastine and vincristine; and RNA
synthesis inhibitors
including actinomycin D, a-amanitine and other fungal amatoxins, cordycepin
(3'-
deoxyadenosine), dichlororibofuranosyl benzimidazole, rifampicine,
streptovaricin and
streptolydigin also can be combined with the compounds of the invention to
provide
pharmaceutical compositions.
In another embodiment, the present invention includes compounds and
compositions in
which a telomerase inhibitor is either combined with or covalently bound to a
cytotoxic agent
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bound to a targeting agent, such as a monoclonal antibody (e.g., a marine or
humanized
monoclonal antibody). It will be appreciated that the latter combination may
allow the
introduction of cytotoxic agents into cancer cells with greater specificity.
Thus, the active form
of the cytotoxic agent (i.e., the free form) will be present only in cells
targeted by the antibody.
Of course, the telomerase inhibitors of the invention may also be combined
with monoclonal
antibodies that have therapeutic activity against cancer.
In addition to the application of the telomerase inhibitors of the present
invention to the
treatment of mammalian diseases characterized by telomerase activity,
telomerase inhibitors
such as those disclosed herein, can be applied to agricultural phytopathogenic
organisms that
are characterized by telomerase activity. These organisms include nematodes
such as
Ceanorhabditis elegans, in which telomerase activity has been found, and in
fungi which are
expected to have telomerase activity based on the determination that the DNA
of the fungus
Ustilago maydis exhibits telomeres having the tandem TTAGGG repeats that are
maintained by
telomerase. Also, protozoans have TTAGGG telomeres and cause human disease.
The
1 S telomerase-inhibiting compounds of the invention can be administered to
plants and soil
infected with phytopathogenic organisms having telomerase activity alone, or
in combination
with other telomerase-inhibiting agents and/or other agents used to control
plant diseases. The
determination of the compositions used to control such phytopathogenic
organisms and the
appropriate modes of delivering such compositions will be known to those
having skill in the
agricultural arts.
The determination that nematodes, protozoans and possibly fungi have
telomerase
activity also indicates that the telomerase inhibitors provided by the present
invention can be
used to treat nematode infections in humans and animals of veterinary interest
such as dogs and
cats. Nematode infection in humans and animals often is in the form of
hookworm or
roundworm infection and leads to a host of deadly secondary illnesses such as
meningitis,
myocarditis, and various neurological diseases. Thus, it will be appreciated
that administration
of the telomerase-inhibiting compounds such as those of the invention, alone,
or in combination
with other telomerase-inhibiting agents and/or other therapeutic agents, can
be used to control
nematode, protozoan and fungal infections in humans and animals.
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In general, a suitable effective dose of a compound of the invention will be
in the range
of 0.001 to 1000 milligram (mg) per kilogram (kg) of body weight of the
recipient per day,
preferably in the range of 0.001 to 100 mg per kg of body weight per day, more
preferably
between about 0. l and 100 mg per kg of body weight per day and still more
preferably in the
range of between 0.1 to 10 mg per kg of body weight per day. The desired
dosage is preferably
presented in one, two, three, four, or more subdoses administered at
appropriate intervals
throughout the day, or by the action of a continuous pump. These subdoses can
be
administered as unit dosage form, for example, containing 5 to 10,000 mg,
preferably 10 to
1000 mg of active ingredient per unit dosage from. Preferably, the dosage is
presented once
per day at a dosing at least equal to TID, or is administered using a
continuous pump delivery
system.
The composition used in these therapies can be in a variety of forms. These
include, for
example, solid, semi-solid, and liquid dosage forms, such as tablets, pills,
powders, liquid
solutions or suspensions, liposomes, and injectable and infusible solutions.
The preferred form
depends on the intended mode of administration and therapeutic application.
The compositions
also preferably include conventional pharmaceutically acceptable carriers and
adjuvants, as is
well known to those of skill in the art. See, e.g., REMINGTON'S PHARMACEUTICAL
SCIENCES, Mack Publishing Co.: Easton, Pa., 17th Ed. (1985). Preferably,
administration
will be by oral or parenteral (including subcutaneous, intramuscular,
intravenous, and
intradennal) routes. More preferably, the route of administration will be
oral. The therapeutic
methods and agents of this invention can of course be used concomitantly or in
combination
with other methods and agents for treating a particular disease or disease
condition.
While it is possible to administer the active ingredient of this invention
alone, it is
preferable to present a therapeutic agent as part of a pharmaceutical
formulation or
composition. The formulations of the present invention comprise at least one
telomerase
activity-inhibiting compound of this invention in a therapeutically or
pharmaceutically effective
dose together with one or more pharmaceutically or therapeutically acceptable
carriers and
optionally other therapeutic ingredients. Various considerations for preparing
such
formulations are described, e.g., in Gilman et al. (eds.) GOODMAN AND
GILMAN'S: THE
PHARMACOLOGICAL BASES OF THERAPEUTICS, 8th Ed., Pergamon Press ( 1990); and
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REMINGTON'S supra. Methods for administration are discussed therein, e.g., for
oral,
intravenous, intraperitoneal, intramuscular, and other forms of
administration. Typically,
methods for administering pharmaceutical compositions will be either topical,
parenteral, or
oral administration methods for prophylactic and/or therapeutic treatment.
Oral administration
is preferred. The pharmaceutical compositions can be administered in a variety
of unit dosage
forms depending upon the method of administration. As noted above, unit dosage
forms
suitable for oral administration include powders, tablets, pills, and
capsules.
One can use topical administration to deliver a compound of the invention by
percutaneous passage of the drug into the systemic circulation of the patient.
The skin sites
include anatomic regions for transdermally administering the drug, such as the
forearm,
abdomen, chest, back, buttock, and mastoidal area. The compound is
administered to the skin
by placing on the skin either a topical formulation comprising the compound or
a transdermal
drug delivery device that administers the compound. In either embodiment, the
delivery
vehicle is designed, shaped, sized, and adapted for easy placement and
comfortable retention on
the skin.
A variety of transdermal drug delivery devices can be employed with the
compounds of
this invention. For example, a simple adhesive patch comprising a backing
material and an
acrylate adhesive can be prepared. The drug and any penetration enhancer can
be formulated
into the adhesive casting solution. The adhesive casting solution can be cast
directly onto the
backing material or can be applied to the skin to form an adherent coating.
See, e.g., U.S. Pat.
Nos. 4,310,509; 4,560,555; and 4,542,012.
In other embodiments, the compound of the invention will be delivered using a
liquid
reservoir system drug delivery device. These systems typically comprise a
backing material, a
membrane, an acrylate based adhesive, and a release liner. The membrane is
sealed to the
backing to form a reservoir. The drug or compound and any vehicles, enhancers,
stabilizers,
gelling agents, and the like are then incorporated into the reservoir. See,
e.g., U.S. Pat. Nos.
4,597,961; 4,485,097; 4,608,249; 4,505,891; 3,843,480; 3,948,254; 3,948,262;
3,053,255; and
3,993,073.
Matrix patches comprising a backing, a drug/penetration enhancer matrix, a
membrane,
and an adhesive can also be employed to deliver a compound of the invention
transdermally.
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The matrix material typically will comprise a polyurethane foam. The drug, any
enhancers,
vehicles, stabilizers, and the like are combined with the foam precursors. The
foam is allowed
to cure to produce a tacky, elastomeric matrix which can be directly affixed
to the backing
material. See, e.g., U.S. Pat. Nos. 4,542,013; 4,460,562; 4,466,953;
4,482,534; and 4,533,540.
Also included within the invention are preparations for topical application to
the skin
comprising a compound of the invention, typically in concentrations in the
range from about
0.001 % to 10%, together with a non-toxic, pharmaceutically acceptable topical
carrier. These
topical preparations can be prepared by combining an active ingredient
according to this
invention with conventional pharmaceutical diluents and carriers commonly used
in topical dry,
liquid, and cream formulations. Ointment and creams may, for example, be
formulated with an
aqueous or oily base with the addition of suitable thickening and/or gelling
agents. Such bases
may include water and/or an oil, such as liquid paraffin or a vegetable oil,
such as peanut oil or
castor oil. Thickening agents that may be used according to the nature of the
base include soft
para~n, aluminum stearate, cetostearyl alcohol, propylene glycol, polyethylene
glycols,
woolfat, hydrogenated lanolin, beeswax, and the like.
Lotions may be formulated with an aqueous or oily base and will, in general,
also
include one or more of the following: stabilizing agents, emulsifying agents,
dispersing agents,
suspending agents, thickening agents, coloring agents, perfumes, and the like.
Powders may be
formed with the aid of any suitable powder base, e.g., talc, lactose, starch,
and the like. Drops
may be formulated with an aqueous base or non-aqueous base also comprising one
or more
dispersing agents, suspending agents, solubilizing agents, and the like.
Topical administration
of compounds of the invention may also be preferred for treating diseases such
as skin cancer
and fungal infections of the skin (pathogenic fungi typically express
telomerase activity).
The topical pharmaceutical compositions according to this invention may also
include
one or more preservatives or bacteriostatic agents, e.g., methyl
hydroxybenzoate, propyl
hydroxybenzoate, chlorocreosol, benzalkonium chlorides, and the like. The
topical
pharmaceutical compositions also can contain other active ingredients such as
antimicrobial
agents, particularly antibiotics, anesthetics, analgesics, and antipruritic
agents.
The compounds of the present invention can also be delivered through mucosal
membranes. Transmucosal (i.e., sublingual, buccal, and vaginal) drug delivery
provides for an
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efficient entry of active substances to systemic circulation and reduces
immediate metabolism
by the liver and intestinal wall flora. Transmucosal drug dosage forms (e.g.,
tablet,
suppository, ointment, pessary, membrane, and powder) are typically held in
contact with the
mucosal membrane and disintegrate and/or dissolve rapidly to allow immediate
systemic
S absorption. Note that certain such routes may be used even where the patient
is unable to
ingest a treatment composition orally. Note also that where delivery of a
telomerase inhibitor
of the invention would be enhanced, one can select a composition for delivery
to a mucosal
membrane, e.g., in cases of colon cancer one can use a suppository to deliver
the telomerase
inhibitor.
For delivery to the buccal or sublingual membranes, typically an oral
formulation, such
as a lozenge, tablet, or capsule, will be used. The method of manufacture of
these formulations
is known in the art, including, but not limited to, the addition of the
pharmacological agent to a
pre-manufactured tablet; cold compression of an inert filler, a binder, and
either a
pharmacological agent or a substance containing the agent (as described in
U.S. Pat. No.
4,806,356); and encapsulation. Another oral formulation is one that can be
applied with an
adhesive, such as the cellulose derivative hydroxypropyl cellulose, to the
oral mucosa, for
example as described in U.S. Pat. No. 4,940,587. This buccal adhesive
formulation, when
applied to the buccal mucosa, allows for controlled release of the
pharmacological agent into
the mouth and through the buccal mucosa.
Parenteral administration is generally characterized by injection, either
subcutaneously,
intramuscularly, or intravenously. Thus, this invention provides compositions
for intravenous
administration that comprise a solution of a compound of the invention
dissolved or suspended
in an acceptable carrier. Injectables can be prepared in conventional forms,
either as liquid
solutions or suspensions, solid forms suitable for solution or suspension in
liquid prior to
injection, or as emulsions. Suitable excipients are, for example, water,
buffered water, saline,
dextrose, glycerol, ethanol, or the like. These compositions will be
sterilized by conventional,
well known sterilization techniques, such as sterile filtration. The resulting
solutions can be
packaged for use as is or lyophilized, the lyophilized preparation being
combined with a sterile
solution prior to administration. In addition, if desired, the pharmaceutical
compositions to be
administered may also contain minor amounts of non-toxic auxiliary substances,
such as
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wetting or emulsifying agents, pH buffering agents and the like, such as for
example, sodium
acetate, sorbitan monolaurate, triethanolamine oleate, etc. Such formulations
will be useful in
treating ovarian cancers.
Another method of parenteral administration employs the implantation of a slow-
release
or sustained-release system, such that a constant level of dosage is
maintained. See, e.g., U.S.
Pat. No. 3,710,795.
Liquid pharmaceutically administrable compositions can, for example, be
prepared by
dissolving, dispersing, etc., an active compound as defined above and optional
pharmaceutical
adjuvants in an excipient, such as, for example, water, saline, aqueous
dextrose, glycerol,
ethanol, olive oil, and other lipophilic solvents, and the like, to form a
solution or suspension.
If desired, the pharmaceutical composition to be administered may also contain
minor amounts
of nontoxic auxiliary substances, such as wetting or emulsifying agents, pH
buffering agents,
and the like, for example, sodium acetate, sorbitan monolaurate,
triethanolamine sodium
acetate, triethanolamine oleate, etc. Actual methods of preparing such dosage
forms are known
and will be apparent to those skilled in this art; for example, see
REMINGTON'S
PHARMACEUTICAL SCIENCES, supra. The composition or formulation to be
administered
will contain an effective amount of an active compound of the invention.
For solid compositions, conventional nontoxic solid carriers can be used and
include,
for example, pharmaceutical grades of mannitol, lactose, starch, magnesium
stearate, sodium
saccharin, talcum, cellulose, glucose, sucrose, magnesium carbonate, and the
like. For oral
administration, a pharmaceutically acceptable nontoxic composition is formed
by incorporating
any of the normally employed excipients, such as those carriers previously
listed, and generally
0.1-95% of active ingredient, preferably about 20%.
The compositions containing the compounds of the invention can be administered
for
prophylactic and/or therapeutic treatments. In therapeutic applications,
compositions are
administered to a patient already suffering from a disease, as described
above, in an amount
sufficient to cure or at least partially arrest the symptoms of the disease
and its complications.
An amount adequate to accomplish this is defined as a "therapeutically
effective amount or
dose." Amounts effective for this use will depend on the severity of the
disease and the weight
and general state of the patient.
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In addition to internal (in vivo) administration, the compounds and
compositions of the
invention may be applied ex vivo to achieve therapeutic effects, as for
example, in the case of a
patient suffering from leukemia. In such an application, cells to be treated,
e.g., blood or bone
marrow cells, are removed from a patient and treated with a pharmaceutically
effective amount
of a compound of the invention. The cells are returned to the patient
following treatment. Such
a procedure can allow for exposure of cells to concentrations of therapeutic
agent for longer
periods or at higher concentrations than otherwise available.
Once improvement of the patient's conditions has occurred, as, for example, by
the
occurrence of remission in the case of a cancer patient, a maintenance dose is
administered if
necessary. Subsequently, the dosage or the frequency of administration, or
both, can be
reduced, as a function of the systems, to a level at which the improved
condition is retained.
When the symptoms have been alleviated to the desired level, treatment can
cease. Patients
can, however, require additional treatment upon any recurrence of the disease
symptoms.
In prophylactic applications (e.g. chemoprevention), compositions containing
the
compounds of the invention are administered to a patient susceptible to or
otherwise at risk of a
particular disease. Such an amount is defined to be a "prophylactically
effective amount or
dose." In this use, the precise amounts again depend on the patient's state of
health and weight.
As will be apparent to those of skill in the art upon reading of this
disclosure, the
present invention provides valuable reagents relating to human and mammalian
telomerase.
The above description of necessity provides a limited and merely illustrative
sampling of
specific compounds, and should not be construed as limiting the scope of the
invention. Other
features and advantages of the invention will be apparent from the following
examples and
claims.
EXAMPLES
The following examples describe specific aspects of the invention to
illustrate the
invention and also provide a description of methods that can be used to
identify and test
compounds that inhibit the activity of telomerase to aid those of skill in the
art in understanding
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and practicing the invention. The examples should not be construed as limiting
the invention in
any manner.
In the following Examples 1 - 24 the following general procedure was employed:
General Procedure 1: Counline 2 4-thiazolidinedione ITZD) to aldehvde
A solution of appropriately substituted aldehyde (1 eq.), 2,4-
thiazolidinedione (1.5 eq.)
and piperidine (1.5 eq.) in EtOH was heated to 90°C for 2-16 h. The
resulting solution was
acidified with aqueous 1N HCI. The resulting solid was either filtered and
washed with water
and/or ether to afford pure product. Alternatively, the acidified solution was
extracted with
chloroform or ethyl acetate, organic phase washed with water, dried over
NaZS04 and
concentrated under reduced pressure to yield crude product as a solid that was
purified either
by column chromatography or recrystallization from appropriate solvent system.
Reactions were generally run on a 0.5 mmolar scale.
Example 1
Preparation of 5-(2-(3,4-Dichlorophenyl)benzylidene)thiazolidine-2,4-dione
CI
Step A: Preparation of aldehyde
To a solution of 2-bromobenzaldehyde (1 eq.) and 3,4-dichlorophenylboronic
acid (1.2
eq.) in acetonitrile was added K2C03 (1.5 eq.) followed by addition of
Pd(PPh3)4 (cat.). The
reaction was heated to 75°C with stirring for 16 h at which time the
reaction was diluted with
EtOAc, washed with water, dried over NaZS04 and concentrated under reduced
pressure to
yield crude aldehyde that was purified by column chromatography.
Sten B. General Procedure 1
General Procedure 1 was then followed to obtain 5-(2-(3,4-Dichlorophenyl)-
benzylidene)thiazolidine-2,4-dione.
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NMR (DMSO-d6, 8): 7.70 (d, 1 H), 7.76 (d, 1 H), 7.59-7.42 (m, SH), 7.28 (dd, 1
H)
MS(ESI) Calcd. 349. Found 348 (M-H)'.
Example 2
Preparation of S-(3-(3,4-Dichlorophenyl)benzylidene)thiazolidine-2,4-dione
w
H
(i
Sten A: Preparation of aldehyde
The requisite aldehyde was prepared from 3,-bromobenzaldehyde using the
procedure of
Example 1, Step A.
Step B. General Procedure 1
General Procedure 1 was then followed to obtain 5-(3-(3,4-Dichlorophenyl)-
benzylidene)-thiazolidine-2,4-dione.
NMR (DMSO-d6, 8): 7.98 (s, 1 H), 7.94 (s, 1 H), 7.84 (s, 1 H), 7.79 (d, 1 H),
7.74-7.66
(m, 2H), 7.63-7.53 (m, 2H)
MS(ESI) Cadcd. 349, found 348 (M-H)'.
Example 3
Preparation of 5-(4-(3,4-Dichlorobenzyloxy)benzylidene)thiazolidine-2,4-dione
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Step A: Preparation of aldehYde
S To a solution of 4-hydroxybenzaldehyde in acetonitrile was added KZC03 (1.5
eq.)
followed by addition of 3,4-dichlorobenzylchloride (3 eq.). The resulting
reaction mixture was
heated to 90 °C for 2-16 h at which time the precipitate was filtered
off: The filtrate was
diluted with EtOAc, washed with water, dried over NazS04 and concentrated
under reduced
pressure to provide crude product. This product was purified by
recrystallization from
CHZCIz/hexane solvent system to yield pure aldehyde.
Step B. General Procedure 1
General Procedure 1 was then followed to obtain 5-(4-(3,4-Dichlorobenzyloxy)-
benzylidene)thiazolidine-2,4-dione.
NMR (DMSO-d6, S): 7.69 (d, 2H), 7.62(d, 1 H), 7.52 (d, 2H), 7.41 (d, I H),
7.12 (d, 2H),
5.20 (s, 2H).
MS (ESI) Calcd. 379. Found 378 (M-H)-.
Example 4
Preparation of 5-(2-(3,4-Dichlorobenzyloxy)benzylidene)thiazolidine-2,4-dione
I
HN ~ I ~ / CI
O O
Step A: Preparation of alde~de
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The requisite aldehyde was prepared from 2-hydroxybenzaldehyde using the
procedure
of Example 3, Step A.
Step B. General Procedure 1
General Procedure 1 was then followed to obtain 5-(2-(3,4-Dichlorobenzyloxy)-
benzylidene)thiazolidine-2,4-dione.
Example 5
Preparation of 5-(4-(3,4-Dichlorobenzamido)benzylidene)thiazolidine-2,4-dione
I
CI
N
HN
O
O
Step A: Preparation of aldehyde
To a solution of 4-nitrobenzaldehyde (1 eq.) in trimethylorthoformate was
added PTSA
(cat.) and the resulting mixture was heated to reflux for 3-5 h at which time
the reaction
mixture was concentrated under reduced pressure. The residue was dissolved in
ether, washed
with NaHC03 followed by water, dried over Na2S04 and concentrated under
reduced pressure
to yield dimethyl acetal.
The crude dimethylacetal was dissolved in EtOH followed by addition of Raney
Ni
(cat). To this mixture at 0°C was added hydrazine hydrate (S eq) drop
wise over 15 min to
keep the effervescence under control. The reaction was warmed to room temp,
stirred for
additional 3 h followed by filtration of catalyst. The filtrate was
concentrated under reduced
pressure. The residue was redissolved in EtOAc, washed with water, dried over
Na2S04 and
concentrated under reduced pressure to yield crude 4-
aminobenzaldehydedimethylacetal.
To a solution of 4-aminobenzaldehydedimethylacetal in CH2C12 at 0°C was
added TEA
(2 eq.) followed by addition of 3,4-dichlorobenzoylchloride (1.5 eq). The
resulting mixture
was stirred for 2-16 h at room temperature at which time it was diluted with
water and EtOAc.
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The organic layer was separated, washed with water followed by concentration
under reduced
pressure. The residue was dissolved in CHCl3 followed by addition of 2N aq.
HCI. This
mixture was stirred for 1 h at which time the organic phase was separated,
washed with sat.
NaHC03, water, dried over Na2S04 followed by concentration under reduced
pressure to yield
crude aldehyde that was purified by either column chromatography or
recrystallization from
appropriate solvent system to yield pure 4-(3,4-
Dichlorobenzamido)benzaldehyde.
Step B. General Procedure 1
General Procedure 1 was then followed to obtain 5-(2-(3,4-Dichlorobenzyloxy)-
benzylidene)thiazolidine-2,4-dione.
NMR (DMSO-d6, 8): 12.50 (br s, 1 H), 10.60 (s, 1 H), 8.18 (d, 1 H), 7.90-7.86
(m, 3H),
7.78 (d, 1 H), 7.70 (s, 1 H), 7.56 (d, 1 H).
MS(ESI) Calcd. 392. Found 391 (M-H)'.
ExamnIe 6
Preparation of S-(4-(N-3,4-Dichlorophenyureido)benzylidene)thiazolidine-2,4-
dione
,~ b
" ~ ~~ o
/ - ~ ci
0
Step A: Preparation of aldehyde
To a solution of 4-nitrobenzaldehyde (1 eq.) in trimethylorthoformate was
added PTSA
(cat.) and the resulting mixture was heated to reflux for 3-5 h at which time
the reaction
mixture was concentrated under reduced pressure. The residue was dissolved in
ether, washed
with NaHC03 followed by water, dried over NaZS04 and concentrated under
reduced pressure
to yield dimethyl acetal.
The crude dimethyl acetal was dissolved in EtOH followed by addition of Raney
Ni
(cat). To this mixture at 0°C was added hydrazine hydrate (5 eq) drop
wise over 1 S min to
keep the effervescence under control. The reaction was warmed to room temp,
stirred for
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additional 3 h followed by filtration of catalyst. The filtrate was
concentrated under reduced
pressure. The residue was redissolved in EtOAc, washed with water, dried over
Na2S04 and
concentrated under reduced pressure to yield crude
aminobenzaldehydedimethylacetal.
To a solution of aminobenzaldehydedimethylacetal.(1 eq.) in acetonitrile was
added
solid 3,4-dichlorphenylisocynate (2 eq.). The resulting mixture was stirred
for 6-16 h at which
time the reaction was diluted with EtOAc, washed with water, dried over Na2S04
followed by
concentration under reduced pressure to yield crude urea. This urea was
dissolved in CHZCIz
followed by addition of 50% aq. TFA. The resulting mixture was stirred for 2 h
at which time
the organic phase was separated , washed with sat aq. NaHC03 and water, arid
dried over
Na2S04 followed by concentration under reduced pressure to yield crude
product.
Step B. General Procedure
General Procedure I was then followed to obtain 5-(4-(N-3,4-
Dichlorophenyureido)-
benzylidene)thiazolidine-2,4-dione.
NMR (DMSO-db, 8): 9.40 (d, 2H), 8.58 (s, IH), 7.84 (d, IH), 7.76 (d, 2H), 7.54
(d, 2H),
7.48 (d, 1 H), 7.30 (dd, 1 H).
MS(ESI) Calcd. 407. Found 406 (M-H)-.
Example 7
Preparation of 5-(2-(N-3,4-Dichlorophenyureido)benzylidene)thiazolidine-2,4-
dione
H
~S
N w I .-
C HN~N ~ CI
CI
Step A: Preparation of aldehyde
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The requisite aldehyde was prepared from 2-nitrobenzaldehyde using the
procedure of
Example 6, Step A.
Step B. General Procedure 1
General Procedure 1 was then followed to obtain 5-(2-(N-3,4-
Dichlorophenyureido)-
benzylidene)thiazolidine-2,4-dione.
NMR (DMS O-d6, 8): 10.10 (Br s, 1 H), 7.70 (d, 1 H), 7.62 (d, 1 H), 7.42 (dd,
1 H), 7.26-
7.22 (m, 2H), 6.94 (t, 1 H), 6.90 (d, 1 H), 6. 70 (d, 1 H), 5.86 (d, 1 H).
Example 8
Preparation of S-(2-(N-3,4-Dichlorophenylcarbamido)benzylidene)thiazolidine-
2,4-dione
HN
i ~. CI
~N \
H
Sten A: Preuaration of 5-(2-carboxvbenzylidene)thiazolidine 2 4 dione
5-(2-carboxybenzylidene)thiazolidine-2,4-dione was prepared by the method of
General
Procedure 1 from 2-carboxybenzaldehyde.
Sten B. Elaboration of carboy group
The 5-(2-carboxybenzylidene)thiazolidine-2,4-dione was dissolved in SOZCl2
followed
by addition of 1-2 drops of DMF. This resulting mixture was heated to ~80
°C for 15-30 min at
which time the reaction was concentrated under reduced pressure. The residue
was dissolved
in THF and added drop wise to a solution of 3,4-dichloroaniline (1.5 eq) and
TEA (2 eq). This
mixture was stirred for additional 1-2 h at which time the solid in the
reaction mixture was
filtered. The filtrate was concentrated under reduced pressure to yield solid
that was washed
with water and ether to afford pure product.
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NMR (DMSO-d6, 8): 10.80 (Br s, 1 H), 8.06 (d, 1 H), 7.93 (s, 1 H), 7.?3-7.54
(m, 6H).
MS(ESI) Calcd. 392. Found 391 (M-H)'
Ex~le 9
Preparation of 5-(3-(N-3,4-Dichlorophenylcarbamido)benzylidene)thiazolidine-
2,4-dione
w
HN
N ~ CI
C
CI
5-(3-(N-3,4-Dichlorophenylcarbamido)benzylidene)thiazolidine-2,4-dione was
prepared
in two steps by the method of Example 8 from 3-carboxybenzaldehyde.
NMR (DMSO-d6,8): 8.12 (d, 1 H), 8.08 (s, 1 H), 7.97 (d, 1 H), 7.82 (s, 1 H),
7.77 (d, 1 H),
7.72-7.64 (m, 2H), 7.60 (d, 2H).
MS(ESI) Calcd. 392. Found 391 (M-H)'.
Example 10
Preparation of 5-(4-(N -3,4-Dichlorophenylcarbamido)benzylidene)thiazolidine-
2,4-dione
S-(4-(N -3,4-Dichlorophenylcarbamido)benzylidene)-thiazolidine-2,4-dione was
prepared by the method of Example 8 from 4-carboxybenzaldehyde.
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NMR (DMSO-d6, 8): 10.60 (Br s, 1 H), 8.12 (s, I H), 8.02 (d, 2H), 7.82 (s, 1
H), 7.71 (d,
3H), 7.58 (d, 1H).
MS(ESI) Calcd. 392. Found 391 (M-H)-.
Example 11
Preparation of 5-(4-(N-3,4-Dichlorophenylcarbamoyloxy)benzylidene)thiazolidine-
2,4-dione
w 0 ~ ~ CI
0
o - - cl
Step A: Preparation of S-(4-hvdroxybenzvlidene)thiazolidine 2 4 dione
5-(4-hydroxybenzylidene)thiazolidine-2,4-dione was prepared by the method of
General
Procedure 1 from 4-hydroxybenzaldehyde.
Step B. Elaboration of hvdroxv ~roun
To a solution of 5-(4-hydroxybenzylidene)thiazolidine-2,4-dione ( 1 eq.) in
acetonitrile
was added K2C03 (xs) followed by addition of solid 3,4-dichlorphenylisocyanate
(2 eq.). The
resulting mixture was stirred for 6-16 h at which time the solid was filtered,
washed with water
to yield pure product.
NMR (DMSO-db, 8): 12.60 (Br s, 1H), 10.60 (br s, 1H), 7.76 (d, 2H), 7.62 (d,
2H), 7.56
dd, 1 H), 7.41 (dd, 1 H), 7.3 7 (dd, 1 H).
MS(ESI) Calcd. 408. Found 407 (M-H)~.
Example 12
Preparation of 5-(4-(3,4-Dichlorophenoxycarbonyl)benzylidene)thiazolidine-2,4-
dione
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5-(4-Carboxybenzylidene)thiazolidine-2,4-dione (from Example 8, Step A) was
dissolved in S02C12 followed by addition of 1-2 drops of DMF. This resulting
mixture was
heated to ~80 °C for 15-30 min at which time the reaction was
concentrated under reduced
pressure. The residue was dissolved in THF and added drop wise to a solution
of 3,4-
dichlorophenol (1.5 ec~ and TEA (2 e~. This resulting mixture was stirred for
additional 1-2 h
at which time the solid in the reaction mixture was filtered off. The filtrate
was concentrated
under reduced pressure to yield solid that was washed with water and ether to
afford pure ester.
Alternatively, to a solution of 5-(4-carboxybenzylidene)thiazolidine-2,4-dione
(I ec~ and 3,4-
dichlorophenol (1 e~ in CH2C12 was added DCC (1 ec~. The resulting reaction
mixture was
stirred for 16 h at which time the reaction was filtered, the filtrate was
washed with water and
concentrated under reduced pressure to provide crude product that was purified
by
recystallization.
NMR (DMSO-db, 8): 8.18 (d, 2H), 7.80 (d, 2H), 7.77-7.68 (m, 3H), 7.38-7.33 (m,
1H).
MS(ESI) Calcd. 393. Found 392 (M-H)'
Example I3
Preparation of 5-(2-(3,4-Dichlorophenoxycarbonyl)benzylidene)thiazolidine-2,4-
dione
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I
HN CI
o ~ ~I
0 0
PCT/US00/18211
5-(2-(3,4-Dichlorophenoxycarbonyl)benzylidene)thiazolidine-2,4-dione was
prepared
by the method of Example 12.
NMR (DMSO-db, S): 8.28 (s, 1 H), 8.22 (d, 1 H), 7.80 (t, 1 H), 7.76 (d, 1 H),
7.73 (d, 1 H),
7.64 (d, 1 H), 7.3 8 (dd, 1 H).
MS(ESI) Calcd. 393. Found 392 (M-H)'.
Example 14
Preparation of 5-(2-(3,4-Dichlorophenylacetoxy)benzylidene)thiazolidine-2,4-
dione
H
CI
C ~CI
Steu A: Preparation of 5-(2-hydroxybenzvlidenelthiazolidine 2 4 dione
5-(2-Hydroxybenzylidene)thiazolidine-2,4-dione was prepared by General
Procedure
from 2-hydroxybenzaldehyde.
Step B. Elaboration of h droxy groin
3,4-Dichlorophenylacetic acid was dissolved in S02C12 followed by addition of
a few
drops of DMF. This resulting mixture was heated to 80 °C for 15-30 min.
followed by
concentration of the reaction mixture under reduced pressure. The residue was
dissolved in
THF and slowly added to a solution of 5-(hydroxybenzylidene)thiazolidine-2,4-
dione (1.5 eq.}
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and TEA (1.5 eq.) in THF. The resulting reaction was stirred for 1-2 h at
which time the solid
was filtered off and filtrate was concentrated under reduced pressure to yield
light yellow solid.
The solid was dissolved in EtOAc followed by washing with sat. aq. K2CO3. The
organic
phase was separated, and dried over Na2S04 followed by concentration under
reduced pressure
to yield pure compound.
NMR (DMSO-db, b): 12.65 (Br s, 1 H), 7.65 (d, 1 H), 7.57 (d, 1 H), 7.55 (s, 1
H), 7.54-7.6
(m, 2H), 7.41 (d, 1 H), 7.37 (dd, 1 H), 7.30 (d, 1 H), 4.00 (s, 2H).
MS(ESI) Calcd. 407. Found 406 (M-H)'.
Example 1 S
Preparation of 5-(3-(3,4-Dichlorophenylacetoxy)benzylidene)thiazolidine-2,4-
dione
CI
" ~
i O ~ CI
O
5-(3-(3,4-Dichlorophenylacetoxy)benzylidene)thiazolidine-2,4-dione was
prepared by
the method of Example 14 from 3-hydroxybenzaldehyde.
NMR (DMSO-db, 8): 12.60 (Br s, 1H), 7.74 (s, 1H), 7.66 (d, 1H), 7.59 (d, 1H),
7.53 (t,
1 H), 7.45 (d, 1 H), 7.38-7.33 (m, 2H), 7.24 (d, 1 H), 4.00 (s, 2H).
MS(ESI) Calcd. 407. Found 406 (M-H)-
Example 16
Preparation of 5-(4-(3,4-Dichlorophenylacetoxy)benzylidene)thiazolidine-2,4-
dione
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CI
H o
CI
O
5-(4-(3,4-Dichlorophenylacetoxy)benzylidene)thiazolidine-2,4-dione was
prepared by
the method of Example 14 from 4-hydroxybenzaldehyde.
NMR (DMSO-d6, S): 7.76 (s, 1H), 7.66 (d, 1H), 7.60 (m, 3H), 7.36 (dd, 1H),
7.28 (d,
2H), 4.00 (s, 2H).
MS(ESI): Calcd. 407. Found 406 (M-H)'
Example 17
Preparation of 5-(2-(3,4-Dichlorobenzoyloxy)benzylidene)thiazolidine-2,4-dione
I
H
i ~ CI
d o ~I
0
SteQ A: Preparation of 5-(2-hvdroxvbenz~lidene~iazolidine 2 4 dione
5-(2-Hydroxybenzylidene)thiazolidine-2,4-dione was prepared by General
Procedure 1
from 2-hydroxybenzaldehyde.
Step B. Acylation of the hydroxy ~roun
A solution of 3,4-dichlorobenzoylchloride in THF was slowly added to a
solution of 5
(hydroxybenzylidene)thiazolidine-2,4-dione (1.5 eq.) and TEA (1.5 eq.) in THF.
The resulting
reaction was stirred for 1-2 h at which time the solid was filtered and
filtrate was concentrated
under reduced pressure to yield light yellow solid. The solid was dissolved in
EtOAc followed
by washing with sat. aq. KZC03. The organic phase was separated, and dried
over Na2S04
followed by concentration under reduced pressure to yield pure compound.
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NMR (DMSO-d6, 8): 8.30 (s, I H), 8.09 (dd, 1 H), 7.90 (d, I H), 7.68-7.62 (m,
1 H), 7.42-
7.28 (m, 3H), 7.18 (s, 1H).
MS (ESI): Calcd. 393. Found 392 (M-H)'
Example 18
Preparation of 5-(3-(3,4-Dichlorobenezoyloxy)benzylidene)thiazolidine-2,4-
dione
w
HN
~ CI
r
CI
5-(3-(3,4-Dichlorobenezoyloxy)benzylidene)thiazolidine-2,4-dione was prepared
by the
method of Example 17 from 3-hydroxybenzaldehyde.
NMR (DMSO-d6, S): 8.27 (s, IH), 8.06 (d, 1H), 7.86 (d, IH), 7.52-7.38 (m, 3H),
7.24
(s, 1 H), 7.20 (d, 1 H).
MS(ESI): Calcd. 393. Found 392 (M-H)'.
Example 19
Preparation of 5-(4-(3,4-Dichlorobenzoyloxy)benzylidene)thiazolidine-2,4-dione
5-(4-(3,4-Dichlorobenzoyloxy)benzylidene)thiazolidine-2,4-dione was prepared
by the
method of Example 17 from 4-hydroxybenzaldehyde.
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NMR (DMSO-D6, 8): 8.26 (d, 1H), 8.04 (dd, 1H), 7.85 (d, 1H), 7.58 (d, 2H),
7.36 (d,
2H), 7.39-7.34 (m, 3H).
MS(ESI): Calcd. 393. Found 392 (M-H)'.
Ex~le 20
Preparation of 5-( 3,4-Bis-(3,4-dichlorobenzyloxy)benzylidine)thiazolidine-2,4-
dione
CI
i
CI
H
O CI
CI
Step A: Preparation of aldehvde
3,4-Dichlorobenzyl chloride (41 S wl, 3 mmol) was added to a mixture of 138 mg
(1
mmol) of 3,4-dihydroxy benzaldehyde and 690 mg of potassium carbonate in DMF.
The
resulting mixture was warmed to 70° C and stirred overnight. The
reaction was then diluted
with 20 mL of water and the mixture was filtered. The resulting white
precipitate was collected
by filtration and air dried to give 426 mg (93%) of the desired product.
1 H NMR (partial) (400 MHz, DMSO-d6) 8 9.9 (s, 1 H), 5.24 (s, 2 H), 5.19 (s, 2
H).
Step B. General Procedure 1
NMR (400MHz, DMSO-d6, 8): 7.68-7.66 (m, 2H), 7.65 (s, 1 H), 7.63-7.59 (m, 2H),
7.41-7.37 (m, 2H), 7.21-7.20 (m, 1H), 7.18-7.15 (m, 2H).
MS(ESI):. Calcd. 553 Found 552 (M-H)'.
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Example 2I
Preparation of 5-(2-(3,4-Dichlorophenoxy)benzylidine)thiazolidine-2,4-dione
Step A: Preparation of aldehyde
A mixture of 2-fluorobenzaldehyde (248.23 mg 210 wL, 2 mmol) and 3,4-
dichlorophenol was stirred with potassium carbonate in 5 mL of
dimethylacetamide at 90°C for
12 hours. The reaction was diluted with 20 mL of water and extracted with 25
mL of ethyl
acetate. The organic layer was washed with saturated aqueous sodium
bicarbonate and
saturated sodium chloride solutions then dried over sodium sulfate and
concentrated in vacuo to
give a brown oil that was taken on without further purification.
Step B. General Procedure 1
NMR (DMSO-d6, 8): 7.73 (s, 1 H), 7.59 (m, 2 H), 7.46 (t, 1 H), 7.34 (m, 2H),
7.02 (m,
2H).
MS(ESI): Calcd. 365. Found 364(M-H)'
Example 22
Preparation of 5-(4-(3,4-Dichlorophenoxy)benzylidine)thiazolidine-2,4-dione
O ~ CI
HN
,- C.
5-(4-(3,4-Dichlorophenoxy)benzylidine)thiazolidine-2,4-dione was prepared by
the
method of 5-(2-(3,4-dichlorophenoxy)benzylidine)thiazolidine-2,4-dione,
(Example 21 ).
MS(ESI): Calcd. 365. Found 364 (M-H)'.
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Example 23
PCT/US00/18211
Preparation of 5-(2,5-Bis-(3,4-dichlorobenzyloxy)benzylidine)thiazolidine-2,4-
dione
5-(2,5-Bis-(3,4-dichlorobenzyloxy)benzylidine)thiazolidine-2,4-dione was
prepared by
the method of 5-(3,4-Bis-(3,4-dichlorobenzyloxy)benzylidine)thiazolidine-2,4-
dione, (Example
20).
NMR (DMSO-d6, 8): 7.88 (s, 1H), 7.70-7.66 (m, 2H), 7.64-7.60 (m, 2H), 7.42-
7.34 (m,
2H), 7.12-7. I 0 (m, 2H), 6.90 (brs, 1 H).
MS(ESI): Calcd. 553. Found 552 (M-H)-.
Example 24
Preparation of 5-(2,4-Bis-(3,4-dichlorobenzyloxy)benzylidine)thiazolidine-2,4-
dione
I ~ CI
CI
O
w I~
0
CI
CI
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v
CI
CI
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5-(2,4-Bis-(3,4-dichlorobenzyloxy)benzylidine)thiazolidine-2,4-dione was
prepared by
the method of 5-(3,4-Bis-(3,4-dichlorobenzyloxy)benzylidine)thiazolidine-2,4-
dione, (Example
20).
MS(ESI): Calcd. 553. Found 552 (M-H)'
In the following Examples 25 - 28, the following General Procedure 2 was
employed:
General Procedure 2~ Counlin~ rhodanine to aldehyde
A solution of appropriately substituted aldehyde ( I eq.), rhodanine ( 1 eq.)
and
ethylenediamine diacetate (I eq.) in methanol was heated to reflux for 1-3 h.
The resulting
precipitate was isolated and washed with methanol, water, 10% aqueous sodium
bisulfate,
saturated aqueous sodium bicarbonate and water and then air dried.
Reactions were generally run on a 0.1 mmolar scale.
Example 25
Preparation of 5-(2-(3,4-Dichlorobenzylthio)-3H-pyrimidin-4-on-6-
ylmethylidene)rhodanine
S
NH
H O ~ N~S I ~ CI
CI
Step A: Preparation of aldehyde
To a suspension of 0.75 g (3.7 mmol) of 6-dimethoxymethyl-2-mercapto-3H-
pyrimidin-
4-one in DMF with 0.66 g of potassium carbonate was added 0.512 mL (3.7 mmol)
of 3,4-
dichlorobenzyl chloride. The suspension was allowed to stand for 2 days. The
mixture was
diluted with 40 mL of ethyl acetate and 40 mL of 10% aqueous sodium bisulfate.
The
precipitate was isolated by filtration and washed with water to give I .0 g of
pure acetal.
A solution of 0.8 g of 6-dimethoxymethyl-2-(3,4-dichlorobenzylthio)-3H-
pyrimidin-4-
one in 70% trifluoroacetic acid in water was allowed to stir for 12 hours. The
solution was
neutralized with solid sodium bicarbonate and extracted with ethyl acetate.
The organic layer
was dried over magnesium sulfate and concentrated. Trituration of the residue
with I :1
ether:hexane provided 600 mg of a pure product.
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Step B. General Procedure 2
PCT/US00/18211
General Procedure 2 was followed to obtain 5-(2-(3,4-
dichlorobenzylthio)pyrimidin-4-
on-6-ylmethylidene)rhodanine.
Example 26
Preparation of 5-(2-(3,4-Dichlorobenzylthio)pyrimidin-4-
ylmethylidene)rhodanine
~N
H
N"S ~ CI
O
CI
Step A' Preparation of aldehvde
A suspension of I .66 g (7.98 mmol) of 4-dimethoxymethylpyrimidine-2-thione
sodium
salt, 2.7g of potassium carbonate and a,3,4-trichlorotoluene was stirred for 2
days. The
mixture was poured into water and extracted with ethyl acetate. The organic
layer was washed
with saturated aqueous sodium chloride solution then dried (MgS04) and
concentrated to give
6S g of product mercapto acetal.
A suspension of 0.8 g of the acetal in 5 mL of concentrated hydrochloric acid
was
refluxed for approximately S minutes until the solution became clear. The
solution was
allowed to cool then diluted with water, neutralized with saturated aqueous
sodium bicarbonate
solution and extracted with ethyl acetate. The organic layer was dried
(anhydrous sodium
sulfate) and concentrated to afford 100 mg of the desired aldehyde.
Step B. General Procedure 2
General Procedure 2 was followed to obtain 5-(2-(3,4-
Dichlorobenzylthio)pyrimidin-4-
ylmethylidene)rhodanine.
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Example 27
Preparation of 5-(2-(3,4-Dichlorobenzylthio)pyrimidin-4-
ylmethylidene)rhodanine
H
O \ ~S \ CI
N
CI
To a stirred suspension of 5-(2-(3,4-dichlorobenzylthio)pyrimidin-4-
ylmethylidene)-
rhodanine (Example 25, 0.3 mmol) in toluene (5 mL) was added diethyl 2,6-
dimethyl-1,4-
dihydro-3,5-pyridine dicarboxylate (109 mg, 0.39 mmol) and 0.3 g of activated
silica gel. The
mixture was heated to 80°C for 20 h then filtered while warm. The
filter cake was rinsed with
ethyl acetate and the combined filtrates were evaporated to dryness. The
residue was
redissolved in ethyl acetate and extracted with 1 N aqueous hydrochloric acid.
The organic
layer was dried (sodium sulfate) and concentrated to give 11 mg of pure
product.
Example 28
Preparation of 5-(3-Cyano-2-(3,4-dichlorobenzylthio)pyridin-6-
yhnethylidene)thiazolidine-2,4
dione
\ CN
HN ~~
\ N"S \ CI
O
~ CI
Step A: Preparation of aldehyde
A suspension of 0.2g ( 1 mmol) of 3-cyano-6-dimethoxymethyl-pyridine-2-thiol,
excess
potassium carbonate and a,3,4-trichlorotoluene (3mmo1) in acetonitrile was
heated at 75° for 10
minutes. Tlc indicated the reaction to be complete. The mixture was poured
into water and
extracted with ethyl acetate. The organic layer was dried (NazS04) and
concentrated to give
the product mercapto acetal as a solid, which was washed with hexane.
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The acetal was dissolved in chloroform (2m1) and 2 mL of 50% aqueous
trifluoroacetic
acid was added. After 16h, TLC indicated the reaction to be nearly complete.
The mixture was
evaporated to dryness and used immediately in the Step B (the NMR spectrum was
consistent
with proposed structure).
Step B. General Procedure 1
General Procedure 1 was followed to obtain 5-(3-Cyano-2-(3,4-
dichlorobenzylthio)-
pyridin-6-ylmethylidene)thiazolidine-2,4-dione.
NMR (DMSO-db, S): 8.12(d, 1H), 7.68(d, 1H), 7.53(d, 1H) 7.48(d, 1H) 7.38-
7.34(m,
1 H) 7.31 (s, 1 H) 4.80(s, 1 H).
Example 29
Preparation of S-(3-(3,4-Dichlorobenzyloxy)benzylidene)thiazolidine-2,4-dione
w
H
O ~ CI
O
CI
Sten A: Preparation of aldehyde
To a solution of 3-hydroxybenzaldehyde in acetonitrile was added KZC03 (1.5
eq.)
followed by addition of 3,4-dichlorobenzylchloride (3 eq.). The resulting
reaction mixture was
heated to 90°C for 2-16 h at which time the precipitate was filtered
off. The filtrate was diluted
with EtOAc, washed with water, dried over Na2S04 and concentrated under
reduced pressure to
provide crude product. This product was purified by recystallization from
CH2Clzlhexane
solvent system to yield pure aldehyde.
Step B. General Procedure 1
General Procedure 1 was followed to obtain 5-(3-(3,4-
Dichlorobenzyloxy}benzylidene)-
thiazolidine-2,4-dione.
NMR (DMSO-d6, 8): 7.69 (d, 2H), 7.61 (d, 1H), 7.43-7.31 (m, 3H), 7.12-7.08 (m,
2H),
6.96 (d, 1 H), 5.11 (s, 2H).
MS (ESl): Calcd. 378.98. Found 378 (M-H)'.
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Example 30
Preparation of Compound 1
2-(4-Methylphenylthio)-5-nitrobenzaldehyde (I.OOg, 3.66 mmol}, 2,4-
thiazolidinedione
( 1.72 g, 14.7 mmol) and piperidine (0.14 mL, 1. 5 mmol) were heated under
reflux for 26 hours
in ethanol (40 mL). The reaction solution was cooled down to room temperature,
and the thus
precipitated crystals were collected by filtration to give Compound 1 (577 mg,
42%).
'H NMR (300 MHz, DMSO-d6) 8 2.38 (s, 3H), 7.07 (d, J = 8.8 Hz, 1H), 7.37 (d, J
= 8.1 Hz, 2H), 7.49 (d, J = 7.9 Hz, 2H), 7.89 (s, I H), 8.14 (dd, J = 8.8, 2.2
Hz, 1 H),
8.22 (d, J = 2.2 Hz, 1 H), 12.8 (br s, 1 H}
FABMS m/z 373 (M+H)+ CI~H~zN2O4S2 = 372
Example 31
Preparation of Compound 2
Compound 1 (200 mg, 0.538 mmol) was dissolved in acetone (30 mL), and the
solution
was mixed with titanium trichloride (20% aqueous solution, 4 mL), followed by
stirring at
room temperature for 30 minutes. To the reaction solution was added an aqueous
saturated
sodium bicarbonate solution, and the mixture was extracted twice with ethyl
acetate. The
organic layer was washed with water and brine, and then dried over anhydrous
sodium sulfate.
The solvent was evaporated under reduced pressure, and the residue was
recrystallized from
ethyl acetate/hexane to give Compound 2 (69 mg, 38%).
'H NMR (300 MHz, CDC13) 82.21 (s, 3H} , 5.86 (br s, 2H),
6.70(dd,J=8.4,2.6 Hz, 1H),6.82(d,J=2.4 Hz,
1 H), 6.89 (d, J = 8.3 Hz, 2H), 7.06 (d, J = 8.1 Hz,
2H) , 7.3 3 (d, J = 8. 4 Hz, 1 H) , 8.06 (s, I H) , 12.5 (br s, I H)
FABMS m/z 342 (M+) C1~H~4N2O2S2 = 342.
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Example 32
Preparation of Compound 3
Compound 2 (20 mg, 0.058 mmol) was dissolved in dimethy 1 formamide ( 1 mL),
and to
the solution were added acetic anhydride ( 1 mL) and triethylamine (0.016 mL,
0.12 mmol),
followed by stirring at room temperature for 50 minutes. To the reaction
solution was added
water, and the mixture was extracted with ethyl acetate. The organic layer was
washed with
water and brine, and then dried over anhydrous sodium sulfate. The solvent was
evaporated
under reduced pressure, and the residue was purified by preparative thin layer
chromatography
(9/1 chloroform/methanol) to give Compound 3 (7.0 mg, 31%).
1H NMR (300 MHz, DMSO-d6) 52.08 (s, 3H), 2.25 (s, 3H),
7.07 (d, J = 8.3 Hz, 2H), 7.14 (d, J = 8.3 Hz, 2H), 7.45 (d, J = 8.6 Hz, 1 H),
7. S 8 (dd, J = 8.4, 2.2 Hz, 1 H), 8.08 (s, 1 H),
8.09 (d, J = 2.4 Hz, 1 H), 10.3 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 385 (M+H)+ C~gH,6N2O3S2 = 384.
Example 33
Preparation of Compound 4
Under ice-cooling, Compound 1 (100 mg, 0.269 mmol) was dissolved in a mixed
solvent of dichloromethane (20 mL} and methanol (4 mL), and the solution was
mixed with m-
chloroperbenzoic acid (50% purity, 100 mg, 0.289 mmol), followed by stirring
at room
temperature for 3 hours. The reaction solution was mixed with a 10% aqueous
sodium
hydrogen sulfite solution and extracted with chloroform. The organic layer was
washed with
an aqueous saturated sodium bicarbonate solution and brine, and then dried
over anhydrous
sodium sulfate. The solvent was evaporated under reduced pressure, and the
residue was
triturated with ethyl acetate to give Compound 4 (86 mg, 82%).
'H NMR (300 MHz, DMSO-d6) 52.30 (s, 3H), 7.32 (d, J
= 8.1 Hz, 2H), 7.48 (d, J = 8.3 Hz, 2H), 7.88 (s, 1H),
8.20 (d, J = 2.2 Hz, 1 H), 8.32 (d, J = 8.6 Hz, 1 H),
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8.50 (dd, J = 8.6, 2.2 Hz, I H), 12.6 (br s, 1 H)
FABMS m/z 387 (M-H)' C,7H,ZN205S2 = 388
Example 34
Preparation of Compound 5
Compound 1 (20 mg, 0.054 mmol) was dissolved in a mixed solvent of
dichlorornethane (5 mL) and methanol (1 mL), and the solution was mixed with
m-chloroperbenzoic acid (50% purity, 187 mg, 0.540 mmol), followed by stirnng
at room
temperature for 1. 5 hours. The reaction solution was mixed with a 10% aqueous
sodium
hydrogen sulf ite solution and extracted four times with chloroform-methanol
(9: I ). The
organic layer was washed with an aqueous saturated sodium bicarbonate solution
and brine,
and then dried over anhydrous sodium sulfate. The solvent was evaporated under
reduced
pressure, and the residue was purified by preparative thin layer
chromatography (6:1
chloroform/acetonitrile) to give Compound 5 ( 10 mg, 46%).
~H NMR (300 MHz, DMSO-d6) 82.36 (s, 3H) , 7.43 (d, J
= 8. 3 Hz, 2H) , 7. 74 (d, J = 8. 4 Hz, 2H) , 8. 06 (s, I H),
8.28 (br s, 1 H), 8.49 (s, 2H), 12.9 (br s, 1 H)
FABMS m/z 403 (M-H)' C,~H,ZNz06S2 = 404
Example 35
Preparation of Compound 6
2-(4-Chlorophenylthio)benzaldehyde (249 mg, 1.00 mmol), 2,4-thiazolidinedione
(176
mg, 1.50 mmol) and piperidine (0.10 mL, 1.0 mmol) were heated under reflux for
3 hours in
ethanol (8 mL). The reaction solution was cooled down to room temperature, and
water and 1
N HC 1 ( 1 mL) were added and the mixture was extracted with ethyl acetate.
The organic layer
was washed with brine, and then dried over anhydrous sodium sulfate. The
solvent was
evaporated under reduced pressure, and the residue was triturated with ethyl
acetate to give
Compound 6 (274 mg, 70%).
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'H NMR (300 MHz, DMSO-d6) 87.24 (d, J = 8.4 Hz, 2H),
7.42 (d, J = 8.6 Hz, 2H), 7.5-7.6 (m, 4H), 8.03 (s,
1 H), 12. 7 (br s, 1 H)
FABMS m/z 348 (M+H)+ C,6H~p35C1NO2S2 = 347
Example 36
Preparation of Compound 7
Under ice-cooling, Compound 6 (20 mg, 0.057 mmol) was suspended in
dichloromethane (5 mL), and the suspension was mixed with m-chloroperbenzoic
acid (50%
purity, 22 mg, 0.063 mmol), followed by stirring for 20 minutes. To the
reaction solution was
added a I O% aqueous sodium hydrogen sulfite solution, and the mixture was
extracted with
chlorofonmmethanol (9:I). The organic layer was washed with an aqueous sodium
bicarbonate
solution, water and brine, and then dried over anhydrous sodium sulfate. The
solvent was
evaporated under reduced pressure, and the residue was triturated with
diisopropyl ether to give
Compound 7 ( 15 mg, 72%).
'H NMR (300 MHz, DMSO-d6) 87.55 (d, J = 9.0 Hz, 2H),
7.59 (d, J = 9.0 Hz, 2H), 7.6-7.8 (m, 2H), 7.9-8.0
(m, 2H), 8.02 (s, 1 H), 12.7 (br s, I H)
FABMS m/z 364 (M+H)+ C16H1035C1NO3Sz = 363
Example 37
Preparation of Compound 8
Under ice-cooling, Compound 6 (20 mg, 0.057 mmol) was suspended in
dichloromethane (5 mL), and the suspension was mixed with m-chloroperbenzoic
acid (50%
purity, 200 mg, 0.57 mmol), followed by stirring for 3 hours. To the reaction
solution was
added a I O% aqueous sodium hydrogen sulfite solution, and the mixture was
extracted with
chloroformmethanol (9: I ). The organic layer was washed with an aqueous
saturated sodium
bicarbonate solution, water and brine, and then dried over anhydrous sodium
sulfate. The
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solvent was evaporated under reduced pressure, and the residue was triturated
with diisopropyl
ether to give Compound 8 ( 16 mg, 74%).
~H NMR (300 MHz, DMSO-db) 57.60 (d, J = 7.5 Hz, 1 H),
7.68 (d, J = 8.8 Hz, 2H), 7.7-7.8 (m, 1 H), 7.81 (d,
J = 8.8 Hz, 2H), 7.87 (td, J = 7.7, 1.3 Hz, 1 H), 8.11
(s, 1 H), 8.27 (dd, J = 7.9, 1.3 Hz, 1 H), 12.8 (br s,
1 H)
FABMS m/z 380 (M+H)+ Cl6Hio35C1NO4S2 = 379
Example 38
Preparation of Compound 9
4-(4-Methylphenylthio)-3-nitrobenzaldehyde (273mg, 1.00 mmol),
2,4-thiazolidinedione (176 mg, 1.50 mmol) and piperidine (0.40 mL, 0.40 mmol)
were heated
under reflux for 19 hours in ethanol (8 mL). The reaction solution was cooled
down to room
temperature, and the thus precipitated crystals were collected by filtration
to give Compound 9
(175 mg, 47%).
~H NMR (300 MHz, DMSO-d6) 82.41 (s, 3H) , 6.92 (d, J
= 8.6 Hz, 1 H), 7.40 (d, J = 7.9 Hz, 2H), 7.54 (d, J
= 8. 3 Hz, 2H) , 7.72 (dd, J = 8.6, 2.2 Hz, 1 H) , 7.73
(s, 1 H}, 8.46 (d, J = 1.8 Hz, 1 H), 12.7 (br s, 1 H)
FABMS m/z 373 (M+H)+ C,~H12N204S2 = 372
Example 39
Preparation of Compound 10
2-Phenoxybenzaldehyde (Synthesis, 28 (1995)) (198 mg, 1.00 mmol),
2,4-thiazolidinedione ( 176 mg, 1.50 mmol) and piperidine (0.10 mL, 1.0 mmol)
were heated
under reflux for 4 hours in ethanol (5 mL). The reaction solution was cooled
down'to room
temperature, water and 1 N HC 1 ( 1 mL) were added, and then the mixture was
extracted with
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ethyl acetate. The organic layer was washed with water and brine, and then
dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced pressure,
and the residue
was triturated with diisopropyl ether to give Compound 10 (199 mg, 67%).
'H NMR (300 MHz, DMSO-d6) 86.95 (d, J = 8.1 Hz, 1H) ,
7.05 (d, J = 7.5 Hz, 2H) , 7.20 (t, J = 7.3 Hz, 1 H) ,
7.32(t,J=7.7 Hz, 1H),7.4-7.5(m,3H),7.58
(dd, J = 7.7, 1.3 Hz, 1 H), 7.95 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 298 (M+H)+ C~6H,1NO3S = 297
Example 40
Preparation of Compound 11
3-Phenoxybenzaldehyde (0.172 mL, 1.00 mmol), 2,4-thiazolidinedione (176 mg,
1.50
mmol) and piperidine (0.10 mL, 1.0 mmol) were heated under reflux for 10 hours
in ethanol (5
mL). The reaction solution was cooled down to room temperature, water and 1 N
HC1 (1 mL)
were added, and then the mixture was extracted with ethyl acetate. The organic
layer was
washed with water and brine, and then dried over anhydrous sodium sulfate. The
solvent was
evaporated under reduced pressure, and the residue was triturated with
diisopropyl ether to give
Compound 1 I ( 141 mg, 47%).
'H NMR (300 MHz, DMSO-d6) 57.0-7.3 (m, SH) , 7.35 (d,
J = 7.9 Hz, 1 H) , 7.44 (dd, J = 8.4, 7.5 Hz, 2H) , 7.54
(t, J = 7.9 Hz, 1 H), 7.82 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 298 (M+H)+ C,6H"N03S = 297
Example 41
Preparation of Compound 12
3-(4-Methylphenoxy)benzaldehyde (0.193 mL, 1.00 mmol), 2,4-thiazolidinedione
(176
mg, 1.50 mmol) and piperidine (0.10 mL, 1.0 mmol) were heated under reflux for
7 hours in
ethanol (S mL). The reaction solution was cooled down to room temperature, and
water and 1
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N HCl (1 mL) were added, and then the mixture was extracted with ethyl
acetate. The organic
layer was washed with water and brine, and then dried over anhydrous sodium
sulfate. The
solvent was evaporated under reduced pressure, and the residue was
recrystallized from
ethanoUethyl acetate to give Compound 12 (98 mg, 32%).
'H NMR (300 MHz, DMSO-db) 82.31 (s, 3H) , 6.9-7.0 (m,
3H), 7.12 (t, J = 2.0 Hz, 1 H), 7.23 (d, J = 8.4 Hz,
2H), 7.29 (br d, J = 7.9 Hz, 1 H), 7.45 (t, J = 7.9
Hz, 1 H), 7.49 (s, 1 H)
FABMS m/z 312 (M+I-~+ C,~H,3N03S = 311
Example 42
Preparation of Compound 13
3-(3,4-Dichlorophenoxy)benzaldehyde (0.198 mL,1.00 mmol), 2,4-
thiazolidinedione
(176 mg, 1.50 mmol) and piperidine (0.10 mL, 1.0 mural) were heated under
reflux for 7 hours
in ethanol (5 mL). The reaction solution was cooled down to room temperature,
water and 1 N
HC 1 ( 1 mL) were added, and then the mixture was extracted with ethyl
acetate. The organic
layer was washed with water and brine, and then dried over anhydrous sodium
sulfate. The
solvent was evaporated under reduced pressure, and the residue was triturated
with ethyl
acetate to give Compound 13 (252 mg, 69%).
'H NMR (300 MHz, DMSO-db) 87.0-7.2 (m, 2H), 7.25 (t,
J = 2.0 Hz, 1 H), 7. 3 8 (d, J = 7. 7 Hz, 1 H), 7.3 8 (d,
J = 2.8 Hz, 1 H), 7.52 (t, J = 8.0 Hz, 1 H), 7.53 (s,
1 H), 7.66 (d, J = 8.8 Hz, 1 H)
FABMS m/z 366 (M+H)+ C,6H9C1N03S = 365
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Example 43
Preparation of Compound 14
4-Phenoxybenzaldehyde (0.175 mL, 1.00 mmol), 2,4-thiazolidinedione ( 176 mg,
1.50
mmol) and piperidine (0.10 mL, 1.0 mmol) were heated under reflux for 6 hours
in ethanol (5
mL). The reaction solution was cooled down to room temperature, and water and
1 N HC1 (1
mL) were added, and then the mixture was extracted with ethyl acetate. The
organic layer was
washed with water and brine, and then dried over anhydrous sodium sulfate. The
solvent was
evaporated under reduced pressure, and the residue was triturated with
hexane/diisopropyl ether
to give Compound 14 (252 mg, 85%).
~H NMR (300 MHz, DMSO-d6) 87.0-7.2 (m, 4H) , 7.23 (t,
J = 7.3 Hz, 1 H), 7.4-7.5 (m, 2H), 7.62 (d, J = 8.8
Hz, 2H), 7.78 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 298 (M+H)+ C,6H11N03S = 297
Example 44
Preparation of Compound 15
In an argon atmosphere, 4-fluorobenzaldehyde (0.53 mL, 5.0 mmol) and p-cresol
(648
mg, 6.0 mmol) were dissolved in dimethylacetamide (8 mL), and to the solution
were added
potassium carbonate (828 mg, 6.0 mmol) and cupric oxide (95 mg, 0.50 mmol) and
the mixture
was heated under reflux for 1.5 hours. The reaction solution was cooled down
to room
temperature, and water was added, and then the mixture was extracted with
ethyl acetate. The organic
layer was washed with a 0.5 N aqueous sodium hydroxide solution, water and
brine, and then
dried over anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure, and
the residue was purified by silica gel column chromatography (6:1 hexane/ethyl
acetate) to
obtain 4-(4methylphenoxy)benzaldehyde (697 mg, 66%).
'H NMR (300 MHz, CDC13) 82.37 (s, 3H) , 6.98 (d, J =
8.4 Hz, 2H) , 7.03 (d, J = 8.6 Hz, 2H) , 7.21 (d, J =
8.6 Hz, 2H) , 7.82 (d, J = 9.0 Hz, 2H) , 9. 91 (s, 1H)
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FABMS m/z 213 (M+H~ C,4H,z02 = 212
The thus obtained 4-(4-methylphenoxy)benzaldehyde (212 mg, 1.00 mmol),
2,4-thiazolidinedione (176 mg, 1.50 mmol) and piperidine (0.10 mL, 1.0 mmol)
were heated
under reflux for 4 hours in ethanol (5 mL). The reaction solution was cooled
down to room
temperature, and water and 1 N Hl (1 mL) were added, and then the mixture was
extracted
with ethyl acetate. The organic layer was washed with water and brine, and
then dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced pressure,
and the residue
was triturated with diisopropyl ether to give Compound 15 (252 mg, 81%).
~H NMR (300 MHz, DMSO-db) 52.31 (s, 3H) , 7. O1 (d, J
= 8.4 Hz, 2H) , 7.06 (d, J = 8.8 Hz, 2H) , 7.25 (d, J
= 8.6 Hz, 2H) , 7. 60 (d, J = 8.8 Hz, 2H) , 7. 76 (s, 1H) ,
12.6 (br s, 1 H)
FABMS m/z 312 (M+H~ C,~H,3N03S = 311.
Example 45
Preparation of Compound 16
(1) In an argon atmosphere, 5-nitrosalicylaldehyde (334 mg, 2.00 mmol) was
dissolved in dimethyformamide (5 mL), and the solution was mixed with benzyl
bromide
(0.238 mL, 2.00 mmol) and sodium hydride (88 mg, 2.4 mmol), followed by
stirnng at 70°C
for 13 hours. The reaction solution was cooled with ice, and water was added,
and then the
mixture was extracted with ethyl acetate. The organic layer was washed with
water and brine,
and then dried over anhydrous sodium sulfate. The solvent was evaporated under
reduced
pressure, and the residue was triturated with diisopropyl ether to obtain 2-
benzyloxy-5-
nitrobenzaldehyde (337 mg, 66%).
'H NMR (300 MHz, CDC,3) 85.33 (s, 2H) , 7.18 (d, J =
9.2 Hz, 1 H) , 7. 4-7. 5 (m, 5 H) , 8. 42 (dd, J =
9. 2, 2. 9 Hz, 1 H), 8.73 (d, J = 2.9 Hz, 1 H), 10.5 (s, 1 H)
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FABMS m/z 258 (M+H)+ C,4H"N04 = 257
(2) The obtained 2-benzyloxy-5-nitrobenzaldehyde (257 mg, 1.00 mmol),
2,4-thiazolidinedione ( 176 mg, 1.50 mmol) and piperidine (0.10 mL, 1.0 mmol}
were heated
under reflux for 11 hours in ethanol (5 mL). The reaction solution was cooled
down to room
temperature, and water and 1 N HCl ( 1 mL) were added, and then the mixture
was extracted
with ethyl acetate. The organic layer was washed with water and brine, and
then dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced pressure,
and the residue
was triturated with diisopropyl ether to give Compound 16 (211 mg, 59%).
~H NMR (300 MHz, DMSO-d6) 85.42 (s, 2H) , 7. 3-7. 6 (m,
6H), 7.91 (s, 1 H), 8.25 (d, J = 2.6 Hz, 1 H), 8.3 5 (dd,
J = 9.2, 2. 8 Hz, 1 H), 12. 7 (br s, 1 H)
FABMS m/z 357 (M+H)+ C,~H12IV2p5S = 356
Example 46
Preparation of Compound 17
( 1 ) In the same manner as described in Example 45 ( 1 ), 2-(3,4-
dichlorobenzyloxy)-
5-nitrobenzaldehyde (482 mg, 74%) was obtained from 5-nitrosalicylaldehyde
(334 mg, 2.00
mmol) and 3,4-dichlorobenzyl chloride (0.305 mL, 2.20 mmol) .
'H NMR (300 MHz, CDC13) 55.27 (s, 2H), 7.14 (d, J =
9.2 Hz, 1 H) , 7.30 (dd, J = 8.3, 2.2 Hz, 1 H) , 7.53 (d,
J = 8.3 Hz, 1 H) , 7.56 (d, J = 2.2 Hz, 1 H} , 8.43 (dd,
J = 9.2, 2.9 Hz, 1 H) , 8.74 (d, J = 2.9 Hz, 1 H) , 10. S
(s, 1 H)
(2) The thus obtained 2-(3,4-dichlorobenzyloxy)-5-nitrobenzaldehyde (326 mg,
1.00 mmol) was treated in the same manner as described in Example 45 (2) to
give Compound
17 (142 mg, 33%).
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'H NMR (300 MHz, DMSO-d6) 85. 42 (s, 2H) , 7.44 (d, J
= 9.4 Hz, 1 H), 7.49 (dd, J = 8.3, 2.0 Hz, 1 H), 7.72
(d, J = 8.3 Hz, 1 H), 7.80 (d, J = 1.8 Hz, 1 H), 7.90
(s, 1 H) , 8.26 (d, J = 2.6Hz 1 H) 8.36 (dd, J = 9.4,
2.8 Hz, 1 H), 12.8 (br s, 1 H)
FABMS m/z 425 (M+H)+ Ci~H,o3sC,2N205S = 424
Example 47
Preparation of Compound 18
( 1 ) In the same manner as described in Example 45 ( 1 ), 2-(4-
methylbenzyloxy)-
5-nitrobenzaldehyde (413 mg, 76%) was obtained from 5-nitrosalicylaldehyde
(334 mg, 2.00
mmol) and 4-methylbenzyl bromide (370 mg, 2.00 mmol).
'H NMR (300 MHz, CDC13) 82. 18 (s, 3H) , 5.28 (s, 2H) ,
7.18 (d, J = 9.4 Hz, 1 H), 7.24 (d, J = 8.1 Hz, 2H),
7.33 (d, J = 8.1 Hz, 2H), 8.40 (dd, J = 9.2, 2.9 Hz,
1 H), 8.72 (d, J = 2.9 Hz, 1 H), 10.5 (s, 1 H)
FABMS m/z272 (M+I-n+ C,sH,3N04 = 271
(2) The thus obtained 2-(4-methylbenzyloxy)-S-nitrobenzaldehyde (271 mg, 1.00
mmol) was treated in the same manner as described in Example 45 (2) to give
Compound 18
(200 mg, 54%).
'H NMR (300 MHz, DMSO-db) 82.32 (s, 3H) , 5.35 (s, 2H),
7.24 (d, J =7.9 Hz, 2H), 7.38 (d, J = 7.9 Hz, 2H),
7.47 (d, J = 9.3 Hz, 1 H), 7.88 (s, 1 H), 8.24 (d, J
= 2.8 Hz, 1 H) , 8.34 (dd, J = 9.2, 2.8 Hz, 1 H) , 12.7
(br s, 1 H)
FABMS m/z 371 (M+H)+ C,8H1aN2OsS = 370
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Example 48
Preparation of Compound 19
( 1 ) In the same manner as described in Example 45 ( 1 ), 3-(4-
methylbenzyloxy)-
4-nitrobenzaldehyde (315 mg, 58%) was obtained from 3-hydroxy-4-
nitrobenzaldehyde (334
mg, 2.00 mmol) and 4-methylbenzyl bromide (370 mg, 2.00 mmol).
1H NMR (300 MHz, CDC13) 82.36 (s, 3H) , 5.27 (s, 2H),
7. 20 (d, J = 7.9 Hz, 2H) , 7.34 (d, J = 8.1 Hz, 2H),
7.53 (dd, J = 8.1, 1.5 Hz, 1 H), 7.64 (d, J =.1.5 Hz,
1 H), 7.92 (d, J = 8.1 Hz, 1 H), 10.0 (s, 1 H)
FABMS m/z 272 (M+H)+ C,SH,3N04 = 271
(2) The obtained 3-(4-methylbenzyloxy)-4-nitrobenzaldehyde (271 mg, 1.00 mmol)
was treated in the same manner as described in Example 45 (2) and
recrystallized from ethyl
acetate/hexane to obtain Compound 19 (95 mg, 26%).
1H NMR (300 MHz, DMSO-d6) 82.31 (s, 3H), 5.32 (s, 2H) ,
7.22 (d, J =7.9 Hz, 2H) , 7.29 (dd, J = 8.4, 1.5 Hz, 1 H),
7.36 (d, J = 8.1 Hz, 2H), 7.64 (d, J = 1.7 Hz, 1 H), 7.81
(s, 1 H), 8.02 (d, J = 8.4 Hz, 1 H), 12.8 (br s, 1 H)
FABMS m/z 371 (M+H)+ C,gH,4NzO5S = 370
Example 49
Preparation of Compound 20
( 1 ) In the same manner as described in Example 45 ( 1 ), 4-(4-
methylbenzyloxy)-
3-nitrobenzaldehyde (365 mg, 67%) was obtained from 4-hydroxy-3-
nitrobenzaldehyde (334
mg, 2.00 mmol) and 4-methylbenzyl bromide (370 mg, 2.00 mmoI).
'H NMR (300 MHz, CDC13) 82. 36 (s, 3H) , 5.31 (s, 2H) ,
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7.21 (d, J = 8.1 Hz, 2H), 7.26 (d, J = 8.8 Hz, 1 H),
7.33 (d, J = 8.1 Hz, 2H) , 8.02 (dd, J = 8.6, 2.0 Hz,
1 H), 8.34 (d, J = 2.0 Hz, 1 H), 9.92 (s, 1 H)
FABMS m/z 272 (M+H)+ C~SH,3N04 = 271
(2) The thus obtained 4-(4-methylbenzyloxy)-3-nitrobenzaldehyde (271 mg, 1.00
mmol) was treated in the same manner as described in Example 45 (2) and
recrystallized from
ethyl acetate/hexane to obtain Compound 20 (123 mg, 33%).
'H NMR (300 MHz, DMSO-db) 82.31 (s, 3H) , 5.34 (s, 2H},
7. 22 (d, J = 7.9 Hz, 2H) , 7.35 (d, J = 8.1 Hz, 2H), 7.60
(d, J = 9. 0 Hz, 1 H) , 7. 81 (s, 1 H) , 7. 84 (dd, J = 9.0,
2.2 Hz, 1 H), 8.15 (d, J = 2.2 Hz, 1 H), 12.7 (br s, 1 H)
FABMS m/z 371 (M+H)+ C~8H,4NZOSS = 370
Example 50
Preparation of Compound 21
( 1 ) In the same manner as described in Example 45 ( 1 ), 5-(4-
methylbenzyloxy)-
2-nitrobenzaldehyde (413 mg, 76%) was obtained from 5-hydroxy-2-
nitrobenzaldehyde (334
mg, 2.00 mmol) and 4-methylbenzyl bromide (370 mg, 2.00 mmol).
'H NMR (300 MHz, CDC13) 82.37 (s, 3H) , 5.16 (s, 2H),
7. 19 (dd, J = 9.0, 2.9 Hz, 1 H) , 7.21 (d, J = 7.5 Hz, 2H),
7.31 (d, J = 8.1 Hz, 2H), 7.41 (d, J = 2.9 Hz, 1 H), 8.1 S
(d, J = 9.2 Hz, 1 H), 10.5 (s, 1 H)
FABMS m/z 272 (M+H)+ C,SH~3N04 = 271
(2) The thus obtained 5-(4-methylbenzyloxy)-2-nitrobenzaldehyde (271 mg, 1.00
mmol), 2,4-thiazolidinedione (176 mg, 1.50 mmol) and piperidine (0.10 mL, 1.0
mmol) were
heated under reflux for 1.5 hours in ethanol (8 mL). The reaction solution was
cooled down to
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room temperature, and water and 1 N HC 1 ( 1 mL) were added, and then the
mixture was
extracted with ethyl acetate. The organic layer was washed with water and
brine, and then
dried over anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure, and
the residue was purified silica gel column chromatography ( 19: I
chloroformlacetonitrile) and
preparative thin layer chromatography (10:1 chloroform/methanol) to give
Compound 21 (30
mg, 8.1 %).
~H NMR (300 MHz, CDC13) b2. 38 (s, 3H) , 5. 16 (s, 2H) ,
7.04 (d, J = 2.4 Hz, 1 H}, 7.09 (dd, J = 9.2, 2.6 Hz,
1 H), 7.22 (d, J = 7.9 Hz, I H), 7.30 (d, J = 7. 8 Hz,
2H), 8.22 (d, J = 9.2 Hz, I H), 8.24 (s, 1 H), 9.00 (br
SR 1 H)
FARMS m/z 371 (M+H)+ C,gH,4N205S = 370
Example 51
Preparation of Compound 22
In an argon atmosphere, salicylaldehyde (0.213 mL, 2.00 mmol) was dissolved in
dimethy 1 formamide (5 mL), and to the solution were added 4-methylbenzyl
bromide (370 mg,
2.00 mmol) and sodium hydride (88 mg, 2.4 mmol), followed by stirring at
70°C for 1.5 hours.
The reaction solution was cooled with ice, and water was added thereto, and
then the mixture
was extracted with ethyl acetate. The organic layer was washed with water and
brine, and then
dried over anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure.
2,4-Thiazolidinedione (176 mg, 1.50 mmol), piperidine (0.10 mL, 1.0 mmol) and
ethanol (5
mL) were added thereto, followed by heating under reflux for 1.5 hours. The
reaction solution
was cooled down to room temperature, and water and 1 N HC1 (1 mL) were added,
and then
the mixture was extracted with ethyl acetate. The organic layer was washed
with water and
brine, and then dried over anhydrous sodium sulfate. The solvent was
evaporated under
reduced pressure, and the residue was triturated with hexane/diisopropyl ether
to give
Compound 22 (542 mg, 83% by two steps).
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~H NMR (300 MHz, DMSO-d6) 82.31 (s, 3H) , 5.19 (s, 2H),
7.10 (t, J = 7.S Hz, 1H), 7.22 (d, J = 7.7 Hz, 2H),
7.24 (d, J = 7.S Hz, 1H), 7.35 (d, J = 8.1 Hz, 2H),
7.4-7. S (m, 2H), 8.01 (s, 1 H), 12.6 (br s, 1 H)
S FABMS m/z 326 (M+H)+ C,BH,SN03S = 32S
Example S2
Preparation of Compound 23
In an argon atmosphere, S-methoxysalicylaldehyde (0.249 mL, 2.00 mmol) was
dissolved in dimethy 1 formamide (S mL), and to the solution were added 4-
methylbenzyl
bromide (370 mg, 2.00 mmol) and sodium hydride (88 mg, 2.4 mmol), followed by
stirring at
70°C for 1.S hours. The reaction solution was cooled with ice, and
water was added, and then
the mixture was extracted with ethyl acetate. The organic layer was washed
with water and
brine, and then dried over anhydrous sodium sulfate. The solvent was
evaporated under
1S reduced pressure. 2,4-Thiazolidinedione {176 mg, 1.50 mmol), piperidine
(0.10 mL, 1.0
mmol) and ethanol (S mL) were added thereto, followed by heating under reflux
for 1.5 hours.
The reaction solution was cooled down to room temperature, and the thus
precipitated crystals
were collected by filtration to give Compound 23 (419 mg, S9% by two steps).
'H NMR (300 MHz, DMSO-d6) 82.31 (s, 3H), 3.75 (s, 3H} ,
S. 12 (s, 2H) , 6.90 (d, J = 2.9 Hz, 1H) , 7.OS (dd, J = 9.0, 2.9 Hz, 1H),
7.18 (d,
J = 9.0 Hz, 1H), 7.20 (d, J = 7.7 Hz, 2H) , 7.32 (d, J = 8.1 Hz, 2H) , 7.95
(s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 3S6 (M+H)+ C,9H1~N04S = 3SS
2S
Example S3
Preparation of Compound 24
In an argon atmosphere, S-chlorosalicylaldehyde (313 mg, 2.00 mmol} was
dissolved in
dimethy 1 forrnamide (S mL), and to the solution were added 4-methylbenzyl
bromide (370 mg,
2.00 mmol) and sodium hydride (88 mg, 2.4 mmol), followed by stirring at
70°C for O.S hour.
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The reaction solution was cooled with ice, and water was added, and then the
mixture was
extracted with ethyl acetate. The organic layer was washed with water and
brine, and then
dried over anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure.
2,4-Thiazolidinedione (176 mg, 1.50 mmol), piperidine (0.10 mL, 1.0 mmol) and
ethanol (15
mL) were added thereto, followed by heating under reflux for 4 hours. The
reaction solution
was cooled down to room temperature, and water and 1 N HC1 (1 mL) were added,
and then
the mixture was extracted with ethyl acetate. The organic layer was washed
with water and
brine, and then dried over anhydrous sodium sulfate. The solvent was
evaporated under
reduced pressure, and the residue was triturated with hexane/diisopropyl ether
to give
Compound 24 (428 mg, 60% by two steps).
'H NMR (300 MHz, DMSO-d6) 82.31 (s, 3H) , 5.19 (s, 2H),
7.21 (d, J = 8.3 Hz, 2H), 7.27 (d, J = 9.0 Hz, 1 H),
7.34 (d, J = 8.4 Hz, 2H), 7.36 (d, J = 2.9 Hz, 1 H),
7.51 (dd, J = 9.0, 2.8 Hz, 1 H), 7. 87 (s, 1 H), 12.6
(br s, 1 H)
FABMS m/z 360 (M+H)+ Ci8H,43sC1NO3S = 359
Example 54
Preparation of Compound 25
In an argon atmosphere, 5-bromosalicylaldehyde (1.00 g, 5.00 mmol) was
dissolved in
dimethylformamide (10 mL), and to the solution were added 4-methylbenzyl
bromide (925 mg,
5.00 mmol) and sodium hydride (220 mg, 5.50 mmol), followed by stirring at
70°C for 0.5
hour. The reaction solution was cooled with ice, and water was added, and then
the mixture
was extracted with ethyl acetate. The organic layer was washed with water and
brine, and then
dried over anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure.
2,4-Thiazolidinedione (702 mg, 6.00 mmol), piperidine (0.50 mL, 5.0 mmol) and
ethanol (40
mL) were added thereto, followed by heating under reflux for 4 hours. The
reaction solution
was cooled down to room temperature, and water and 1 N HC 1 (5 mL) were added,
and then
the mixture was extracted with ethyl acetate. The organic layer was washed
with water and
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brine, and then dried over anhydrous sodium sulfate. The solvent was
evaporated under
reduced pressure, and the residue was triturated with hexane/diisopropyl ether
to give
Compound 25 (1.27 mg, 63% by two steps).
'H NMR (300 MHz, DMSO-d6) 52.31 (s, 3H), 5.19 (s, 2H),
7.21 (d, J = 8.8 Hz, 3H), 7.34 (d, J = 8.0 Hz, 2H),
7.48 (d, J = 2.4 Hz, IH), 7.62 (dd, J = 8.8, 2.4 Hz,
1 H), 7.86 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 406, 404 (M+H)+ C,8H14'9BrN03S = 403
Example SS
Preparation of Compound 26
4-Diphenylaminobenzaldehyde (273 mg, 1.00 mmol), 2,4-thiazolidinedione (176
mg,
1.50 mmol) and piperidine (0.10 mL, 1.0 mmol) were heated under reflux for 4
hours in ethanol
(8 mL). The reaction solution was cooled down to room temperature, and water
and 1 N HC 1
( 1 mL) were added, and then the mixture was extracted with ethyl acetate. The
organic layer
was washed with water and brine, and then dried over anhydrous sodium sulfate.
The solvent
was evaporated under reduced pressure, and the residue was triturated with
ethyl acetate to give
Compound 26 (293 mg, 79%).
'H NMR (300 MHz, DMSO-db) 86.93 (d, J = 7.2 Hz, 2H) ,
7.1-7.2 (m, 6H), 7.3-7.5 (m, 6H), 7.67 (s, 1 H), 12.5 (br s, 1 H)
FABMS m/z 373 (M+H)+ C22H'6NZOZS = 372
Example 56
Preparation of Compound 27
2-Phenylbenzaldehyde (Tetrahedron Lett., 38(32):5575 (1997)) (273mg,
I.SOmmo1),
2,4-thiazolidinedione (263 mg, 2.25 mmol) and piperidine (0.15 mL, 1.5 mmol)
were heated
under reflux for 3 hours in ethanol (8 mL). The reaction solution was cooled
down to room
temperature, and water and 1 N HC 1 (2 mL) were added, and then the mixture
was extracted
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with ethyl acetate. The organic layer was washed with water and brine, and
then dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced pressure,
and the residue
was triturated with diisopropyl ether to give Compound 27 (344 mg, 82%).
~H NMR (300 MHz, DMSO-db) 57.3-7.4 (m, 2H), 7.4-7.7
(m, 8H), 12.6 (br s, 1 H)
FABMS m/z 282 (M+H)+ C,6H"NOZS = 281
Example 57
Preparation of Compound 28
3-Phenylbenzaldehyde (Tetrahedron Lett., 38(32):5575 (1997)) (269 mg, 1.48
mmol),
2,4-thiazolidinedione (259 mg, 2.22 mmol) and piperidine (0.15 mL, 1.5 mmol)
were heated
under reflux for 3 hours in ethanol (8 mL). The reaction solution was cooled
down to room
temperature, and water and 1 N HC 1 (2 mL) were added, and then the mixture
was extracted
1 S with ethyl acetate. The organic layer was washed with water and brine, and
then dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced pressure,
and the residue
was triturated with diisopropyl ether to give Compound 28 (355 mg, 85%).
'H NMR (300 MHz, DMSO-d6) 87.3-7.8 (m, 8H), 7.89 (s,
2H), 12.6 (br s, IH)
FABMS m/z 282 (M+H)+ C,6Hi iNO2S = 281
Example 58
Preparation of Compound 29
4-Phenylbenzaldehyde (182 mg, 1.00 mmol), 2,4-thiazolidinedione (176 mg, 1.5
mmol) and piperidine (0.10 mL, 1.0 mmol) were heated under reflux for 3 hours
in ethanol (6
mL). The reaction solution was cooled down to room temperature, and water and
1 N HC 1 ( 1
mL) were added, and then the mixture was extracted with ethyl acetate. The
organic layer was
washed with water and brine, and then dried over anhydrous sodium sulfate. The
solvent was
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evaporated under reduced pressure, and the residue was triturated with
diisopropyl ether to give
Compound 29 ( 187 mg, 67%).
'H NMR (300 MHz, DMSO-d6) 87.4-7.6 (m, 3H) , 7.70 (d,
J = 8.4 Hz, 2H), 7.75 (d, J = 7.2 Hz, 2H), 7.85 (s,
1 H), 7.86 (d, J = 8.1 Hz, 2H), 12.6 (br s, IH)
FABMS m/z 282 (M+H)+ Cl6Hi,N02S = 281
Example 59
Preparation of Compound 30
4-(a-Hydroxybenzyl)benzaldehyde (.l. Org. Chem., 62: 4643 (1997)) (1.35 g,
6.37
mmol), 2,4-thiazolidinedione (894 mg, 7.64 mmol) and piperidine (0.64 mL, 6.4
mmol) were
heated under reflux for 10 hours in ethanol (6 mL). The reaction solution was
cooled down to
room temperature, and water and 1 N HC1 (7 mL) were-added, and then the
mixture was
extracted with ethyl acetate. The organic layer was washed with water and
brine, and then dried
over anhydrous sodium sulf ate. The solvent was evaporated under reduced
pressure, and the
residue was triturated with diisopropyl ether to give Compound 30 (1.73 g,
87%).
1H NMR (300 MHz, DMSO-d6) 85.75 (d, J = 2.7 Hz, 1H) ,
6.03 (d, J = 2.7 Hz, 1 H), 7.21 (tt, J = 7.2, 1.5 Hz,
1 H), 7.31 (t, J= 7.2 Hz, 2H), 7.38 (d, J = 7.3 Hz,
2H), 7.53 (s, 4H), 7.75 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 312 (M+H)+ C,~H,3N03S = 311
Example 60
Preparation of Compound 31
Compound 30 (622 mg, 2.00 mmol) was dissolved in acetonitrile (80 mL), and to
the
solution was added manganese dioxide (2.61 g) and the mixture was heated under
reflux for 4.5
hours. The reaction solution was cooled down to room temperature and filtered
through celite,
and then the solvent was evaporated under reduced pressure. The residue was
purified by silica
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gel column chromatography (9:1 chlorofonm/acetonitrile) and triturated with
diisopropyl ether
to give Compound 31 (73 mg, 12%).
~H NMR (300 MHz, DMSO-d6) 87.58 (t, J = 7.5 Hz, 2H),
7.6-7.8 (m,'SH), 7.86 (d, J = 7.9 Hz, 2H), 7.87 (s, 1H), 12.7 (br s, 1H)
FABMS m/z 310 (M+H)+ C,~H, ~N03S = 309.
Exam lp a 61
Preparation of Compound 32
Compound 30 (115 mg, 0.39 mmol) was dissolved in dichloromethane (15 mL), and
to
the solution were added trifluoroacetic acid (0.30 mL, 3.9 mmol) and
triethylsilane (0.81 mL,
S.1 mmol) and the mixture was heated under reflux for 12 hours. The solvent
was evaporated
under reduced pressure, and the residue was recrystallized from acetone/hexane
to give
Compound 32 (70 mg, 61%).
'H NMR (300 MHz, DMSO-d6) 84.00 (s, 2H) , 7.1- 7.3 (m,
SH),7.39(d,J=8.1Hz,2H),7.52(d,J=8.lHz,
2H), 7.75 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 296 (M+H)+ Ci~H~3N02S = 295
Example 62
Preparation of Compound 33
4-Formyltrityl alcohol (J. Org. Chem., 63:9924 (1998)) (576 mg, 2.00 mmol),
2,4-thiazolidinedione (281 mg, 2.40 mmol) and piperidine (0.20 mL, 2.0 mmol)
were heated
under reflux for 9 hours in ethanol ( 15 mL). The reaction solution was cooled
down to room
temperature, and water and 1 N HC 1 (2 mL) were added, and then the mixture
was extracted
with ethyl acetate. The organic layer was washed with water and brine, and
then dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced pressure,
and the
residue was triturated with ethyl acetate/ diisopropyl ether to give Compound
33 (684 mg,
78%).
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'H NMR (300 MHz, DMSO-d6) 86.61 (s, 1H), 7.2-7.4 (m,
l OH) , 7.37 {d, J = 8 .4 Hz, 2H) , 7. 55 (d, J = 8. 4 Hz,
2H), 7.76 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 388 (M+H)+ C23H1~NO3S = 387
Example 63
Preparation of Compound 34
Compound 33 (219 mg, 0.566 mmol) was dissolved in dichloromethane (15 mL), and
to
the solution were added trifluoroacetic acid (0.385 mL, 0.50 mmol) and
triethylsilane (0.80 mL,
0.50 mmol), followed by stirring at room temperature for 10 minutes. The
solvent was
evaporated under reduced pressure, and the residue was triturated with hexane
to give
Compound 34 ( 198 mg, 94%).
'H NMR {300 MHz, DMSO-db) 85.70 (s, 1 H), 7.1-7.4 (m,
12H), 7.55 (d, J = 8.3 Hz, 2H}, 7.75 (s, 1 H), I 2.6 (br s, 1 H)
FABMS m/z 372 (M+H)+ C23H1~NOZS = 371
Example 64
Preparation of Compound 35
In an argon atmosphere, diphenylamine (338 mg, 2.00 mmol) and 4-bromobenzyl
bromide (500 mg, 2.00 mmol) were dissolved in dimethylformamide (8 mL), and to
the
solution was added sodium hydride (88 mg, 2.2 mmol) under ice -cooling,
followed by stirnng
at room temperature for 4 hours. To the reaction solution was added water, and
the mixture was
extracted with ethyl acetate. The organic layer was washed with water and
brine, and then dried
over anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure, and the
residue was purified by silica gel column chromatography ( 14:1
hexane/acetone) to obtain
N-(4-bromobenzyl)diphenylamine (478 mg, 71 %).
'H NMR (300 MHz, CDC13) 84.93 (s, 2H) , 6.94 (tt, J =
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7.3, 1.1 Hz, 2H), 7.03 (dd, J = 8.8, 1.1 Hz, 4H),
7.2-7.3 (m, 6H), 7.42 (d, J = 8.6 Hz, 2H)
FABMS m/z 339, 337 (M+) C,9H16~9BrN = 337
In an argon atmosphere, the thus obtained N-(4-bromobenzyl)diphenylamine (440
mg,
1.30 mmol) was dissolved in tetrahydrofuran (6 mL) and cooled to -78°C.
A 1.6 M n-butyl
lithium hexane solution (1.3 mL, 2.0 mmol) was added thereto, and, 5 minutes
thereafter,
dimethylformamide (0.20 mI,, 2.6 mmol) was further added thereto, followed by
stirring for 5
minutes. To the reaction solution was added water, and the mixture was
extracted with ethyl
acetate. The organic layer was washed with brine, and then dried over
anhydrous sodium
sulfate. The solvent was evaporated under reduced pressure, and the residue
was purified by
silica gel column chromatography (9:1 hexane/ethyl acetate) to obtain
4-(N,N-diphenylaminomethyl)benzaldehyde (213 mg, 57%).
' H NMR (300 MHz, CDC 13) 85.06 (s, 2H), 6.96 (t, J =
7.3 Hz, 2H), 7.03 (d, J = 8.6 Hz, 4H), 7.25 (dd, J =
8.6, 7.3 Hz, 4H), 7.52 (d, J = 8.4 Hz, 2H), 7.83
(d, J = 8.3 Hz, 2H), 9.97 (s, 1 H)
FABMS m/z 287 (M~ C2oH,~NO = 287
The obtained 4-(N,N-diphenylaminomethyl)benzaldehyde (198 mg, 0.690 mmol),
2,4-thiazolidinedione (117 mg, 1.10 mmol) and piperidine (0.068 mL, 0.69 mmol)
were heated
under reflux for S hours in ethanol (6 mL). The reaction solution was cooled
down to room
temperature, and the precipitated crystals were collected by filtration to
give Compound 35
(240 mg, 90%).
'H NMR (300 MHz, DMSO-d6) 85.06 (s, 2H) , 6.92 (t, J
= 7.2 Hz, 2H), 7.05 (d, J = 8.4 Hz, 4H), 7.25 (t, J
= 7.7 Hz, 4H), 7.48 (d, J = 8.3 Hz, 2H), 7.55 (d, J
= 8.4 Hz, 2H), 7.75 (s, 1 H), 12.6 (br s, 1 H)
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FABMS m/z 386 (M+) C23H,8N202S = 386
Example 65
Preparation of Compound 36
In an argon atmosphere, 4 -bromoaniline (344 mg, 2.00 mmol) was dissolved in
dimethylformamide (5 mL), and to the solution were added sodium hydride (200
mg, 5.00
mmol) and benzyl bromide (0.52 mL, 4.4 mmol) under ice-cooling, followed by
stirring at
room temperature for 11 hours. To the reaction solution was added water, and
the mixture was
extracted with ethyl acetate. The organic layer was washed with water and
brine, and then dried
over anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure, and the
residue was recrystallized from ethyl acetate/hexane to obtain 4-bromo-N,N-
dibenzylaniline
(442 mg, 63%).
~H NMR (300 MHz, CDC13) 84.63 (s, 4H) , 6.59 (d, J =9.0, 2H), 7.2-7.4 (m, 12H)
FABMS m/z 353, 351 (M+) C2oH,8~9BrN = 351
In an argon atmosphere, the obtained 4-bromo-N,N-dibenzylaniline (425 mg, 1.21
mmol) was dissolved in tetrahydrofuran (S mL) and cooled to -78"C. A 1.6 M n-
butyl lithium
hexane solution (1.1 mL, 1.8 mmol) was added thereto, and, 5 minutes
thereafter,
dimethylformamide (0.186 mL, 2.4 mmol) was further added thereto, followed by
stirring for 5
minutes: To the reaction solution was added water, and the mixture was
extracted with ethyl
acetate. The organic layer was washed with a saturated brine, and then dried
over anhydrous
sodium sulfate. The solvent was evaporated under reduced pressure, and the
residue was
purified by silica gel column chromatography (9:1 hexane/ethyl acetate) to
obtain
4-(dibenzylamino)benzaldehyde (192 mg, 53%).
iH NMR (300 MHz, CDC13) 84.75 (s, 4H) , 6.79 (d, J =9.0 Hz, 2H),
7.2 - 7.4 (m, 1 OH) , 7.69 (d, J = 9.0, 2H), 9.73 (s, 1 H)
FABMS m/z302 (M+H)+ C2,H,9N0 = 301
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The obtained 4-(dibenzylamino) benzaldehyde (181 mg, 0.601 mmol),
2,4-thiazolidinedione (105 mg, 0.900 mmol) and piperidine (0.059 mL, 0.60
mmol) were
heated under reflux for S hours in ethanol (7 mL). The reaction solution was
cooled down to
room temperature, and water and 1 N HC 1 (0.6 mL) were added, and then the
mixture was
extracted with ethyl acetate. The organic layer was washed with water and
brine, and then dried
over anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure, and the
residue was triturated with diisopropyl ether to give Compound 36 (207 mg,
86%).
'H NMR (300 MHz, DMSO-d6) 84.81 (s, 4H), 6.81 (d, J
= 8.8 Hz, 2H), 7.2-7.4 (m, 12H), 7.61 (s, 1 H), 12.3 (br s, 1 H)
FABMS m/z 400 (M+) CZqH20N202s = 400.
Example 66
Preparation of Compound 39
5-Nitro-2-[(4-trifluoromethyl)phenoxy]benzaldehyde (311 mg, 1. 00 mmol),
2,4-thiazolidinedione (234 mg, 2.00 mmol) and piperidine (0.10 mL, 1.0 mmol)
were heated
under reflux for 13 hours in ethanol (8 mL). The reaction solution was cooled
down to room
temperature, and water and 1 N HC 1 ( 1 mL) were added, and then the mixture
was extracted
with ethyl acetate. The organic layer was washed with water and brine, and
then dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced pressure,
and the residue
was purified by silica gel column chromatography (15:1
chloroform/acetonitrile), and then
triturated with diisopropyl ether to give Compound 39 (146 mg, 36%).
1H NMR (300 MHz, DMSO-d6) 87.16 (d, J = 9.2 Hz, 1H),
7. 45 (d, J = 8. 4 Hz, 2H), 7. 89 (d, J = 8. 4 Hz, 2H), 7.93 (s, 1 H),
8.30 (dd, J = 9.2, 2.8 Hz, 1 H), 8.39 (d, J = 2.2 Hz, 1 H), 12.8 (br s, 1 H)
FABMS m/z 411 (M+H)+ C,~H9F3N205S = 410.
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Example 6?
Preparation of Compound 40
In an argon atmosphere, 2-bromo-5-hydroxybenzaldehyde (201 mg, 1.00 mmol) was
dissolved in dimethylformamide (3 mL), and to the solution were added 4-
methylbenzyl
bromide (185 mg, 1.00 mmol} and sodium hydride (48 mg, 1.2 mmol), followed by
stirring at
70°C for 3 hours. The reaction solution was cooled with ice, and water
was added, and then the
mixture was extracted with ethyl acetate. The organic layer was washed with
water and brine,
and then dried over anhydrous sodium sulfate. The solvent was evaporated under
reduced
pressure. 2,4-Thiazolidinedione (176 mg, 1.5 mmol) , piperidine (0.10 mL, 1.0
mmol) and
ethanol (6 mL) were added thereto, followed by heating under reflux for 13
hours. The
reaction solution was cooled down to room temperature, and water and 1 N HC1
(5 mL) were
added, and then the mixture was extracted with ethyl acetate. The organic
layer was washed
with water and brine, and then dried over anhydrous sodium sulfate. The
solvent was
evaporated under reduced pressure, and the residue was purified by silica gel
column
chromatography (20:1 chloroform/acetonitrile) and triturated with diisopropyl
ether to give
Compound 40 (142 mg, 35% by two steps).
'H NMR (300 MHz, DMSO-db) 82.31 (s, 3H), 5.13 (s, 2H),
7.0-7.1 (m, 2H), 7.21 (d, J = 7.7 Hz, 2H), 7.34 (d, J = 7.7 Hz, 1 H),
7.69 (d, J = 8.6 Hz, 1 H), 7.79 (s, 1 H), 12.7 (br s, 1 H)
FABMS m/z 406, 404 (M+H)+ Ci8H14~9BrN03S = 403.
Example 68
Preparation of Compound 41
In an argon atmosphere, 2,5-dihydroxybenzaldehyde (138 mg, 1.00 mmol) was
dissolved in dimethy 1 formamide (5 mL), and to the solution were added 4-
methylbenzyl
bromide (370 mg, 2.00 mmol) and sodium hydride (88 mg, 2.2 mmol), followed by
stirring at
70°C for 2 hours. The reaction solution was cooled with ice, and water
was added, and then the
mixture was extracted with ethyl acetate. The organic layer was washed with
water and brine,
and then dried over anhydrous sodium sulfate. The solvent was evaporated under
reduced
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pressure. 2,4-Thiazolidinedione (176mg, l.Smmo1), piperidine (0. 10 mL, 1.0
mmol) and
ethanol (8 mL) were added thereto, followed by heating under reflex for 2
hour. The reaction
solution was cooled down to room temperature, and water and 1 N HC 1 (5 mL)
were added,
and then the mixture was extracted with ethyl acetate. The organic layer was
washed with water
S and brine, and then dried over anhydrous sodium sulfate. The solvent was
evaporated under
reduced pressure, and the residue was triturated with hexane/diisopropyl ether
to give
Compound 41 (340 mg, 76% by two steps).
~H NMR (300 MHz, DMSO-d6) 82.30 (s, 6H), 5.05 (s, 2H), 5.11 (s, 2H),
6.93 (br s, 1H), 7.1-7.2 (m, 2H), 7.20 (d, J = 7.7 Hz, 4H), 7.32 (d, J = 7.9
Hz,
4H), 7.93 (s, 1 H), 12.6 Hz, 1 H)
FABMS m/z 446 (M+H)+ C26H23N04S = 445
Example 69
Preparation of Compound 42
In an argon atmosphere, 5'-bromo-2'-hydroxyacetophenone (215 mg, 1.00 mmol)
was
dissolved in dimethylformamide (5 mL), and to the solution were mixed with 4-
methylbenzyl
bromide (185 mg, 1.00 mmol) and potassium carbonate (152 mg, 1.1 mmol),
followed by
stirring at 70°C for S hours. The reaction solution was cooled with
ice, and water was added,
and then the mixture was extracted with ethyl acetate. The organic layer was
washed with a 0.1
N aqueous sodium hydroxide solution, water and brine, and then dried over
anhydrous sodium
sulfate. The solvent was evaporated under reduced pressure. 2,4-
Thiazolidinedione ( 176 mg,
1.5 mmol) and sodium acetate (123 mg, 1.0 mmol) were added thereto, followed
by heating at
190°C for 3-5 hours. The reaction solution was cooled down to room
temperature, and water
~ and 1 N HC 1 ( 1 mL) were added, and then the mixture was extracted with
ethyl acetate. The
organic layer was washed with water and a saturated brine, and then dried over
anhydrous
sodium sulfate. The solvent was evaporated under reduced pressure, and the
residue was
purified by silica gel column chromatography (30:1 chloroform/acetonitrile) to
give Compound
42 ( 102 mg, 24% by two steps).
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IH NMR (300 MHz, DMSO-d6) 52.35 (s, 3H), 2.61 (s, 3H), 6.88 (d, J = 9.0 Hz,
IH),
7.1-7.3 (m, SH), 7.41 (dd, J = 8.8, 2.6 Hz, 1 H), 8.14 (br s, 1 H)
FABMS m/z 419, 418 (M+H)+ C,9Hi6'9BrNO3S = 417.
Example 70
Preparation of Compound 43
5-Bromo-2-(4-chlorophenylthio)thiophene-3-carboxyaldehyde (167 mg, 0.500
mmol),
2,4-thiazolidinedione (88 mg, 0.75 mmol) and piperidine (0.049 mL, 0.5 mmol)
were heated
under reflux for 6 hours in ethanol (S rnL). The reaction solution was cooled
down to room
temperature, water and 1 N HC I (0.5 mL) were added, and then the mixture was
extracted with
chloroform. The organic layer was washed with water and brine, and then dried
over anhydrous
sodium sulfate. The solvent was evaporated under reduced pressure, and the
residue was
triturated with diisopropyl ether/ethyl acetate to give Compound 43 (I38 mg,
64%).
'H NMR (300 MHz, DMSO-db) 57.32 (d, J = 8.6 Hz, 2H), 7.47 (d, J = 8.6 Hz, 2H),
7.82
(s, 1 H), 7.98 (s, 1 H), 12.7 (br s, IH).
FABMS m/z 433, 43I (M+) C~4H~'9Br35C1NO2S3 = 431.
Example 71
Preparation of Compound 44
In an argon atmosphere, Compound 43 (22 mg, 0.051 mmol) was dissolved in
dimethoxyethane (2 mL), and to the solution were added
tetrakistriphenylphosphine palladium
(6 mg, 10 mol%), sodium carbonate aqueous solution (0.5 M, 0.6 mL) and phenyl
boronic acid
(31 mg, 0.25 mmol) and heated under reflux for 12 hours. The reaction solution
was cooled
down to room temperature, and water was added, and then the mixture was
extracted with ethyl
acetate. The organic layer was washed with brine, and then dried over
anhydrous sodium
sulfate. The solvent was evaporated under reduced pressure, and the residue
was purified by
preparative thin layer chromatography ( 10:1 chloroform/acetone) to give
Compound 44 (8.3
mg, 38%).
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~H NMR (300 MHz, DMSO-d6) 57.17 (d, J = 8.4 Hz, 2H),
7.2-7.5 (m, 8H), 7.91 (s, IH), 8.16 (br s, 1H)
FABMS m/z 429 (M~ CZOHi23sC1N02S3 = 429.
Example 72
Preparation of Compound 45
In an argon atmosphere, Compound 43 (32 mg, 0.075 mmol) was dissolved in
tetrahydrofuran (5 mL) and cooled down to - 78°C. n-Butyl lithium (a
1.6 mol/L hexane
solution, 0.3 mL) was added thereto, followed by stirring for 15 minutes. The
reaction solution
was mixed with water and extracted with ethyl acetate. The organic layer was
washed with
brine, and then dried over anhydrous sodium sulfate. The solvent was
evaporated under
reduced pressure, and the residue was purified by preparative thin layer
chromatography ( 10:1
chloroform/acetone) to give Compound 45 (3.8 mg, 14%).
~H NMR (300 MHz, DMSO-d6) 87.24 (d, J = 4.2 Hz, 1H), 7.27 (d, J = 9.0 Hz, 2H),
7.30
(d, J = 3.9 Hz, 1 H), 7.31 (d, J = 9.2 Hz, 2H), 7.91 (s, 1 H), 8.42 (br s, 1
H)
FABMS m/z 353 (M~ C~4Hg35C1NO2S3 = 353.
Example 73
Preparation of Compound 46
In an argon atmosphere, tris(4-bromophenyl)amine (964 mg, 2.00 mmol) was
dissolved
in tetrahydrofuran (10 mL) and cooled to -78°C. n-Butyl lithium (a 1.6
mol/L hexane solution,
1.5 mL, 2.4 mmol) and dimethylformamide (0.19 mL, 2.4 mmol) was dripped
thereto at a
system temperature of -60°C or less, followed by stirring for 10
minutes. To the reaction liquid
was added water and the product was extracted with ethyl acetate. The organic
layer was
washed with water and brine, and then dried over anhydrous sodium sulfate. The
solvent was
evaporated under reduced pressure, and the residue was purified by silica gel
column
chromatography (elution solvent: hexane/ethyl acetate = 8/1) to give
4-[bis(4-bromophenyl)amino]benzaldehyde (377 mg, 44%).
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~H NMR (300 MHz, CDC13) 8 (ppm) 7.02 (d, J = 8.8 Hz, 4H), 7.04 (d, J = 8.6 Hz,
2H),
7.45 (d, J = 8.8 Hz, 4H), 7.71 (d, J = 8.8 Hz, 2H), 9.84 (s, 1 H)
FABMS m/z 433, 431, 429 (M~ CigH~3~9BR2NO = 429
The 4-[bis(4-bromophenyl)amino]benzaldehyde (356 mg, 0.826 mmol),
2,4-thiazolidinedione (145 mg, 1,24 mmol), and piperidine (0.083 mL, 0.83
mmol) were heated
under reflux for four hours in ethanol (8 mL). The reaction liquid was cooled
to room
temperature, mixed with water and 1 mol/L HCl (1 mL), and was extracted with
chloroform.
The organic layer was washed with water and brine, and then dried over
anhydrous sodium
sulfate. The solvent was evaporated under reduced pressure, the residue was
purified by silica
gel column chromatography (elution solvents chloroform/acetonitrile = 15/1)
and was triturated
with hexane to give Compound 46 (388 mg, 89%).
1H NMR (300 MHz, DMSO-d6) 8 (ppm) 7.03 (d, J = 8.8 Hz, 2H), 7.06 (d, J = 8.8
Hz, 4H), 7.51 (d, J = 8.8 Hz, 2H), 7.54 (d, J = 8.8 Hz, 4H), 7.70 (s, 1 H),
12.5 (br s, 1 H)
FABMS m/z 532, 530, 528 (M~ CZZH,4~9BRZNzOZS = 528
Example 74
Preparation of Compound 47
In an argon atmosphere, Compound 25 (40 mg, 0.10 mmol) was dissolved in
1,2-dimethoxyethane, (4 mL). Phenyl boric acid (24 mg, 0.20 mmol), a 2 mol/L
sodium
carbonate aqueous solution (0.15 mL), water (0.5 mL), and
tetrakis(triphenylphosphine)
palladium (6 mg, 5 mol%) were added and the product was heated under reflux
for 8 hours.
The reaction liquid was cooled to room temperature, mixed with water and 1
mol/L HC1, and
was extracted with ethyl acetate. The organic layer was washed with water and
brine, and then
diied over anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure, and
the residue was purified by preparative thin layer chromatography (development
solvent:
chloroform/acetonitrile = 12/1) and was triturated with isopropyl ether to
give Compound 47
(27 mg, 67%).
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'H NMR (300 MHz, DMSO-d6) b (ppm) 2.32 (s, 3H), 5.25 2H), 7.23 (d, J = 8.1 Hz,
2H), 7.3-7.4 (m, 4H), 7.49 (t, J = 7.6 Hz, 2H), 7.63 (d, J = 8.6 Hz, 2H), 7.64
(d, J = 1.7
Hz, 1 H), 7.76 (dd, J = 8.6, 2.2 Hz, 1 H), 8.03 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 401 (M~ C24H,9NO3S = 401
Example 75
Preparation of Compound 48
2-Thienyl boric acid (26 mg, 0.20 mmol) was used instead of phenyl boric acid
to
obtain Compound 48 (7.5 mg, 18%) from Compound 25 (40 mg, 0.10 mmol), using
the same
method as Example 74.
~H NMR (300 MHz, DMSO-db) b (ppm) 2.32 (s, 3H), 5.23 (s, 2H), 7.14 (dd, J =
5.0, 3.7
Hz, 1 H), 7.22 (d, J = 7.9 Hz, 2H), 7.30 (d, J = 8.8 Hz, 1H), 7.36 (d, J = 8.1
Hz, 2H),
7.44 (d, J = 3 . 5 Hz; 1 H), 7.5 3 (d, J = 4.8 Hz, 1 H), 7.62 (d, J = 2.0 Hz,
1 H), 7.75 (dd, J =
8.8, 2.0 Hz, 1 H), 7.98 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 407 (M+) C22H»NO3S2 = 407.
Example 76
Preparation of Compound 49
In an argon atmosphere, tris(4-bromophenyl)amine (7.23 g, 15.0 mmol) was
dissolved
in tetrahydrofuran (100 mL) and cooled to -78°C. n-Butyl lithium (a 1.6
mol/L hexane solution,
34 mL, 54 mmol) and dimethylformamide (4.6 mL, 60 mmol) was dripped thereto at
a system
temperature of -60°C or less, followed by stirring for 10 minutes at
the same temperature.
Water was added to the reaction liquid and the product was extracted with
ethyl acetate. The
organic layer was washed with water and brine, and then dried over anhydrous
sodium sulfate.
The solvent was evaporated under reduced pressure, and the residue was
purified by silica gel
column chromatography (elution solvent: hexane/ethyl acetate = 4/1 followed by
chloroform/acetonitrile = 30/1) to give tris(4-formylphenyl)amine (2.97 g,
60%).
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1H NMR (300 MHz, CDC13) 8 (ppm) 7.25 (d, J = 8.8 Hz, 6H), 7.85 (d, J = 8.8 Hz,
6H),
9.95 (s, 3H).
FABMS m/z 330 (M+H)+ Cz~H,5N03 = 329.
The tris(4-formylphenyl)amine (165 mg, 0.502mmol) was dissolved in methyl
alcohol
(8 mL) and chloroform (S mL). To the solution sodium borohydride (9.5 mg, 0.25
mmol) was
added under ice-cooling, followed by stirring at room temperature for 15
minutes. Water was
added to the reaction liquid and the product was extracted with chloroform:
The organic layer
was washed with brine, and then dried over anhydrous sodium sulfate. The
solvent was
evaporated under reduced pressure, and the residue was purified by silica gel
column
chromatography (elution solvent: chloroform/acetonitrile = 20/1-10/1) to give
4-[[bis(4-
hydroxymethyl)phenyl]amino]benzaldehyde ( 107 mg, 64%).
'H NMR (300 MHz, CDC13) 8 (ppm) 2.46 (br s, 2H), 4.66 (s, 4H), 6.99 (d, J =
8.8 Hz,
2H), 7.13 (d, J = 8.4 Hz, 4H), 7.32 (d, J = 8.4 Hz, 4H), 7.64 (d, J = 8.8 Hz,
2H), 9.75 (s,
1 H)
FABMS m/z 333 (M+) CziH19NO3 = 333.
The 4-[[bis(4-hydroxymethyl)phenyl]amino]benzadehyde (100 mg, 0.300
mmol)°,
2,4-thiazolidinedione (53 mg, 0.45 mmol), and piperidine (0.030 mL, 0.30 mmol)
were heated
under reflux for three hours in ethanol (5 mL). The reaction liquid was cooled
to room
temperature, mixed with water and 1 mol/L HC1 (1 mL), and the product was
extracted with
ethyl acetate. The organic layer was washed with water and brine, and then
dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced pressure,
and triturated
with hexane to give Compound 49 (113 mg, 87%).
'H NMR (300 MHz, DMSO-db) b (ppm) 4.48 (d, J = 4.4 Hz, 4H), 5.16 (br t, J =
5.1 Hz,
2H), 6.89 (d, J = 8.8 Hz, 2H), 7.09 (d, J = 8.3 Hz, 4H), 7.32 (d, J = 8.5 Hz,
4H), 7.44
(d,J = 9.0 Hz, 2H), 7.66 (s, 1 H), 12.4 (br s, 1 H)
FABMS m/z 432 (M+) Cz4Hz°N204S = 432.
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Example 77
Preparation of Compound 50
In an argon atmosphere, N-(4-bromobenzyl)diphenylamine (169 mg, 0.500 mmol)
obtained in Example 64 was dissolved in dimethylformamide ( 1.5 mL), and
phosphorus
oxychloride (0.116 mL, 1.25 mmol) was added thereto, followed by stirring at
100°C for 30
minutes. The reaction liquid was cooled to room temperature, poured into
saturated sodium
acetate aqueous solution, and was extracted with ethyl acetate. The organic
layer was washed
with brine, and then dried over anhydrous sodium sulfate. The solvent was
evaporated under
reduced pressure, and the residue was purified by silica gel column
chromatography (elution
solvent: hexane/ethyl acetate = 4/1) to give 4-[N-(4-bromobenzyl)-N-
phenylamino]-
benzaldehyde (159 mg, 87%).
'H NMR (300 MHz, CDC13) b (ppm) 4.98 (s, 2H), 6.78 (d, J = 8.8 Hz, 2H), 7.18
(d, J =
8.3 Hz, 2H), 7.26 (m, 1 H), 7.27 (d, J = 8.4 Hz, 2H), 7.42 (m, 2H), 7.45 (d, J
= 8.4 Hz,
2H), 7.65 (d, J = 8.8 Hz, 2H), 9.?5 (s, 1 H)
FABMS m/z 368, 366 (M+H)+ C2oH16~9BrN0 = 365.
The 4-[N-(4-bromobenzyl)-N-phenylamino]benzaldehyde (153 mg, 0.418 mmol),
2,4-thiazolidinedione (73 mg, 0.63. mmol), and piperidine (0.042 mL, 0.42
mmol) were heated
under reflux for three hours in ethanol (5 mL). The reaction liquid was cooled
to room
temperature, mixed with water and 1 mol/L HC1 (1 mL), and was extracted with
ethyl acetate.
The organic layer was washed with water and brine, and then dried over
anhydrous sodium
sulfate. The solvent was evaporated under reduced pressure, and triturated
with
hexane/isopropyl ether to give Compound 50 (150 mg, 87%).
~H NMR (300 MHz, DMSO-d6) 8 (ppm) 5.06 (s, 2H), 6.88 (d, J = 8.8 Hz, 2H), 7.21
(t, J
= 7.3 Hz, 1H), 7.27 (d, J = 8.3 Hz, 2H), 7.32 (d, J = 7.5 Hz, 2H), 7.40 (d, J
= 8.8 Hz,
2H), 7.42 (t, J = 7.3 Hz, 2H), 7.51 (d, J = 8.3 Hz, 2H), 7.64 (s, 1 H), I 2.4
(br s, 1 H)
FABMS m/z 466, 464 (M+} Cz3Hi~79BrN202S = 464.
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Exam~Ie 78
Preparation of Compound S 1
Under ice-cooling, Compound 43 (50 mg, 0.12 mmol) was dissolved in a mixed
solvent
of dichloromethane (8 mL) and methanol (2 mL). m-Chloroperbenzoic acid (50%
purity, 65
mg, 0.19 mmol) was added thereto, followed by stirring at room temperature for
18 hours. To
the reaction liquid a 5% sodium hydrogen sulfite aqueous solution was added
and the product
was extracted with chloroform. The organic layer was washed, with sodium bi-
carbonate
aqueous solution and brine, and then dried over anhydrous sodium sulfate. The
solvent was
evaporated under reduced pressure, and the residue was triturated with
methanol to give
Compound 51 (30 mg, 56%).
IH NMR (300 MHz, DMSO-d6) 8 (ppm) 7.73 (d, J = 8.4 Hz, 2H), 7.75 (s, 1H), 7.84
(d,
J = 8.6 Hz, 2H), 7.98 (s, 1 H), 12.8 (br s, 1 H)
FABMS m/z 450, 448 (M-H)' C,4H~~9Br35CliNO3S3 = 449.
Example 79
Preparation of Compound 52
Under ice-cooling, Compound 43 (50 mg, 0.12. mmol) was dissolved in a mixed
solvent of dichloromethane (8 mL) and methanol (2 mL). m-Chloroperbanzoic acid
(50%
purity, 398 mg, 1.2 nunol) was added thereto, followed by stirring at room
temperature for 17
hours. To the reaction liquid a 5% sodium hydrogen sulfite aqueous solution
was added and the
product was extracted with chloroform. The organic layer was washed with
sodium
hydrogencarbonate aqueous solution and brine, and then dried over anhydrous
sodium sulfate.
The solvent was evaporated under reduced pressure, and triturated with
methanol to give
Compound 52 (23 mg, 41 %).
'H NMR (300 MHz, DMSO-db) 8 (ppm) 7.71 (d, J = 8.6 Hz, 2H), 7.78 (s, 1 H),
8.00 (s,
1 H), 8.05 (d, J = 8.6 Hz, 2H), 12.9 (br s, 1 H)
FABMS m/z 466, 464 (M-H)' C,4H7~9Br35CiNO4S3 = 465.
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Example 80
Preparation of Compound S3
Commercially available S-bromo-2-(4-chlorophenyl-thio)thiophene-3-
carboxaldehyde
(1.00 g, 2.99 mmol) (MAYBRIDGE, Catalog Number: KMOS476) was dissolved in
methanol
(7S mL). p-Toluenesulfonic acid (S2 mg, 0.30 mol) was added thereto, and the
mixture was
heated under reflux for 1.S hours. The solvent was evaporated under reduced
pressure until the
total volume becomes about 30 mL. Water and saturated sodium bicarbonate
aqueous solution
were added thereto, and the product was extracted with ethyl acetate. The
organic layer was
washed with water and brine, and then dried over anhydrous sodium sulfate. The
solvent was
evaporated under reduced pressure to give S-bromo-2-(4-
chlorophenylthio)-3-(dimethoxymethyl)-thiophene (1.12g, 100%).
'H NMR (300 MHz, CDC13) S (ppm) 3.36 (s, 6H), S.SS (s, 1H), 7.07 (s, 1H), 7.15
(d, _
1S J-8.6Hz,2H),7.24(d,J=8.8Hz,2H)
FABMS m/z 380, 378 (M~ C13H12~9Br35C1O2S2 = 378.
In an argon atmosphere, S-bromo-2-(4-chlorophenylthio)-3-(dimethoxymethyl)
thiophene ( 1.04 g, 2.75 mmol) was dissolved in tetrahydrofuran ( 1 S mL) and
the mixture was
cooled to -78°C. n-Butyl lithium (1.6 mol/L hexane solution; 2.S mL,
4.1 mmol) was added
thereto, followed by stirnng for S minutes. Dry ice (about 1 g) was added
thereto, followed by
stirring for 10 minutes. Water and 1 moUL sodium hydroxide aqueous solution
was added to
the reaction liquid so that pH becomes 10, and the mixture was washed with
ether. 1 mol/L
Hydrochloric acid was added to the aqueous layer so that pH becomes 3, and the
aqueous layer
was extracted with ethyl acetate. The organic layer was washed with water and
brine and then
dried over anhydrous sodium sulfate, and the solvent was evaporated under
reduced pressure.
The residue was dissolved in tetrahydrofuran (10 mL). 1 mol/L hydrochloric
acid (2 mL) was
added thereto, followed by stirring at room temperature for 2 hours. Water was
added to the
reaction liquid and the product was extracted with ethyl acetate. The organic
layer was washed
with water and brine, and then dried over anhydrous sodium sulfate. The
solvent was
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evaporated under reduced pressure, and the residue was triturated with
isopropyl ether to give
2-(4-chlorophenylthio}-3-formyl-5- thiophenecarboxylic acid (439 mg, 54%).
~H NMR (300 MHz, CDC13) 8 (ppm) 7.50 (d, J = 8.6 Hz, 2H), 7.62 (d, J = 8.6 Hz,
2H),
8.06 (s, 1 H), 9.68 (s, 1 H)
FABMS m/z 299 (M+H)+ C,ZH~35C 1 O3Sz = 298.
2-(4-Chlorophenylthio)-3-formyl-5-thiophensecarboxylic acid (400 mg, 1.34
mmol}, 2,4-
thiazolidinedione (188 mg, 1.01 mmol}, and piperidine (0.133 mL, 1.34 mmol)
were heated
under reflux for 3.5 hours in ethanol ( 12 mL). The reaction mixture was
cooled to room
temperature. 1 moUL hydrochloric acid (1.5 mL) was added thereto, and the thus
precipitated
crystals were collected by filtration to give Compound 53 (411 mg, 77%).
'H NMR (300 MHz, DMSO-d6) 8 (ppm) 7.66 (d, J = 8.4 Hz, 2H), 7.77 (d, J = 8.4
Hz,
2H), 7.83 (s, 1 H), 7.90 (s, 1 H), 12.6 {br s, 1 H), 13.4 (br s, 1 H)
FABMS m/z 396 (M-H)' Ci5Hg35C1N04S3 = 397.
Example 81
Preparation of Compound 54
Compound 53 (80 mg, 0.20 mmol) was dissolved in dimethylformamide (3 mL).
1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (77 mg, 0.40 mmol)
and
diethylamine (0.041 mL, 0.40 mmol) were added thereto, followed by stirnng at
room
temperature for 3 hours. Water was added to the reaction liquid and the
product was extracted
with ethyl acetate. The organic layer was washed with water and brine, and
then dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced pressure,
and the residue
was purified by preparative thin layer chromatography (9:1
chloroform/methanol) and was
triturated with hexane to give Compound 54 (22 mg, 24%).
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~H NMR (300 MHz, DMSO-d6) 8 (ppm) 1.06 (br s, 6H), 3.13 (br s, 2H), 3.41 (br
s, 2H),
7.41 (d, J = 8.6 Hz, 2H), 7.47 (d, J = 8.6 Hz, 2H), 7.69 (s, 1 H), 8.00 (s, 1
H), 12.7 (br s,
1 H)
FABMS m/z 451 (M-H)' C,9H1~35C1N2O3S3 = 452.
Example 82
Preparation of Compound 55
Aniline (0.037 mL, 0.40 mmol) was used instead of diethylamine to obtain
Compound
SS (37 mg, 39%), using the same method as Example 81.
'H NMR (300 MHz, DMSO-d6) S (ppm) 7.13 (t, J = 7.3 Hz, 1 H), 7.37 (t, J = 7.7
Hz,
2H), 7.60 (d, J = 8.4 Hz, 2H), 7.68 (d, J = 8.6 Hz, 2H), 7.71 (d, J = 7.7 Hz,
2H), 7.84(s,
1 H), 8.04 (s, 1 H), 10.3 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 471(M-H)' C2iH,3C1N2O3S3 = 472.
Example 83
Preparation of Compound 56
1-Methylpiperazine (0.044 mL, 0.40 mmol) was used instead of diethylamine to
obtain
Compound 56 { 11 mg, 12%), using the same method as Example 81. Compound 56
was
obtained in the form of hydrochloride.
'H NMR (300 MHz, DMSO-d6, hydrochloride) 8 (ppm) 2.80 (s, 3H), 3.25 (m, 8H),
7.46
(d, J = 8.8 Hz, 2H), 7.49 (d, J = 8.8 Hz, 2H), 7.69 (s, 1 H), 7.98 (s, 1 H)
FABMS m/z 478 (M-H)' C2oH18CIN3O3S3 = 479.
Example 84
Preparation of Compound 57
Morpholine (0.035 mL, 0.40 mmol) was used instead of diethylamine to obtain
Compound 57 (22 mg, 24%), using the same method as Example 81.
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'H NMR (300 MHz, DMSO-d6) S (ppm) 3.32 (m, 4H), 3.55 (m, 4H), 7.43 (d, J = 8.6
Hz, 2H), 7.47 (d, J = 8.6 Hz, 2H), 7.69 (s, 1 H), 7.98 (s, 1 H), 12.7 (br s, 1
H)
FABMS m/z 465 (M-H)' C,9H,SCINZO4S3 = 466
Example 85
Preparation of Compound 58
Compound 53 (36 mg, 0.090 mmol) was dissolved in dichloromethane (2 mL) and
tetrahydrofuran (2 mL). Thionyl chloride (0.032 mL, 0.36 mmol) was added to
the solution and
the mixture was heated under reflux for 2 hours. The reaction liquid was
cooled with ice.
Methanol (1 mL) and triethylamine (0.038 mL, 0.27 mmol) were added thereto,
followed by
stirring for 10 minutes. Water was added to the reaction liquid and the
product was extracted
with ethyl acetate. The organic layer was washed with water and brine, and
then dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced pressure,
and the residue
was purified by preparative thin layer chromatography (9:1
chloroform/methanol) and was
triturated with isopropyl ether to give Compound 58 (15 mg, 40%).
'H NMR (300 MHz, DMSO-db) 8 (ppm) 3.85 (s, 3H), 7.67 (d, J = 8.6 Hz, 2H),
7.77 (d, J = 8.4 Hz, 2H), 7.88 (s, 1 H), 7.92 (s, 1 H), 12.6 (br s, 1 H)
FABMS m/z 410 (M-H)- Ci6H,o SC1NOQS3 = 411
Example 86
Preparation of Compound 59
Compound 59 (260 mg, 86%) was obtained from commercially available
3-phenoxythiophene-2-carboxaldehyde (204 mg, 1.00 mmol) (MAYBRIDGE, Catalog
Number: KM05428), 2,4-thiazolidinedione (176 mg, 1.5 mmol), and piperidine
(0.099 mL, 1.0
mmol), using the same method as Example 70.
'H NMR (300 MHz, DMSO-db) 8 (ppm) 6.93 (d, J = 5.5 Hz, 1H), 7.02 (dd, J = 8.6,
1.1
Hz, 2H), 7.15 (t, J = 7.5 Hz, 1H), 7.39 (dd, J = 8.6, 7.5 Hz, 2H), 7.61 (s,
1H), 7.91.(d, J
= 5.5 Hz, 1 H)
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FABMS m/z 302 (M-H)' Ci4H9NO3S2 = 303.
Example 87
Preparation of Compound 60
4-Bromothiophene-2-carboxaldehyde (2.00 g, 10.5 mmol) was dissolved in
methanol
(80 mL). p-Toluenesulfonic acid (181 mg, 1.05 mmol) was added thereto,
followed by stirring
for 3 hours. The solvent was evaporated under reduced pressure until the total
volume becomes
about 20 mL. Sodium hydrogencarbonate aqueous solution was added thereto, and
the product
was extracted with ether. The organic layer was washed with water and brine,
and then dried
over anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure to give 4-
bromo-2- dimethoxymethylthiophene (2.36g, 100%).
'H NMR (300 MHz, CDC13) 8 (ppm) 3.36 (s, 6H), 5.58 (s, 1H), 7.00 (t, J = 1.0
Hz,
1 H), 7.20 (d, J = 1.3 Hz, 1 H)
In an argon atmosphere, 4-bromo-2-dimethoxymethylthiophene (236 mg, 1.00 mmol)
was dissolved in tetrahydrofuran (4 mL) and cooled to -78°C. n-Butyl
lithium (1.6 moUL
hexane solution, 0.81 mL, 1.3 mmol), and tetrahydrofuran (1.5 mL) solution of
4,4'-dichlorodiphenyl disulfide (287 mg, 1.0 mmol) was added thereto, followed
by stirring for
15 minutes. Water was added to the reaction liquid and the product was
extracted with ether.
The organic layer was washed with water and brine and then dried over
anhydrous sodium
sulfate, and the solvent was evaporated under reduced pressure. The residue
was dissolved in
tetrahydrofuran (6 mL) and 1 mol/L hydrochloric acid was added thereto,
followed by stirring
at room temperature for 1 hour. Sodium hydrogencarbonate aqueous solution was
added to the
reaction liquid and the product was extracted with ethyl acetate. The organic
layer was washed
with water and brine, and then dried over anhydrous sodium sulfate. The
solvent was
evaporated under reduced pressure, and the residue was purified by preparative
thin layer
chromatography (9:1 hexane/ethyl acetate) to give
4-(4-chlorophenylthio)thiophene-2-carboxaldehyde (71 mg, 28%),
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2,3-bis(4-chlorophenylthio)thiophene-5-carboxaldehyde (29 mg, 7.3%), and 5-(4-
chlorophenyl-
thio)thiophene-2-carboxaldehyde (53 mg, 21%).
4-(4-chlorophenylthio)thiophene-2-carboxaldshyde:
~ H NMR (300 MHz, CDC 13) 8 (ppm) 7.21 (d, J = 8.8 Hz, 2H), 7.28 (d, J = 9.0
Hz,
2H), 7.65 (d, J = 1.5 Hz, 1 H), 7.69 (t, J = 1.4 Hz, 1 H), 9.87 (d, J = 1.3
Hz, 1 H)
FABMS m/z 254 (M+) C"H~35CI OSZ = 254
2,3-bis(4-chlorophenylthio)thiophene-5-carboxaldehyde:
~H NMR (300 MHz, CDC13) S (ppm) 7.19 (d, J = 8.4 Hz, 2H), 7.29 (d, J = 8.4 Hz,
2H),
7.37 (d, J = 8.6 Hz, 2H), 7.43 (d, J = 8.6 Hz, 2H), 7.53 (s, 1H), 9.67 (s, 1H)
FABMS m/z 397 (M+H)+ C,~HIO35C120S3 = 396
5-(4-chlorophenylthio)thiophenes-2-carboxaldehyde:
~H NMR (300 MHz, CDC13) 8 (ppm) 7.13 (d, J = 3.9 Hz, 1H), 7.33 (d, J = 8.8 Hz,
2H), 7.37 (d, J = 9.0 Hz, 2H), 7.63 (d, J = 3.9 Hz, 1 H), 9.78 (s, 1 H)
FABMS m/z 255 (M+H)+ C,1H~35CIOS2 = 254
4-(4-chlorophenylthio)thiophene-2-carboxyaldehyde (70 mg, 0.28 mmol),
2,4-thiazolidinedione (39 mg, 0.33 mmol), and piperidine (0.028 mL, 0.28 mmol)
were heated
under reflux for 4 hours in ethanol (4 mL). The reaction liquid was cooled to
room temperature,
mixed with water and 1 mol/L hydrochloric acid (1 mL), and the product was
extracted with
ethyl acetate. The organic layer was washed with water and brine, and then
dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced pressure
and the residue
was recrystallized from ethyl acetate to give Compound 60 (57 mg, 58%).
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1H NMR (300 MHz, DMSO-d6) S (ppm) 7.25 (d, J = 8.6 Hz, 2H), 7.41 (d, J = 8.6
Hz, 2H),
7.65 (m, 1 H), 8.02 (s, 1 H), 8.12 (m, 1 H), 12.6 (br s, 1 H)
FABMS m/z 352 (M-H)' C"Hg35C1NO2S2 = 353.
Example 88
Preparation of Compound 61
Compound 61 (19 mg, 54%) was obtained from 2,3-bis(4-
chlorophenylthio)thiophene-
5-carboxyaldehyde (28 mg, 0.071 rnmol) obtained in Example 87, 2,4-
thiazolidinedione (10
mg, 0.085 mmol), and piperidine (0.007 mL, 0.07 mmol), using the same method
as Example
87.
1NMR (300 MHz, DMSO-db) 8 (ppm) 7.30 (d, J = 8.6 Hz, 2H), 7.35 (d, J = 8.6 Hz,
2H},
7.42 (d, J = 8.6 HZ, ZH), 7.46 (d, J = 8.6 Hz, 2H), 7.56 (s, 1 H), 7.96 (s, 1
H), 12.7 (br s,
1 H)
FABMS m/z 494 (M-H)' C~gH"35C12NOZS4 = 495.
Example 89
Preparation of Compound 62
Compound 62 ( 113 mg, 62%) was obtained from 5-(4-chlorophenylthio)thiophene-
2-carboxyaldehyde (132 mg, 0.520 mmol) obtained in Example 87, 2,4-
thiazolidinedione (73
mg, 0.62 mmol), and piperidine (0.052 mL, 0.52 mmol), using the same method as
Example
87.
IH NMR (300 MHz, DMSO-d6) 8 (ppm) 7.33 (d, J = 8.6 Hz, 2H), 7.44 (d, J = 8.6
Hz,
2H), 7.54 (d, J = 3.9 Hz, 1 H), 7.68 (d, J = 3.9 Hz, 1 H), 8.01 (s, 1 H), 12.6
(br s, 1 H}
FABMS m/z 352 (M-H)' C,3H835CINOZSz = 353.
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Example 90
Preparation of Compound 63
In an argon atmosphere, a tetrahydrofuran (3 mL) solution of diisopropylamine
(0.21
mL, 1.5 mol) was cooled with ice, n-butyl lithium (1.6 mol/L hexane solution,
0.81 ml, 1.3
mmol) was added thereto, and the mixture was cooled to -78°C. A
tetrahydrofuran (1 mL)
solution of 4-bromo-2-dimethoxymethylthiophene (236 mg. 1.00 mmol) obtained in
Example
87 was added thereto, followed by stirring for 30 minutes. Then, a
tetrahydrofuran (1 mL)
solution of 4,4'-dichlorodiphenyl disulfide (287 mg, 1.0 mmol) was added
thereto, followed by
stirring for 10 minutes. To the reaction liquid water was added, and the
product was extracted
with ether. The organic layer was washed with water and brine, dried over
anhydrous sodium
sulfate and then the solvent was evaporated under reduced pressure to give 3-
bromo-2-(4-
chlorophenylthio)-5-dimetoxymethylthiophen (355 mg, 94%).
~H NMR (300 MHz, CDC 13) 8 (ppm.) 3.36 (s, 6H), 5.55 (d, J = 0.7 Hz, 1 H),
7.07 (d,
J = 0.9 Hz, 1 H), 7.16 (d, J = 8.4 Hz, 2H), 7.24 (d, J = 8.6 Hz, 2H)
FABMS m/z 380, 378 (M+) Cl;Hi2~9Br35C1OzS2 = 378
3-Bromo-2-(4-chlorophenylthio)-5-dimetoxymethylthiophen (170 mg, 0.449 mmol)
was dissolved in tetrahydrofuran (4 mL) and 1 mol/L hydrochloric acid (0.5 mL)
was added
thereto, followed by stirring at room temperature for 4.5 hours. To the
reaction liquid was
added a sodium hydrogencarbonate aqueous solution and the product was
extracted with ethyl
acetate. The organic layer was washed with water and brine, and then dried
over anhydrous
sodium sulfate. The solvent was evaporated under reduced pressure, and the
residue was
purified by silica gel column chromatography (9:1 hexane/ethyl acetate) to
give
3-bromo-2-(4-chlorophenylthio)thiophan-5-carboxaldehyde (126 mg, 85%).
'H NMR (300 MHz, CDC13) b (ppm) 7.38 (d, J = 8.4 HZ, 2H), 7.43 (d, J = 8.6 Hz,
2H),
7.63 (s, 1 H), 9.71 (s, 1 H)
FABMS m/z 335, 333 (M+H)+ C, iH6'9Br35C1OS2 = 332.
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Compound 63 (73 mg, 49%) was obtained from 3-bromo-
2-(4-chlorophenylthio)thiophen-5-carboxaldehyde ( 114 mg, 0.342 mmol),
2,4-thiazolidinedione (48 mg, 0.41 mmol), and piperidine (0.034 mL, 0.34 mmol)
using the
same method as Example 87.
~H NMR (300 MHz, DMSO-d6) b (ppm) 7.31 (d, J = 8.8 Hz, 2H), 7.47 (d, J = 8.6
Hz,
2H), 7.81 (s, 1 H), 7.96 (s, 1 H), 12.7 (br s, 1 H)
FABMS m/z 432, 430 (M-H)' Ci4H~~9Br35CINO2S3 = 431
Example 91
Preparation of Compound 64
Compound 64 (14 mg, 714) was obtained from Compound 63 (19 mg, 0.044 mmol) and
m-chloroperbenzoic acid (50% purity, 23 mg, 0.066 mmol), using the same method
as Example
78.
1H NMR (300 MHz, DMSO-d6) b (ppm) 7.73 (d, J = 9.0 Hz, 2H), 7.75 (s, 1 H),
7.84 (d,
J = 8.6 Hz, 2H), 7.98 (s, 1 H), 12.8 (br s, 1 H)
FABMS m/z 448, 446 (M-H)' C,4H~~9Br35C1NO3S3 = 447
Example 92
Preparation of Compound 65
In an argon atmosphere, 3-bromo-2-(4-chlorophenylthio)-5-
dimetoxymethylthiophene
(500 mg, 1.32 mmol) obtained in Example 90 was dissolved in tetrahydrofuran (6
mL) and
cooled to -78°C. n-Butyl lithium (a 1.6 moUL hexane solution, 0.81 mL,
1.3 mmol) and
tetrahydrofuran (1 mL) solution of 4-chloro-N-methoxy-N-methylbenzamide (528
mg, 2.64
mmol) were added thereto, followed by stirring for 10 minutes. Water was added
to the
reaction liquid and the product was extracted with ethyl acetate. The organic
layer was washed
with water and brine, and then dried over anhydrous sodium sulfate. The
solvent was
evaporated under reduced pressure, and the residue was purified by silica gel
column
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chromatography ( 1 S:1 hexane/ethyl acetate) to give 3-(4-chlorobenzoyl)-2-
(4-chlorophenylthio)-5-dimetoxymethylthiophene (345 mg, 60%).
~H NMR (300 MHz, CDC 13) 8 (ppm) 3.31 (s, 6H), 5.45 1 H), 7.15 (d, J = 1.1 Hz,
114),
7.39 (d, J = 8.8 Hz, 2H), 7.46 (d, J = 8.6 Hz, 2H), 7.53 (d, J = 8.8 Hz, 2H),
7.73 (d, J =
8.6 Hz, 2H)
FABMS m/z 439 (M+H)+ C2pH1635C12~3S2 = 438
3-(4-Chlorobenzoyl)-2-(4-chlorophenylthio)-5-dimetoxymethylthiopene (335 mg,
0.736
mmol) was dissolved in tetrahydrofuran (6 mL). 1 mol/L Hydrochloric acid ( 1
mL) was added
thereto, followed by stirring at room temperature for 1.5 hours. Sodium
hydrogencarbonate
aqueous solution was added to the reaction liquid and the product was
extracted with ethyl
acetate. The organic layer was washed with water and brine, and then dried
over anhydrous
sodium sulfate. The solvent was evaporated under reduced pressure, and the
residue was
purified by silica gel column chromatography (5:1 hexane/ethyl acetate) and
recrystallized
from ethyl acetate/hexane to give 3-(4-chlorobenzoyl)-2-(4-
chlorophenylthio)thiophen-
5-carboxyaldehyde (203 mg, 68%).
1H NMR (300 MHz, CDC13) 8 (ppm) 7.50 (d, J = 8.4 Hz, 2H), 7.52 (d, J = 8.4 Hz,
2H);
7.62 (d, J = 8. 8 Hz, 2H), 7.74 (d, J = 8.8 Hz, 2H), 7.83 (s, 1 H), 9.65 (s, 1
H)
FABMS m/z 393 (M+H)+ C~6H1p35C12OZS2 = 392
Compound 65 (211 mg, 88%) was obtained from 3-(4-chlorobenzoyl)-2-
(4-chlorophenylthio)thiophen-5-carboxyaldehyde (193 mg, 0.490 mmol), 2,4-
thiazolidinedione
(69 mg, 0.59 mmol), and piperidine (0.049 mL, 0.49 mmol), using the same
method as
Example 70.
~H NMR (300 MHz, DMSO-db) 8 (ppm) 7.64 (d, J = 8.6 Hz, 2H), 7.65 (d,J = 8.6
Hz,
2H), 7.73 (d, J = 8.4 Hz, 2H), 7.81 (d, J = 8.4 Hz, 2H), 7.87 (s, 1 H), 7.94
(s, 1 H), 12.6
(br s, 1 H)
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FABMS m/z 490 (M-H)' C2~H"35C1ZNO3S3 = 491.
Example 93
Preparation of Compound 66
Compound 66 (14 mg, 28%) was obtained from Compound 65 (50 mg, 0.10 mmol) and
m-chloroperbenzoic acid (50% purity, 53 mg, 0.15 mol), using the same method
as Example
78.
'H NMR (300 MHz, DMSO-d6) S (ppm) 7.60 (d, J = 8.4 Hz, 2H), 7.71 (d, J = 8.4
Hz,
2H), 7.72 (d, J = 8.8 Hz, 2H), 7.82 (s, 1 H), 7.89 (d, J = 8.8 Hz, 2H), 8.04
(s, 1 H), 12.8
(br s, 1 H)
FABMS m/z 506 (M-H)- C2,H~135C12NO4S3 = 507
Exam~e 94
Preparation of Compound 67
Commercially available 2-[(4-chlorophenyl}thioJ-5-nitrobenzaldehyde (0.3 g,
1.0
mmol) (MAYBRIDGE, Catalog Number: XAX00154) was dissolved in ethanol (8 mL),
and
thiazolidinedione (0.36 g, 3.0 mmol) and piperidine (0.1 mL, 1.0 mmol) were
added thereto,
and the mixture was heated under reflux for S.5 hours in a flask equipped with
a reflux
condenser and drying tube (CaCl2), the temperature was reduced to room
temperature, and a 1
M HC 1 aqueous solution was added thereto. After a conventional treatment, the
residue was
purified by silica gel column chromatography (chloroform-chloroform:methanol =
99:1 ), and
purified by recrystallization from ethyl acetate and hexane, to obtain
Compound 67 (0.109 g,
27.9%).
1H NMR(300MHz, DMSO-d6) b (ppm) 7.24 (d, J = 8.8 Hz, 1H), 7.59(s, 4H), 7.88
(s,
1 H), 8.17 (dd, J = 2.6, 8.8 Hz, 1 H}, 8.24(d, J = 2.6Hz, 1 H), 12.84 (br s, 1
H)
FABMS m/z 393 (M+H)+ Ci6H935C1NZO4S2 = 392.
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Example 95
Preparation of Compound 68
Compound 68 (0.0137 g, 65.7%} was obtained by using Compound 67 (0.02 g, 0.051
mmol), using the same method as Example 78.
1H-NMR(300MHz, DMSO-d6) S (ppm) 7.61(d, J = 2.6Hz, 4H), 7.67(s, 1H), 8.25(d,
J = 8.8 Hz, 1 H), 8.35(d, J = 2.3Hz, 1 H), 8.40 (dd, J = 2.3, 8.8 Hz, 1 H), NH
is not found
EIMS m/z 409 (M+H}+C~6H935C1N2OSS2 = 408.
Example 96
Preparation of Compound 69
Compound 67 (0.03 g, 0.077 mmol) was dissolved in dichloromethane (5 mL} and
methanol ( 1 mL), and m-chloroperbenzoic acid (0.03 g, 0.0092 mmol) was added
thereto,
followed by stirring at room temperature for 1 hour. A 10% aqueous sodium
hydrogen sulfite
solution was added, and a conventional treatment was performed, after which
the residue was
purified by thin layer column chromatography (chloroform: methanol = 12:1 ),
followed by
another purification by thin layer chromatography (chloroform:acetonitryl =
6:1), to obtain
Compound 69 (0.014 g, 15.9%).
1H-NMR(300MHz, DMSO-d6) ( (ppm) 7.71 (dt, J = 2.0, 8.8 Hz, 2H), 7.87 (dt, J =
2.0,
8.8 Hz, 2H), 7.98 (s, 1 H), 8.31 (s, 1 H), 8.49 (d, J = 2.0 Hz, 2H), NH is not
found
EIMS m/z 423(M-H)- C16H935C1N2O6S2 = 424.
Example 97
Preparation of Compound 70
A 2.5 mol/L sodium hydroxide aqueous solution (1.2 mL, 3.1 mmol) and
tetrabutylammonium bromide (0.012 g, 0.031 mmol) were added to 3-
chlorobenzenethiol (0.11
g, 0.73 mmol), followed by stirnng at 25°C for 10 minutes. A toluene
(1.2 mL) solution of
2-fluoro-5-nitrobenzaldehyde (0.12 g, 0.73 mmol) was added to the reaction
liquid, followed by
stirring at 110°C for 1.5 hours. After the conventional post-reaction
treatment, the residue was
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purified by silica gel chromatography (eluted by chloroform) to give 2-[(3-
chlorophenyl)thio]-5-nitrobenzaldehyde (72 mg, 34%).
~H NMR (300 MHz, CDC13) ( (ppm) 7. O1 (d, J = 8.8 Hz, 1H), 7.44-7.54 (m, 3H),
7.58
(br s, 1 H), 8.17 (dd, J = 2.4, 8.8 Hz, 1 H}, 8.69 (d, J = 2.6 Hz, 1 H), 10.29
(s, 1 H)
FABMS m/z 294 (M+H)+C,3Hg3sC1N03S = 293.
2-[(3-Chlorophenyl)thiol-5-nitrobenzaldehyde (70 mg, 0.24 mmol) was dissolved
in
toluene (3.5 mL). 2,4-Thiazolidinedione (0.11 g, 0.95 mmol), piperidine
(0.0094 mL, 0.095
mmol), acetic acid (0.0054 mL, 0.095 mmol) and molecular sieves 4A (0.35 g)
were added
thereto, followed by stirring at 110°C for 3 hours. After the
conventional post-reaction
treatment, the residue was purified by thin-layer chromatography (developed
with
chloroform/acetonitrile = 10/I ) to give Compound 70 (41 mg, 440).
'H NMR (300 MHz, DMSO-d6) 8 (ppm) 7.34 (d, J = 8.8 Hz, 1 H), 7.60-7.47 (m,
3H),
7.63 (br s, 1 H), 7.8 8 (s, 1 H), 8.20 (dd, J = 8.8, 2.6 Hz, 1 H), 8.26 (d, J
= 2.6 Hz, 1 H),
12.83 (m, 1 H)
FABMS m/z 391 (M-H)- C16H9ssC1NzO4Sz = 392
Example 98
Preparation of Compound 71
A 2.5 mol/L sodium hydroxide aqueous solution (1.7 mL, 4.4 mmol) and
tetrabutylammonium bromide (0.017 g, 0.051 mmol) were added to 2-
chlorobenzenethiol (0.17
g, 1.0 mmol), followed by stirring at 25°C for 10 minutes. To the
reaction liquid was added a
toluene ( 1.7 mL) solution of 2-fluoro-5-nitrobenzaldehyde (0.18 g, 1.0 mmol),
followed by
stirring at 110°C for 2 hours. After the conventional post-reaction
treatment, the residue was
purified by silica gel chromatography (eluted by chloroform) to give 2-[(2-
chlorophenyl)thin]-5-nitrobenzaldehyde (0.25 g, 83%).
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'H NMR (300 MHz, CDCl3) 8 (ppm) 6.88 (d, J = 8.8 Hz, 1H), 7.42 (ddd, J = 7.5,
7.5,
1.4 Hz, 1 H), 7.51 (ddd, J = 7.3, 7.3, 1.6 Hz, 1 H), 7.62 (dd, J = 8.1, 1.5
Hz, 1 H), 7.69
(dd, J =
7.5. 1.7 Hz, 1 H), 8.16 (dd, J = 8.8, 2.6 Hz, 1 H), 8.70 (d, J = 2.5 Hz, 1 H),
10.32 (s, 1 H)
FABMS m/z 293 (M+) C,3Hg35C1NO3S = 293
2-[(2-Chlorophenyl)thin]-5-nitrobenzaldehyde (0.14 g, 0.49 mmol) was dissolved
in
ethanol (5.8 mL). 2,4-Thiazolidinedione (0.23 g, 2.0 mmol} and piperidine
(0.039 mL, 0.39
mmol) were added thereto, followed by stirring at 80°C for 3 hours.
After the conventional
post-reaction treatment, the residue was purified by thin-layer chromatography
(developed with
chloroform/acetonitrile = 10/1) to give Compound 71 (24 mg, 13%).
'H NMR (300 MHz, DMSO-d6) 8 (ppm) 7.20 (d, J = 8.8 Hz, 1H), 7.43-7.62 (m, 3H),
7.71 (m, 1 H), 7.89 (s, 1 H), 8.20 (dd, J = 8.8, 2.6 Hz, 1 H), 8.28 (m, 1 H),
12.82 (m, 1 H)
FABMS m/z 391 (M-H)- Ci6H935C1N2O4S2 = 392
Example 99
Preparation of Compound 72
2-[(3,4-Dichlorophenyl)thio]-5-nitrobenzaldehyde (0.16 g, 79%) was obtained
from
3,4-dichlorobenzenethiol (0.12 g, 0.68 mmol), a 2.5 mol/L sodium hydroxide
aqueous solution
(1.2 mL, 2.9 mmol), tetrabutylammonium bromide (0.011 g, 0.034 mmol) and a
toluene (1.2
mL) solution of 2-fluoro-5-nitrobenzaldehyde (0.12 g, 0.68 rnmol) using the
same method as
Example 97.
'H NMR (300 MHz, CDC13) S (ppm) 7.02 (d, J = 8.8 Hz, 1H), 7.41 (dd, J = 8.2,
2.0 Hz,
1 H), 7.60 (d, J = 8.2 Hz, 1 H), 7.69 (d, J = 2.0 Hz, 1 H), 8.18 (dd, J = 8.8,
2.5 Hz, 1 H),
8.69 (d, J = 2.6 Hz, 1 H), 10.28 (s, 1 H)
FABMS m/z 328 (M+H)+ C,3H~3sC1zN03S = 327.
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Compound 72 (74 mg, 49%) was obtained from 2-[(3,4-dichlorophenyl)thio]-S-
nitrobenzaldehyde (0.12 g, 0.35 mmol), toluene (5.8 mL), 2,4-thiazolidinedione
(0.16 g, 1.4
mmol), piperidine (0.014 mL, 0.14 mmol), acetic acid (0.0080 mL, 0.14 mmol)
and molecular
sieves 4A (0.58 g) using the same method as Example 97.
'H NMR (300 MHz, DMSO-d6) 8 (ppm) 7.39 (d, J = 8.8 Hz, 1 H), 7.50 (dd, J =
8.4,
1.8 Hz, 1 H), 7.75 (d, J = 8.5 Hz, 1 H), 7.88-7.83 (m, 2H), 8.18 (d, J = 8.6,
2.2 Hz, 1 H),
8.25 (m, 1 H), 12.82 (m, 1 H)
FABMS m/z 425 (M-H)' C16Hg3sC 12N204S2 = 426
Example 100
Preparation of Compound 73
2-[(4-Bromophenyl)thio]-5-nitrobenzaldehyde (0.28 g, 82%) was obtained from
4-bromobenzenethiol (0.19 g, 1.0 mmol), a 2.5 mol/L sodium hydroxide aqueous
solution (1.7
mL, 4.3 mmol), tetrabutylammonium bromide (0.016 g, 0.051 mmol) and a toluene
( 1.7 mL)
solution of 2-fluoro-5-nitrobenzaldehyde (0.17 g, 1.0 mmol) using the same
method as
Example 97.
'H NMR (300 MHz, CDC13) 8 (ppm) 6.98 (d, J = 8.9 Hz, 1H), 7.41-7.46 (m, 2H),
7.63-7.69 (m, 2H), 8.13 (dd, J = 8.8, 2.4 Hz, 1 H), 8.68 (d, J = 2.6 Hz, 1 H),
10.29 (s, 1 H)
FABMS m/z 338 (M+H)+ C,3Hg~9BrN03S = 337.
Compound 73 (26 mg, 66%) was obtained from 2-[(4-bromophenyl)thio]-
5-nitrobenzaldehyde (0.031 g, 0.091 mmol), toluene (1.5 mL), 2.4-
thiazolidinedlone (0.043 g,
0.36 mmol), piperidine (0.0036 mL, 0.036 mmol), acetic acid (0.0025 mL, 0.036
mmol) and
molecular sieves 4A (0-092 g) using the same method an Example 97.
Compound 73: 'H NMR (300 MHz, DMSO-d6) S (ppm) 7.25 (d, J = 8.8 Hz, 1H),
7.48-7.52 (m, 2H), 7.20-7.25 (m, 2H), 7.88 (s, 1 H), 8.18 (dd, J = 8.8, 2.6
Hz, 1 H), 8.25
(d, J = 2.4 Hz, 1 H), 12.85 (br s, 1 H)
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FABMS m/z 435 (M-H)' C16H9~9BrN2OoS2 = 436.
Example 101
Preparation of Compound 74
2-[(4-Methoxyphenyl)thio]-5-nitrobenzaldehyde (0.22 g, 76%) was obtained from
4-methoxybenzenethiol (0.14 g, 0.99 mmol), a 2.5 mol/L sodium hydroxide
aqueous solution
(1.8 mL, 4.4 mmol), tetrabutylammonium bromide (0.016 g, 0.050 mmol) and a
toluene (1.8
mL) solution of 2-fluoro-5-nitrobenzaldehyde (0.17 g. 0.99 mmol) using the
same method
as Example 98.
~ H NMR (300 MHz, CDC 13) b (ppm) 3.89 (s, 3H), 6.92 (d, J = 8.9 Hz, 1 H),
7.03 (d, J =
8.6 Hz, 2H), 7.49 (d, J = 8.6 Hz, 2H), 8.08 (dd, J = 9.0, 2.4 Hz, 1 H), 8.63
(d, J = 2.2 Hz,
1 H), 10.29 (s, 1 H)
FABMS m/z 289 (M+) C,4H"N204 = 289.
Compound 74 (26 mg, 18%) was obtained from 2-[(4-methoxyphenyl)thio]-5
nitrobenzaldehyde (0.11 g, 0.39 mmol) ethanol (4.4 mL), 2,4-thiazolidinedione
(0.18 g, 1.5
mmol), and piperidine (0.015 mL, 0.15 mmol) using the same method as Example
98.
1H NMR (300 MHz, DMSO-d6) 8 (ppm) 3.34 (s, 3H), 6.98 (d, J = 8.8 Hz, 1 H),
7.13 (d,
J = 9.0 Hz, 2H), 7.57 (d, J = 8.6 Hz, 2H), 7.89 (s, 1 H), 8.14 (dd, J = 8.6,
2.4 Hz, 1 H),
8.21 (d, J = 2.4 Hz, 1 H)
FABMS m/z 387 (M-H)~ C,~H,ZN205Sz = 388
EXamDle 102
Preparation of Compound 75
2-[(4-Ethylphenyl)thio]-5-nitrobenzaldehyde (0.22 g, 82%) was obtained from
4-ethylbenzenethiol (0.13 g, 0.95 mmol), a 2.5 mol/L sodium hydroxide aqueous
solution (1.6
mL, 4.0 mmol), tetrabutylammonium bromide (0.015 g, 0.047 mmol) and a toluene
( 1.6 mL)
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solution of 2-fluoro-5-nitrobenzaldehyde (0.16 g, 0.95 mmol) using the same
method as
Example 97.
~H NMR (300 MHz, CDC13) 8 (ppm) 1.30 (t, J = 7.6 Hz, 3H), 2.74 (q, J = 7.5 Hz,
2H),
6.96 (d, J = 9.0 Hz, 1H), 7.36 (d, J = 7.5 Hz, 2H), 7.48 (d, J = 8.1 Hz, 2H),
8.10 (dd, J =
9Ø 2.2 Hz, 1 H), 8.66 (d, J = 2.6 Hz, 1 H), 10.31 (s, 1 H)
FABMS m/z 288 (M+H)+ C,SH,3N03S = 287.
Compound 75 (0.14 g, 51 %} was obtained from 2-[(4-ethylphenyl)thio]-5-
nitrobenzaldehyde (0.20 g, 0.69 mmol), toluene (9.9 mL), 2,4-thiazolidinedione
(0.32 g, 2.7
mmol), piperidine (0.027 mL, 0.27 mmol), acetic acid (0.016 mL, 0.27 mmol) and
a molecular
sieve 4A (0.99 g) using the same method as Example 97.
Compound 75: 1H NMR (300 MHz, DMSO-d6) 8 (ppm) 1.21 (t, J = 7.7 Hz, 3H), 2.68
(q, J = 7.7 Hz, 2H), 7.09 (d, J = 8.8 Hz, 1H), 7.39 (d, J = 8.1 Hz, 2H), 7.51
(d, J = 8.1
Hz, 2H), 7.39 (s, 1 H), 8.16 (dd, J = 8.8, 2.3 Hz, 1 H), 8.22 (d, J = 2.0 Hz,
1 H), 12.82 (m,
1 H)
FABMS m/z 385 (M-H)' C,8H14N2O4S2 = 386.
Example 103
Preparation of Compound 76
Compound 76 (0.134 g, 35.9%) was obtained by using commerciallyavailable
(MAYBRIDGE, Catalog Number: XAX00131 ) 2-benzylthio-5-nitrobenzaldehyde (0.27
g, 1.0
mmol) using the same method as Example 94.
'H-NMR(300MHz, DMSO-d6) 8 (ppm) 4.51 (s, 2H), 7.31 (m, 3H),7.46 (dt, J = 1.7,
6.6Hz, 2H), 7.78 (s, 1 H), 7.81 (d, J = 8.9Hz, 1 H), 8.17 (d, J = 2.6Hz, 1 H),
8.22 (dd,
J = 2.6, 8.9Hz, 1 H), 12.82 (br s, 1 H)
EIMS m/z 371(M-H)' Ci~HizN2O4S2 = 372.
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Example 104
Preparation of Compound 77
Compound 77 (0.0055 g, 17.6%) was obtained by using Compound 76 (0.03 g, 0.081
mmol) using the same method as Example 96.
'H-NMR, (300 MHz, DMSO-d6) S (ppm) 4.24 (d, J = 6.8Hz, 2H), 6.94 (dd, J = 2:2,
S.OHz, 2H}, 7.20 (dd, J = 2.2, 5.0 Hz, 3H), 7.4 (s, 1H), 7.89 (d, J = 8.7Hz,
1H), 8.21
d, J = 2.2 Hz, 1 H), 8.38 (dd, J = 2.2, 8.7Hz, 1 H), NH is not found
EIMS m/z 369(M+H)' C17H,ZN2OSS2 = 388.
Example 105
Preparation of Compound 78
Compound 78 (0.0192 g, 59.0%) was obtained by using Compound 76 (0.03 g, 0.081
mmol) using the same method as Example 96.
~H-NMR (300 MHz, DMSO-d6) 8 (ppm) 4.78 (s, 2H), 7.10 (dd, J = 1.8, 4.5 Hz,
2H),
7.27 (dd, J = 1.8, 4.5 Hz, 3H), 8.06 (d, J = 8.8 Hz, 1H), 8.08 (s, 1H), 8.36
(dd, J = 2.5,
8.8 Hz, 1 H), 8.39 (d, J = 2.5 Hz, 1 H), NH is not found
EIMS m/z 403(M-H)' C,~H,2NZO6S2 = 404.
Example 106
Preparation of Compound 79
2-[(4-Chlorobenzyl)thio]-5-nitrobenzaldehyde (0.17 g, 67%) was obtained from
4-chlorobenzylthiol (0.11 g, 0.83 mmol), a 2.5 moUL sodium hydroxide aqueous
solution ( 1.4
mL, 3.5 mmol), tetrabutylammonium bromide (0.013 g, 0.041 mmol) and a toluene
(1.4 mL)
solution of 2-fluoro-5-nitrobenzaldehyde (0.14 g, 0.83 mmol) using the same
method as
Example 97. Compound 79 (0.11 g, 70%) was obtained from
2-[(4-chlorobenzyl)thio]-5-nitrobenzaldehyde (0.12 g, 0.39 mmol), toluene (6.1
mL),
2,4-thiazolidinedione (0.18 g, 1.6 mmol), piperidine (0.016 mL, 0.16 mmol),
acetic acid
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{0.0090 mL, 0.16 mmol) and molecular sieves 4A (0.61 g) using the same method
as Example
97.
2-[{4-chlorobenzyl)thio]-5-nitrobenzaldehyde:
'H NMR (300 MHz, CDC13) b (ppm) 4.26 (s, 2H), 7.34 (s, 4H), 7.49 (d, J = 8.8
Hz,
1 H), 8.29 (dd, J = 8.8, 2.6 Hz, 1 H), 8.65 (d, J= 2.5 Hz, 1 H), 10.23 (s, 1
H)
FABMS m/z 307 {M~ C,aHio35C1N03S = 307.
Compound 79:
~H NMR (300 MHz, DMSO-d6) b (ppm) 4.50 (s, 2H), 7.39 (d, J = 8.5 Hz, 2H), 7.46
(d,
J = 8.4 Hz, 2H), 7.77 (s, 1 H), 7.80 (d, J = 8.8 Hz, 1 H), 8.17 (d, J = 2.4
Hz, 1 H), 8.21
(dd, J = 9.0, 2.6 Hz, 1 H), 12.81 (m, 1 H)
FABMS m/z 405 (M-H)' C,7H,~35C1N2OqS2 = 406.
Example 107
Preparation of Compound 80
Commercially available (MAYBRIDGE, Catalog Number: XAX00146)
4-[(4-bromophenyl)thio]-3-nitrobenzaldehyde (0.12 g, 0.35 mmol) was dissolved
in toluene
(5.9 mL). 2,4-Thiazolidinedione (0.16 g, 1.4 mmol), piperidine (0.014 mL, 0.14
mmol), acetic
acid (0.0080mL, 0.14 mmol) and molecular sieves 4A (0.59 g) were added
thereto, followed by
stirring at 110°C for 3 hours. After the conventional post-reaction
treatment, the residue was
triturated by using ethanol to give Compound 80 (21 mg, 14%).
'H NMR (300 MHz, DMSO-d6) 8 (ppm) 7.01 (d, J = 8.6 Hz, 1H), 7.58-7.62 (m, 2H),
7.73-7.82 (m, 4H), 8.50 (d, J = 2.0 Hz, 1 H), 12.71 (m, 1 H)
FABMS m/z 435 (M-H)' C~6Hg~9BrNzO4S2 = 436.
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Example 108
Preparation of Compound 81
Commercially available (MAYBRIDGE, Catalog Number: NRB00117)
4-[(4-chlorophenyl)thio]-3-nitrobenzaldehyde (0.31 g, 1.0 mmol) was dissolved
in ethanol ( 12
mL). 2,4-Thiazolidinedione (0.16 g, 1.4 mmol) and piperidine (0.014 mL, 0.14
mmol) were
added thereto, followed by stirring at 80°C for 3 hours. The reaction
liquid was cooled to 25°C,
and the precipitated crystals were collected by filtration to give Compound 81
(0.15 g, 36%).
~H NMR (300 MHz, DMSO-d6) 8 (ppm) 6.98 (d, J = 8.5 Hz, 1H), 7.08 (br s, 1H),
7.70-7.59 (m, SH), 7.73 (dd, J = 8.6, 2.0 Hz, 1H), 8.45 (d, J = 2.0 Hz, 1H)
FABMS m/z 391 (M-H)' C,6H9ssC1NZO4S2 = 392.
Example 109
Preparation of Compaund 82
Compound 82 (0.0195 g, 62.0%) was obtained by using Compound 9 (0.03 g, 0.081
mmol) using the same method as Example 96.
'H-NMR(300MHz, DMSO-db) b (ppm) 2.49 (s, 3H), 7.31 (d, J = 8.2 Hz, 2H), 7.56
(d, J
= 8.2 Hz, 2H), 7.84 (s, 1 H), 8.28 (dd, J = 1.8, 8.4 Hz, 1 H), 8.46 (s, 1 H),
8.49 (t, J =
1.8 Hz, 1 H), NH is not found
EIMS m/z 389 (M+H)+ C,~H,ZN205S2 = 388.
Example 110
Preparation of Compound 83
Compound 9 (0.07 g, 0.19 mmol) was dissolved in dichloromethane (12 mL),
methanol
(2.3 mL) and m-chloroperbenzoic acid (0.13 g, 0.38 mmol) was added thereto,
followed by
stirnng at room temperature for 1 hour. A 10% sodium hydrogen sulfite aqueous
solution was
added thereto. After the conventional treatment, the residue was purified by
thin-layer
chromatography (chloroform: methanol = 12:1) followed by trituration with
chlorofrom to give
Compound 83 (0.019 g, 25.4%) by triturating with chloroform.
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'H-NMR (300 MHz, DMSO-d6) 8 (ppm) 2.41(s, 3H), 7.50 (d, J = 8.3 Hz, 2H), 7.84
(s, 1 H), 7.86 (d, J = 8.1 Hz, 2H), 8.03 (d, J = 8.3 Hz, 1 H), 8.23 (d, J =
1.7 Hz, IH), 8.44
(d, J = 8.3 Hz, 1 H) , NH is not found
EIMS m/z 405(M+H)+ C,~H,2NZO6S2 = 404.
Example 111
Preparation of Compound 84
Compound 84 (0.152 g, 41.8%) was obtained by using commercially available
(MAYBRIDGE, Catalog Number: XAX00135) 2-(cyclohexylthio)-5-nitrobenzaldehyde
(0.26
g, 1.0 mmol) using the same method as Example 94.
'H-NMR (300 MHz, DMSO-d6) 8 (ppm) 1.30 (m, 1H), 1.46 (m, 4H), 1.61 (m, 1H),
1.73 (m, 2H), 2.00 (m, 2H), 3.67 (m, 1 H), 7.79 (d, J = 8.5 Hz, 1 H), 7.83 (s,
1 H), 8.19 (d,
J = 2.4 Hz, 1 H), 8.22 (dd, J = 2.4, 8. S Hz, 1 H), 12.82 (br s, 1 H)
FABMS m/z 363(M-H}- C16Hi6N204Sz = 364.
Example 112
Preparation of Compound 85
Compound 85 (0.0149 g, 71.3%) was obtained by using Compound 84 (0.02 g, 0.055
mmol) using the same method as Example 96.
'H-NMR (300 MHz, DMSO-db) 8 (ppm) 1.30 (m, 8H), 1.77 (m, 2H), 2.73(m, 1H),
7.60
(s, 1 H), 8.24 (s, 1 H), 8.05 (d, J = 8.6 Hz, 1 H}, 8.36 (d, J = 2.2Hz, 1 H),
8.43 (dd, J = 2.2,
8.6 Hz, 1 H), NH is not found
FABMS m/z 381(M+H)' Ci6H,6N2O5S2 = 380.
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Example 113
Preparation of Compound 86
Compound 86 (0.0034 g, 10.9%) was obtained by using Compound 84 (0.03 g,
0.,082
mmol) using the same method as Example 96.
S
1H-NMR (300 MHz, DMSO-db) 8 (ppm) 1.27 (m, 6H), 1.59 (m, 1H), 1.69 (m, 4H),
8.23
(d, J = 8.4 Hz, 1 H), 8.24 (s, 1 H), 8.44 (s, 1 H), 8.46 (d, J = 8.4 Hz, 1 H),
NH is not found
EIMS m/z 395 (M-H)' Ci6Hi6NZO6S2 = 396.
Example 114
Preparation of Compound 87
p-Toluenethiol (0.12 g, 1.0 mmol) was dissolved in a 10% sodium hydroxide
aqueous
solution (1.7 mL), and tetrabutylammonium bromide (0.016 g, 0.05 mmol) was
added thereto,
followed by stirring at room temperature for 5 minutes. 5-Bromo-2-
fluorobenzaldehyde (0.2 g,
1.0 mmol) dissolved in toluene (1.7 mL) was added thereto by dropping,
followed by stirring at
room temperature for 4 hours. After the conventional treatment, and was
recrystallized and
purified with ethanol and hexane, to give 5-bromo-2-[(4-
methylphenyl)thio]benzaldehyde
(0.23 g, 74.1 %).
IHNMR (300 MHz, CDC13) 8 (ppm) 2.39 (s, 3H), 6.90 (d, J = 8.4 Hz, 1H), 7.22
(d,
J = 7.9 Hz, 2H), 7.35 (dd, J =1.7, 8.1 Hz, 2H), 7.46 (dd, J = 2.4, 8.4 Hz,
1H), 7.96 (d, J =
2.2 Hz, 1 H), 10.32 (s, 1 H).
FABMS m/z 308(M+I-n+ C,4I-Iil~9grOS = 307.
Compound 87 (0.021 g, 52%) was obtained by using 5-bromo-2-[(4-methylphenyl)-
thio]benzaldehyde (0.03 g, 0.1 mmol) using the same method as Example 94.
'H-NMR {300 MHz, DMSO-d6) b (ppm) 2.31 (m, 3H), 7.14 (d, J = 8.8 Hz, 1H), 8.23
(d,
J = 8.3 Hz, 2H), 8.28 {d, J = 8.3 Hz, 2H), 7.60 (s, 1 H), 7.62 (dd, J = 2.2,
8.8 Hz, 1 H),
7.90 (s, 1 H), 12.82 (br s, 1 H)
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EIMS m/z 407(M+H)+ C,7H,z'9grN02Sz = 406.
Example 115
Preparation of Compound 88
3-Bromo-4-[(4-methylphenyl)thio]benzaldehyde (0.19 g, 63.6%) was obtained by
using
3-bromo-4-fluorobenzaldehyde (0.2 g, 0.99 mmol) using the same method as
Example 114
(synthesis of 5-bromo-2-[(4-methylphenyl)thio]benzaldehyde).
'H-NMR (300 MHz, CDC13) 8 (ppm) 2.44 (s, 3H), 6.73 (d, J = 8.3 Hz, 1H), 7.30
(d, J =
8.4 Hz, 2H), 7.47 (d, J = 8.1 Hz, 2H), 7.54 (dd, J = 1.7, 8.4 Hz, 1 H), 8.00
(d, J = 1.7 Hz,
1 H), 9.84 (s, 1 H)
FABMS m/z 307(M~ C,4H"BrOS = 307.
Compound 88 (0.087 g, 43.9%) was obtained by using 3-bromo-4-[(4-methylphenyl)-
thin]benzaldehyde (0.15 g, 0.49 mmol) using the same method as Example 94.
~H-NMR (300 MHz, DMSO-d6) 8 (ppm) 2.49 (s, 3H), 6.74 (d, J = 8.3 Hz, 1H), 7.37
(d,
J = 8.1 Hz, 2H), 7.44 (dd, J = 2.0, 8.5 Hz, 1 H), 7.48 (d, J = 8.1 Hz, 2H),
7.70 (s, 1 H),
7.89 (d, J = 2.0 Hz, 1H), 12.64 (br s, 1 H)
EIMS m/z 407(M+H)+C~?H~2'9BrNO2S2 = 406.
Example 116
Preparation of Compound 89
The reaction was performed in accordance with a document (Tetrahedron Lett.
Vol. 36,
No. S0, pp. 9085-9088,.1995) and the product was treated by using 5-bromo-2-
[(4-
methylphenyl)thio]-benzaldehyde (0.06 g, 0.2 mmol) and 2-pyridyl
trifluoromethanesulfonate
(0.03 g, 0.2 mmol). The residue was purified by thin-layer chromatography
(hexane: ethyl
acetate = 8:1)) to obtain 2-[(4-methylphenyl)thio)-5-(2-pyridyl)benzaldehyde
(0.024 g, 38.5%}.
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'H-NMR (300 MHz,CDC 13) 8 (ppm) 2.41 (s, 3H), 7.09 (d, J = 8.4 Hz, 1 H), 7.24
(m,
3H), 7.42 (d, J = 8.4 Hz, 2H), 7.76 (dd, J = 1.7, 6.6 Hz, 2H), 8.01 (dd, J =
2.2, 8.5 Hz,
1 H), 8.50 (d, J = 2.2 Hz, 1 H), 8.69 (d, J = 4.6 Hz, 1 H), 10.43 (s, 1 H)
EIMS m/z 306(M+H)+ C,9H,sNOS = 305.
Compound 89 (0.026 g, 84.4%) was obtained by using 2-[(4-methylphenyl)thio]-
5-(2-pyridyl)benzaldehyde (0.024 g, 0.08 mmol) using the same method as
Example 94.
'H-NMR (300 MHz, DMSO-db) 8 (ppm) 2.32 (s, 3H), 7.26 (d, J = 8.4 Hz, 2H), 7.30
(d, J = 8.4 Hz, 2H),7.33 (d, J = 8.3 Hz, 2H), 7.41 (m, 1 H), 7.92 (m, 1 H),
7.97 (d, J = 7.7
Hz, 1 H), 8.08 (s, 1 H), 8.07 (dd, J = 2.0, 7.7 Hz, 1 H), 8.32 (d, J = 1.7 Hz,
1 H), 8.71
(d, J = 4.8 Hz, 1 H), 12.70 (br s, 1 H)
EIMS m/z 405 (M-H)- Cz2Hi6N2OZS2 = 406.
1 S Example 117
Preparation of Compound 90
Tri(dibenzylideneacetone)-dipalladium (0.18 g, 0.2 mmol), and
triphenylphosphine
(0.21 g, 0.8 mmol) were dissolved in tetrahydrofuran (60 mL), followed by
stirring at room
temperature for 30 minutes. Then, 5-brorno-2-fluorobenzaldehyde (0.4 g, 2.0
mmol) and
2-(tributylstannyl)-furan ( 1.25 mL, 4. 0 mmol) were added thereto, followed
by heating under
reflux for 10 hours. The mixture was cooled to room temperature, and after the
conventional
treatment, the residue was purified by silica gel column chromatography
(hexane: ethyl acetate
= 8:1), to give 2-fluoro-5-(2-furyl)benzaldehyde (0.38 g, 100%).
'H-NMR (300 MHz, CDC13) S (ppm) 6.49 (dd, J = 1.8, 3.5 Hz, 1 H), 6.69 (d, J =
3.3Hz,
1 H), 7.21 (t, J = 9.9 Hz, 1 H), 6.49 (d, J = 1.8 Hz, 1 H), 7.90 (m, 1 H),
8.14 (dd, J = 2.4,
6.6 Hz, 1 H), 10.39 (s, 1 H) .
FABMS m/z 190(M+) C"H~I9F02 = 190
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2-[(4-Methylphenyl)thio]-5-(2-furyl)benzaldehyde (0.14 g, 87.8%) was obtained
by
using 2-fluoro-5-(2-furyl)benzaldehyde (0.1 g, 0.53 mmol) using the same
method as Example
114 (synthesis of 5-bromo-2-[(4-methylphenyl)thio]benzaldehyde).
'H-NMR (300 MHz, CDCI~) 8 (ppm) 2.39 (s, 3H), 6.59 (d, J = l.BHz, 1H), 6.69
(d, J
= 3.3 Hz, 1 H), 7.07 (d, J = 8.4 Hz, 1 H), 7.21 (d, J = 8.3 Hz, 2H), 7.36 (d,
J = 8.1 Hz, 2H)
7.48 (s, 1 H), 7.66 (dd, J = 2.0, 8.3 Hz, 1 H), 8.14 (d, J = 2.0 Hz, 1
H),10.41 (s, 1 H)
FABMS m/z 294(M+) ClgH,4O2S = 294
Compound 90 (0.17 g, 92.8%) was obtained by using 2-[(4-methylphenyl)thio]-5
(2-furyl)benzaldehyde (0.14 g, 0.47 mmol) using the same method as Example 94.
'H-NMR (300 MHz, DMSO-d6) b (ppm) 2.50 (s, 3H), 6.65 (m, 1H), 7.04 (d, J = 3.5
Hz,
1 H), 7.22 (d, J = 8.4 Hz, 2H), 7.26 (d, J = 8.4Hz, 2H), 7.32 (d, J = 8.4 Hz,
1 H),7.74 (d, J
= 8.2 Hz, 1 H), 7.81 (d, J = 7.7 Hz, 1 H), 8.03 (s, 1 H), 12.71 (br s, 1 H)
EIMS m/z 392 (M-H)' C2,HISN03Sz = 393.
Example 118
Preparation of Compound 91
Compound 91 (0.069 g, 66.5%) was obtained by using compound 74 (0.1 g, 0.25
mmol)
using the same method as Example 96.
1H-NMR (300 MHz, DMSO-d6) S (ppm) 2.30 (s, 3H), 6.67 (dd, J = 1.8, 3.5 Hz,
1H),
7.17 (d, J = 8.6 Hz, 2H), 7.31 (d, J = 8.6 Hz, 2H), 7.45 (d, J = 8.3 Hz, 2H),
7.76 (s, 1 H),
7.87 (d, J = 1.8 Hz, 1 H) , 8.01 (d, J = 5.9 Hz, 2H}, 8.02 (s, 1 H), 12.77 (br
s, 1 H)
EIMS m/z 410 (M+H)+, C2,H,SN04S2 = 409.
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Example 119
Preparation of Compound 92
2-Fluoro-5-(2-thienyl)benzaldehyde (0.2 g, 100%) was obtained by using
2-(triutylstannyl)thiophene (0.63 mL, 2.0 mmol) using the same method as
Example 117
(synthesis of 2-fluoro-5-(2-furyl)benzaldehyde).
1H-NMR (300 MHz, CDC 13) 8 (ppm) 7.10 (t, J = 4.4 Hz, 1 H}, 7.21 (dd, J = 8.8,
9.9 Hz,
1 H), 7. 3 3 (d, J = 4.4 Hz, 2H), 7. 83 (m, 1 H), 8.08 (dd, J = 2.6, 6. 5 Hz,
1 H}, 10.40 (s, 1 H)
FABMS m/z 206 (M~ C~,H~~9FOS = 206.
2-[(4-Methylphenyl)thio]-5-(2-thienyl)benzaldehyde (0.12 g, 77.4%) was
obtained by
using 2-fluoro-5-(2-thienyl)benzaldehyde (0.1 g, 0.49 mmol) using the same
method as
Example 114 (synthesis of 5-bromo-2-[(4-methylphenyl)thioJbenzaldehyde).
~H-NMR (300 MHz, CDC13) 8 (ppm) 2.39 (s, 3H), 7.06 (d, J = 8.3 Hz, 1H), 7.10
(m,
1 H), 7.24 (d, J = 8.3 Hz, 2H), 7.33 (m, 4H), 7.37 (d, J = 8.1 Hz, 2H) , 7.60
(dd, J = 2.4,
8.3 Hz, 1 H), 8.07 (d, J = 2.4 Hz, 1 H),10.42 (s, 1 H)
FABMS m/z 310 (M+) Ci8H,40S2 = 310.
Compound 92 (0.14 g, 91.3%) was obtained by using 2-[(4-methylphenyl)thiol-5-
(2-thienyl)benzaldehyde (0.12 g, 0.38 mmol) using the same method as Example
94.
~H-NMR (300 MHz, DMSO-d6) 8 (ppm) 2.30 (s, 3H), 7.18 (t, J = 3.8 Hz, 1H), 7.24
(d,
J = 4.8 Hz, 4H), 7.56 (d, J = 7.7 Hz, 1 H), 7.63 (d, J = 5.1 Hz, 1 H), 7.72
(s, 1 H), 7.73 (d,
J = 7.5 Hz, 1 H), 8.03 (s, 1 H) , 12.71 (br s, 1 H)
EIMS m/z 408 (M-H)- C2iH,sNOZS3 = 409.
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Example 120
Preparation of Compound 93
Compound 93 (0.051 g, 49.4%) was obtained by using Compound 92 (0.1 g, 0.24 .
mmol) using the same method as Example 96.
1H-NMR (300 MHz, DMSO-d6) 8 (ppm) 2.30 (s, 3H), 7.20 (dd, J = 3.7, 5.0 Hz,
1H),
7.32 (d, J = 8.1 Hz, 2H), 7.46 (d, J = 8.3 Hz, 2H), 7.66 (d, J = 3.7 Hz, 1 H),
7.69 (d, J =
3.7 Hz, 2H), 8.01 (m, 3H), 12.78 (br s, 1H)
EIMS m/z 426 (M+H)+ CZ~H15N03S3 = 425.
Example 121
Preparation of Compound 94
Diisopropylamine (0.35 mL, 2.5 mmol) was dissolved in tetrahydrofuran (3.5 mL)
and
the temperature was adjusted to 0°C. Then, n-butyllithium (hexane
solution) (1.24 mL, 2.0
mmol) was dropped thereto, followed by stirring for 10 minutes. Thereafter the
reaction
temperature was brought to -78°C. 4-Fluorobenzonitile (0.2 g, 1.65
mmol) was added thereto,
stirnng for 1 hour. Then, dimethylformamide (0.19 mL, 2.5 mmol) was dropped
thereto,
followed by stirring for 20 minutes and then the conventional treatment was
performed. The
residue was purified by thin-layer chromatography (hexane: ethyl acetate = 8:1
), to give
2-fluoro-5-cyanobenzaldehyde (0.11 g, 43.4%).
' H-NMR (300 MHz, CDC 13) 8 (ppm) 7.46 (t, J = 8.8 Hz, 1 H), 7.90 (m, 1 H),
7.71 (dd, J
= 2.2, 6.2 Hz, 1 H), 10.3 6 (s, 1 H)
EIMS m/z 148(M-H) CgH4~9FNO = 149
5-Cyano-2-[(4-methylphenyl)thio]benzaldehyde (0.15 g, 86.9%) was obtained by
using
2-fluoro-5-cyanobenzaldehyde (0.1 g, 0.67 mmol) using the same method as
Example 114
(synthesis of 5-bromo-2-[(4-methylphenyl)thio]benzaldehyde).
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'H-NMR (300 MHz, CDC 13) 8 (ppm) 2.44 (s, 3H), 6.92 (d, J = 8.6 Hz, 1 H), 7.31
(d,
J = 7.9 Hz, 2H), 7.44 (d, J = 8.3 Hz, 2H), 7.49 (dd, J = 1.8, 8.4 Hz, 1 H),
8.07 (d, J =
2.0 Hz, 1 H), 10.27 (s, 1 H)
EIMS m/z 253(M~ C,SH,1NOS = 253.
Compound 94 (0.09 g, 64.5%) was obtained by using 5-cyano-2-[(4-methylphenyl)-
thio]-benzaldehyde (0.1 g, 0.4 mmol) using the same method as Example 94.
'H-NMR (300 MHz, DMSO-d6) b (ppm) 2.37 (s, 3H), 7.00 (d, J = 8.3 Hz, 1H), 7.34
(d,
J = 8.3 Hz, 2H), 7.46 (d, J = 8.3 Hz, 2H), 7.76 (dd, J = 1.7, 8.4 Hz, 1 H),
7.80 (s, 1 H), .
7.84 (s, 1 H), 12.01 (br s, 1 H)
EIMS m/z 353 (M+H)+ C,gH~2N20zS2 = 352.
Example 122
Preparation of Compound 95
3-Cyano-4-[(4-methylphenyl)thio]benzaldehyde (0.3 g, 89.9%) was obtained by
using
2-fluoro-5-formylbenzonnitrile (0.2 g, 1.34 mmol) using the same method as
Example 114
(synthesis of S-bromo-2-[(4-methylphenyl)thio]benzaldehyde).
'H-NMR (300 MHz, CDC13) 8 (ppm) 2.44 (s, 3H), 6.94 (d, J = 8.4 Hz, 1H), 7.31
(d, J =
7.9 Hz, 2H), 7.47 (dd, J = 1.8, 8.1 Hz, 2H), 7.78 (dd, J = 1.8, 8.4 Hz, 1 H),
8.07 (d, J =
S Hz, 1 H), 9.90 (s, 1 H)
FABMS m/z 254 (M+H)+ CISH,~NOS = 253.
Compound 95 (0.126 g, 45.3%) was obtained by using 3-cyano-4-[(4-methylphanyl)-
thio]benzaldehyde (0.2 g, 0.79 mmol) using the same method as Example 94.
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1H-NMR (300 MHz, DMSO-d6) S (ppm) 2.50 (s, 3H), 7.06 (d, J = 8.6 Hz, 1H), 7.36
(d,
J = 8.4 Hz, 2H), 7.49 (d, J = 8.3 Hz, 2H), 7.66 (s, 1 H), 7.71 (dd, J = 2.0,
8.5 Hz, 1 H),
8.08 (d, J = 2.0 Hz,1H), NH is not found
EIMS m/z 352 (M+) CigH~2NZO2S2 = 352.
Example 123
Preparation of Compound 96
3-Bromo-4-[(4-methylphenyl}thio]benzaldehyde (0.18 g, 59.0%) was obtained by
using
3-bromo-4-fuluorobenzaldehyde (0.12 g, 1.0 mmol) using the same method an
Example 114
(synthesis of 5-bromo-2-[(4-methylphenyl)thio]benzaldehyde).
~ H-NMR (300 MHz, CDC 13) 8 {ppm) 2.44 (s, 3H), 6.73 (d, J = 8.3 Hz, 1 H),
7.30 (d, J =
7.9 Hz, 2H), 7.47 (d, J = 8.1 Hz, 2H), 7.54 (dd, J = 1.7, 8.3 Hz, 1 H), 7.99
(d, J = 1.7 Hz,
1 H), 9.84 (s, 1 H)
EIMS m/z 307(M+) C14H1179BrOS = 307.
3-Bromo-4-[(4-methylphenyl)thio]benzaldehyde (0.2 g, 0.65 mmol) was dissolved
in
methanol (6.0 mL) and dichloromethane (6.0 mL). Sodium borohydride (0.025 g,
0.65 mmol)
was added thereto, followed by stirring for 15 minutes and the conventional
treatment was
performed. The solvent was removed by using a vacuum dryer. The residue was
dissolved in
dichloromethane (7.0 mL). Tert-butyldimethylsilylchloride (0.12 g, 0.78 mmol)
and imidazole
(0.053 g, 0.65 mmol} were added thereto, followed by stirring for 2 hours and
then the
conventional treatment was performed. The product was purified by silica gel
chromatography
(hexane: ethyl acetate = 16:1), to give
3-bromo-4-[(4-methylpheny)thio]benzyl(tent-butyldimethylsilyl}ether (0.27 g,
96.4%).
1H-NMR (300 MHz, CDC13) ( 0.62 (s, 6H), 0.84 (s, 9H), 2.29 (s, 3H), 4.56 (s,
2H),
6.76 (d, J = 8.3 Hz, 1 H), 7.00 (dd, J = 1.8, 8.3 Hz, l H), 7.11 (d, J = 8.4
Hz, 2H), 7.26 (d,
J = 1.8 Hz, 2H}, 7.43 (d, J = 1.7 Hz, 1 H)
EIMS m/z 424 (M+H)+ C20H2779BrOSSi = 423.
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In an argon atmosphere, hexane ( 1.4 mL) and diethyl ether ( 10.4 mL) was
cooled to
-78°C. n-Butyl lihlum (hexane solution) (2.17 mL, 3.47 mmol) and
tetramethylethylene-
diamine (0.52 mL, 3.47 mmol) were added thereto, followed by stirring for 15
minutes.
3-Bromo-4-[4-methylphenyl)thio]benzyl tert-butyldimethylsilyl ether (0.37 mL,
0.87 mmol)
was added thereto, further followed by stirring for 30 minutes. Then,
dimethylformamide (0.3
mL, 3.47 mmol) dissolved in diethylether (4.3 mL) was added thereto by
dropping, followed by
stirnng for 45 minutes. After heating the mixture up to room temperature, the
conventional
treatment was performed. The product was purified by thin-layer chromatography
(hexane:
ethyl acetate = 16:1), to give 5-hydroxymethyl-2-[(4-
methylphenyl)thioJbenzaldehyde (0.58 g,
66.7%).
1H-NMR (300 MHz, CDC13) ( (ppm) 0.62 (s, 6H), 0.84 (s, 9H), 2.28 ~(s, 3H);
4.64 (s,
2H), 6.95 (d, J = 8.1 Hz, 1 H), 7.09 (d, J = 7.9 Hz, 2H), 7.23 (d, J = 8.3 Hz,
2H), 7.28
(dd, J = 2.2, 8.2 Hz, 1 H), 7.70 (d, J = 1.8 Hz, 1 H), 10.29 (9, 1 H)
EIMS m/z 373(M+H)+ C2,H2g02SSi = 372.
Compound 96 (0.067 g, 70.4%) was obtained by using 5-hydroxymethyl-2-[(4-
methyl-
phenyl)thio]benzaldehyde (0.1 g, 0.27 mmol) using the same method as Example
94.
~H-NMR (300 MHz, DMSO-d6) b (ppm) 2.27 (s, 3H), 4.53 (d, J = 5.5 Hz, 2H), 5.37
(t,
J = 5.5 Hz, 1 H), 7.16 (d, J = 3.5 Hz, 4H), 7.31 (s, 1 H), 7. S 8 (s, 1 H),
7.98 (s, 1 H), 12.00
(br s, 1 H)
EIMS m/z 356(M-H)- CIgH,5N03Sz = 357.
Example 124
Preparation of Compound 97
In an argon atmosphere, 5-bromo-2-fluorobenzaldehyde (0.2 g, 1.0 mmol) was
dissolved in toluene (2 mL). Tributyl(1-ethoxyvlnyltin) (0:37 mL, 1.1 mmol)
and
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bis(triphenylphosphine)palladiumchloride (0.007 g, 0.01 mmol) were added
thereto, followed
by heating at 100°C for 10 hours. Then the conventional treatment was
performed when the
temperature of the residue fell to room temperature. The product was purified
by thin-layer
chromatography (hexane: ethyl acetate = 8:1), to give 5-acetyl-2-
fuluorobenzaldehyde (0.19 g,
58.3%).
'H-NMR (300 MHz, CDC 13) 8 (ppm) 2.65 (s, 3H), 7.31 (d, J = 9.3 Hz, 1 H), 8.27
(m, 1 H),
8.45 (dd, J = 2.4, 6.6 Hz, 1 H), 10.40 (s, 1 H)
CIMS m/z 167(M+H)+ C9H~'9F02 = 166.
5-Acetyl-2-[(4-methylphenyl)thio]benzaldehyde (0.043 g, 26.5%) was obtained by
using 5-acetyl-2-fuluorobenzaldehyde (O.I g, 0.6 mmol) using the same method
as Example
114 (synthesis of 5-bromo-2-[(4-methylphenyl)thin]benzaldehyde).
'H NMR (300 MHz, CDC13) S (ppm) 2.43 (s, 3H), 2.60 (s, 3H), 6.93 (d, J = 8.6
Hz,
1 H), 7.29 (d, J = 7.9 Hz, 2H), 7.44 (d, J = 8.3 Hz, 2H), 7.86 (dd, J = 8.5,
2.0 Hz, 1 H),
8.38 (d, J = 2.0 Hz, 1 H), 10.34 (s, 1 H)
FABMS m/z 271 (M+H)+ C16H1402S = 270.
Compound 97 (0.059 g,99.8%) was obtained by using 5-acetyl-2-[(4-methylphenyl)-
thio]benzaldehyde (0.043 g, 0.16 mmol) using the same method as Example 94.
'H-NMR (300 MHz, DMSO-d6) b (ppm) 2.49 (s, 3H), 2.51 (s, 3H), 7.09 (d, J = 8.3
Hz,
1 H), 7.31 (d, J = 8.3 Hz, 2H), 7.41 (d, J = 6.8 Hz, 2H), 7.89 (d, J = 8.6 Hz,
1 H), 7.95 (s,
1 H), 8.01 (s, 1 H), 12.00 (br s, 1 H)
EIMS m/z 370 (M+H)+ Ci9Hi5N03S2 = 369.
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Example 125
Preparation of Compound 98
Compound 98 (0.040 g, 49.8%) was obtained by using Compound 97 (0.083 g, 0.21
mmol) using the same method as Example 96.
'H-NMR (300 MHz, DMSO-d6) 8 (ppm) 2.29 (s, 3H), 2.50 (s, 3H}, 7.30 (d, J = 8.3
Hz,
2H), 7.47 (d, J = 7.9 Hz, 2H), 7.78 (s, 1 H), 7.88 (s, 1 H), 8.04 (s, 1 H),
8.16 (s, 1 H), 8.17
(d, J = 3.3 Hz, 1 H), NH is not found
EIMS mlz 386(M+H)+ C,9H1sN04S2 = 385.
Example 126
Preparation of Compound 98
2-[(4-Methylphenyl)thio]-6-(trifluoromethyl)benzaldehyde (0.44 g, 100%) was
obtained
by using 2-fluoro-6-(trifluoromethyl)benzaldehyde (0.19 g, 1.5 mmol) using the
same method
as Example 114 (synthesis of 5-bromo-2-[(4-methylphenyl)thio]benzaldehyde).
'H-NMR (300 MHz, CDC13) 8 (ppm) 2.42 (s, 3H), 7.08 (d, J = 8.3 Hz, 1H), 7.27
(d, J =
7.3 Hz, 2H), 7.33 (t, J = 7.7 Hz, 1 H), 7.43 (d, J = 8.1 Hz, 2H), 7.50 (d, J =
7.7 Hz, 1 H),
10.53 (q, J = 2.2 Hz, 1 H)
EIMS m/z 296 (M+) C~sH"19F30S = 296.
Compound 99 (0.11 g, 81.2%) was obtained by using 2-[(4-methylphenyl)thio]-6-
(trifluoromethyl)benzaldehyde (0.1 g, 0.3 mmol) using the same method as
Example 94
(synthesis of Compound 67).
~H-NMR (300 MHz, DMSO-d6) 8 (ppm) 2.34 (s, 3H), 7.23 (d, J = 7.7 Hz, 1H), 7.29
(d,
J = 7.9 Hz, 2H), 7.37 (d, J = 8.3 Hz, 2H), 7.51 (t, J = 7.7 Hz, 1 H), 7.66 (d,
J = 7.7 Hz,
1 H), 7.70 (s, 1 H), 12.00 (br s, 1 H)
EIMS m/z 394 (M-H)' CiBH,z~9FNOzS2 = 395.
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Example 127
Preparation of Compound 100
2-[(4-Methylphenyl)thio]-S-(trifluoromethyl)benzaldehyde (0.45 g, 86.4%) was
obtained by using 2-fluoro-5(trifluoromethyl)benzaldehyde (0.1 g , 0.5 mmol)
using the same
method as Example 114 (synthesis of 5-bromo-2-[(4-
methylphenyl)thioJbenzaldehyde).
~H-NMR (300 MHz, CDC13) 8 (ppm) 2.43 (s, 3H), 6.99 (d, J = 8.6 Hz, 1H), 7.28
(d, J
=11.0 Hz, 2H), 7.43 (d, J = 8.1 Hz, 2H), 7.52 (d, J = 8.4 Hz, 1 H), 8.07 (s, 1
H), 10.34 (s, 1 H)
FABMS m/z 296 (M~ CASH" ~9F30S = 296
Compound 100 (0.068 g, 50.9%) was obtained by using 2-[(4-methylphenyl)thio]-
5-(trifluoromethyl)benzaldehyde (0.1 g, 0.34 mmol) using the same method as
Example 94.
'H-NMR (300 MHz, DMSO-db) 8 (ppm) 2.35 (s, 3H), 7.15 (d, J = 8.8 Hz, 1H), 7.32
(d,
J = 7.7 Hz, 2H), 7.43 (d, J = 7.7 Hz, 2H), 7.70 (s, 1 H),7.71 (d, J = 7.0 Hz,
1 H), 7.92 (s,
1 H), 12.80 (br s, 1 H)
EIMS m/z 394(M-H)' CigH12~9F3NO2Sz= 395.
Example 128
Preparation of Compound 101
2-[(4-Methylphenyl)thio]-4-(trifluoromethyl)benzaldehyde (0.47 g, 89.5%) was
obtained by using 2-fluoro-4(trifluoromethyl)benzaldehyde (0.1 g, 0.5 mmol)
using the same
method as Example 114 (synthesis of 5-bromo-2-[(4-
methylphenyl)thioJbenzaldehyde).
'H-NMR (300 MHz, CDC 13) 8 (ppm) 2.41 (s, 3H), 7.23 (d, J = 4.6 Hz, 2H), 7.24
(m,
1H), 7.38 (d, J = 8.1 Hz, 2H), 7.50 (d, J = 8.1 Hz, 1H) , 7.95 {d, J = 7.9 Hz,
1 H), 10.41
(s, 1 H)
FABMS m/z 296 (M+) C,SH"'9F30S = 296
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Compound 101 (0.082 g, 60.9%) was obtained by using 2-[(4-methylphenyl)thio]-4-
(trifluoromethyl)benzaldehyde (0.1 g, 0.34 mmol) using the same method as
Example 94.
'H-NMR (300 MHz, DMSO-d6) b (ppm) 2.33 (s, 3H), 7.28 (d, J = 8.1 Hz, 2H), 7.35
(d,
J = 8.3 Hz, 2H), 7.3 8 (s, 1 H), 7.71 (d, J = 8.1 Hz, 1 H), 7.77 (d, J = 8.8
Hz, 1 H), 7.96 (s,
1 H), 12.78 (br s, 1 H)
EIMS m/z 394 (M-H)' C~6H,2'9F3NOZSz = 395.
Example 129
Preparation of Compound 102
2-[(4-methylphenyl)thioJ-3-(trifluoromethyl)benzaldehyde (0.44 g, 100%) was
obtained
by using 2-fluoro-3(trifluoromethyl)benzaldehyde (0.19 g, 1.5 mmol) using the
same method
as Example 114 (synthesis of S-bromo-2-[(4-methylphenyl)thin]benzaldehyde).
'H-NMR (300 MHz, CDC 13) 8 (ppm) 2.26 (s, 3H), 6.97 (d, J = 2.2 Hz, 2H), 7.03
(d, J =
8.1 Hz, 2H), 7.64 (t, J = 7.9 Hz, 1 H), 8.02 (d, J = 7.9 Hz, 1 H), 8.09 (d, J
= 7.7 Hz, 1 H),
10.60 (d, J = 0.7 Hz, 1 H)
EIMS m/z 296 (M~ C,SH"'9F30S = 296.
Compound 102 (0.13 g, 97.5%) was obtained by using 2-[(4-methylphenyl)thio]-3-
(trifluoromethyl)benzaldehyde (0.1 g, 0.3 mmol) using the same method as
Example 94.
'H-NMR (300 MHz, DMSO-ds) b (ppm) 2.19 (s, 3H), 6.89 (d, J = 8.3 Hz, 2H), 7.05
(d,
J = 7.9 Hz, 2H), 7.80 (s, 1 H), 7.81 (d, J = 4.0 Hz, 1 H), 7.87 (s, 1 H) ,
8.00 (dd, J = 2.8,
6.6 Hz, 1H), 12.01 (br s, 1H)
EIMS m/z 394 (M-H)- C,gHi2'9F3NOaS2 = 395.
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Example 130
Preparation of Compound 103
Compound 103 (0.043 g, 41.7%) was obtained by using Compound 102 (0.1 g, 0.25
mmol) using the same method as Example 96.
'H-NMR (300 MHz, DMSO-d6) S (ppm) 2.27 (s, 3H), 7.23 (d, J = 8.3 Hz, 2H), 7.27
(d,
J = 8.8 Hz, 2H), 7.79 (d, J = 7.7 Hz, 1 H), 7.93 (t, J = 7.7 Hz, 1 H), 8.07
(d, J = 7.7 Hz,
1 H), 8.18 (s, 1 H), 12.62 (br s, 1 H)
EIMS m/z 412 (M+H)+ C,gH,2'9F3NO3S2 = 411.
Example 131
Preparation of Compound 104
By using 2-fluoro-S-methoxybenzaldehyde (0.25 g, 2.0 mmol), 2-[(4-
methylphenyl)-
thio]-5-methoxybenzaldehyde (0.05 g, 10.1 %) was obtained through a reaction
as described in
Example 114 (synthesis of 5-bromo-2-[(4-methylphenyl)thioJ-benzaldehyde), and
a
purification by thin layer chromatography (hexane: ethyl acetate = 8:1 ),
followed by another
purification with thin layer chromatography (hexane: ethyl acetate = 24:1 ).
'H-NMR (300 MHz, CDC13) b (ppm) 2.32 (s, 3H), 3.86 (s, 3H), 7.07 (dd, J = 3.1,
8.6 Hz,
2H), 7.13 (m, 4H), 7.32 (d, J = 8.6 Hz, 1 H), 7.44 (d, J = 3.1 Hz, 1 H), 10.51
(s, 1 H)
EIMS m/z 258(M+) Ct5H,40zS = 258.
Compound 104 (0.07 g, 97.1 %)was obtained by using 2-[(4-methylphenyl)thio]-
5-methoxybenzaldehyde (0.05 g, 0.2 mmol) using the same method as Example 94.
'H-NMR (300 MHz, DMSO-d6) 8 (ppm) 2.24 (s, 3H), 3.84 (s, 3H), 7.02 (d, J = 8.3
Hz,
2H), 7.09 (m, 2H), 7.12 (d, J = 8.1 Hz, 2H), 7.52 (d, J = 8.6 Hz, 1 H), 8.04
(s, 1 H), 12.01
(br s, 1 H)
EIMS m/z 356 (M-H)' C,gH,5N03Sz = 357.
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Example 132
Preparation of Compound 105
2-[(4-Methylphenyl)thio]-4-methoxybenzaldehyde (0.078 g, 46.2%) was obtained
by using
2-fluoro-4-methoxybenzaldehyde (0.1 g, 0.65 mmol) using the same method as
Example 114
(synthesis of 5-bromo-2-[(4-methylphenyl)thio]benzaldehyde).
'H-NMR (300 MHz, CDC13) b (ppm) 2.39 (s, 3H), 3.70 (s, 3H), 6.45 (d, J = 2.4
Hz,
1 H), 6.76 (dd, J = 2.4, 8.6 Hz, 1 H), 7.22 (d, J = 7.7 Hz, 2H), 7.39 (d, J =
8.1 Hz, 2H)
7.80 (d, J = 8.6 Hz, 1 H), 10.20 (s, 1 H)
FABMS m/z 259 (M+H)+ C,SH,40zS = 258
Compound 105 (0.091 g, 85.2%) was obtained by using 2-[(4-methylphenyl)thio]-
4-methoxybenzaldehyde (0.078 g, 0.3 mmol) using the same method as Example 94
(synthesis
of Compound 67).
'H-NMR (300 MHz, DMSO-d6) 8 (ppm) 2.49 (s, 3H), 3.74 (s, 3H), 6.75 (d, J = 2.7
Hz,
I H), 7.07 (dd, J = 2.7, 8.6 Hz, 1 H), 7.23 (d, J = 8.3 Hz, 2H), 7.49 (d, J =
8.8 Hz, 1 H),
8.04 (s, 1 H), 12.59 (br s, 1 H)
EIMS m/z 358 (M+H)+ Ct6H~5NO3S2 = 357
Example 133
Preparation of Compound 106
5-Chloro-2-fluorobenzaldehyde (0.55 g, 77.6%) was obtained by using
4-chlorofluorobenzene (0.48 mL, 4.5 mmol) using the same method an Example 121
(synthesis
of 2-fluoro-5-cyanobenzaldehyde).
'H-NMR (300 MHz, CDC13) S (ppm) 7.16 (t, J = 9.4 Hz, 1H), 7.56 (m, 1H), 7.84
(dd, J
= 2.8, 5.9 Hz, 1 H), 10.32 (s, 1 H)
EIMS m1z 157(M-H) C~H435C119FN0 = I58.
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5-Chloro-2-[(4-methylphenyl)thio]benzaldehyde (0.21 g, 85.1 %) was obtained by
using 5-chloro-2-fluorobenzaldehyde (0.15 g, 0.95 mmol) using the same method
as Example
114 (synthesis of 5-bromo-2-[(4-methylphenyl)thio]benzaldehyde).
'H-NMR (300 MHz, CDC13) 8 (ppm) 2.38 (s, 3H), 6.99 (d, J = 8.6 Hz, 1H), 7.21
(d, J =
7.9 Hz, 2H), 7.33 (m, 1H) 7.35 (d, J = 8.3 Hz, 2H) 7.82 (d, J = 2.4 Hz, 1H),
10.34 (s,
1 H)
FABMS m/z 262(M+) C,4H~,35C10S = 262.
Compound 106 (0.096 g,70.1%) was obtained by using S-chloro-2-[(4-
methylphenyl)-
thio]benzaldehyde (0.1 g, 0.38 mmol) using the same method as Example 94.
1H-NMR (300 MHz, DMSO-d6) 8 (ppm) 2.30 (s, 3H), 7.23 (d, J = 8.6 Hz, 2H), 7.25
(m,
1 H), 7.27 (d, J = 8.4 Hz, 2H), 7.48 (s, 1 H), 7.49 (d, J = 7.0 Hz, 1 H), 7.91
(s, 1 H) 12.01
(br s, 1 H)
EIMS m/z 362 (M+H)+ C1~H1235C1NOzS2 = 361.
Example 134
Preparation of Compound 107
Compound 107 (0.094 g, 90.0%) was obtained by using Compound 106 (0.1 g, 0.28
mmol) using the same method as Example 96.
~H-NMR (300 MHz, DMSO-db) S (ppm) 2.30 (s, 3K), 7.31 (d, J = 8.4 Hz, 2H), 7.43
(d,
J = 8.1 Hz, 2H), 7.49 (d, J = 2.0 Hz, 1 H), 7.79 (dd, J = 2.0, 8.5 Hz, 1 H),
7.88 (s, 1 H),
8.02 (d, J = 8.4 Hz, 1H), NH is not found
EIMS m/z 378 (M+H)+ C,~H,z3sC1N03Sz = 377.
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Example 135
Preparation of Compound 108
4-Chloro-2-[(4-methylphenyl)thio)benzaldehyde (0.32 g, 60.4%) was obtained by
using
4-chloro-2-fluorobenzaldehyde (0.32 g, 2.0 mmol) using the same method as
Example 114
(synthesis of 5-bromo-2-[(4-methylphenyl)thio]benzaldehyde).
~ H-NMR (300 MHz, CDC 13) 8 (ppm) 2.41 (s, 3H), 6.90 (d, J = 1.8 Hz, 1 H),
7.24 (m,
3 H), 7.40 (d, J = 8.1 Hz, 2H), 7.76 (d, J = 8.3 Hz, 1 H), 10.29 (s, 1 H)
EIMS m/z 262 (M+) C(4H"35C1OS = 262
Compound 108 (0.052 g, 75.1 %) was obtained by using 4-chloro-2-[(4-
methylphenyl)thioJbenzaldehyde (0.05 g, 0.2 mmol) using the same method as
Example 94.
IH-NMR (300 MHz, DMSO-d6) S (ppm) 2.33 (s, 3H), 7.11 (d, J = 1.5 Hz, 1H), 7.28
(d,
1 S J = 8.3 Hz, 2H), 7.35 (d,J = 8.3 Hz, 2H), 7.50 (d, J = 2.2 Hz, 1 H), 7.51
(s, 1 H) 7.95 (s,
1 H),12.72 (br s, 1 H)
EIMS m/z 326 (M+) C,~H~ZNOzS2 = 326.
Example 136
Preparation of Compound 109
3-Chloro-4-[(4-methylphenyl)thio]benzaldehyde (0.29 g, 87.9%) was obtained by
using 3-chloro-4-fluorobenzaldehyde (0.2 g, 1.26 mmol) using the same method
as Example
114 (synthesis of 5-bromo-2-[(4-methylphenyl)thin]benzaldehyde).
'H-NMR (300 MHz, CDC13) 8 (ppm) 2.44 (s, 3H), 6.77 (d, J = 8.3 Hz, 1H), 7.30
(d, J
= 8.4 Hz, 2H), 7.46 (d, J = 8.1 Hz, 2H), 7.50 (dd, J = 1.8, 8.4 Hz, 1H), 7.82
(d, J = 1.7
Hz, 1 H), 9.85 (s, 1 H)
FABMS m/z 262 (M~ C,4H"35C 1 OS = 262.
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Compound 109 (0.13 g, 65.1 %) was obtained by using 3-chloro-4-[(4-
methylphenyl)-
thin]benzaldehyde (0.15 g, 0.57 mmol) using the same method as Example 94.
'H-NMR (300 MHz, DMSO-db) 8 (ppm) 2.38 (s, 3H), 6.79 (d, J = 8.4 Hz, 1H), 7.37
(d,
J = 7.9 Hz, 2H), 7.41 (dd, J = 2.2, 8.7 Hz, 1 H), 7.48 (d, J = 8.1 Hz, 2H),
7.71 (s, 1 H),
7.75 (d, J = 1.8 Hz, 1 H), NH is not found
EIMS m/z 361(M~ C,~H,235C1N02S2 = 361
Example 137
Preparation of Compound 110
2-Fluoro-5-nitrobenzaldehyde (0.055 g, 0.33 mmol) was dissolved in N,N-
dimethyl-
formamide (2.8 mL), and phenol (0.077 g, 0.82 mmol) and potassium carbonate
(0.11 g, 0.82
mmol) were added thereto, followed by stirring at 25°C for 1 hour.
After a conventional
treatment, the residue was purified by silica gel chromatography (eluted with
chloroform), to
obtain 5-vitro-2-phenoxybenzaldehyde (77 mg, 98%).
~H NMR (300 MHz, CDC13) 8 (ppm) 6.91 (d, J = 9.4 Hz, 1H), 7.20-7.14 (m, 2H),
7.35
(dd, J = 7.2 Hz, 7.2 Hz, 1 H) 7.47-7.55 (m, 2H), 8.31 (dd, J = 9.2, 2.8 Hz, 1
H), 8.79 (d, J
= 3.0 Hz, 1 H), 10.60 (s, 1 H)
FABMS m/z 244 (M+H)+ C13H9NO4 = 243
5-Nitro-2-phenoxybenzaldehyde (77 mg, 0.32 mmol) was dissolved in toluene (3.9
mL), and 2,4-thiazolidinedione (0.15 g, 1.3 mmol), piperidine (0.013 mL, 0.13
mmol), acetic
acid (0.0073 mL, 0.13 mmol) and molecular sieves 4A (0.39 g) were added
thereto, followed
by stirnng at 110°C for 3 hours. After the conventional post-reaction
treatment, the residue was
purified by thin layer chromatography (developed with chloroform/acetonitrile
= 10/1 ), to
obtain Compound 110 (67 mg, 61 %).
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IH NMR (300 MHz, DMSO-d6) 8 (ppm) 6.94 (d, J = 9.2 Hz,1H), 7.27 (d, J = 7.7
Hz,
2H), 7.36 (d, J = 7.1, 7.1 Hz, 1H), ?.SO-7.58 (m, 2H), 7.99 (s, 1H), 8.28 (dd,
J = 2.7, 9.2
Hz, 1 H), 8.3 8 (d, J = 2. 8 Hz, 1 H), 12.81 (m, 1 H)
FABMS m/z 341 (M-H)' C,6H,oN205 = 342.
Example 138
Preparation of Compound 111
2-Fluoro-5-nitrobenzaldehyde (0.13 g, 0.79 mmol) was dissolved in N,N-
dimethylformamide (6.7 mL), and p-cresol (0.22 g, 2.0 mmol) and potassium
carbonate (0.27
g, 2.0 mmol) were added thereto, followed by stirring at 25°C for 1.5
hours. After a
conventional treatment, the residue was purified by silica gel chromatography
(eluted with
chloroform), to obtain 5-nitro-2-(4-methylphenoxy)benzaldehyde (0.20 g, 98%).
~H NMR (300 MHz, CDC13) 8 (ppm) 2.41 (s, 3H), 6.89 (d, J = 9.4 Hz, 1H), 7.04
(d, J =
8.5 Hz, 2H), 7.25-7.32 (m, 2H), 8.39 (dd, J = 9.2, 3.0 Hz, 1 H), 8.79 (d, J =
2.9 Hz, 1 H),
10.60 (s, 1 H)
FABMS m/z 257 (M+) CiaHi iNO4S = 257.
5-Nitro-2-(4-methylphenoxy)benzaldehyde (0.11 g, 0.41 mmol) was dissolved in
toluene (5.3 mL), and 2,4-thiazolidinedione (0.19 g, 1.6 mmol), piperidine
(0.016 mL, 0.16
mmol), acetic acid (0.0094 mL, 0.16 mmol) and molecular sieves 4A (0.53 g)
were added
thereto, followed by stirnng at 110°C for 2 hours. After the
conventional post-reaction
treatment, the residue was triturated with ethanol, to obtain Compound 111 (60
mg, 41 %).
'H NMR (300 MHz, DMSO-db) 8 (ppm) 2.36 (s, 3H), 6.90 (d, J = 9.2 Hz, 1H), 7.15
(d,
J = 8.2 Hz, 2H), 7.33 (d, J = 8.6 Hz, 2H), 7.99 (s, 1 H), 8.25 (dd, J = 9.2,
1.8 Hz, 1 H),
8.36 (d, J = 2.6 Hz, 1 H), 12.80 (m, 1 H)
FABMS m/z 355 (M-H)'Ci~H,2N205S = 356.
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Example 139
Preparation of Compound 112
Commercially available 2-[4-(2,2-dimethylethyl)phenaxy]-5-nitrobenzaldehyde
(0.12 g,
0.41 mmol) (MAYBRIDGE, Catalog Number: XAX00137) was dissolved in ethanol (4.9
mL),
and 2,4-thiazolidinedione (0.19 g, 1.6 mmol) and piperidine (0.016 mL, 0.16
mmol) were
added thereto, followed by stirring at 80°C for 2 hours. After the
conventional post-reaction
treatment, the residue was purified by thin layer chromatography (developed
with
chloroform/acetonitrile = 10/1), to obtain Compound 112 (49 mg, 30%).
~H NMR (300 MHz, DMSO-d6) 8 (ppm) 1.31 (s, 9H), 6.92 (d, J = 9.2 Hz, 1H), 7.18
(d,
J = 8.6 Hz, 2H), 7.53 (d, J = 8.6 Hz, 2H), 7.98 (s, 1 H), 8.27 (dd, J = 9.2,
2.4 Hz, 1 H),
8. 3 5 (d, J = 2.2 Hz, I H), 12.78 (br s, 1 H)
FABMS m/z 397 (M-H)- CZOH,8NZO5S = 398.
Example 140
Preparation of Compound 113
Compound 113 (0.079 g, 71.1 %) was obtained by using Compound 100 (0.1 I g,
0.27
mmol) using the same method as Example 96.
~H-NMR (300 MHz, DMSO-d6} b (ppm) 2.29 (s, 3H), 7.31 (d, J = 8.3 Hz, 2H), 7.47
(d,
J = 8.1 Hz, 2H), 7.74 (s, 1 H), 7.77 (s, I H), 8.03 (d, J = 7.9 Hz, 1 H), 8.24
(d, J = 7.9Hz,
1 H), NH is not found
EIMS m/z 412 (M+H)+ C,gH12~9F3NO3S2 = 411.
Example 141
Preparation of Compound 114
Compound 114 (0.046 g, 43.5%) was obtained by using Compound 94 (0.1 g, 0.28
mmol) using the same method as Example 96.
'H-NMR (300 MHz, DMSO-db) 8 (ppm) 2.29 (s, 3H), 7.29 (d, J = 8.1 Hz, 2H), 7.47
(d,
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J = 8.3 Hz, 2H), 7.83 (s, 1 H), 7.86 (d, J = 7.9 Hz, 1 H), 8.01 (d, J = 8.1
Hz, 1 H), 8.15 (d,
J = 8.3 Hz, 1 H), NH is not found
EIMS mlz 369 (M+H)+C,6H,ZN2O3Sz = 368.
Example 142
Preparation of Compound 115
2-[(4-Trifluoromethylphenyl)thio]-5-nitrobenzaldehyde (0.13 g, 93%) was
obtained
from 4-trifluoromethylbenzenethiol (74 mg, 0.42 mmol), a 2.5 mol/L sodium
hydroxide
aqueous solution (0.71 mL, 1.8 mmol), tetrabutylammonium bromide (6.7 mg,
0.021 mmol)
and a toluene (0.71 mL) solution of 2-fluoro-5-nitrobenzaldehyde (70 mg, 0.41
mmol) using
the same method as Example 97.
1H NMR (300 MHz, CDC13) 8 (ppm) 7.04 (d, J = 8.8 Hz, 1H), 7.70 (d, J = 8.1 Hz,
2H),
7.77 (d, J = 8.6 Hz, 2H), 8.18 (dd, J = 8.8, 2.5 Hz, 1 H), 8.71 (d, J = 2.5
Hz, 1 H), 10.31
(s, 1 H)
FABMS m/z 327 (M-) C,4Hg19F3NO3S = 327.
Compound 115 (44 mg, 51 %) was obtained from 2-[(4-trifluoromethylphenyl)-
thio]-5-nitrobenzaldehyde (0.12 g,0.35 mmol), toluene(5.8 mL), 2,4-
thiazolidinedione (0.17 g,
1.4 mmol), piperidine (0.014 mL, 0.14 mmol), acetic acid (0.0080 mL, 0.14
mmol), and
molecular sieves 4A (0.58 g) using the same method as Example 97.
~ H NMR (300 MHz, DMSO-d6) 8 (ppm) 7.52 (d, J = 8.7 Hz, 1 H), 7.65 (d, J = 8.3
Hz,
2H), 7.81 (d, J = 8.2 Hz, 2H), 7.88 (s, 1 H), 8.24 (dd, J = 8.6, 2.5 Hz, 1 H),
8.29 (d, J =
2.4 Hz, 1 H), 12.82 (m, 1 H)
FABMS m/z 425 (M-H)- Cl7Hg'9F3N2O4SZ = 426.
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Example 143
Preparation of Compound 116
Compound 74 (57 mg, 0.15 mmol) obtained in Example 101 was dissolved in
dichloromethane (11 mL) and methanol (2.3 mL), and m-chloroperbenzoic acid (SS
mg, 0.16
mmol) was added thereto, followed by stirring at 25°C for 1.5 hours.
After the conventional
post-reaction treatment, the residue was purified by thin layer chromatography
(developed with
chloroform/methanol = 15/ 1 ), to obtain Compound 116 (34 mg, 57%).
'H NMR (300 MHz, DMSO-d6) 8 (ppm) 3.76 (s, 3H), 7.02-7.08 (m, 2H), 7.49-7.55
(m,
2H),' 7.81 (s, 1 H), 8.19 (d, J = 2.0 Hz, 1 H), 8.33 (d, J = 8.6 Hz, 1 H),
8.52 (dd, J = 8.6,
2.2 Hz, 1 H), 12. 86 (m, 1 H)
FABMS m/z 403 (M-H)' C,~H,ZN2O6S2 = 404.
Example 144
Preparation of Compound 117
Compound 117 (37 mg, 68%) was obtained from Compound 71 (52 mg, 0.13 mmol)
obtained in Example 98, dichloromethane (10 mL), methanol (2.1 mL) and
m-chloroperbenzoic acid (50 mg, 0.15 mmol) using the same method as Example
143.
'H NMR (300 MHz, DMSO-d6) S (ppm) 7.55-7.66 (m, 3H), 7.80-7.85 (m, 1H), 8.08
(s,
1 H), 8.15 (d, J = 8.6 Hz, 1 H), 8.27 (m, 1 H), 8.45 (dd, J = 8.6, 2.2 Hz, 1
H), 12.88 (m,
1 H)
FABMS m/z 407 (M-H)- C~6H935C1N2OSS2 = 408.
Example 145
Preparation of Compound 118
3-bromo-4-[(4-methylphenyl)thio]benzaldehyde (0.2 g, 0.65 mmol) was dissolved
in
acetone (3 mL), cooled to O°C, and Jones reagent (0.082 mL) was added
thereto, followed by
stirring for 3.5 hours. Then, isopropyl alcohol (0.1 mL) was added thereto.
After a conventional
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treatment, the residue was recrystallized and purified with hexane and ethyl
acetate, to obtain
3-bromo-4-[(4-methylphenyl)thio]benzoic (0.18 g, 84.7%).
'H-NMR (300 MHz, DMSO-d6) S (ppm) 2.39 (s, 3H), 6.70 (d, J = 8.4 Hz, 1H), 7.37
(d,
J = 8.3 Hz, 2H), 7.49 (d, J = 7.9 Hz, 2H), 7.76 (dd, J = 1.1, 8.7 Hz, 1 H),
8.07 (d, J = 1.1
Hz, I H), C02H is not found
FABMS m/z 322 (M-H)' C,aH"79BrOS = 323
S-Carboxy-2-[(4-methylphenyl)thio]benzaldehyde (0.031 g, 36.7%) was obtained
by
using 3-bromo-4-[(4-methylphenyl)thio]benzoic acid (0.1 g, 0.31 mmol) using
the same
method as Example 115 (synthesis of 3-bromo-4-[(4-
methylphenyl)thio]benzaldehyde).
'H-NMR (300 MHz, DMSO-d6) 8 (ppm) 2.50 (s, 3H), 6.84 (d, J = 8.4 Hz, 1H), 7.36
{d,
J = 8.1 Hz, 2H), 7.46 (d, J = 8.1 Hz, 2H), 7.93 (dd, J = 2.0, 8.4 Hz, 1 H),
8.46 (d, J = 2.0
Hz, 1 H), 10.22 (s, 1 H), COZH is not found
FABMS m/z 271 (M-H)' C~SH12~3s = 272
Compound 118 (0.032 g, 25.3%) was obtained by using 5-carboxy-2-[(4-methyl-
phenyl)thio]benzaldehyde (0.094 g, 0.34 mmol) using the same method as
described in
Example 94.
'H-NMR (300 MHz, DMSO-d6) 8 (ppm} 2.34 (s, 3H), 6.96 (d, J = 8.6 Hz, 1 H),
7.29 (d,
J = 8.1 Hz, 2H), 7.33 (s, IH), 7.37 (d, J = 7.2 Hz, 2H), 7.64 (s, 1H), 7.69
(d, J = 8.4 Hz,
1H), 8.20 (s, 1H), NH is not found
FABMS m/z 370 (M-H)' C,gH~3NOqS2 = 371.
Example 146
Preparation of Compound 1 I 9
Compound 119 (0.120 g, 33.5%) was obtained by using commercially available
5-vitro-2-(pyrid-2-ylthio)benzaldehyde (0.26 g, 1.0 mmol) (MAYBRIDGE, Catalog
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Number: XAX00153) using the same method an Example 94.
~H-NMR (300 MHz, DMSO-d6) b (ppm) 7.28 (dd, J = 4.8, 7.7 Hz, 1H), 7.40 (dd, J
=
0.9, 7.7 Hz, 1 H), 7.78 (dd, J = I .3, 7.7 Hz, I H), 7.86 (s, 1 H), 7.87 (d, J
= 8.4 Hz, 1 H),
8.28 (dd, J = 2.4, 8.4Hz, I H), 8.32 (d, J = 2.4 Hz, I H), 8.42 (dd, J = 0.9,
4.8 Hz, 1 H),
I 2.78 (br s, I H)
EIMS m/z = 360 (M+H)+ Cl5H9N3~4S2 = 359.
Example 147
Preparation of Compound 120
Compound 120 (0.0044 g, 21.2%) was obtained by using Compound 119 (0.02 g,
0.056
mrnol) obtained in Example 146 using the same method as Example 96.
~H-NMR (300 MHz, DMSO-d6) 8 (ppm) 7.49 (m, 1H), 7.87 (s, 1H), 7.98 (d, J = 7.9
Hz,
1 H), 8.07 (m, 1 H), 8.08 (d, J = 8.5 Hz, 1 H), 8.26 (dd, J = 2.2, 8.5 Hz, 1
H), 8.43 (d, J =
2.2 Hz, 1 H), 8.54 (d, J = 4.8 Hz, 1 H), NH is not found
EIMS m/z = 376 (M+H)+ C,SH9N305S2 = 375.
Example 148
Preparation of Compound 121
N,N-diphenylbenzylamine (935 mg, 3.60 mmol) was suspended in acetic acid (20
mL),
and hexamethylenetriamine (1.12 g, 7.96 mmol) was added thereto, followed by
stirnng at
90°C for 12 hours. The reaction liquid was cooled to room temperature,
a 6 mol/L sodium
hydroxide aqueous solution and water were added thereto, and then the product
was extracted
with chloroform. The organic layer was washed with brine, and then dried over
anhydrous
sodium sulfate. The solvent was evaporated under reduced pressure, and the
residue was
purified by silica gel column chromatography (elution solvent: hexane/ethyl
acetate = 20/I -~
10/1), to obtain 4-(N-phenylbenzylamino)benzaldehyde (742 mg, 72%).
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'H NMR (270 MHz, CDC 13} 8 (ppm) 5.04 (s, 2H), 6.80 (d, J = 8.6 Hz, 2H), 7.2-
7.45
{m, l OH), 7.63 (d, J = 8.6 Hz, 2H), 9.73 (s, 1 H).
4-(N-phenylbenzylamino)benzaldehyde (109 mg, 0.378 mmol), 2,4-
thiazolidinedione
(59.9 mg, 0.511 mmol) and piperidine (0.045 mL, 0.46 mmol) were heated under
reflux for 6
hours in ethanol (5 mL). The reaction liquid was cooled to room temperature, 1
mol/L HC1
was added thereto, and then the product was extracted with chloroform. The
organic layer was
washed with brine, and then dried over anhydrous sodium sulfate. The solvent
was evaporated
under reduced pressure, and the residue was purified by preparative thin layer
chromatography
(chloroform/methanol = 20/1), to obtain Compound 121 (123 mg, 84%).
'H NMR (270 MHz, CDC13) 8 (ppm) 5.08 (s, 2H), 6.88 (d, J = 8.6 Hz, 2H), 7.15-
7.45
(m, 12H), 7.61 (s, 2H), 12.38 (br s, 1H).
Example 149
Preparation of Compound 129
2-Fluoro-5-nitrobenzaldehyde(3 I mg, 0.18 mmol) was dissolved in N,N-
dimethylformamide (3.1 mL), and 4-mercaptobenzoic acid (85 mg, 0.55 mmol) and
triethylamine (0.13 mL, 0.92 mmol) were added thereto, followed by stirnng at
25 °C for 20
minutes. After the conventional post-reaction treatment, the product was
purified by thin layer
chromatography (developed with chloroform/methanol = 10/1) to give 5-vitro-2-
[(4-
carboxylphenyl)thio]benzaldehyde (60 mg, 100%).
5-Nitro-2-[(4-carboxylphenyl)thin]benzaldehyde (60 mg, 0.20 mmol) was
dissolved in
ethanol (2.4 mL), and 2,4-thiazolidinedione (92 mg, 0.79 mmol), piperidine
(0.027 mL, 0.28
mmol), acetic acid (0.0045 mL, 0.079 mmol) and molecular sieves 4A (0.30 g)
were added
thereto, followed by stirring at 80 °C for 2.5 hours. After the
conventional post-reaction
treatment, thin layer chromatography (developed with chloroform/methanol =
5/1), to give
Compound 129 ( 12 mg, 15%).
5-Nitro-2-[(4-carboxyiphenyl)thin]benzaldehyde:
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'H NMR (300 MHz, CDC13) 8(ppm) 7.04 (d, J = 8.8 Hz, 1 H), 7.74 (d, J = 8.0 Hz,
2H),
8.09 (d, J = 8.2 Hz, 2H), 8.28 (dd, J = 8.8, 2.5 Hz, 1 H), 8.87 (d, J = 2.5
Hz, 1 H), 10.28 (s, 1 H),
13.10 (m, 1 H)
Compound 129:
'H NMR (300 MHz, DMSO-d6) 8(ppm) 7.46 (d, J = 8.8 Hz, 1H), 7.55 (d, J = 8.3
Hz,
2H), 7.87 (s, 1 H), 7.98 (d, J = 8.7 Hz, 2H), 8.21 (dd, J = 8.7, 2.4 Hz, 1 H),
8.30 (d, J = 2.4 Hz,
1H), 12.57-13.31 (m, 2H)
FABMS m/z 401 (M-H)' C,~HipN2O6S2 = 402
Exam~Ie 1 SO
Preparation of Compound 130
5-Nitro-2-[(4-carboxylphenyl)thio]benzaldehyde (71 mg, 0.23 mmol) obtained in
Example 149 was dissolved in N,N-dimethylformamide (7.1 mL), and a 2.0 mol/L
dimethylamine methanol solution (0.23 mL, 0.47 mmol), 1-ethyl-3-(3-
dimethylaminopropyl)carbodiimide hydrochloride (90 mg, 0.47 mmol), and 1-
hydroxybenzotriazole hydrate (0.11 g, 0.94 mmol) were added thereto, followed
by stirring at
°C for 30 minutes. After the conventional post-reaction treatment, the
product was purified
by silica gel chromatography (eluted by chloroform) to give N,N-dimethyl-4-[(2-
formyl-4-
nitrophenyl)thio]benzamido (59 mg, 75%).
20 N,N-Dimethyl-4-[(2-formyl-4-nitrophenyl)thio]benzamido (0.12 g, 0.36 mmol)
was
dissolved in toluene (6.0 mL), and 2,4-thiazolidinedione (0.17 g, 1.5 mmol),
piperidine (0.014
mL, 0.15 mmol), acetic acid (0.0083 mL, 0.15 mmol) and molecular sieves 4A
(0.60 g) were
added thereto, followed by stirring at 110 °C for 4.5 hours. After the
conventional post-
reaction treatment, thin layer chromatography (developed with
chloroform/acetonitrile = 3/1),
25 to give Compound 130 (48 mg, 31 %).
N,N-dimethyl-4-[(2-formyl-4-nitrophenyl)thio]benzamido:
1H NMR (300 MHz, CDC13) 8(ppm) 3.02 (br s, 3H), 3.16 (br s, 3H), 7.02 (d, J =
9.0 Hz,
1 H), 7.53-7.64 (m, 4H), 8.13 (dd, J = 8.8, 2.6 Hz, 1 H), 8.69 (d, J = 2.6 Hz,
1 H), 10.31 (s, 1 H) ,
FABMS m/z 331 (M+H)+ C,6H,qN3O4S = 330
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Compound 130:
'H NMR (300 MHz, DMSO-db) S(ppm) 2.92 (br s, 3H), 2.99 (br s, 3H), 7.36 (d, J
= 8.8
Hz, 1 H), 7.50 (d, J = 8.2 Hz, 2H), 7.57 (d, J = 8.0 Hz, 2H), 7.89 (s, 1 H),
8.21 (dd, J = 8.8, 2.2
Hz, 1 H), 5.27 (d, J = 2.2 Hz, 1 H), 12.83 (m, 1 H)
FABMS mlz 428 (M-H)' C,9H,SN305S2 = 429
Example 151
Preparation of Compound 131
4-{4-[(2-Formyl-4-nitrophenyl)thio]benzoyl}morpholine (0.13 g, 78%) was
obtained
from 5-vitro-2-[(4-carboxylphenyl)thio]benzaldehyde (0.13 g, 0.42 mmol)
obtained in
Example 149, N,N-dimethylformamide (7.1 mL), morpholine (0.073 mL, 0.83 mmol),
1-ethyl-
3-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.16 g, 0.83 mmol), and 1-
hydroxybenzotriazole monohydrate (0.11 g, 1.7 mmol), using the same method as
Example 150.
Compound 131 (48 mg, 31%) was obtained from 4-{4-[(2-formyl-4-
nitrophenyl)thio]benzoyl}morpholine (0.13 g, 0.34 mmol), toluene (6.3 mL), 2,4-
tltiazolidinedione (0.16 g, 1.3 mmol), piperidine (0.013 mL, 0.13 mmol),
acetic acid (0.0077
mL, 0.13 mmol) and molecular sieves 4A (0.63 g).
4-{4-[(2-formyl-4-nitrophenyl)thio]benzoyl }morpholine:
1H NMR (300 MHz, CDCl3) 8(ppm) 3.39-3.93 (m, 8H), 7.04 (d, J = 9.0 Hz, 1H),
7.56
(d, J = 8.2 Hz, 2H), 7.62 (d, J = 8.3 Hz, 2H), 8.13 (dd, J = 8.8, 2.5 Hz, 1
H), 8.69 (d, J = 2.6 Hz,
1 H), 10.30 (s, 1 H)
FABMS m/z 373 (M+H)+ C~8H,6N205S = 372
Compound 131:
'H NMR (300 MHz, DMSO-db) 8(ppm) 3.27-3.45 (m, 4H), 3.61 (br s, 4H), 7.39 (d,
J
=8.9 Hz, 1 H), 7.51 (d, 3 = 8.4 Hz, 2H), 7.57 (d, J = 8.4 Hz, 2H), 7.89 (s, 1
H), 8.21 (dd, J = 8.8,
2.6 Hz, 1 H), 8.27 (d, J = 2.2 Hz, 1 H), 12.83 (m, 1 H)
FABMS m/z 470 (M-H)' CZ,Hi7N3O6S3 = 471
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Example 152
Preparation of Compound 132
A 2.5 mol/L sodium hydroxide aqueous solution (1.6 mL, 4.0 mmol) and
tetrabutylammonium bromide (15 mg, 0.047 mmol) were added to 4-
(methylthio)benzenethiol
(0.15 mL, 0.93 mmol), followed by stirring at 25 °C for 10 minutes. A
toluene solution (1.6
mL) of 2-fluoro-5-nitrobenzaldehyde (0.16 g, 0.93 mmol) was added to the
reaction liquid,
followed by stirring at 110 °C for 1.5 hours. After the conventional
post-reaction treatment, the
product was purified by silica gel chromatography (eluted by chloroform) to
give S-nitro-2-[(4-
methylthio)phenyl]thiobenzaldehyde (0.25 g, 89%).
5-nitro-2-[4-(methylthio)phenyl]thiobenzaldehyde (0.23 g, 0.76 mmol) was
dissolved in
toluene (12 mL), and 2,4-thiazolidinedione (0.35 mg, 3.0 mmol), piperidine
(0.030 mL, 0.30
mmol), acetic acid (0.017 mL, 0.30 mmol) and molecular sieves 4A (1.2 g) were
added thereto,
followed by stirring at 110 °C for 4 hours. After the conventional post-
reaction treatment, the
product was triturated by using ethanol to give Compound 132 (12 mg, 15%).
S-nitro-2-[4-(methylthio)phenyl]thiobenzaldehyde:
'H NMR (300 MHz, CDC13} S(ppm) 2.55 (s, 3H), 6.98 (d, J = 9.0 Hz, 1H), 7.32-
7.38
(m, 2H), 7.44-7.50 (m, 2H), 8.11 (dd, J = 9.0, 2.5 Hz, 1 H), 8.66 (d, J = 2.4
Hz, 1 H), 10.30 (s,
1 H)
FABMS m/z 306 (M+H)+ Cl4Ht,N03Sz = 305
Compound 132:
~H NMR (300 MHz, DMSO-d6) 8(ppm) 2.52 (s, 3H), 7.09 (d, J = 9.2 Hz, 1H), 7.41
(d, J
= 8.6 Hz, 2H), 7.52 (d, J = 8.3 Hz, 2H), 7.38 (s, 1 H), 8.16 (dd, J = 8.8, 2.6
Hz, 1 H), 8.23 (d, J
=2.4 Hz, 1 H), 12.83 (m, 1 H)
FABMS m/z 403 (M-H)' CmH12N2O4S3 = 464
Example 153
Preparation of Compound 133
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Compound 133 (0.12 g, 100%) was obtained from Compound 75 (0.11 g, 0.27 mmol)
obtained in Example 102, dichloromethane (21 mL), methanol (4.2 mL) and m-
chloroperbenzoic acid (0.10 g, 0.30 mmol), using the same method as Example
134.
Compound 133:
iH NMR (300 MHz, DMSO-db) 8(pprn) 2.46-2.55 (m, 3H), 3.93-4.04 (m, 2H), 8.73
(br
d, J = 7.7 Hz, 2H), 8.88 (br d, J =7.5 Hz, 2H), 9.27 (s, 1 H), 9.58 (s, 1 H),
9.72 (d, J = 8.6 Hz,
1 H), 9.91 (br d, J = 8.3 Hz, 1 H), 14.21 (m, 1 H)
FABMS m/z 401 (M-H)' C,$H,4N205S2 = 402
. Example 154
Preparation of Compound 134
Compound 134 (90 mg, 76%) was obtained from Compound 70 (0.11 g, 0.29 mmol)
obtained in Example 97, dichloromethane (23 mL), methanol (4.5 mL) and m-
chloroperbenzoic
acid (0.11 g, 0.32 mmol), using the same method as Example 134.
Compound 134:
'H NMR (300 MHz, DMSO-db) 8(ppm) 7.49-7.62 (m, 3H), 7.66 (s, 1H), 7.99 (s,
1H),
8.21 {d, J = 1.7 Hz, 1 H), 8.3 0 (d, J = 8.9 Hz, 1 H), 8.49 (dd, J = 8.4, 2.0
Hz, 1 H), 12. 89 (m, 1 H)
FABMS m/z 407 (M-H)' C~6H935C1N2OSS2 = 408
Example 1 SS
Preparation of Compound 135
Compound 135 (32 mg, 37%) was obtained from Compound 72 (83 mg, 0.19 mmol)
obtained in Example 99, dichloromethane ( 17 mL), methanol (3.3 mL) and m-
chloroperbenzoic
acid (74 mg, 0.21 mmol), using the same method as Example 134.
Compound 135:
'H NMR (300 MHz, DMSO-ds) 8(ppm) 7.52 (dd, J = 8.4, 1.8 Hz, 1H), 7.79 (d, J =
8.4
Hz, 1 H), 7.86 (d, J = 2.1 Hz, 1 H), 7.99 (s, 1 H), 8.22 (d, J = 2.0 Hz, 1 H),
8.30 (d, J = 8.6 Hz,
1 H), 8.48 (dd, J = 8.6, 2.0 Hz, 1 H), 12.88 {m, 1 H)
FABMS m/z 441 (M-H)' C,6Hg35C12N2O5S2 = 442
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Example 156
Preparation of Compound 136
Compound 80 (60 mg, 0.14 mmol) obtained in Example 107 was dissolved in N,N-
dimethylformamide (3.0 mL), and m-chloroperbenzoic acid (0.11 g, 0.33 mmol)
was added
thereto, followed by stirring at 25 °C for 1 hour. After the
conventional post-reaction
treatment, thin layer chromatography (developed with chloroform/methanol =
20/1), to give
Compound 136 (20 mg, 33%).
Compound 136:
~H NMR (300 MHz, DMSO-d6) S(ppm) 7.66 (d, J = 8.6 Hz, 2H), 7.73 (d, J = 8.4
Hz,
2H), 7.99 (s, 1 H), 8.27-8.32 (m, 1 H), 8.48 (d, J = 8.2 Hz, 1 H), 8.54 (d, J
= 1.8 Hz, 1 H), 12.83
(m, 1 H)
FABMS mlz 451 (M-H)' Ci6H9~9BrN2O5S2 = 452
Example 157
Preparation of Compound 137
A reaction similar to the synthesis of 5-bromo-2-[(4-
methylphenyl)thio]benzaldehyde in
Example 114 was performed using 4-fluoro-3-phenoxybenzaldehyde (0.20 g, 0.93
mmol),
followed by the conventional treatment.
The solvent was removed from the obtained mixture of the starting material and
the
product by using a vacuum dryer, and, using the whole quantity, Compound 137
(0.1 I g, 45%)
was obtained in a manner similar to the synthesis of Compound 67.
Compound 137:
'H-NMR(300MHz, DMSO-db) 8(ppm) 2.50(s, 3H), 6.90(d, J = 8.3Hz, 1H), 7.06(d, J
=
9.0Hz, 2H), 7.07(m, 1 H), 7.21 (t, J = 7.3Hz, 1 H), 7.30(m, 1 H), 7.32(d, J =
8.1 Hz, 2H) , 7.43(d, J
= 8.1 Hz, 4H) , 7.63(s, 1 H) , 12.55(br s, 1 H)
EI-MS m/z 419(M+), C23HI~N03Sz=419
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Example 158
Preparation of Compound 138
A reaction similar to the synthesis of S-bromo-2-[(4-
methylphenyl)thio]benzaldehyde in
Example 114 was performed using 4-fluoro-3-metoxybenzaldehyde (0.20 g, 1.3
mmol),
followed by the conventional treatment.
The solvent was removed from the obtained mixture of the starting material and
the
product by using a vacuum dryer, and, using the whole quantity, Compound 138
(0.13 g, 44%)
was obtained in a manner similar to the synthesis of Compound 67.
Compound 138:
IH-NMR(300MHz, DMSO-d6) 8(ppm) 2.36(s, 3H), 3.90(s, 3H), 6.71 (d, J = 8.1 Hz,
1 H),
7.06(d, J = 8.1 Hz, 1 H), 7.24(d, J = 1.3Hz, 1 H), 7.31 (d, J=8.3Hz, 2H),
7.39(d, J = 8.1 Hz, 2H) ,
7.74(s, 1H) , 12.57(br s, 1H)
EI-MS m/z 357(M'), C,BH,SN03S2=357
Example 159
Preparation of Compound 139
Compound 118 (0.33 g, 0.89 mmol) obtained in Example 145 was dissolved in
dimethylformamide ( 15 mL), potassium carbonate (0.12 g, 0.89 mmol) and benzyl
bromide
(0.13 mL, 1.1 mmol) were added thereto, followed by stirring at room
temperature for 10
hours. Then the conventional treatment was performed, and the product was
purified with thin
layer chromatography (hexane/ etyhl acetate = 8/1) to give Compound 139 (0.077
g, 50%).
Compound 139:
1H-NMR(300MHz, DMSO-d6) S(ppm) 2.33(s, 3H), 5.37(s, 2H), 7.11(d, J=8.4Hz, 1H),
7.31(d, J = 8.3Hz, 2H), 7.38(d, J = 8.3Hz, 2H), 7.39(m, 1H), 7.41(d, J =
8.3Hz, 2H) , 7.46(d, J
= 7.3Hz, 2H) , 7.91 (dd, J = 1.8, 8.4Hz, 1 H), 7.96(s, 1 H) , 8.08(d, J = 1.
SHz, 1 H), 12.76(br s,
1 H)
EI-MS m/z 461(M'), C25H,9N04S2=461
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Example 160
Preparation of Compound 140
Compound 140 (0.067 g, 64.0%) was obtained by using Compound 95 (0.1 g,
0.28 mmol) using the same method as Example 96 (synthesis of Compound 69).
Compound 140:
IH NMR (300 MHz, DMSO-d6) 8(ppm) 2.34 (s, 3H), 7.40 (d, J = 8.4 Hz, 2H), 7.63
(d, J
= 8.1 Hz, 2H), 7.80 (s, 1 H), 8.09 (dd, J = 1.7, 8.4 Hz, 1 H), 8.19 (d, J = 1.
S Hz, 1 H) , 8.22 (d, J =
8.3 Hz, 1 H), 12. 80 (br s, 1 H}
FABMS m/z 369 (M+H)+ C,sH,ZN203S2 = 368
Example 161
Preparation of Compound 141
Compound 141 (0.094 g, 89.9%) was obtained by using Compound 109 (0.1 g,
0.28 mmol) using the same method as Example 96 (synthesis of Compound 69).
Compound 141:
~H NMR (300 MHz, DMSO-d6 8(ppm) 2.49 (s, 3H), 7.36 (d, J = 8.4 Hz, 2H), 7.62
(d, J
= 8.1 Hz, 2H), 7.76 (s, 1 H), 7.79 (s, 1 H), 7.82 (d, J = 8.3 Hz, 1 H) , 8.10
(d, J = 8.3 Hz, 1 H),
12.77 (br s, 1 H)
FABMS m/z 377 (M-H)' C,~H,23sC1N03Sz = 378
Example 162
Preparation of Compound 142
3-Bromo-4-[(4-methylphenyl)thio]benzaldehyde (0.1 g, 0.33 mmol) obtained in
Example 123 was dissolved in methanol (5 mL), p-toluenesulfonic acid
monohydrate (0.0006
g, 0.003 mmol) and orthoformic acid methyl (0.035 mL, 0.33 mmol) were added
thereto, and
the product was heated under reflux for 3 hours. After the conventional
treatment, the solvent
was removed by using a vacuum dryer, and 3-bromo-4-[(4-
methylphenyl)thio]benzaldehydedimethylacetal was obtained in the form of the
crude product.
Using the whole quantity of the crude product of 3-bromo-4-[(4-
methylphenyl)thio]benzaldehydedimethylacetal, 3-formyl-4-[(4-
methylphenyl)thio]
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benzaldehydedimethylacetal (0.09 g, yield 90.3%) was obtained in a manner
similar to the
synthesis of 5-[hydroxymethyl-2-[(4-methylphenyl)thio)benzaldehyde in Example
123.
Using 3-formyl-4-[(4-methylphenyl)thio]benzaldehydedimethylacetal (0.2 g, 0.67
mmol), Compound 142 (0.21 g, 76.3%) was obtained using the same method as
Example 94
(synthesis of Compound 67).
3-Formyl-4-[(4-methylphenyl)thio)benzaldehydedimethylacetal:
1H-NMR(300MHz, CDC13) 8(ppm) 2.39 (s, 3H), 3.32 (s, 6H), 5.39 (s, 1H), 6.99
(d,
J=8.3Hz, 1H), 7.22(d, J=7.9Hz, 2H), 7.38(d, J=8.3Hz, 2H), 7.43(dd, J=2.0,
8.3Hz, IH), 7.94(d,
J=2.OHz, 1 H), 10.3 5 (s, 1 H)
FAB-MS m/z=302(M-), Ci~H1803S=302
Compound I42:
~H NMR (300 MHz, DMSO-d6) 8(ppm) 2.49 (s, 3H), 3.27 (s, 6H) , 5.44 (s, 1H),
7.26
(d, J = 2.6 Hz, 4H), 7.40 (d, J = 8.4 Hz, 1 H), 7.54 (s, 1 H), 8.04 (s, 1 H),
12.68 (br s, 1 H)
FABMS m/z 401 (M-H)' CZpH~9NO4S2 = 402
Example 163
Preparation of Compound 143
Using Compound 88 (0.07 g, 0.17 mmol), Compound 143 (0.049 g, 69.4%) was
obtained using the same method as Example 96 (synthesis of Compound 69).
Compound 143:
'H NMR (300 MHz, DMSO-db) 8(ppm) 2.33 (s, 3H), 7.35 (d, J = 8.1 Hz, 2H), 7.63
(d, J
= 8.3 Hz, 2H), 7.79 (s, 1 H), 7.86 (d, J = 8.4 Hz, 1 H), 7.91 (s, I H), 8.08
(d, J = 8.3 Hz, I H),
12.76 (br s, 1 H)
FABMS m/z 422 (M+) C~7H,2~9BrNO3S2 = 422
Example 164
Preparation of Compound 144
Compound I42 (0.17 g, 0.43 mmol) was dissolved in
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dichloromethane ( 14 mL) and methanol (2.6 mL}, and a 1 mol/L hydrogen
chloride aqueous
solution (0.5 mL) was added thereto. After heating under the reflux for 2
hours, the product
was cooled to room temperature, and extracted with ethyl acetate. The solvent
was evaporated
under reduced pressure and the residue was recrystallized from ethyl acetate
and hexane to
obtain Compound 144 (0.15 g, 98.2%).
Compound 144:
'H NMR (300 MHz, DMSO-d6) 8(ppm) 2.49 (s, 3H), 7.12 (d, J = 8.4 Hz, IH), 7.34
(d, J
= 8.3 Hz, 2H), 7.45 (d, J = 8.1 Hz, 2H), 7.83 (d, J = 8.3 Hz, 1 H), 7.95 (s, 1
H), 7.97 (s, 1 H), 9.97
(s, 1 H), 12.78 (br s, 1 H)
FABMS m/z 356 (M+H)+ C,gH,3NO3S2 = 355
Example 165
Preparation of Compound 145
Compound 144 (0.05 g, 0.14 mmol) was dissolved in chloroform (4 mL), and
((tert-
butoxycarbonyl)methylene]triphenylphosphine (0.13 g, 0.35 mmol) was added
thereto. After
heating under the reflux for 2 hours, the product was cooled to room
temperature, and extracted
with ethyl acetate. The solvent was evaporated under reduced pressure, and the
residue was
purified by preparative thin layer chromatography (development solvent:
chloroform/
acetonitrile = 18/1). Then the product was recrystallized from ethyl acetate
and hexane to give
Compound 145 (0.038 g, 59.8%).
Compound 145:
1H NMR (300 MHz, DMSO-db) 8(ppm) 1.48 (s, 9H), 2.33 (s, 3H), 6.49 (d, J = 16.1
Hz,
1H), 7.08 (d, J = 8.3 Hz, 1H}, 7.27 (d, J = 8.6 Hz, 2H), 7.35 (d, J = 8.3 Hz,
2H), 7.56 (d, J =
16.1 Hz, 1 H), 7.69 (s, 1 H) , 7.72 (d, J = 8.3 Hz, 1 H), 7.95 (s, 1 H), 12.70
(br s, 1 H)
FABMS m/z 453 (M-H)' C24HzsNOaS2 = 454
Example I66
Preparation of Compound 146
Compound 145 (0.02 g, 0.04 mmol) was dissolved in dichloromethane (4 mL), and
trifluoroacetic acid ( 1 mL) was added thereto, followed by stirring at room
temperature for 1
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hour. Then the conventional treatment was performed, the solvent was
evaporated under
reduced pressure, and the residue was triturated with ethyl acetate to give
Compound 146
(0.015 g, 87.5%).
Compound 146:
~H NMR (300 MHz, DMSO-d6) S(ppm) 2.50 (s, 3H), 6.51 (d, J = 16.0 Hz, 1 H),
7.11 (d,
J = 8.1 Hz, 1 H), 7.27 (d, J = 8.4 Hz, 2H), 7.34 (d, J = 8.1 Hz, 2H), 7.60 (d,
J = 16.0 Hz, 1 H),
7.70 (s, 1 H) , 7.72 (d, J = 8.4 Hz, 1 H), 7.96 (s, 1 H), 12.50 (br s, 1 H) ,
12.71 (br s, 1 H)
FABMS m/z 396 (M-H)' CzoHisNOasz = 397
Example 167
Preparation of Compound 147
Using 4-fluoro-5-(trifluoromethyl)benzaldehyde (0.20 g, 1.0 mmol), 4-[(4-
methylphenyl)thio]-3-(trifluoromethyl)benzaldehyde (0.30 g, 100%) was obtained
in a manner
similar to the synthesis of 5-bromo-2-[(4-methylphenyl)thio]benzaldehyde.
Compound 147 (0.13 g, 50.2%) was obtained by using 4-[(4-methylphenyl)thio]-3-
(trifluoromethyl)benzaldehyde (0.2 g, 0.7 mmol) using the same method as
Example 94
(synthesis of Compound 67).
4-[(4-methylphenyl)thio]-3-(trifluoromethyl)benzaldehyde:
~H NMR (300 MHz, CDC13) 8(ppm) 2.43 (s, 3H), 7.01 (d, J = 8.3 Hz, 1H), 7.29
(d, J =
7.9 Hz, 2H), 7.46 (d, J = 8.1 Hz, 2H), 7.73 (d, J = 8.4 Hz, 1 H), 8.11 (s, 1
H), 9.93 (s, 1 H)
FABMS m/z 296 (M+ ) CISH"F30S = 296
Compound 147:
'H NMR (300 MHz, DMSO-d6) 8(ppm) 2.49 (s, 3H), 7.08 (d, J = 8.3 Hz, 1 H), 7.35
(d, J
= 8.1 Hz, 2H), 7.46 (d, J = 8.1 Hz, 2H), 7.67 (d, J = 8.4 Hz, 1 H), 7.82 (s, 1
H), 8.02 (s, 1 H) ,
12.68 (br s, 1 H)
FABMS m/z 395 (M+) C,gH,z~9F3NOZSz = 395
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Example 168
Preparation of Compound 148
Compound 148 (0.047 g, 64.3%) was obtained by using Compound 147 (0.07 g, 0.18
mmol) using the same method as Example 96 (synthesis of Compound 69).
Compound 148:
~H NMR (300 MHz, DMSO-db) 8(ppm) 2.49 (s, 3H), 7.37 (d, J = 8.4 Hz, 2H), 7.53
(d, J
= 8.3 Hz, 2H), 7.93 (s, 1 H), 8.10 (d, J = 7.7 Hz, 1 H), 8.11 (s, 1 H), 8.3 5
(d, J = 8.3 Hz, 1 H) ,
12.80 (br s, 1 H)
FABMS m/z 412 (M+H)+ CigH,2~9F3NO3S2 = 411
Example 169
Preparation of Compound 149
A reaction similar to Example 81 was performed using 5-carboxy-2-[(4-
methylphenyl)thio]benzaldehyde (0.10 g, 0.37 mmol) to obtain 5-(N,N-
diethylaminocarbonyl)-
2-[(4-methylphenyl)thio]benzaldehyde (0.046 g, 36.7%).
Compound 149 (0.043 g, 76.1 %) was obtained by using 5-(N,N-
diethylaminocarbonyl)-2-[(4-
methylphenyl)thio]benzaldehyde (0.046 g, 0.13 rnmol) using the same method as
Example 94
(synthesis of Compound 67).
5-(N,N-diethylaminocarbonyl)-2-[(4-methylphenyl)thio]benzaldehyde:
1H NMR (300 MHz, CDC13) S(ppm) 1.19 (m, 3H), 1.53 (m, 4H), 2.41 (s, 3H}, 6.98
(d, J
= 8.3 Hz, 1 H), 7.23 (d, J = 8.4 Hz, 2H), 7.36 (dd, J = 2.0, 8.3 Hz, 1 H),
7.40 (d, J = 8.3 Hz, 2H),
7. 86 (d, J = 2.0 Hz, 1 H), 10.3 5 (s, 1 H)
FABMS m/z 328 (M+H)+ C,9H2,NOZS = 327
Compound 149:
~H NMR (300 MHz, DMSO-db) S(ppm) 1.1 (m, 6H), 2.50 (s, 3H), 3.38 (m, 4H), 7.16
(d, J = 8.3 Hz, 1H), 7.27 (d, J = 8.3 Hz, 2H) , 7.34 (d, J = 8.4 Hz, 2H) ,
7.37 (d, J = 8.1 Hz, 1H)
7.39 (s, 1 H), 7.97 (s, 1 H), 12.03 (br s, 1 H)
FABMS m/z 427 (M)+ CZZH2zN203S2 = 427
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Example 170
Preparation of Compound 150
Compound 150 (0.024 g, 26.2%) was obtained by using Compound 149 (0.09 g, 0.21
mmol) using the same method as Example 96 (synthesis of Compound 69}.
Compound 150:
'H NMR (300 MHz, DMSO-db) 8(ppm) 1.1 (m, 6H}, 2.29 (s, 3H), 2.49 (m, 4H), 7.31
(d, J = 8.4 Hz, 2H), 7.40 (s, 1 H} , 7.46 (d, J = 8.1 Hz, 2H) , 7.63 (d, J =
7.9 Hz, 1 H) , 7.84 (s,
1 H), 8.05 (d, J = 8.1 Hz, 1 H), NH not found
FABMS m/z 443 (M)+ C22H22N2~4S2 = 443
Example 171
Preparation of Compound 151
4-{3-Formyl-4-[(4-methylphenyl)thioJbenzoyl}-morpholine (0.092g, 24.4%) was
obtained by using 5-carboxy-2-[(4-methylphenyl)thio]benzaldehyde (0.3 g, 1.1
mmol) and
using morpholine (0.19 mL, 2.2 mmol) instead of diethylamine with the same
method as
Example 81.
Compound 151 (0.066 g, 55.8%) was obtained by using 4-{3-formyl-4-[(4
methylphenyl)thio]benzoyl}-rnorpholine (0.092 g, 0.27 mmol) using the same
method as
Example 94 (synthesis of Compound 67).
4-{3-Formyl-4-[(4-methylphenyl)thioJbenzoyl}-morpholine:
'H NMR (300 MHz, CDC13} s(ppm) 2.41 (s, 3H), 2.89 (s, 4H), 2.97 (s, 4H), 6.97
(d, J =
8.3 Hz, 1 H), 7.22 (m, 1 H), 7.26 (d, J = 7.9 Hz, 2H), 7.41 (d, J = 8.1 Hz,
2H), 7.89 (d, J = 1.8
Hz, 1 H), 10.34 (s, 1 H)
FABMS m/z 342 (M+H)+ C,9H,9N03S = 341
Compound 151:
'H NMR (300 MHz, DMSO-d6) 8(ppm) 2.30 (s, 3H), 3.40 (s, 4H), 3.60 (s, 4H},
7.13 (d,
J = 8.3 Hz, 1 H), 7.27 (d, J = 8.4 Hz, 2H) , 7.35 (d, J = 7.7 Hz, 2H) , 7.39
(d, J = 8.3 Hz, 1 H) ,
7.92 (s, 1 H), 8.29 (d, J = 3.9 Hz, 1 H), 12.02 (br s, 1 H)
FABMS m/z 441 (M)+ C22HZaNZO4S2 = 441
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Example 172
Preparation of Compound 152
3-Acetyl-4-fluorobenzaldehyde (0.38 g, 47.0%) was obtained by using 5-bromo-4-
fluorobenzaldehyde (1.0 g, 4.9 mmol) in a manner similar to the synthesis of 5-
acetyl-2-
fluorobenzaldehyde in Example 124.
3-Acetyl-4-[(3,4-dichlorophenyl)thio]benzaldehyde (0.10 g, 13.7%) was obtained
by
using 3-acetyl-4-fluorobenzaldehyde (0.38 g, 2.3 mmoI) and using 3,4-
dichlorothiophenol
instead of 4-methylthiophenol in a manner similar to the synthesis of 5-bromo-
2-[(4-
methylphenyl)thio]benzaldehyde in Example 114.
Compound 152 (0.044 g, 32.5%) was obtained by using 3-acetyl-4-[(3,4-
dichlorophenyl)thio]benzaldehyde (0.1 g, 0.32 mmol} using the same method as
Example 94
(synthesis of Compound 67).
3-acetyl-4-fluorobenzaldehyde:
~H NMR (300 MHz, CDCl3} 8(ppm) 2.64 (s, 3H), 7.25 (dd, J = 8.4, 10.4 Hz, 1H),
8.Oi
(m, I H), 8.35 (dd, J = 2.4, 7.1 Hz, 1 H), 9.95 (s, 1 H)
CIMS m/z 167 (M+H)+ C9H~F0z = 166
3-Acetyl-4-[(3,4-dichlorophenyl)thio]benzaldehyde:
~H NMR (300 MHz, CDCI3) 8(ppm) 2.72 (s, 3H), 6.95 (d, J = 2.0 Hz, I H), 7.37
(dd, J =
2.0, 8.1 Hz, 1 H), 7.53 (d, J = 2.0 Hz, 1 H), 7.62 (d, J = 2.0 Hz, 1 H), 7.72
(dd, J = 1.8, 8.3 Hz,
1 H) , 8.3 5 (d, J = 1.8 Hz, 1 H), 9.95 (s, 1 H)
FABMS m/z 325 (M~ C,SHioC120zS = 325
Compound 152:
1H NMR (300 MHz, DMSO-d6) S(ppm) 2.69 (s, 3H), 6.96 (d, J = 8.4 Hz, IH), 7.53
(dd,
J = 2.0, 8.3 Hz, 1 H) , 7.66 (dd, J = 1.8, 8.5 Hz, 1 H) , 7.74 (s, 1 H) , 7.78
(d, J = 8.3 Hz, 1 H),
7.86 (d, J = 2.0 Hz, 1 H), 8.32 (d, J = 1. 8 Hz, 1 H), NH not found
FABMS m/z 424 (M+) CIgH"C1zN03S2 = 424
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Example 173
Preparation of Compound 153
Compound 153 (0.094 g, 89.9%) was obtained by using Compound 152 (0.03 g,
0.071
mmol) using the same method as Example 96 (synthesis of Compound 69).
Compound 153:
~H NMR (300 MHz, DMSO-db) S(ppm) 2.50 (s, 3H), 7.65 (dd, J = 2.0, 8.3 Hz, 1H),
7.74 (d, J = 8.6 Hz, 1 H) , 7.93 (d, J = 1.7 Hz, 1 H) , 7.96 (s, 1 H) , 8.11
(dd, J = 1.8, 8.3 Hz, 1 H),
8.43 (s, 1 H) , 8.46 (d, J = 8.8 Hz, 1 H), 12.77 (br s, 1 H)
FABMS m/z 440 (M~) CIgH" C1zN04S2 = 440
Example 174
Preparation of Compound 154
2-[(2,3-Dichlorophenyl)thio]-5-nitrobenzaldehyde (0.57 g, 97.9%) was obtained
by
using 5-nitro-2-fluorobenzaldehyde (0.30g, 1.77 mmol) and using 2,3-
dichlorothiophenol
instead of p-toluenethiol in a manner similar to the synthesis of 5-bromo-2-
[(4-methylphenyl)-
thio]benzaldehyde in Example 114.
Compound 154 (0.088 g, 22.5%} was obtained by using 2-[(2,3-
dichlorophenyl}thio]-5-
nitrobenzaldehyde (0.3 g, 0.91 mmol) using the same method as Example 96
(synthesis of
Compound 69).
2-[(2,3-Dichlorophenyl)thio]-5-nitrobenzaldehyde:
~H NMR (300 MHz, CDCl3) 8(ppm) 6.91 (d, J = 8.8 Hz, 1H), 7.34 (d, J = 8.1 Hz,
1H),
7. 5 8 (dd, J = 1. 5, 7.7 Hz, 1 H), 7.66 (dd, J = 1.7, 8.1 Hz, 1 H), 8.19 (dd,
J = 2.6, 8.9 Hz, 1 H) ,
8.72 (d, J = 2.6 Hz, 1 H), 10. 31 (s, 1 H)
FABMS m/z 328 (M+) C,3H7C12N03S = 328
Compound 153:
1H NMR (300 MHz, DMSO-d6) S(ppm) 7.25 (d, J = 8.6 Hz, 1H), 7.55 (d, J = 2.0
Hz,
1 H), 7.57 (s, 1 H) , 7.81 (s, 1 H) , 7.92 (d, J = 2.0 Hz, 1 H} , 8.16 (dd, J
= 2.4, 8.7 Hz, 1 H), 8.3 0
(d, J = 2.4 Hz, 1 H}, NH not found
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FABMS m/z 425 (M-H)' C~6Hg35CIZN2O4S2 = 426
Example 175
Preparation of Compound 155
2-[(2,4-Dichlorophenyl)thin]-5-nitrobenzaldehyde (0.53 g, 91.7%) was obtained
by
using S-vitro-2-fluorobenzaldehyde (0.30 g, 1.77 mmol) and using 2,4-
dichlorothiophenol
instead of p-toluenethiol in a manner similar to the synthesis of 5-bromo-2-
[(4-methylphenyl)-
thio]benzaldehyde in Example 114.
Compound 155 (0.13 g, 32.4%) was obtained by using 2-[(2,4-
dichlorophenyl)thio]-5-
nitrobenzaldehyde (0.3 g, 0.91 mmol) using the same method as Example 96
(synthesis of
Compound 69).
2-[(2,4-Dichlorophenyl)thio]-5-nitrobenzaldehyde:
'H NMR (300 MHz, CDCl3) 8(ppm} 6.86 (d, J = 8.8 Hz, 1 H), 7.40 (dd, J = 2.2,
8.3 Hz,
1 H), 7.61 (s, 1 H), 7.64 (t, J = 2.6 Hz, 1 H), 8.17 (dd, J = 2.6, 8.8 Hz, 1
H) , 8.71 (d, J = 2.6 Hz,
1 H), 10.30 (s, 1 H)
FABMS m/z 328 (M+H)+ C13H~35C12NO3S = 327
Compound 155:
~H NMR (300 MHz, DMSO-d6) 8(ppm) 7.38 (d, J = 8.8 Hz, 1H), 7.41 (s, 1H), 7.43
(d,
J = 1.3 Hz, 1 H) , 7.75 (dd, J = 3.9, 5.7 Hz, 1 H) , 7.80 (s, 1 H) , 8.20 (dd,
J = 2.4, 8.8 Hz, 1 H),
8.32 (d, J = 2.4 Hz, 1H), NH not found
FABMS m/z 425 (M-H)' C,6Hg35C12NzO4S2 = 426
Example 176
Preparation of Compound 156
Compound 156 (0.016 g, 43.1%) was obtained by using Compound 154 (0.037 g,
0.085
mmol) using the same method as Example 96 (synthesis of Compound 69).
Compound 156:
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'H NMR (300 MHz, DMSO-d6) S(ppm) 7.74 (dd, J = 2.0, 8.4 Hz, 1H), 7.77 (s, 1H),
7.79 (s, 1 H) , 7.82 (t, J = 2.2 Hz, 1 H) , 8.00 (d, J = 8.6 Hz, 1 H) , 8.30
(dd, J = 2.2, 8.7 Hz, 1 H),
8.42 (d, J = 2.2 Hz, 1 H), NH not found
FABMS m/z 441 (M-H)- Ci6H835CI2NzO5S2 = 442
Example 177
Preparation of Compound 157
Compound 157 (0.030g, 44.1%) was obtained by using Compound 155 (0.066 g,
0. l5mmol) using the same method as Example 96 (synthesis of Compound 69).
Compound 157:
'H NMR (300 MHz, DMSO-d6) 8(ppm) 7.67 (t, J = 7.7 Hz, 1H), 7.85 (d, J = 0.9
Hz,
1 H), 7.88 (s, 1 H) , 8.10 (d, J = 8.8 Hz, 1 H) , 8.14 (s, 1 H) , 8.28 (d, J =
2.2 Hz, 1 H), 8.42 (d, J =
2.4 Hz, 1 H), NH not found
FABMS m/z 441 (M-H)' C16H835C12NZOSS2 = 442
Example 178
Preparation of Compound 158
Compound 158 (0.0085 g, 26.4%) was obtained by using Compound 154 (0.03 g,
0.070
mmol) using the same method as Example 96 (synthesis of Compound 69).
Compound 158:
'H NMR (300 MHz, DMSO-db) 8(ppm) 7.19 (d, J = 8.8 Hz, 1H), 7.55 (d, J = 1.7
Hz,
1 H), 7.55 (s, 1 H) , 7.70 (s, 1 H) , 7.92 (s, 1 H) , 8.11 (dd, J = 2.6, 8.8
Hz, 1 H), 8.36 (d, J = 2.4
Hz, 1 H), NH not found
Example 179
Preparation of Compound 159
Compound 159 (0.013 g, 38.9%) was obtained by using Compound 155 (0.03 g,
0.070
mmol) using the same method as Example 96 (synthesis of Compound 69).
Compound 159:
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'H NMR (300 MHz, DMSO-d6) 8(ppm) 7.35 (d, J = 8.8 Hz, 1H), 7.40 (s, 1H), 7.42
(d, J
= 4.4 Hz, 1 H) , 7.75 (dd, J = 2.4, 6.7 Hz, 1 H) , 7.76 (s, 1 H) , 8.18 (dd, J
= 2.6, 8.8 Hz, 1 H), 8.35
(d, J = 2.6 Hz, 1 H), NH not found
Exam In a 180
Preparation of Compound 160
2-Fluoro-S-nitrobenzaldehyde (0.16 g, 0.95 mmol) was dissolved in N,N-
dimethylformamide (8.1 mL), and 4-mercaptophenol (0.11 g, 0.86 mmol) and
triethylamine
(0.27 mL, 1.9 mmol) were added thereto, followed by stirring at 25°C
for 1 hour. After the
conventional treatment, the product was purified by silica gel column
chromatography (eluted
by n-hexane/acetone = 4/1-1/1) to give 5-nitro-2-[(4-
hydroxyphenyl)thio]benzaldehyde (0.18 g,
70%).
5-Nitro-2-[(4-hydroxyphenyl)thio]benzaldehyde (0.17 g, 0.61 mmol) was
dissolved in
toluene (8.4 mL), and 2,4-thiazolidinedione (0.29 mg, 2.4 mmol), piperidine
(0.024 mL, 0.24
mmol), acetic acid (0.014 mL, 0.24 mmol) and molecular sieves 4A (0.84 g) were
added
thereto, followed by stirring at 110 °C for 2 hours. After the
conventional post-reaction
treatment, thin layer chromatography (developed with chloroform/methanol =
10/1 ) to give
Compound 160 (34 mg, 15%).
5-Nitro-2-[(4-hydroxyphenyl)thio]benzaldehyde:
iH NMR (300 MHz, CDCl3) 8(ppm) 6.97 (d, J = 9.0 Hz, 1H), 7.03 (d, J = 8.6 Hz,
2H),
7.40 (d, J = 8.8 Hz, 2H), 8.06 (m, 1 H), 8.61 (br s, 1 H), 8.63 (d, J = 2.3
Hz, 1 H), 10.29 (s, 1 H)
Compound 160:
~H NMR (300 MHz, DMSO-db) S(ppm) 6.94 (d, J = 8.8 Hz, 3H), 7.45 (d, J = 8.6
Hz,
2H), 7.87 (s, 1 H), 8.13 (dd, J = 2.4, 8.8 Hz, 1 H), 8.20 (d, J = 2.2 Hz, 1
H), 10.20 (s, 1 H), 12.86
(br s, 1 H)
FABMS m/z 373 (M-H)' C~6H1oN205S2 = 374
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Example 181
Preparation of Compound 161
2-[(3,4-Dimethylphenyl)thio]-5-nitrobenzaldehyde (0.16 g, 45%) was obtained
from
3,4-dimethylbenzenethiol (0.16 mL, 1.2 mmol), a 2.5 mol/L sodium hydroxide
aqueous
solution (2.1 mL, 5.2 mmol), tetrabutylammonium bromide (0.020 mg, 0.61 mmol)
and a
toluene solution (2.1 mL) of 2-fluoro-5-nitrobenzaldehyde (0.21 g, 1.2 mmol)
using the same
method as Example 97.
Compound 161 (0.13 g, 64%) was obtained from 2-[(3,4-dimethylphenyl)thio]-S-
nitrobenzaldehyde (0.16 g, 0.54 mmol), toluene (7.8 mL), 2,4-thiazolidinedione
(0.26 mg, 2.2
mmol), piperidine (0.022 mL, 0.22 mmol), acetic acid (0.012 mL, 0.22 mmol) and
molecular
sieves 4A (0.78 g).
2-[(3,4-DimethylphenyI)thio]-S-nitrobenzaldehyde:
'H NMR (300 MHz, CDC13) 8(ppm) 2.31 (s, 3H), 2.36 (s, 3H), 6.97 (d, J = 9.0
Hz, 1H),
7.24-7.36 (m, 3H), 8.09 (dd, J = 9.0, 2.5 Hz, 1 H), 8.67 (d, J = 2.6 Hz, 1 H),
10.31 (s, 1 H)
FABMS m/z 287 (M)+ C,SHi3N03S = 287
Compound 169:
'H NMR (300 MHz, DMSO-d6) S(ppm) 2.25 (s, 3H), 2.28 (s, 3H), 7.06 (d, J = 9.0
Hz,
1 H), 7.33 (s, 2H), 7.40 (s, 1 H), 7.88 (s, 1 H), 8.15 (m, 1 H), 8.21 (m, 1
H), 12.86 (br s, 1 H)
FABMS m/z 385 (M-H)- ClgH~4N2O4Sz = 386
Example 182
Preparation of Compound 162
Compound 162 (0.60 mg, 0.9%) was obtained from Compound 72 (0.061 g, 0.14
mmol)
obtained in Example 56, dichloromethane (6.1 mL), methanol (1.2 mL) and m-
chloroperbenzoic acid (0.75 g, 2.1 mmol) using the same method as Example 134.
Compound 162:
1H NMR (300 MHz, DMSO-db) 8(ppm) 7.96 (d, J = 8.4 Hz, 1 H), 8.02 (dd, J = 8.7,
2.2
Hz, 1 H), 8.17 (s, 1 H), 8.28 (s, 1 H), 8.32 (d, J = 2.2 Hz, 1 H), 8.44 (d, J
= 1.3 Hz, 2H)
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Example 183
Preparation of Compound 163
Compound 163 (0.034 g, 32%) was obtained from Compound 81 (0.10 g, 0.26 mmol)
obtained in Example 108, N,N-dimethylformamide (5.1 mL) and m-chloroperbenzoic
acid
(0.14 g, 0.82 mmol) using the same method as Example 156.
Compound 163:
1H NMR (300 MHz, DMSO-d6) 8(ppm) 7.58 (d, J = 8.4 Hz, 2H), 7.74 (d, J = 8.4
Hz,
2H), 8.00 (s, I H), 8.30 (dd, J = 8.2, 1.4 Hz, 1 H), 8.48 (d, J = 8.2 Hz, 1
H), 8.53 (d, J = 1.4 Hz,
1 H), 12. 83 (br s, 1 H)
FABMS m/z 407 {M-H)' Ci6H935C1N2OSS2 = 408
Example 184
Preparation of Compound 164
A 2.5 mol/L sodium hydroxide aqueous solution (2.0 mL, 4.9 mmol) and
tetrabutylammonium bromide (0.019 g, O.S8 mmol) were added to 4-
ethylbenzenethiol (0.20 g,
1.2 mmol), followed by stirring at 2S °C for S minutes. A toluene
solution (2.0 mL) of 4
fluoro-3-nitrobenzaldehyde (0.20 g, 1.2 mmol) was added to the reaction
liquid, followed by
stirring at 2S°C for 12 hours. After the conventional post-reaction
treatment, the product was
purified by silica gel column chromatography (eluted by chloroform) to give 4-
[(4-
ethylphenyl)thio]-3-nitrobenzaldehyde (0.25 g, 75%).
4-[(4-Ethylphenyl)thin]-3-nitrobenzaldehyde (0.25 g, 0.86 mmol) was dissolved
in
ethanol (9.9 mL), and 2,4-thiazolidinedione (0.40 g, 3.4 mmol) and piperidine
(0.034 mL, 0.34
mmol) were added thereto, followed by stirring at 80°C for 7 hours. The
precipitated crystals
were collected by filtration to give Compound 164 (0.061 g, 19%).
4-[(4-ethylphenyl)thio]-3-nitrobenzaldehyde:
'H NMR (300 MHz, CDCl3) 8(ppm) 1.31 (t, J = 7.7 Hz, 3H), 2.76 (q, J = 7.S Hz,
2H),
7.01 (d, J = 8.4 Hz, 1 H), 7.37 (d, J = 8.2 Hz, 2H), 7.50 (d, J = 8.2 Hz, 2H),
7.81 (dd, J = 8.4, 1.8
Hz, 1 H), 8.69 (d, J = 1.6 Hz, 1 H), 9.97 (s, 1 H)
FABMS mlz 287 (M)+ ClsH,3N03S = 287
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Compound 164:
1H NMR (300 MHz, DMSO-db) 8(ppm) 1.24(t, J = 7.5 Hz, 3H), 2.70 (q, J = 7.3 Hz,
2H), 6.92 (d, J = 8.6 Hz, 1 H), 7.08 (br s, 1 H), 7.43 (d, J = 8.1 Hz, 2H),
7.56 (d, J = 8.0 Hz, 2H),
7.58 (s, 1H), 7.72 (dd, J = 8.6, 2.0 Hz, 1H), 8.44 (d, J = 1.8 Hz, 1 H)
FABMS m/z 385 (M-H)' C,$H,4NzO4S2 = 386
Example 185
Preparation of Compound 165 and Compound 166
Compound 165 (0.014 g, 26%) and Compound 166 (O.OI 6 g, 26%) were obtained
from
Compound 164 (0.052 g, 0.14 mmol) obtained in Example 184, N,N-
dimethylformamide (2.6
mL) and m-chloroperbenzoic acid (0.44 g, 1.3 mmol) using the same method as
Example 156.
Compound 165:
1H NMR (300 MHz, DMSO-d6) 8(ppm) 1.13 (t, J = 7.5 Hz, 3H), 2.61 (q, J = 7.5
Hz,
2H), 7.34 (d, J = 8.3 Hz, 2H), 7.51-7.56 (m, 2H), 7.95 (s, 1H), 8.30 (dd, J =
8.4, 1.6 Hz, 1H),
8.48-8.52 (m, 2H), 12.87 (m, 1 H)
FABMS m/z 401 (M-H)' ClgH~4N2O5S2 = 402
Compound 166:
~H NMR (300 MHz, DMSO-db) 8(ppm) 1.19 (t, J = 7.7 Hz, 3H), 2.71 (q, J = 8.4
Hz,
2H), 7.51-7.60 (m, 2H), 7. 80 (s, 1 H), 7.86-7.95 (m, 2H), 8.03 (dd, J = 8.8,
1.5 Hz, 1 H), 8.21 (d,
J = 1.1 Hz, 1 H), 8.45 (d, J = 8.2 Hz, 1 H), 12.90 (m, 1 H)
FABMS m/z 417 (M-H)' C,gH,4Nz06S2 = 418
Example 186
Preparation of Compound 167
4-[(3,4-Dichlorophenyl)thioJ-3-nitrobenzaldehyde (0.29 g, 84%) was obtained
from
3,4-dichlorobenzenethiol (0.13 mL, 1.0 mmol), a 2.5 moUL sodium hydroxide
aqueous solution
(1.8 mL, 4.4 mmol), tetrabutylammonium bromide (0.017 g, 0.52 mmol) and a
toluene solution
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(1.8 mL) of 4-fluoro-3-nitrobenzaldehyde (0.18 g, 1.0 mmol) using the same
method as
Example 184.
Compound 167 (0.077 g, 15%) was obtained from 4-[(3,4-dichlorophenyl)thio]-3-
nitrobenzaldehyde (0.40 g, 1.2 mmol), ethanol (16 mL), 2,4-thiazolidinedione
(0.57 g, 4.9
mmol) and piperidine (0.048 mL, 0.49 mmol).
4-[(3,4-dichlorophenyl)thio]-3-nitrobenzaldehyde:
'H NMR (300 MHz, CDC13) 8(ppm) 7.03 (d, J = 8.4 Hz, 1H), 7.44 (dd, J = 8.3,
2.0 Hz,
1 H), 7.63 (d, J = 8.3 Hz, 1 H), 7.72 (d, J = 2.0 Hz, 1 H), 7.88 (dd, ~J =
8.4, 1.8 Hz, 1 H), 8.73 (d, J
= 1.8 Hz, 1 H), 10.00 (s, 1 H)
FABMS m/z 328 (M+H)+ C13H735C12NO3S = 327
Compound 167:
'H NMR (300 MHz, DMSO-d6) 8(ppm) 7.11 (d, J = 8.6 Hz, 1H), 7.65 (dd, J = 8.3,
2.2
Hz, 1 H), 7.77 (dd, J = 2.0, 8.6 Hz, 1 H), 7.85 (d, J = 8.2 Hz, 1 H), 7.87 (s,
1 H), 8.02' (d, J = 2.0
Hz, 1 H), 8.52 (d, J = 2.0 Hz, 1 H), 12.74 (br s, 1 H)
FABMS m/z 425 (M-H)- C,6Hg35C12N2O4S2 = 426
Example 187
Preparation of Compound 168
Compound 168 (0.017 g, 54%) was obtained from Compound 167 (0.031 g, 0.073
mmol) obtained in Example 186, N,N-dimethylformamide (1.6 mL) and m-
chloroperbenzoic
acid (0.035 g, 0.10 mmol) using the same method as Example 156.
'H NMR (300 MHz, DMSO-d6) S(ppm) 7.69 (m, 1H), 7.78 (d, J = 8.5 Hz, 1H), 8.00
(s,
2H), 8.28 (d, J =8.1 Hz, 1 H), 8.48 (d, J = 8.9 Hz, 1 H), 8.53 (s, 1 H), 12.84
(br s, 1 H)
FABMS m/z 441 (M-H)' C,6Hg35C12N2O5S2 = 442
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Example 188
Preparation of Compound 169 and Compound 170
Compound 169 (0.080 g, 75%) and Compound 170 (2.5 mg, 2.3%) were obtained from
Compound 161 (0.10 g, 0.27 mmol) obtained in Example 181, N,N-
dimethylformamide (5.2
mL) and m-chloroperbenzoic acid (0.46 g, 4.0 mmol) using the same method as
Example 156.
Compound 169: FABMS m/z 401 (M-H)' C,gH,4N205S2 = 402
Compound 170: FABMS m/z 417 (M-H)' C,gH~qN2O6S2 = 418
Example 189
Preparation of N (4-Chlorophenyl)-4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-
benzenesulfonamide
ci
0
H
Step A: General Procedure 1.
General Procedure 1 with benzaldehyde was followed to obtain 5-benzylidene-
thiazolidine-2,4-
dione.
NMR (DMSO-db): 7.75 (s, 1 H), 7.6-7.4 (m, SH)
MS (ESI) 205. Found 204 (M-H).
Step B. Preparation of 4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-
benzenesulfonyl chloride.
To a flask charged with chlorosulfonic acid (10 eq.) cooled in an ice bath was
added 5-
benzylidene-thiazolidine-2,4-dione. The solution was stirred at 0 °C
for one hour, warmed to
room temperature and stirred overnight. The reaction was then poured carefully
onto ice and
the resulting precipitate was filtered and air dried to give pure product.
NMR (DMSO-d6): 7.72 (s, 1 H), 7.66 (d, 2H), 7.51 (d, 2H)
MS (ESI) 303. Found 302 (M-H).
Step C. Sulfonylation Reaction.
The sulfonyl chloride and 4-chloro-phenylamine were stirred together in
pyridine heated at
60°C for 12 h. The solution was diluted with EtOAc and washed with 10%
aq. NaHS04
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followed by sat aq. NaCI. The organic phase was separated, dried over NaZS04
and
concentrated under reduced pressure to give pure compound.
MS (ESI) 394. Found 393 (M-H).
Example 190
Preparation of 4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-N p-tolyl-
benzenesulfonamide
i
4~~p \
H
S
4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-Np-tolyl-benzenesulfonamide was
prepared
by the method of Example 189 from p-toluidine.
MS (ESI) 374. Found 373 (M-H).
Example 191
Preparation of 4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-N (4-methoxy-phenyl}-
benzenesulfonamide
I0.
H
4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl}-N (4-methoxy-phenyl)-
benzenesulfonamide was
prepared by the method of Example 189from p-anisidine.
MS (ESI) 390. Found 389 (M-H).
Example 192
Preparation of 4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-N (4-trifluoromethyl-
phenyl)-
benzenesulfonamide
~CF3
O Q. ..O I
\ S. \
\ I~ H
4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-N (4-trifluoromethyl-phenyl)-
benzenesulfonamide
was prepared by the method of Example I 89from 4-(trifluoromethyl)aniline.
MS (ESI) 428. Found 427 (M-H).
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Example 193
Preparation of 4,5-Dichloro-N [3-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-
phenyl]-phthalamic
acid
0
ozH
s
H
CI
CI
Step A: Coupling 2,4-thiazolidinedione (TZD) to aldehyde under acidic
conditions.
3-Nitro benzaldehyde, TZD ( 1.25 eq.), NaOAc (2 eq.), and acetic anhydride ( 1
eq.) were
dissolved in acetic acid and heated to reflux for 12 hours. The reaction was
allowed to cool and
a precipitate was collected, washed and air dried to give pure product. The
filtrate was poured
into water and the resulting precipitate was filtered to afford an additional
yield of product.
MS (ESI) 250. Found 249 (M-H).
Step B: Reduction of the nitro group.
5-(3-Nitro-benzylidene)-thiazolidine-2,4-dione was dissolved in a large volume
of acetic acid.
A small amount of methanol was added as needed to completely dissolve the
starting material.
The solution was warmed gently and iron powder (5 eq.) was added. After 4
hours the mixture
was filtered to remove iron then diluted with an equal volume of water and
extracted with
EtOAc. The organic phase was concentrated under vacuum to give a brown solid.
T'he residue
was dissolved in methanol and impurities were remove by filtration. The
filtrate was
concentrated and the resulting residue was triturated with hexane and
collected by filtration to
give product.
MS (ESI) 220. Found 219 (M-H).
Step C: Condensation with Anhydride
S-(3-Amino-benzylidene)-thiazolidine-2,4-dione was dissolved in THF and
treated with a
solution of 4,5-dichlorophthalic anhydride and a catalytic amount of DMAP. The
solution was
stirred for 8 hours, then filtered and concentrated under vacuum. The residue
was dissolved in
EtOAc and washed with water. The organic layer was concentrated to an oil and
triturated with
hexane to give the product as a solid which was collected by filtration.
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MS (ESI) 436. Found 435 (M-H).
Example 194
Preparation of 3,6-Dichloro-N [3-(2,4-dioxo-thiazolidin-S-ylidenemethyl)-
phenyl]-phthalamic
acid
O OH
I ~ CI
I H I~
CI
H
O
3,6-Dichloro-N [3-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenyl]-phthalamic
acid was
prepared by the method of Example 193 from 3,6-dichlorophthalic anhydride.
MS (ESI) 436. Found 435 (M-H).
Example 195
Preparation of 4-tert-Butyl-N [3-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-
phenyl]-phthalamic
acid
0
N I ,
4-tent-Butyl-N [3-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenyl]-phthalamic
acid was
prepared by the method of Example 193 from 4-tert-butylphthalic anhydride and
isolated as a
64:36 mixture of regioisomers.
MS (ESI) 425. Found 423 (M-2H).
Example 196
Preparation of N [3-(2,4-Dioxo-thiazolidin-5-ylidenemethyl}-phenyl]-3-hydroxy-
phthalamic
acid
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O
_ I
\ H I\
HO
H
O
N [3-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenyl]-3-hydroxy-phthalamic acid
was
prepared by the method of Example 193 from 3-hydroxyphthalic anhydride to give
a 80:20
mixture of regioisomers.
MS (ESI} 384. Found 383 (M-H).
Example 197
Preparation of 3-[3-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenylcarbamoyl]-
pyrazine-2
carboxylic acid
0
I H
H~o
3-[3-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenylcarbamoyl]-pyrazine-2-
carboxylic acid
was prepared by the method of Example 193 from 2,3-pyrazinedicarboxylic
anhydride
MS (ESI) 370. Found 369 (M-H).
1 S Example 198
Preparation of 3-[3-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenylcarbamoyl]-
isonicotinic
acid
\ I O
H O C
li
3-[3-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenylcarbamoyl]-isonicotinic
acid was prepared
by the method of Example 193 from pyridine-3,4-dicarboxylic anhydride as a
70:30 mixture of
isomers.
MS (ESI) 369. Found 368 (M-H).
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Example 199
Preparation of 3-[3-(2,4-Dioxo-thiazolidin-S-ylidenemethyl)-phenylcarbamoyl]-
pyridine-2
carboxylic acid
H O H O
w
H I ~"
3-[3-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenylcarbamoyl]-pyridine-2-
carboxylic acid
was prepared by the method of Example 193 from 2,3-pyridinedicarboxylic
anhydride and
isolated as a mixture of isomers.
MS (ESI) 369. Found 368 (M-H}.
Example 200
Preparation of 4-(4-Chloro-benzenesulfonylamino)-N [3-(2,4-dioxo-thiazolidin-5
ylidenemethyl)-phenyl]-benzamide
0
s ~ I ~ w
~ ~s,o
I~
ci
Step A: Condensation with Acid Chloride
5-(3-Amino-benzylidene)-thiazolidine-2,4-dione prepared in was dissolved in
pyridine and
treated with 4-nitrobenzoyl chloride and a catalytic amount of DMAP. The
reaction was stirred
for 20 minutes then a precipitate was collected by filtration. The solid was
washed repeatedly
with water, saturated aq. NaHC03, and 10% aq HCl solution. The resulting solid
was air dried.
MS (ESI) 369. Found 368 (M-H).
Step B: Reduction of the Nitro Groun
4-Nitro-N [3-(2,4-dioxo-thiazolidin-S-ylidenemethyl)-phenyl]-benzamide was
suspeded in
isopropanol. A small amount of amonium chloride dissolved in water was added
and the
mixture was heated to 70°C for several hours. Iron powder was added and
stirring continued for
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four hours. The solids were removed by filtration and the filtrate
concentrated under vacuum.
The residue was dissolved in EtOAc and washed with water. The organic layer
was
concentrated to an oil and the product precipitated with the addition of
hexanes.
MS (ESI) 339. Found 338 (M-H).
Step C: Condensation with sulfonyl chlorides
4-Amino-N [3-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenyl]-benzamide was
dissolved in
pyridine and treated with 4-chloro-benzenesulfonyl chloride in pyridine at
60°C for 12 h. The
solution was cooled and diluted with EtOAc then washed with 10% aq. NaHS04
followed by
saturated aq. NaCI. The organic phase was separated, dried over NaZS04 and
concentrated
under reduced pressure to give an oil. Trituration with hexane gave pure
product.
MS (ESI) 513. Found S 12 (M-H).
Example 201
Preparation of N [3-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenyl]-4-(toluene-
4-
sulfonylamino)-benzamide
~ "o
NH ~ I NH~
'~J~H
O S
N [3-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenyl]-4-(toluene-4-
sulfonylamino)-benzamide
was prepared by the method of Example 200 from p-toluenesulfonyl chloride.
MS (ESI) 493. Found 492 (M-H).
Example 202
Preparation of N [3-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenylJ-4-(4-
methoxy
benzenesulfonylamino)-benzamide
0
H
O S H I / ~ ~~O
H
O
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N [3-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenyl]-4-(4-methoxy-
benzenesulfonylamino)-
benzamide was prepared by the method of Example 200 from 4-
methoxybenzenesulfonyl
chloride.
MS (ESI) 509. Found 508 (M-H).
Exam In a 203
Preparation of N [3-(2,4-Dioxo-thiazolidin-5-ylidenernethyl)-phenyl]-4-(4-
trifluoromethoxy
benzenesulfonylamino)-benzamide
_ o.. ..o
O ~ ~ NH ~ ~ NH
N CF3
O
N [3-(2,4-Dioxo-thiazolidin-S-ylidenemethyl)-phenyl]-4-(4-trifluoromethoxy-
benzenesulfonylamino)-benzamide was prepared by the method of Example 200 from
4-
(trifluoromethoxy)benzenesulphonyl chloride
MS (ESI) 563. Found 562 (M-H).
Example 204
Preparation of 4-Benzenesulfonylamino-N [3-(2,4-dioxo-thiazolidin-5-
ylidenemethyl)-phenyl]
benzamide
~ "o
NH ~ ~ NH ~ /
H
O S
4-Benzenesulfonylamino-N [3-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenyl]-
benzamide
was prepared by the method of Example 200 from benzenesulfonyl chloride
MS (ESI) 479. Found 478 (M-H).
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Example 205
Preparation of N (4-Chloro-phenyl)-4-[4-(2,4-dioxo-thiazolidin-5-
ylidenemethyl)-phenoxy)
benzenesulfonamide
O
H ~ I I , H ~ ~ CI
a o
Step A: Preparation of 4-Phenoxy-benzaldehyde
A suspension of potasium carbonate (1 eq.), 4-flurobenzaldehyde, and phenol
(1.2 eq.) was
heated to 150 °C and stirred for two days. The reaction was allowed to
cool to room
temperatrure and poured into saturated aq. sodium bicarbonate and ice. The
solution was
extracted with ether. The combined organic layers were washed with water and
dried with
Na2S04. Concentration under vacuum gave pure product as an orange oil.
Step B: Condensation with TZD to give 5-(4-Phenoxy-benzylidene)-thiazolidine-
2,4-dione
A solution of 4-Phenoxy-benzaldehyde, TZD (1.5 eq.) and piperidine (2 eq.) in
EtOH was
heated at 70°C and stirred overnight. The reaction was cooled to room
temperature and poured
into 10% aq. HCI. The resulting precipitate was filtered, washed with water
and allowed to air
dry.
MS (ESI) 297. Found 296 (M-H).
Step C: Chlorosulfonylation to give 4-[4-(2,4-Dioxo-thiazolidin-5-
ylidenemethyl)-phenoxy)-
benzenesulfonyl chloride
5-(4-Phenoxy-benzylidene)-thiazolidine-2,4-dione was dissolved in
chlorosulfonic acid and
stirred at 0°C for 30 minutes. The solution was then poured onto ice
and the precipitate was
collected by filtration and air dried to give crude product.
MS (ESI) 395. Found 394 (M-H).
Step D: Condensation with 4-chloroaniline to$ive the sulfonamide
A solution of 4-[4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenoxy)-
benzenesulfonyl chloride
and 4-chloroaniline ( 1.1 eq.) was stirred in pyridine with gentle warming.
Pyridine was then
removed under vacuum and the residue dissolved in EtOAc. The organic phase was
washed
with 10% aq HCl solution, aq. sodium bicarbonate and aq. sodium chloride
solution then
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concentrated under vacuum. The resulting residue was triturated with hexane to
give pure
product.
MS (ESI) 486. Found 485 (M-H).
Example 206
Preparation of 4-[4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N p-tolyl-
benzenesulfonamide
i
~ H~-
0 0
4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-Np-tolyl-
benzenesulfonamide was
prepared by the method of Example 205 from p-toluidine.
MS (ESI) 466. Found 465 (M-H).
Example 207
Preparation of 4-[4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N (4-
trifluoromethoxy-
phenyl)-benzenesulfonamide
o _
~ I I ~ H \ ~ OCF3
S ~I~O
O
4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N {4-trifluoromethoxy-
phenyl)-
benzenesulfonamide was prepared by the method of Example 205 from 4-
(trifluoromethoxy)aniline.
MS (ESI) 536. Found 535 (M-H).
E_xamnle 208
Preparation of 4-[4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N (4-
methoxy-phenyl)
benzenesulfonamide
o ~ _
w wl li \/
0
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4-[4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenoxyJ-N (4-methoxy-phenyl)-
benzenesulfonamide was prepared by the method of Example 205 from p-anisidine.
MS (ESI) 482. Found 481 (M-H).
Example 209
Preparation of 4-[4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N phenyl-
benzenesulfonamide
H O
I
O O
4-[4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N phenyl-
benzenesulfonamide was
prepared by the method of Example 205 from aniline.
MS (ESI) 452. Found 451 (M-H).
Example 210
Preparation of 4-[4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenoxyJ-N (3,4,5-
trimethoxy-
phenyl)-benzenesulfonamide
H
I °~ ~ a
o ~ o-
4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxyJ-N (3,4,5-trimethoxy-
phenyl)-
benzenesulfonamide was prepared by the method of Example 205 from 3,4,5-
trimethoxyaniline.
MS (ESI) 542. Found 541 (M-H).
Example 211
Preparation of 4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N (4-
morpholin-4-yl-
phenyl)-benzenesulfonamide
o n- ~~~ ~ _
S ~ I ~H~~O
0
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4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N (4-morpholin-4-yl-
phenyl)-
benzenesulfonamide was prepared by the method of Example 205 from 4-
morpholinoaniline.
MS (ESI) 537. Found 536 (M-H).
Example 212
Preparation of 4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N (4-
isopropyl-phenyl)-
benzenesulfonamide
H O
~Hfr--
O O
4-[4-(2,4-Dioxo-thiazolidin-S-ylidenemethyl)-phenoxy]-N (4-isopropyl-phenyl)-
benzenesulfonamide was prepared by the method of Example 205 from 4-
isopropylaniline.
MS (ESI) 494. Found 493 (M-H).
Exam In a 213
Preparation of N (2-Chloro-phenyl)-4-[4-(2,4-dioxo-thiazolidin-5-
ylidenemethyl)-phenoxy]-
benzenesulfonamide
o c
H
p ~ I I~
O O
N (2-Chloro-phenyl)-4-[4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-
benzenesulfonamide was prepared by the method of Example 205 from 2-
chloroaniline.
MS (ESI) 486. Found 485 (M-H).
Example 214
Preparation of N (3-Chloro-phenyl)-4-[4-(2,4-dioxo-thiazolidin-5-
ylidenemethyl)-phenoxy]-
benzenesulfonamide
ci
H ~ I
O O
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N (3-Chloro-phenyl)-4-[4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenoxyJ-
benzenesulfonamide was prepared by the method of Example 205 from 3-
chloroaniline.
MS (ESI) 486. Found 485 (M-H).
Example 21 S
Preparation of 4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N (4-
hydroxy-phenyl)-
benzenesulfonamide
0
\ \ I I ~ ~ ~ off
0 0
4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N (4-hydroxy-phenyl)-
benzenesulfonamide was prepared by the method of Example 205 from 4-
aminophenol.
MS (ESI) 468. Found 467 (M-H).
Example 216
Preparation of 4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N (2-
hydroxy-phenyl)-
benzenesulfonamide
H
\ I I
O O
4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N (2-hydroxy-phenyl)-
benzenesulfonamide was prepared by the method of Example 205 from 2-
aminophenol.
MS (ESI) 468. Found 467 (M-H).
Example 217
Preparation of N (2-tert-Butyl-phenyl)-4-[4-(2,4-dioxo-thiazolidin-5-
ylidenemethyl}-phenoxy]-
benzenesulfonamide
o \
I~ H
0 0
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N (2-tent-Butyl-phenyl)-4-[4-(2,4-dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-
benzenesulfonamide was prepared by the method of Example 205 from 2-tert-
butylaniline.
MS (ESI) 508. Found 507 (M-H).
Example 218
Preparation of 4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N
isopropyl-N phenyl-
benzenesulfonamide
H O
\ /
0 o
4-(4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-phenoxy]-N isopropyl-N phenyl-
benzenesulfonamide was prepared by the method of Example 205 from N-
isopropylaniline.
MS (ESI) 494. Found 493 (M-H).
Example 219
Preparation of 4-[4-(2,4-Dioxo-thiazolidin-S-ylidenemethyl)-phenoxy]-N (3-
hydroxy-phenyl)-
benzenesulfonamide
H
H O / \
/ H \ /
O O
4-[4-(2,4-Dioxo-thiazolidin-S-ylidenemethyl)-phenoxy]-N (3-hydroxy-phenyl)-
benzenesulfonamide was prepared by the method of Example 205 from 3-
aminophenol.
MS (ESI) 468. Found 467 (M-H).
Example 220
Preparation of 5-[2-(3,4-Dichloro-benzylsulfanyl)-pyrimidin-4-ylmethyl]-2-
thioxo-thiazolidin-
4-one
H O
S
~CI
CI
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Step A: Alkylation of a p~rrimidine thiol with a benzyl halide
To a suspension of the sodium salt of 4-Dimethoxymethyl-pyrimidine-2-thiol in
DMF with
potassium carbonate (2 eq.) was added alpha, 3,4-trichlorotoluene (1 eq.). the
suspension was
stirred for 2 days. The reaction mixture was diluted with water and extracted
with EtOAc. The
organic layer was washed with water, aq. sodium chloride, dried over MgS04 and
concentrated
in vacuum.
Step B: Deprotection of the acetal
A suspension of 2-(3,4-Dichloro-benzylsulfanyl)-4-dimethoxymethyl-pyrimidine
in
concentrated HCl was refluxed for 5 minutes. The reaction was cooled and
poured into water.
Saturated aqueous sodium bicarbonate was added and the neutralized solution
was extracted
with EtOAc. The organic phase was dried over Na2S04 and concentrated under
vacuum to give
the product.
Step C: Condensation with rhodanine
A solution of rhodanine ( 1 eq.), ethylenediamine diacetate ( 1 eq.) and 2-
(3,4-Dichloro-
benzylsulfanyl)-pyrimidine-4-carbaldehyde in methanol was stirred reflux for 1
hour. The
resulting precipitate was collected by filtration and washed with methanol,
water, aq. sodium
bisulfate and aq. sodium bicarbonate. The solid was air dried.
Step D: Reduction of the double bond
A suspension of 5-[2-(3,4-dichloro-benzylsulfanyl)-pyrimidin-4-ylmethylene]-2-
thioxo-
thiazolidin-4-one in toluene was heated to 80 °C in the presence of 2,6-
dimethyl-1,4-dihydro-
3,5-pyridine carboxylate (1.1 eq.) and activated silica gel. The suspension
was filtered and
rinsed with EtOAc. The filtrate was evaporated in vacuum and the resulting
residue was
redissolved in EtOAc and washed with 1 N aq. HCl solution. The organic layer
was dried over
Na2S04 and concentrated under vacuum to give the crude product. Flash
chromatography
afforded pure product.
MS (ESI) 415. Found 414 (M-H).
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Example 221
5-[3-Amino-2-(2,4-dichloro-benzoy 1)-thieno [2, 3-b]pyridin-6-ylmethylene]-
thiazolidine-2,4-
dione
N2
~ O
CI
S ~
CI
Step A: Alkylation and cyclization of 6-dimethoxymethyl-2-mercanto-
nicotinonitrile
A mixture of 6-dimethoxymethyl-2-mercapto-nicotinonitrile and potassium
carbonate ( 1.52
eq.) was treated with 2,4-dibromoacetopheneone (3.1 eq.) and stirred
overnight. The alkylated
and cyclized product was isolated as a solid precipitate.
Step B: Deprotection of the acetal.
A solution of (3-amino-6-dimethoxymethyl-thieno[2,3-b]pyridin-2-yl)-(2,4-
dichloro-phenyl)-
methanone in a mixture of TFA and H20 was stirred at room temperature until
TLC indicated
the reaction was complete. The reaction was neutralized with cold aq. NaHC03
and extracted
with EtOAc. The organic layer was washed with aq. sodium chloride, then
partially
concentrated under vacuum and stored at 0 °C overnight. The resulting
precipitate was
collected to give the product as a pure yellow powder.
Step C: Condensation with TZD
The aldehyde was heated with TZD and piperidine in ethanol at 90 °C for
3 days. The reaction
was allowed to cool, then poured into 10% aq. HCl solution. Pure product was
isolted by
filtration as a yellow solid.
MS (ESI) 449. Found 448 (M-H).
Example 222
Preparation of 2-Chloro-3-(2,4-dioxothiazolidin-5-ylidene-methyl)quinoline.
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Step A: Coupling 2,4-thiazolidinedione (TZD) to aldehyde under basic
conditions.
2-Chloro-3-quinolinecarboxaldehyde, TZD (1.5 eq.), piperidine (1.5 eq) were
dissolved in
ethanol and heated to reflux for 5 hours. The reaction mixture was allowed to
cool to RT and
poured into ethanol, IN HCl was added and a yellow precipitate was collected,
washed several
times with ether, air dried for 24 hrs at RT to give pure product (Yield-45%).
~H NMR (DMSO-d6) 8: 12.8 (br.s, 1H, NH), 8.5 (s, 1H), 8.2 (d, 1H), 8.0-7.8 (m,
3H), 7.7 (t,
1H).
MS (ESI) 290. Found 289 (M-H). HPLC: 92% pure.
Example 223
Preparation of 2-Phenylthio-3-(2,4-dioxothiazolidin-5-ylidene-
methyl)quinoline.
Step A: 2-Chloro-3-(2,4-dioxothiazolidin-5-ylidene-methyl)quinoline and
thiophenol (1.2 eq)
were mixed with ethoxyethanol and heated to reflux under NZ for 2 hours. The
reaction
mixture was allowed to cool to RT, ether was added and bright-yellow
precipitate was filtered
off, washed several times with ether, air dried for 24 hrs at RT to give pure
product ( Yield-
36%).
1H NMR (DMSO-d6) S (br.s , 1 H, NH), 8.3 (s, 1 H),'8.1 (d, 1 H), 7.7 (t, 1 H),
7.6-7.5 (m, 4H),
7.5-7.4 (m, 3H).
MS (ESI) 364. Found 363 (M-H). HPLC: 88% pure.
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Example 224
Preparation of 2-(4-Chlorophenylthio)-3-(2,4-dioxothiazolidin-S-ylidene-
methyl)quinoline.
The title compound was prepared by the method of Example 223.Yield-69%.
1H NMR (DMSO-d6) 8 12.8 (br,s, 1 H, NH), 8.3 (s, 1 H), 8.00 (d, 2H), 7.9 (s, 1
H), 7.7 (t, 1 H),
7.6-7.4 (m, 6H).
MS(ESI) 398. Found 397 (M-H); 399 (M+H). HPLC: 92% pure.
EXamDle 225
Preparation of 2-(3,4-Dichlorophenylthio)-3-(2,4-dioxothiazolidin-5-ylidene-
methyl)quinoline.
The title compound was prepared by the method of Example 223, using ethanol as
a solvent.
Yield-72%.
~H NMR (DMSO-d6) S 12.8 (br.s, 1H, NH), 8.34 (s, 1H), 8.05 (d, 1H, J= 8), 7.87
(d, 1H,
J=1.6), 7.83 (s, 1 H), 7.7I -7.53 (m, 6H).
MS(ESI) 433. Found 431;432; 433 (M-H). 433; 435; 436 (M+H), HPLC: 96% pure..
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Example 226
Preparation of 2-(4-Fluorophenylthio)-3-(2,4-dioxothiazolidin-5-ylidene-
methyl)quinoline.
The title compound was prepared by the method of Example 223. Yield-23%.
~ H NMR (DMSO-db) b 12.6 (br.s 1 H, NH), 8.3 (s, 1 H), 8.00 (d, 1 H), 7.9 (s,
1 H), 7.7-7.4 (m,
SH), 7.3 (t, 2H).
MS(ESI) 382. Found 381 (M-H). HPLC: 100% pure.
Example 227
Preparation of 2-(4-Methylphenylthio)-3-(2,4-dioxothiazolidin-5-ylidene-
methyl)quinoline.
The title compound was prepared by the method of Example 223. Yield-24%. 'H
NMR
(DMSO-d6) 8 12.8 (br.s, 1 H, NH), 8.3 (s, 1 H), 8.00 (d, 1 H), 7.9 (s, 1 H),
7.7 (t, 1 H), 7.6-7.5 (m,
2H), 7.4 (d, 2H), 7.3 (d, 2H), 2.1 (s, 3H).
MS (ESI) 378. Found 377 (M-H). HPLC: 86% pure.
Example 228
Preparation of 2-(4-Methoxyphenylthio)-3-(2,4-dioxothiazolidin-5-ylidene-
methyl)quinoline.
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The title compound was prepared by the method of Example 223. Yield-35%.
~ H NMR (DMSO-d6) 8 12.8 (br.s, 1 H, NH), 8.2 (s, 1 H), 8.0 (d, 1 H), 7.9 (s,
1 H), 7.8-7.4 (m,
SH), 7.0 (d, 2H), 3.8 (s, 3H).
MS (ESI) 394. Found 393 (M-H). HPLC: 98% pure.
EXample 229
Preparation of 2-[4-Trifluoromethylphenylthio]-3-(2,4-dioxothiazolidin-5-
ylidene-
methyl)quinoline.
The title compound was prepared by the method of Example 223. Yield-SS%
1H NMR (DMSO-d6) 8 12.8 ( br.s. 1 H, NH), 8.3 (s, 1 H), 8.1 (d, 1 H), 7.9 (s,
1 H), 7.8-7.6 (m,
SH), 7.5 (t, 2H).
MS (ESI) 432. Found 431 (M-H). HPLC: 100% pure.
Example 230
Preparation of 2-(4-Chlorophenylsulfinyl)-3-(2,4-dioxothiazolidin-5-ylidene-
methyl)quinoline.
Step A.
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The compound prepared in Example 224 was dissolved in CHZC12/CH30H (5:1 ), and
to this
solution was added 3-chloroperoxybenzoic acid (2eq. of 77% max. purity). The
mixture was
stirring at RT for 4 hrs before TLC analysis showed the disappearance of the
starting
compound. The reaction mixture was stirring for an additionally 2 hrs. At the
end of the
reaction, white crystalline product precipitated. The product was collected by
filteration,
washed several times with ether, and air dried for 48 hrs to provide the final
product with a
yield of 30%.
1H NMR (DMSO-db) 8 12.8 (br. s, 1 H, NH), 8.5 (s, 1 H), 8.4 (s, 1 H), 8.2 (d,
1 H), 8.1 (d, 1 H),
7.9 (t, 1 H), 7.7 (t, 1 H), 7.6-7.4 (m, 4H).
MS (ESI) 414. Found 413 (M-H); 415 (M+H). HPLC: 100% purity.
Reference Example 1
Preparation of Compound 122
Compound 5 (23 mg, 0.056 mmol) obtained in Example 5 was dissolved in dimethyl
sulfoxide (2.0 mL) and a 0.04 mol/L buffer solution (pH 7.2) (4.1 mL), and 2-
mercaptoethanol
(0.012 mL, 0.17 mmol) was added thereto, followed by stirring at 25°C
for 40 minutes. After
the conventional post-reaction treatment, the residue was purified by thin
layer chromatography
(developed with chloroform/methanol =10/1), to obtain a coarse purified
product. The product
was further purified with preparative HPLC [ODS column; eluted with an
acetonitrile/disodium
hydrogenphosphate-potassium dihydrogen phosphate buffer solution (0.04 mol/L;
pH 7.2) _
30/70], to obtain Compound 122 (5.7 mg, 31 %).
~H NMR (300 MHz, DMSO-db) 8 (ppm) 3.26-3.39 (m, 2H), 3.65-3.73 (m, 2H), 5.12
(m, 1 H), 7.73 (d, J = 8.6 Hz, 1 H), 7.82 (s, 1 H), 8.16-8.24 (m, 2H), 12.80
(m, 1 H)
FABMS m/z 325 (M-H)' C,2H1pN2O5S2 = 326
Reference Example 2
Preparation of Compound 123
2-Fluoro-5-nitrobenzaldehyde (48 mg, 0.28 mmol) was dissolved in N,N-
dimethylformamide (2.4 mL), and di-n-propylamine (0.15 mL, 1.1 mmol) and
potassium
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carbonate (0.16 g, 1.1 mmol) were added thereto, followed by stirring at
25°C for lhour. After
the conventional post-reaction treatment, the residue was purified by silica
gel
chromatography(eluted with chloroform), to obtain 2-(N,N-di-n-propylamino)-5-
nitrobenzaldehyde (49 mg, 69%).
'H NMR (300 MHz, CDC13) 8 (ppm) 0.89 (t, J = 7.6 Hz, 6H), 1.66 (tq, J = 7.6,
7.6 Hz,
4H), 3.38 (m, 4H), 7.03 (d, J = 9.1 Hz, 1H), 8.21 (dd, J = 9.1, 2.7 Hz, 1H),
8.60 (d, J =
3.0 Hz, 1 H), 10.01 (s, 1 H)
FABMS m/z 251 (M+H)+ C,3HigO3Nz = 250
2-(N,N-di-n-propylamino)-5-nitrobenzaldehyde (46 mg, 0.18 mmol) was dissolved
in
ethanol ( 1.8 mL), and 2,4-thiazolidinediione (86 mg, 0.74 mmol) and
piperidine, (0.0073 mL,
0.073 mmol) were added thereto, followed by stirring at 80°C for 1
hour. After the
conventional post-reaction treatment, the residue was purified by thin layer
column
chromatography (developed with chloroform/acetonitrile = 10/1), to obtain
Compound 123 (53
mg,82%).
'H NMR (300 MHz, DMSO-d6) S (ppm} 0.81 (t, J = 7.3 Hz, 6H), 1.48-1.61 (m, 4H),
3.21 (t, J = 7.2 Hz, 4H),7.24 (d, J = 9.4 Hz, 1 H), 7.62 (s, 1 H), 8.13 (m, 1
H), 8.20 (m,
1 H), 12.63 (br s, 1 H)
FABMS m/z 348 (M-H)- Cl6H~gO4N3S = 349
Reference Example 3
Preparation of Compound 124
2-Fluoro-5-nitrobenzaldeh yde (0.11 g, 0.63.mmo1) was dissolved in ethanol
(4.2 mL),
and 2,4-thiazolidinedione (0.29 mg, 2.5 mmol) and piperidine (0.025 mL, 0.25
mmol) were
added thereto, followed by stirring at 80°C for 6.5 hours. After the
conventional post-reaction
treatment, the residue was purified by thin layer column chromatography
(eluted with
chloroform/acetonitrile = 10/1), to obtain Compound 124 (24 mg, 12%).
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'H NMR (300 MHz, DMSO-db) S (ppm) 1.57-1.72 (m, 6H), 3.08-3.14 (m, 4H), 7.26
(d,
J = 8.6 Hz, 1 H), 7.65 (s, 1 H), 8.20 (m, 1 H), 8.21 (s, 1 H), 12.66 (m, 1 H)
FABMS m/z 332 (M-H)' C,SH,5N304S = 333
Reference Example 4
Preparation of Compound 125
Commercially available 2-morpholino-S-nitrobenzaldehyde (0.11 g, 0.44 mmol)
(MAYBRIDGE, Catalog Number: RH01290) was dissolved in ethanol (4.2 mL), and
2,4-
thiazolidinedione (0.21 g, 1.8 mmol) and piperidine (0.018 mL, 0.18 mmol) were
added thereto,
followed by stirring at 80°C for 6 hours. The reaction liquid was
cooled to 25°C, and the
precipitated crystals were collected by filtration to give Compound 125 (0.12
g, 82%).
'H NMR (300 MHz, DMSO-d6) 8 (ppm) 3.10-3.16 (m, 4H), 3.73-3.80 (m, 4H), 7.30
(m, 1 H), 7.67 (br s, l H), 8.221-8.28 (m, 2H), 12.65 (m, 1 H)
FABMS m/z 334 (M-H)-C,4H~3N305S = 335
Reference Example 5
Preparation of Compound 126
Compound 126 (85 mg, 73%) was obtained from 4-hydroxy-2-methoxybenzaldehyde
(71 mg, 0.47 mmol), ethanol (2.8 mL), 2,4-thiazolidinedione (0.22 g, 1.9 mmol)
and piperidine
(0.019 mL, 0.19 mmol) using the same method as Reference Example 4.
'H NMR (300 MHz, DMSO-db) b (ppm) 3.82 (s, 3H), 6.51 (s, 1 H), 6.53 (dd, J =
11.7,
2.2 Hz, 1 H), 7.25 (d, J = 8. S Hz, 1 H), 7.93 (s, 1 H), 10.3 9 (s, 1 H), 12.3
8 (br s, 1 H)
FABMS m/z 250 (M-H)' C"H9N04S = 251
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Reference Example 6
Preparation of Compound 127
Compound 127 (53 mg, 47%) was obtained from 2-trifluoromethoxybenzaldehyde (73
mg. 0.38 mmol), ethanol (2.9 mL), 2,4-thiazolidinedione (0.18 g, 1.5 mmol) and
piperidine
(0.015 mL, 0.1 S mmol) using the same method as Reference Example 3.
'H NMR (300 MHz, DMSO-d6) S (ppm) 7.53-7.68 (m, 4H), 7.80 (s, 1H)
FABMS m/z 288 (M-H)- C"H619N03S = 289
Reference Example 7
Preparation of Compound 128
Compound 128 (34 mg, 20%) was obtained from 10-[(4-
chlorophenyl)thio]anthracene-
9-carboxaldehyde (0.13 g, 0.38 mmol), ethanol (5.4 mL), 2,4-thiazolidiedione
(0.18 g, 1.5
mmol) and piperidine (0.01 S mL, 0.1 S mmol) using the same method as
Reference Example 4.
'H NMR (300 MHz, DMSO-db) 8 (ppm) 6.92 (d, J = 8.6 Hz, 2H), 7.25 (d, J = 8.4
Hz,
2H), 7.64-7.77 (m, 4H), 8.18 (d, J =7.9 Hz, 2H), 8.65 (s, 1 H), 8.78 (d, J =
8.1 Hz, 2H),
12.68 (m, 1 H)
FABMS m/z 446 (M-H)- Cz4H,435C1NOZS2 = 447
Reference Example 8
Preparation of Compound 171
A 2.5 mol/L sodium hydroxide aqueous solution (2.1 mL, 5.2 mmol) and
tetrabutylammonium bromide (0.020 g, 0.061 mmol) were added to 2-
naphthalenethiol (0.20 g,
1.2 mmol), follwed by stirring at 25°C for 15 minutes. A toluene (2.1
mL) solution of 2-fluoro-
5-nitrobenzaldehyde (0.21 g, 1.2 mmol) was added to the reaction liquid,
followed by stirring at
110 °C for 3.5 hours. The conventional post-reaction treatment was
performed to give 2-[(2-
formyl-4-nitrophenyl)thio]naphthalene. 2-[(2-Formyl-4-
nitrophenyl)thio]naphthalene (0.30 g,
0.98 mmol) was dissolved in toluene (15 mL), and 2,4-thiazolidinedione (0.46
g, 3.9 mmol),
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piperidine (0.039 mL, 0.39 mmol), acetic acid (0.022 mL, 0.039 mmol) and
molecular sieves
4A ( 1.5 g) were added thereto, followed by stirring at 110 °C for 2.5
hours. After the
conventional post-reaction treatment, the product was triturated with ethanol
to give
Compound 171 (0.13 g, yield 33%).
2-[(2-formyl-4-nitrophenyl)thio]naphthalene:
~H NMR (300 MHz, CDC13) 8(ppm) 6.99 (d, J = 8.8 Hz, 1H), 7.52 (dd, J = 8.4,
1.7 Hz,
1H), 7.56-7.68 (m, 2H), 7.85-7.98 (m, 3H), 8.05 (dd, J = 2.4, 8.8 Hz, 1H),
8.15 (s, 1H), 8.68 (d,
J = 2.4 Hz, 1 H), 10.3 3 (s, 1 H)
FABMS m/z 310 (M+H)+ C,~H"N03S = 309
Compound 171:
'H NMR (300 MHz, DMSO-d6) S(ppm) 7.19 (d, J = 8.8 Hz, 1H), 7.57 (br d, J = 8.5
Hz,
1 H), 7.59-7.68 (m, 2H), 7.95 (s, 1 H), 7.97-8.04 (m, 2H), 8.08 (d, J = 8.8
Hz, 1 H), 8.13 (dd, J =
8.4, 1.8 Hz, 1 H), 8.28 (br s, 2H), 12.82 (m, 1 H)
FABMS m/z 407 (M-H)+ C2oH,2N20aS2 = 464
Example 231
Preparation of Affinity Purified Extract
Extracts used for screening telomerase inhibitors were routinely prepared from
293 cells
over-expressing the protein catalytic subunit of telomerase (hTERT). 'These
cells were found
to have 2-5 fold more telomerase activity than parental 293 cells. 200 ml of
packed cells
(harvested from about 100 liters of culture) were resuspended in an equal
volume of hypotonic
buffer (10 mM Hepes pH 7.9, 1 mM MgCl2, 1 mM DTT, 20 mM KCI, 1 mM PMSF) and
lysed
using a dounce homogenizer. The glycerol concentration was adjusted to 10% and
NaCI was
slowly added to give a final concentration of 0.3 M. The lysed cells were
stirred for 30 min
and then pelleted at 100,000 x g for 1 hr. Solid ammonium sulfate was added to
the S 100
supernatant to reach 42% saturation. The material was centrifuged; the pellet
was resuspended
in one fifth of the original volume and dialyzed against Buffer 'A' containing
50 mM NaCI.
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After dialysis the extract was centrifuged for 30 min at 25,000 x g. Prior to
affinity
chromatography, Triton X-100 (0.5 %}, KCI (0.3 M) and tRNA (50 pg/ml) were
added.
Affinity oligo (5' biotinTEG-biotinTEG-biotinTEG-GTA GAC CTG TTA CCA guu agg
guu
ag 3'; lower case represents 2' O-methyl ribonucleotides and upper case
represents
deoxynucleotides) was added to the extract ( 1 nmol per 10 ml of extract).
After an incubation
of 10 min at 30 °C, Neutravidin beads (Pierce; 250 wl of a 50%
suspension) were added and the
mixture was rotated overnight at 4 °C. The beads were pelleted and
washed three times with
Buffer 'B' containing 0.3 M KCI, twice with Buffer 'B' containing 0.6 M KCI,
and twice more
with Buffer B containing 0.3 M KCI. Telomerase was eluted in Buffer 'B'
containing 0.3 M
KCI, 0.15% Triton X-100 and a 2.5 molar excess of displacement oligo (5'-CTA
ACC CTA
ACT GGT AAC AGG TCT AC-3' at 0.5 ml per 125 pl of packed Neutravidin beads)
for 30
min. at room temperature. A second elution was performed and pooled with the
first. Purified
extracts typically had specific activities of 10 fmol nucleotides
incorporated/min/~l extract, or
200 nucleotides/min/mg total protein.
Buffer 'A' Buffer 'B'
mM Hepes pH 20 mM Hepes pH 7.9
7.9
I mM MgCl2 1 mM EDTA
1 mM DTT 1 mM DTT
1 mM EGTA 10% glycerol
10 % glycerol 0.5 Triton
Example 232
20 . Telomerase Specific Activity Determination
Three separate 100 pl telomerase assays are set up with the following buffer
solutions:
50 mM Tris acetate, pH 8.2, 1 mM DTT, I mM EGTA, 1 mM MgClz, 100 mM K acetate,
500
pM dATP, 500 p,M TTP, l OpM 32P-dGTP (25 Ci/mmol), and a00 nM d(TTAGGG)3. To
the
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individual reactions 2.5, 5 or 10 ~1 of affinity-purified telomerase (see
Example 231 ) is added
and the reactions are incubated at 37 °C. At 45 and 90 minutes, 40 ~1
aliquots are removed
from each reaction and added to 160 ~1 of Stop Buffer ( 1 OOmM NaCI, 10 mM Na
pyrophosphate, 0.2% SDS, 2 mM EDTA, 100 ~g/ml tRNA). 10 p,l trichloroacetic
acid (TCA)
(100%) is added and the sample is incubated on ice for 30 minutes. The sample
is pelleted in a
microcentrifuge (12000 x g force) for 15 minutes. The pellet is washed with 1
ml 95% ethanol
and pelleted again in the microcentrifuge ( 12000 x g force) for 5 minutes.
The pellet is
resuspended in 50 p,l dHzO and transferred to a 12 x 75 glass test tube
containing 2.5 ml of ice
cold solution of 5% TCA and 10 mM Na pyrophosphate. The sample is incubated on
ice for 30
minutes. The sample is filtered through a 2.5 cm wet (dH20) GFC membrane (S&S)
on a
vaccum filtration manifold. The filter is washed three times under vacuum with
5 ml ice cold
1% TCA, and once with 5 ml 95% ethanol. The filter is dried and counted in a
scintillation
counter using scintillation fluid. The fmol of nucleotide incorporated is
determined from the
specific activity of radioactive tracer. The activity of extract is calculated
based on the dNTP
incorporated and is expressed as fmol dNTP/min/pl extract.
Exam.".ple 233
Telomerase Activity Assay
Bio-Tel FlashPlate Assav
An assay is provided for the detection and/or measurement of telomerase
activity by
measuring the addition of TTAGGG telomeric repeats to a biotinylated
telomerase substrate
primer; a reaction catalyzed by telomerase. The biotinylated products are
captured in
streptavidin-coated microtiter plates. An oligonucleotide probe complementary
to 3.5 telomere
repeats labeled with [33P] is used for measuring telomerase products, as
described below.
Unbound probe is removed by washing and the amount of probe annealing to the
captured
telomerase products is determined by scintillation counting.
Method:
1. Compounds are stored as concentrated stocks and dissolved in 100
dimethylsulfoxide (DMSO).
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2. For testing, the compounds are diluted to a 15X working stock in 50% DMSO
and
2 pl is dispensed into two wells of a 96-well microtiter dish (assayed in
duplicate).
3. Telomerase extract is diluted to a specific activity of 0.04 - 0.09 fmol
dNTP
incorporated/min./~.1 in Telomerase Dilution Buffer and 18 ul added to each
sample well to
preincubate with compound for 30 minutes at room temperature.
4. The telornerase reaction is initiated by addition of 10 ~1 Master Mix to
the wells
containing telomerase extract and compound. The plates are sealed and
incubated at 37°C for
90 min.
5. The reaction is stopped by the addition of 10 ~1 HCS.
6. 25 pl of the reaction mixture is transferred to a 96-well streptavidin-
coated
FlashPlate (NEN) and incubated for 2 hours at room temperature with mild
agitation.
7. The wells are washed three times with 180 ~l 2X SSC without any incubation.
8. The counts of probe annealed to biotinylated telomerase products are
detected on a
scintillation counter.
Buffers:
Telomerase Dilution Buffer
50 mM Tris-acetate, pH 8.2
I mM DTT
1 mM EGTA
1 mM MgCl2
830 nM BSA
Master Mix (MM)
50 mM Tris-acetate, pH 8.2
1 mM DTT
1 mM EGTA .
1 mM MgCl2
150 mM K acetate
10 ~M dATP
20 ~.M dGTP
120 ~.M dTTP
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100 nM biotinylated primer (5'-biotin-AATCCGTCGAGCAGAGTT-3')
5.4 nM labeled probe [5'-CCCTAACCCTAACCCTAACCC-(33P} A~_so - 3']; specific
activity approximately 109 cpm/ug or higher
Hybridization Capture Solution (HCS~
12X SSC (1X = 150 mM NaCI/30 mM Na3Citrate)
40 mM EDTA
40 mM Tris-HCI, pH 7.0
Using the foregoing assay, the compounds of Examples 1-29 were shown to have
telomerase ICso values below 100 ~M.
Example 234
Anti-tumor Activity
Ex vivo Studies
a. Reduction of Telomere Length in Tumor Cells
Colonies of the tumor cell lines, such as the ovarian tumor cell lines OVCAR-5
and SK-
OV-3, and normal human cells used as a control (e.g., normal human BJ cells}
are prepared
using standard methods and materials. In one test, the colonies are prepared
by seeding 15-
centimeter dishes with about 106 cells in each dish. The dishes are incubated
to allow the cell
colonies to grow to about 80% confluence, at which time each of the colonies
are divided into
two groups. One group is exposed to a subacute dose of a compound of the
invention at a
predetermined concentration (e.g., between about 5 pM and about 20 ~M) for a
period of about
4-8 hours after plating following the split; the other group is exposed to a
control (e.g., DMSO}.
Each group is then allowed to continue to divide, and the groups are split
evenly again
(near confluence). The same number of cells are seeded for continued growth.
The compound
or control is added every fourth day to the samples at the same concentration
delivered initially.
Remaining cells are analyzed for telomere length. As the untested cell
cultures near confluence,
the samples are split again as just described. This sequence of cell doubling
and splitting is
continued for about 20 to 25 doublings. Thus, a determination of telomere
length as a function
of cell doublings is obtained.
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Telomere length is determined by digesting the DNA of the cells using
restriction
enzymes specific for sequences other than the repetitive T2 AG3 sequence of
human telomeres
(TRF analysis). The digested DNA is separated by size using standard
techniques of gel
electrophoresis to determine the lengths of the telomeric repeats, which
appear, after probing
with a telomere DNA probe, on the gel as a smear of high-molecular weight DNA
(approximately 2 Kb-15 Kb).
The results of the telomere length analysis are expected to indicate that the
compounds
of the invention have no affect on the rate of decrease in telomere length for
control cells as a
function of progressive cell doublings. With respect to the tumor cell lines,
however,
measurable decreases in telomere length are expected to be determined for
tumor cells exposed
to the compounds of the invention. Tumor cells exposed to the control are
expected to
maintain steady telomere lengths. Thus, the compounds of the invention are
expected to cause
resumption of the normal loss of telomere length as a function of cell
division in tumor cells.
In another experiment, HEK-293 cells are incubated with a compound of the
invention
and a control at concentrations between about 1 pM and about 20 pM using the
protocol just
described. Cells are expected to enter crisis (i.e., the cessation of cell
function) within several
weeks following administration of the test compound of the invention. In
addition, TRF
analysis of the cells using standard methodology is expected to show that the
test compounds of
the invention are effective in reducing telomere length. In addition to the
HEK-293 cells
described above, this assay can be performed with any telomerase-positive cell
line, such as
HeLa cells.
b. Specificity
Compounds of the invention are screened for activity (ICso) against telomerase
and
several enzymes having nucleic acid binding or modifying activities related to
telomerase using
standard techniques. The enzymes being screened include Telomerase, DNA
Polymerase I,
HeLa RNA Polymerase II, T3 RNA Polymerase, MMLV Reverse Transcriptase,
Topoisomerase I, Topoisomerase II, Terminal Transferase and Single-Stranded
DNA Binding
Protein (SSB). The specificity of a compound of the invention for telomerase
is determined by
comparing the ICSO of the compound with respect to telomerase with the ICso
values of the
compound for each of the enzymes being screened. The compound is determined to
have high
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specificity for telomerase if the ICso for telomerase of the compound is lower
than the ICso
vaules for each of the enzymes being screened.
Alternatively, telomerase inhibitory activity of the compounds was measured in
accordance with a known method (U.S. Patent 5,760,062). That is, a dimethyl
sulfoxide
(DMSO) solution of each agent was mixed with partially purified telomerase
from a nuclear
extract of HEK293 cells and incubated in the presence of an
oligodeoxynucleotide to be used
as the substrate and deoxynucleotide triphosphate. The obtained reacted
product (DNA having
a telomere sequence) was adsorbed on a membrane, and hybridization was earned
out using a
labeled oligonucleotide probe having a sequence complementary to the telomere
sequence.
The inhibition ratio was calculated based on the ratio of the signal of label
on the membrane in
the presence of the agent to the signal of label in the absence of the agent
(control). Also,
concentration of each agent which inhibits SO% of the enzyme activity based on
the control
was used as ICso. Results of the measurement of inhibition activity of
selected compounds are
shown in Table 7.
Table 7
In vitro telomerase inhibition activity
Compound ICso (~M)
1 21
4 30
5 23
9 8.3
12 5.8
13 5.9
14 4.0
16 26
17 10
18 11
19 70
9.0
21 4.3
23 14
24 5.3
4.2
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26 4.2
31 15
32 21
33 5.5
35 3.9
36 5.1
37 6.0
43 2.6
44 1.9
47 4.1
48 1.6
49 4.4
53 0.77
54 7.4
55 5.3
56 7.9
57 5.4
58 56%
59 2.1
60 4.5
61 0.33
62 7.8
63 6.5
64 9.7
65 6.2
66 3.7
67 1.0
68 2.7
69 0.48
70 4.8
71 2.7
_
72 4.3
73 7.0
74 3.7
76 3.9
78 S.5
79 3.5
80 5.5
82 51%
83 58%
84 3.9
86 8.0
87 6.9
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89 . ~56%)
90 3.7
91 _
61%
92 5.6
93 55%
94 7.8
96 62%
97 5.6
99 6.9
100 4.4
101 62%
104 61%
106 4.5
108 6.2
110 53%
113 68%
115 2.7
120 9.3
121 4.0
122 7,7
123 6.9
124 54%
125 1.6
126 1.3
127 5.7
128 2.6
129 g, l
132 2.6
133 9.1
134 59%
135 4.5
137 5.5
138 4.3
139 4.4
141 64%
143 62%
153 53%
171 7.2
Tro litazone 16
Pio litazone 83
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In parenthesis, residual activity of in vitro telomerase activity is shown in
the presence of the
compound at a concentration of 10 pm.
c. Cytotoxicity
The XTT assay for cytotoxicity is performed using HeLa cells. The cell lines
used in
the assay are exposed to a compound of the invention for 72 hours at
concentrations ranging
from about 1 pM to about 1,000 pM. During this period, the optical density
(OD) of the
samples is determined for light at 540 nanometers (nm}. No significant
cytotoxic effects are
expected to be observed at concentrations less than about 5 pM. It will be
appreciated that
other tumor cells lines such as the ovarian tumor cell lines OVCAR-5 and SK-OV-
3 can be
used to determine cytotoxicity in addition to control cell lines such as
normal human BJ cells.
Other assays for cytotoxicity such as the MTT assay (see Berridge et al.,
1996, Biochemica
4:14-19) and the alamarBlueTM assay (U.S. Patent No. 5,501,959) can be used as
well.
Some compounds may induce G2 arrest at concentrations above about 5 pM (i.e.,
at
10 pM-20 p.M concentrations or higher). Preferably, to observe any telomerase
inhibiting
effects the compounds should be administered at a concentration below the
level of
cytotoxicity. Nevertheless, since the effectiveness of many cancer
chemotherapeutics derives
from their cytotoxic effects, it is within the scope of the present invention
that the compounds
of the present invention be administered at any dose for which
chemotherapeutic effects are
observed.
In vivo Studies
A human tumor xenograft model in which OVCAR-5 tumor cells are grafted into
nude
mice can be constructed using standard techniques and materials. The mice are
divided into
two groups. One group is treated intraperitoneally with a compound of the
invention. The
other group is treated with a control comprising a mixture of either DMSO or
ethanol and
emulphor (oil) and phosphate buffer solution (PBS). The average tumor mass for
mice in each
group is determined periodically following the xenograft using standard
methods and materials.
In the group treated with a compound of the invention, the average tumor mass
is
expected to increase following the initial treatment for a period of time,
after which time the
tumor mass is expected to stabilize and then begin to decline. Tumor masses in
the control
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group are expected to increase throughout the study. Thus, the compounds of
the invention are
expected to lessen dramatically the rate of tumor growth and ultimately induce
reduction in
tumor size and elimination of the tumor.
In another experiment, each agent was allowed to contact with human renal
carcinoma
cell line ACHN for 3 days, and then a cell extract was prepared by a known
method (U.S.
Patent 5,629,154) to measure the enzyme activity. That is, a cell extract was
prepared using a
buffer solution containing 0.5% CHAPS. Using the extract, TRAP (Telomeric
Repeat
Amplification Protocol) assay was carried out in vitro (TRAPEZETM ELISA
Telomerase
Detection Kit, manufactured by Intergen). The ratio (%) of the enzyme activity
in the extract
from agent-treated cells to the enzyme activity in the extract from agent-
untreated cells was
calculated. The results are shown in Table 8.
Table 8
In vivo telomerase inhibition activity
Compound Concentration (~tM) Residual Enzyme Activity
(%)
1 30 11
3 100 0
4 10 16
5 10 16
11 100 50
12 100 26
14 100 47
.
16 100 0
21 30 35
22 100 33
37 30 39
38 30 36
54 30 50
56 30 26
57 30 24
66 30 0
68 10 18
69 10 16
78 10 25
82 3 23
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83 10 32
86 10 44
89 30 2
92 30 41
97 30 41
113 30 37
120 10 40
133 10 11
134 10 4
135 3 14
141 30 26
143 30 36
Thus, the present invention provides novel compounds, compositions and methods
for
inhibiting telomerase activity and treating disease states in which telomerase
activity has
deleterious effects, especially cancer. The compounds of the invention provide
a highly
selective and effective treatment for malignant cells that require telomerase
activity to remain
immortal; yet, without affecting non-malignant cells.
While the prefer ed embodiment of the invention has been illustrated and
described, it
will be appreciated that various changes can be made therein without departing
from the spirit
and scope of the invention.
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