U.S. patent application number 10/115550 was filed with the patent office on 2002-10-17 for ppar gamma ligands.
Invention is credited to Collins, Jon Loren, Holmes, Christopher Patrick, Lenhard, James Martin, Willson, Timothy Mark.
Application Number | 20020151569 10/115550 |
Document ID | / |
Family ID | 10842180 |
Filed Date | 2002-10-17 |
United States Patent
Application |
20020151569 |
Kind Code |
A1 |
Collins, Jon Loren ; et
al. |
October 17, 2002 |
PPAR gamma ligands
Abstract
The present invention discloses a method for treating
osteoporosis by administration of a PPAR gamma antagonist.
Inventors: |
Collins, Jon Loren; (Durham,
NC) ; Holmes, Christopher Patrick; (Saratoga, CA)
; Lenhard, James Martin; (Raleigh, NC) ; Willson,
Timothy Mark; (Durham, NC) |
Correspondence
Address: |
DAVID J LEVY, CORPORATE INTELLECTUAL PROPERTY
GLAXOSMITHKLINE
FIVE MOORE DR., PO BOX 13398
RESEARCH TRIANGLE PARK
NC
27709-3398
US
|
Family ID: |
10842180 |
Appl. No.: |
10/115550 |
Filed: |
April 3, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10115550 |
Apr 3, 2002 |
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09831672 |
May 11, 2001 |
|
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09831672 |
May 11, 2001 |
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PCT/EP99/08477 |
Nov 9, 1999 |
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Current U.S.
Class: |
514/342 ;
514/369; 546/269.7; 548/187 |
Current CPC
Class: |
A61P 35/00 20180101;
A61K 31/4439 20130101; A61P 3/10 20180101; G01N 2333/70567
20130101; A61P 3/00 20180101; A61P 9/00 20180101; G01N 33/566
20130101; A61K 31/443 20130101; A61P 29/00 20180101; A61P 19/10
20180101; A61K 31/426 20130101; A61P 43/00 20180101; A61P 3/04
20180101; C07D 277/14 20130101 |
Class at
Publication: |
514/342 ;
546/269.7; 514/369; 548/187 |
International
Class: |
A61K 031/4439; C07D
417/02; A61K 031/427 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 11, 1998 |
GB |
98246143 |
Claims
What is claimed is:
1. A compound of the following formula 2or a pharmaceutically
acceptable salt or solvate thereof, where n is2,3, or 4, R.sub.1 is
hexyl, heptyl, or C.sub.4--6 alkyl-phenyl, R.sub.2 is butyl or
benzyl optionally substituted with 1 or 2 halogen, R.sub.3 is
butyl, benzyl optionally substituted with a trifluoromethyl group
or with 1 to 3 halogen, --C.sub.4H.sub.8OH, p-pyridyl, o-pyridyl,
ethylpropionate, propyl, ethyl acetate, o-thiophenmethyl,
2,3-methylenedioxobenzyl, 2-thiazolemethyl, 2-furfuryl, R.sub.4 is
--COOH, --NHC(O)NH.sub.2, --NHS(CH.sub.3)O.sub.2,
--S(NH.sub.2)O.sub.2, hydantoin, --OH, --OCH.sub.2CO.sub.2H,
--OCH.sub.2CONH.sub.2, --OCH.sub.3, R.sub.5 is hydrogen or R.sub.5
and R.sub.4 are bonded together to form a methylenedioxo ring.
2. The compound of claim 1 wherein R.sub.3 is butyl, benzyl
optionally substituted with 1 or 2 halogen, or p-pyridyl.
3. A compound of claims 1 or 2 wherein the stereochemistry around
the 2 and 5 carbon atoms is such that the compound is the trans
pair of the (2S, 5S) enantiomer and the (2R, 5R) enantiomer.
4. A compound of claim 3 wherein the stereochemistry around the 2
and 5 carbon atoms is such that the compound is the (2S,5S)
enantiomer.
5. The compound of claim 1 wherein said compound is selected from
the group consisting of
4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazol- idine
N,N-dibenzylacetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-hept-
yl-4-oxo-5-thiazolidine N,N-dibenzylacetamide,
(2S,5S)-4-0(4-(4-carboxyphe-
nyl)butyl)-2-heptyl-4-oxo-5-thiazolidine N,N-dibenzyacetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-hexyl-4-oxo-5-thiazolidine
N,N-dibutylacetamide,
(2S*,5S*)-4-(2-(4-carboxyphenyl)ethyl)-2-octyl-4-ox-
o-5-thiazolidine N,N-dibenzylacetamide,
(2R*,5S*)-4(2-(4-carboxyphenyl)eth-
yl)-2-octyl-4-oxo-5-thiazolidine N,N-dibenzylacetamide,
(2S*,5S*)-4-(2-(4-carboxyphenyl)ethyl)-2-octyl-4-oxo-5-thiazolidine
N,N-di-(3-iodo)benzylacetamide,
(2S*,5S*)-4-(3-(4-carboxyphenyl)propyl)-2-
-heptyl-4-oxo-5-thiazolidine N,N-benzylacetamide,
(2S*,5S*)-4-(4-(4carboxy-
phenyl)butyl)-2-heptyl-4-oxo-5-thiazolidine
N,N-di-(3-benzylacetamide,
(2S*,5S*)-4-(2-(4-carboxyphenyl)ethyl)-2-(6-phenylhexyl)-4-oxo-5-thiazoli-
dine N,N-dibenzyiacetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-(6-ph-
enylhexyl)-4-oxo-5-thiazolidine N,N-dibenzylacetamide,
4-(4-(4-carboxyphenyl)butyl)-2-(4-phenylbutyl)-4-oxo-5-thiazolidine
N,N-dibenzylacetamide,
(2S*,5S*)-4-(2-(4-ureidophenyl)ethyl)-2-octyl-4oxo- -5-thiazolidine
N,N-dibenzylacetamide, (2S*,5S*)-4-(2-(4-methylsuffonamido-
phenyl)ethyl)-2-octyl-4-oxo-5-thiazolidine N,N-dibenzylacetamide,
(2S*,5S*)-4-(2-(4-aminosuffonylphenylethyl)-2-octyl-4-oxo-5-thiazolidine
N,N-dibenzylacetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-hepyl-4-o-
xo-5-thiazolidine N-benzyl-N-(4-trifluorobenzyl)acetamide,
(2R*,S*)-4-(4-(4-carboxyphenyl)butyl)-2-hepyl-4-oxo-5-thiazolidine
N-benzyl-N-(4-trifluorobenzyl)acetamide,
(2S*,5S*)-4-(2-(4-(3-hydantoino)-
phenyl)ethyl)-2-octyl-4-oxo-5-thiazolidine N,N-dibenzylacetamide,
(2S,5S*)-4-(2-(3,4-dioxomethylenephenyl)ethyl)-2-heptyl-4-oxo-5-thiazolid-
ine N,N-dibenzylacetamide,
(2S*,5S*)-4-(4-(4-arboxyphenyl)butyl)-2-octyl-4-
-oxo-5-thiazolidine N-benzyl-N-(4-hydroxybutyl)acetamide,
(2S*,5S*)-4-(2-(3,4dioxomethylenephenyl)ethyl)-2octyl-4-oxo-5-thiazolidin-
e N,N-dibenzylacetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-
-oxo-5-thiazolidine N-benzyl-N-(4-pyridyl)acetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidine
N-benzyl-N-(2-pyridyl)acetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-
-heptyl-4-oxo-5-thiazolidine
N-benzyl-N-(2-ethoxycarboxyethyl)acetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidine
N-benzyl-N-butylacetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-hepty-
l-4-oxo-5-thiazolidine N-benzyl-N-isopropylacetamide,
(2S*,5S*)-4-(2-(4-hydroxyphenyl)ethyl)-2-heptyl-4-oxo-5-thiazolidine
N,N-dibenzylacetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4--
oxo-5thiazoildne N-benzyl-N-ethoxycarboxymethylacetamide,
(2S*,5S*)-4-(4-(4arboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidine
N,N-di-(4-fluorobenzyl)acetamide,
(2S*,5S*)-4-(2-(4-carboxymethoxyphenyl)-
ethyl)-2-heptyl-4-oxo-5-thiazolidine N,N-dibenzylacetamide,
(2S*,5S*)-4-(2-(4-carboxyamidomethoxyphenyl)ethyl)-2-heptyl-4-oxo-5-thiaz-
olidine N,N-dibenzylacetamide,
(2S*,5S*)-4-(2-(4-methoxyphenyl)ethyl)-2-he-
ptyl-4-oxo-5-thiazolidine N,N-dibenzylacetamide,
(2S*,5S*)-4-(4-(4-carboxy-
phenyl)butyl)-2-heptyl-4-oxo-5-thiazolidine
N-benzyl-N-(2-thienylmethyl)ac- etamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolid-
ine N-benzyl-N-(2,3-dioxomethylenebenzyl)acetamide,
(2S*,S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazoidine
N-benzyl-N-(2-thiazolemethyl)acetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)b-
utyl)-2-heptyl-4-oxo-5-thiazolidine
N-benzyl-N-(2-furfuryl)acetamide, and pharmaceutically acceptable
salts and solvates thereof.
6. The compound of claim 1 wherein said compound is selected from
the group consisting of
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-
-5-thiazolidine N,N-dibenzylacetamide,
(2S,5S)-4-(4-(4-carboxyphenyl)butyl-
)-2-heptyl-4-oxo-5-thiazolidine N,N-dibenzylacetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-hexyl-4-oxo-5-thiazolidine
N,N-dibutylacetamide,
(2S*,5S*)-4-(4(4-carboxyphenyl)butyl)-2octyl-4-oxo-- 5-thiazolidine
N-benzyl-N-(4-hydroxybutyl)acetamide,
(2S*,5S*)-4-(4-(4-carboxyphenylbutyl)-2-heptyloxo-5thiazolidine
N-benzyl-N-(4-pyridyl)acetamide,
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-
-heptyl-4-oxo-5-thiazolidine
N-benzyl-N-(2-thiazolemethyl)acetamide, and pharmaceutically
acceptable salts and solvates thereof.
7. A pharmaceutical composition comprising a compound according to
any of claims 1-6.
8. A pharmaceutical composition according to claim 6 further
comprising a pharmaceutically acceptable diluent or carrier.
9. A compound according to any of claims 1-6 for use in
therapy.
10. A method for the prevention or treatment of a PPARgamma
mediated disease or condition comprising administration of a
therapeutically effective amount of a compound of any of claims
1-6.
11. The method of claim 10 wherein said disease or condition is
diabetes, obesity, dyslipidemia, metabolic syndrome, osteoporosis,
acne, cardiovascular disease, inflammation, or cancer.
12. The method of claim 10 wherein said disease or condition is
osteoporosis.
13. A method for the prevention or treatment of osteoporosis
comprising administration of a therapeutically effective amount of
a PPARgamma antagonist.
14. The method of claim 13 wherein said antagonist is a compound of
any of claims 1-6.
15. Use of a compound of any of claims 1-6 for the manufacture of a
medicament for the prevention or treatment of a PPARgamma mediated
disease or condition.
16. Use according to claim 15 wherein said disease or condition is
diabetes, obesity, dyslipidemia, metabolic syndrome, osteoporosis,
acne, cardiovascular disease, inflammation, or cancer.
17. Use according to claim 16 wherein said disease or condition is
osteoporosis.
18. Use of a PPARgamma antagonist compound for the manufacture of a
medicament for the prevention or treatment of osteoporosis.
19. Use according to claim 18 wherein said antagonist is a compound
according to any of claims 1-6.
20. A method for identifying compounds that will be useful for the
treatment of a PPAR gamma mediated disease or condition, comprising
the step of binding a compound of claim 1 to PPAR gamma.
21. The method of claim 20 wherein said disease or condition is
diabetes, impaired glucose tolerance, obesity, or a cardiovascular
disorder.
22. A method for treating a PPAR gamma mediated disease or
condition comprising administration of a therapeutically effective
amount of a compound that was identified as useful for such
treatment by the method of claim 20.
23. The method of claim 22 wherein said disease or condition is
diabetes, impaired glucose tolerance, obesity, or a cardiovascular
disorder.
24. Use of a compound identified by the method of claim 20 for the
manufacture of a medicament for use in the treatment of a PPAR
mediated disease or condition.
25. Use according to claim 24 wherein the PPAR mediated disease or
condition is diabetes, impaired glucose tolerance, obesity or a
cardiovascular disorder.
26. A method for identifying compounds which would be useful in the
treatment of osteoporosis comprising the step of determining
whether a compound antagonises PPAR gamma.
Description
[0001] The present invention relates to compounds that bind to and
affect PPAR gamma. In another aspect, the present invention relates
to methods for prevention or treatment of PPARgamma mediated
diseases and conditions and to a method for prevention or treatment
of osteoporosis.
[0002] Peroxisome Proliferator Activated Receptors (PPARs) are
orphan receptors belonging to the steroid/retinoid receptor
superfamily of ligand-activated transcription factors. See, for
example, Willson, T. M. and Wahli, W., Curr. Opin. Chem. Biol.,
(1997), Vol. 1, pp 235-241.
[0003] Three mammalian PPARs have been identified which are termed
PPAR-alpha, PPAR-gamma, and PPAR-delta. PPARs regulate expression
of target genes by binding to DNA response elements as heterodimers
with the retinoid X receptor. These DNA response elements (PPRE)
have been identified in the regulatory regions of a number of genes
encoding proteins involved in lipid metabolism and energy balance.
The biological role of the PPARs in the regulation of lipid
metabolism and storage has been recently reviewed. See, for
example, Spiegelman, B. M., Diabetes, (1998), Vol. 47, pp 507-514,
Schoonjans, K., Martin, G., Staels, B., and Auwerx, J., Curr. Opin.
Lipidol., (1997), Vol. 8, pp 159-166, and Brun, R. P., Kim, J. B.,
Hu, E., and Spiegelman, B. M., Curr. Opin. Lipidol., (1997), Vol.
8, pp 212-218.
[0004] PPAR-gamma ligands of the thiazolidinedione class (TZD)
enhance the actions of insulin in man and reduce circulating
glucose levels in rodent models of diabetes. The PPAR-gamma
receptor is expressed in adipose tissue and plays a pivotal role
the regulation of adipocyte differentiation in vitro. TZD such as
rosiglitazone induce adipocyte differentiation in vitro through
activation of the PPAR-gamma receptor. Thus, although there are
clearly therapeutic uses for PPAR-gamma ligands in the treatment of
diseases of lipid metabolism and energy balance, it is possible
that there will be side effects of these drugs. For example,
PPAR-gamma ligands that promote adipocyte differentiation in vivo
could lead to increased fat accumulation and weight gain. This side
effect might offset the beneficial effects of a PPAR-gamma ligand
in the treatment of diabetes or other diseases where obesity is a
risk factor. See, for example, the Spiegelman and Brun articles
cited above.
[0005] Essential dietary fatty acids and certain of their
eicosanoid metabolites are naturally occurring hormones for the
PPAR-gamma receptor. These hormones can promote adipogenesis
through activation of the PPAR-gamma receptor. See, for example,
Kliewer, S. A., et al., Proc. Natl. Acad. Sci. USA, (1997), Vol.
94, pp 4318-4323, and Kliewer, S. A., et al., Cell, (1995), Vol.
83, pp 813-819. Molecules that inhibit the adipogenic effects of
endogenous PPAR-gamma hormones may be useful in the treatment of
diseases caused by increased fat accumulation or lipid storage.
See, for example, Tontonoz, P., Hu, E., and Spiegelman, B. M.,
Curr. Opin. Genet. Dev., (1995), Vol. 5, pp 571-576. Examples of
these diseases are obesity, osteoporosis, and acne. For example, it
has also been noted that TZD promote adipogenesis in bone marrow
and inhibit expression of markers of the osteoblast phenotype such
as alkaline phosphatase. See, for example, Paulik, M. A. and
Lenhard, J. M., Cell Tissue Res., (1997), Vol. 290, pp 79-87. These
effects may lead to low bone mineral density and osteoporosis.
Compounds that promote osteogenesis activity may be useful in the
treatment of osteoporosis. Similarly, it is known that the TZDs can
promote lipid accumulation in sebocytes. See, for example,
Rosenfield, R. L., Deplewski, D., Kentsis, A., and Ciletti, N.
Dermatology, (1998), Vol. 196, pp 43-46. These effects may lead to
sebocyte differentiation and acne formation. Thus, molecules that
block adipogenesis in adipocytes, pre-adipocytes, bone marrow, or
sebocytes may have beneficial effects in the treatment of obesity,
osteoporosis, or acne.
[0006] The PPARgamma receptor has been found in tissues other than
adipose, and it is believed that synthetic PPAR-gamma ligands and
natural PPAR-gamma hormones (natural ligands) may have beneficial
effects in many other diseases including cardiovascular disease,
inflammation, and cancer. See, for example, the Schoonjans article
cited above, Ricote, M. et al., Nature, (1998), Vol. 391, pp 79-82,
and Mueller, E. et al., Mot. Cell, (1998), Vol. 1, pp 465-470.
[0007] There is precedent among other member of the
steroid/retinoid receptor superfamily that synthetic ligands can be
identified which mimic many of the beneficial effects but inhibit
some of the detrimental side effects of the natural hormones. See,
for example, McDonnell, D. P., Biochem. Soc. Trans., (1998), Vol.
26, pp 54-60. These synthetic ligands have been given various
labels, including antagonists, anti-hormones, partial agonists,
selective receptor modulators, tissue selective ligands, and
others. See, for example, Katzenellenbogen, J. A., O'Malley, B. W.,
and Katzenellenbogen, B. S., Mol. Endocinol., (1996), Vol. 10, pp
119-131.
[0008] As used herein, a "PPARgamma ligand" is a compound that
binds to human PPARgamma with a pKi of greater than 5 when tested
in the binding assay described below. As used herein a "PPARgamma
antagonist" is a PPARgamma ligand that gives greater than 50%
inhibition of lipogenesis when tested in the adipocyte
differentiation assay described below and greater than 50%
inhibition of transactivation by rosiglitazone when tested in the
cell-based reporter assay described below.
[0009] Briefly, in one aspect, the present invention discloses
compounds of Formula (1) and pharmaceutically acceptable salts and
solvates thereof, 1
[0010] where n is 2, 3, or 4,
[0011] R.sub.1 is hexyl, heptyl, or C.sub.4--6alkyl-phenyl,
[0012] R.sub.2 is butyl or benzyl optionally substituted with 1 or
2 halogen,
[0013] R.sub.3 is butyl, benzyl optionally substituted with a
trifluoromethyl group or with 1 to 3 halogen, --C.sub.4H.sub.8OH,
p-pyridyl, o-pyridyl, ethylpropionate, propyl, ethyl acetate,
o-thiophenmethyl, 2,3-methylenedioxobenzyl, 2-thiazolemethyl,
2-furfuryl,
[0014] R.sub.4 is --COOH, --NHC(O)NH.sub.2, --NHS(CH.sub.3)O.sub.2,
--S(NH.sub.2)O.sub.2, hydantoin, --OH, --OCH.sub.2CO.sub.2H,
--OCH.sub.2CONH.sub.2, --OCH.sub.3,
[0015] R.sub.5 is hydrogen or R.sub.5 and R.sub.4 are bonded
together to form a methylenedioxo ring.
[0016] In another aspect, the present invention discloses a method
for prevention or treatment of a PPARgamma mediated disease or
condition comprising administration of a therapeutically effective
amount of a compound of this invention. As used herein, "a compound
of the invention" means a compound of formula (I) or a
pharmaceutically acceptable salt, or solvates thereof.
[0017] In another aspect, the present invention provides
pharmaceutical compositions comprising a compound of the invention,
preferably in association with a pharmaceutically acceptable
diluent or carrier.
[0018] In another aspect, the present invention discloses a method
for prevention or treatment of osteoporosis comprising
administration of a therapeutically effective amount of a PPARgamma
antagonist.
[0019] In another aspect, the present invention provides a compound
of the invention for use in therapy, and in particular, in human
medicine.
[0020] In another aspect, the present invention provides the use of
a compound of the invention for the manufacture of a medicament for
the treatment of a PPARgamma mediated disease or condition.
[0021] In another aspect, the present invention provides the use of
a PPARgamma antagonist for the manufacture of a medicament for the
treatment of osteoporosis.
[0022] In another aspect, the present invention provides a method
for identifying compounds that will be useful for the treatment of
a PPAR gamma mediated disease or condition, comprising the step of
binding a compound of this invention to PPAR gamma.
[0023] In another aspect, the present invention provides a method
for treating a PPAR gamma mediated disease or condition comprising
administration of a therapeutically effective amount of a compound
that was identified as useful for such treatment by the above
method ( in other words, by a method comprising the step of binding
a compound of this invention to PPAR gamma).
[0024] In another aspect, the present invention provides the use of
a compound that was identified as useful for treating a PPAR gamma
mediated disease or condition by the above method (in other words,
by a method comprising the step of binding a compound of this
invention to PPAR gamma), for the manufacture of a medicament for
the treatment of a PPAR gamma mediated disease or condition.
[0025] In another aspect, the invention provides a method for
identifying compounds which will be useful in treatment of
osteoporosis comprising the step of determining whether a compound
antagonises PPAR gamma.
[0026] Preferably, when any of the R groups in Formula (I) are
alkyl, they are straight chain alkyl.
[0027] Preferably, R.sub.3 is butyl, benzyl optionally substituted
with 1 or 2 halogen, or p-pyridyl.
[0028] Suitable compounds of the present invention include:
[0029] 4-(4-(4-carboxyphenyl)butyl)2-heptyl4-oxo-5-thiazolidine
N,N-dibenzylacetamide,
[0030]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N,N-dibenzylacetamide,
[0031]
(2S,5S)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5thiazolidine
N,N-dibenzylacetamide,
[0032]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)2-hexyl)-4-oxo-5-thiazolidine
N,N-dibutylacetamide,
[0033]
(2S*,5S*)-4-(2-(4-carboxyphenyl)ethyl)-2-octyl-4-oxo-5-thiazolidine
N,N-dibenzylacetamide,
[0034]
(2R*,5S*)-4-(2-(4-carboxyphenyl)ethyl)-2-octyl-4-oxo-5-thiazolidine
N,N-dibenzylacetamide,
[0035]
(2S*,5S*)-4-(2-(4-carboxyphenyl)ethyl)-2-octyl-4-oxo-5-thiazolidine
N,N-di-(3-iodo)benzylacetamide,
[0036]
(2S*,5S*)-4-(3-(4-carboxyphenyl)propyl)-2-heptyl-4-oxo-5thiazolidin-
e N,N-benzylacetamide,
[0037]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl-2-heptyl-4-oxo-5-thiazolidine
N,N-di-(3-benzylacetamide,
[0038]
(2S*,5S*)-4-(2-(4-carboxyphenyl)ethyl)-2-(6-phenylhexyl)-4-oxo-5-th-
iazolidine N,N-dibenzylacetamide,
[0039]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-(6phenylhexyl)-4-oxo-5-thi-
azolidine N,N-dibenzylacetamide,
[0040]
4-(4-(4-carboxyphenyl)butyl)-2-(4-phenylbutyl)-4-oxo-5-thiazolidine
N,N-dibenzylacetamide,
[0041]
(2S*,5S*)-4-(2-(4-ureidophenyl)ethyl)-2-octyl-4-oxo-5-thiazolidine
N,N-dibenzylacetamide,
[0042]
(2S*,5S*)-4-(2-(4-methylsulfonamidophenyl)ethyl)-2-octyl-4-oxo-5-th-
iazolidine N,N-dibenzylacetamide,
[0043]
(2S*,5S*)-4-(2-(4-aminosulfonylphenyl)ethyl)-2-octyl-4-oxo-5-thiazo-
lidine N,N-dibenzylacetamide,
[0044]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-hepyl-4-oxo-5-thiazolidine
N-benzyl-N-(4-trifluorobenzyl)acetamide,
[0045]
(2R*,5S*)-4-(4(4-carboxyphenyl)butyl)-2-hepyl-4-oxo-5-thiazolidine
N-benzyl-N-(4-trifluorobenzyl)acetamide,
[0046]
(2S*,5S*)-4-(2-(4-(3-hydantoino)phenyl)ethyl)-2-octyl-4-oxo-5-thiaz-
olidine N,N-dibenzylacetamide,
[0047]
(2S*,5S*)-4-(2-(3,4-dioxomethylenephenyl)ethyl)-2-heptyl4-oxo-5-thi-
azolidine N,N-dibenzylacetamide,
[0048]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-octyl-4oxo-5-thiazolidine
N-benzyl-N-(4-hydroxybutyl)acetamide,
[0049]
(2S*,5S*)-4-(2-(3,4-dioxomethylenephenyl)ethyl)-2-octyl-4-oxo-5-thi-
azolidine N,N-dibenzylacetamide,
[0050]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N-benzyl-N-(4-pyridyl)acetamide,
[0051]
(2S*,5S*)-4-(4carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidine
N-benzyl-N-(2-pyridyl)acetamide,
[0052]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N-benzyl-N-(2-ethoxycarboxyethyl)acetamide,
[0053]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N-benzyl-N-butylacetamide,
[0054]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl4-oxo-thiazolidine
N-benzyl-N-isopropylacetamide,
[0055]
(2S*,5S*)-4-(2-(4-hydroxyphenyl)ethyl)-2-heptyl-4-oxo-5-thiazolidin-
e N,N-dibenzylacetamide,
[0056]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N-enzyl-N-ethoxycarboxymethylacetamide,
[0057]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N,N-di-(4-fluorobenzyl)acetamide,
[0058]
(2S*,5S*)-4-(2-(4-arboxymethoxyphenyl)ethyl)-2-heptyl-4-oxo-5-thiaz-
olidine N,N-dibenzylacetamide,
[0059]
(2S*,5S*)-4-(2-(4-carboxyamidomethoxyphenyl)ethyl)-2-heptyl-4-oxo-5-
-thiazolidine N,N-dibenzylacetamide,
[0060]
(2S*,5S*)-4-(2-(4-methoxyphenyl)ethyl)-2-heptyl-4-oxo-5-thiazolidin-
e N,N-dibenzylacetamide,
[0061]
(2S*,5S*)-4-(4-(4arboxyphenyl)butyl)-2-heptyl4-oxo-thiazolidine
N-benzyl-N-(2-thienylmethyl)acetamide,
[0062]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N-benzyl-N-(2,3-dioxomethylenebenzyl)acetamide,
[0063]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N-benzyl-N-(2-thiazolemethyl)acetamide,
[0064]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N-benzyl-N-(2-furfuryl)acetamide,
[0065] and pharmaceutically acceptable salts and solvates
thereof.
[0066] Particularly preferred compounds of the present invention
include:
[0067]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N,N-dibenzylacetamide,
[0068]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N,N-dibenzylacetamide,
[0069]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-hexyl-4-oxo-thiazolidine
N,N-dibutylacetamide,
[0070]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-octyl-4-oxo-5-thiazolidine
N-benzyl-N-(4-hydroxybutyl)acetamide,
[0071]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N-benzyl-N-(4-pyridyl)acetamide,
[0072]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N-benzyl-N-(2-thiazolemethyl)acetamide,
[0073] and pharmaceutically acceptable salts and solvates
thereof.
[0074] Those skilled in the art will recognise that stereocenters
exist in compounds of Formula (I). Accordingly, the present
invention includes all possible stereoisomers and geometric isomers
of formula (I) and includes not only racemic compounds but also the
optically active isomers as well. When a compound of formula (I) is
desired as a single enantiomer, it may be obtained either by
resolution of the final product or by stereospecific synthesis from
either isomerically pure starting material or any convenient
intermediate. Resolution of the final product, an intermediate or a
starting material may be effected by any suitable method known in
the art. See, for example, Stereochemistry of Carbon Compounds by
E. L. Eliel (Mcgraw Hill, 1962) and Tables of Resolving Agents by
S. H. Wilen. Additionally, in situations where tautomers of the
compounds of formula (I) are possible, the present invention is
intended to include all tautomeric forms of the compounds.
[0075] While the compounds of this invention include all
enantiomers and diastereomers, the trans pair of diastereomers is
preferred. This pair consists of the (2S, 5S) enantiomer and the
(2R, 5R) enantiomer. This diastereomer pair will be abbreviated as
(2S*,5S*). Most preferred are the (2S,5S) enantiomers.
[0076] It will also be appreciated by those skilled in the art that
the compounds of the present invention may also be utilised in the
form of a pharmaceutically acceptable salt or solvate thereof. The
physiologically acceptable salts of the compounds of formula (i)
include conventional salts formed from pharmaceutically acceptable
inorganic or organic acids or bases as well as quaternary ammonium
acid addition salts. More specific examples of suitable acid salts
include hydrochloric, hydrobromic, sulfuric, phosphoric, nitric,
perchloric, fumaric, acetic, propionic, succinic, glycolic, formic,
lactic, maleic, tartaric, citric, pamoic, malonic, hydroxymaleic,
phenylacetic, glutamic, benzoic, salicylic, fumaric,
toluenesulfonic, methanesulfonic, naphthalene-2-sulfonic,
benzenesulfonic hydroxynaphthoic, hydroiodic, malic, steroic,
tannic and the like. Other acids such as oxalic, while not in
themselves pharmaceutically acceptable, may be useful in the
preparation of salts useful as intermediates in obtaining the
compounds of the invention and their pharmaceutically acceptable
salts. More specific examples of suitable basic salts include
sodium, lithium, potassium, magnesium, aluminium, calcium, zinc,
N,N'-dibenzylethylenediam- ine, chloroprocaine, choline,
diethanolamine, ethylenediamine, N-methylglucamine and procaine
salts. References hereinafter to a compound according to the
invention include both compounds of Formula (I) and their
pharmaceutically acceptable salts and solvates.
[0077] It will be appreciated by those skilled in the art that
reference herein to treatment extends to prophylaxis as well as the
treatment of established diseases or symptoms. Moreover, it will be
appreciated that the amount of a compound of the invention required
for use in treatment will vary with the nature of the condition
being treated and the age and the condition of the patient and will
be ultimately at the discretion of the attendant physician or
veterinarian. In general, however, doses employed for adult human
treatment will typically be in the range of 0.02-5000 mg per day,
preferably 1-1500 mg per day. The desired dose may conveniently be
presented in a single dose or as divided doses administered at
appropriate intervals, for example as two, three, four or more
sub-doses per day.
[0078] While it is possible that compounds of the present invention
may be therapeutically administered as the raw chemical, it is
preferable to present the active ingredient as a pharmaceutical
formulation. Accordingly, the present invention further provides
for a pharmaceutical formulation comprising a compound of Formula
(I) or a pharmaceutically acceptable salt or solvate thereof,
preferably together with one or more pharmaceutically acceptable
carriers therefor and, optionally, other therapeutic and/or
prophylactic ingredients.
[0079] Formulations of the present invention include those
especially formulated for oral, buccal, parenteral, transdermal,
inhalation, intranasal, transmucosal, implant, or rectal
administration, however, oral administration is preferred. For
buccal administration, the formulation may take the form of tablets
or lozenges formulated in conventional manner. Tablets and capsules
for oral administration may contain conventional excipients such as
binding agents, (for example, syrup, acacia, gelatin, sorbitol,
tragacanth, mucilage of starch or polyvinylpyrrolidone), fillers
(for example, lactose, sugar, microcrystalline cellulose,
maize-starch, calcium phosphate or sorbitol), lubricants (for
example, magnesium stearate, stearic acid, talc, polyethylene
glycol or silica), disintegrants (for example, potato starch or
sodium starch glycollate) or wetting agents, such as sodium lauryl
sulfate. The tablets may be coated according to methods well known
in the art.
[0080] Alternatively, the compounds of the present invention may be
incorporated into oral liquid preparations such as aqueous or oily
suspensions, solutions, emulsions, syrups or elixirs, for example.
Moreover, formulations containing these compounds may be presented
as a dry product for constitution with water or other suitable
vehicle before use. Such liquid preparations may contain
conventional additives such as suspending agents such as sorbitol
syrup, methyl cellulose, glucose/sugar syrup, gelatin,
hydroxyethylcellulose, carboxymethyl cellulose, aluminum stearate
gel or hydrogenated edible fats; emulsifying agents such as
lecithin, sorbitan mono-oleate or acacia; non-aqueous vehicles
(which may include edible oils) such as almond oil, fractionated
coconut oil, oily esters, propylene glycol or ethyl alcohol; and
preservatives such as methyl or propyl p-hydroxybenzoates or sorbic
acid. Such preparations may also be formulated as suppositories,
e.g., containing conventional suppository bases such as cocoa
butter or other glycerides. Additionally, formulations of the
present invention may be formulated for parenteral administration
by injection or continuous infusion. Formulations for injection may
take such forms as suspensions, solutions, or emulsions in oily or
aqueous vehicles, and may contain formulatory agents such as
suspending, stabilising and/or dispersing agents Alternatively, the
active ingredient may be in powder form for constitution with a
suitable vehicle (e.g., sterile, pyrogen-free water) before
use.
[0081] The formulations according to the invention may also be
formulated as a depot preparation. Such long acting formulations
may be administered by implantation (for example, subcutaneously or
intramuscularly) or by intramuscular injection. Accordingly, the
compounds of the invention may be formulated with suitable
polymeric or hydrophobic materials (as an emulsion in an acceptable
oil, for example), ion exchange resins or as sparingly soluble
derivatives as a sparingly soluble salt, for example.
[0082] The formulations according to the invention may contain
between 0.1-99% of the active ingredient, conveniently from 30-95%
for tablets and capsules and 3- 50% for liquid preparations.
[0083] The novel thiazolidine acetamides of this invention can be
used to inhibit adipogenesis. Surprisingly, it has been found that
PPARgamma ligands that inhibit adipogenesis also stimulate alkaline
phosphatase activity, which is a surrogate marker for stimulation
of osteogenesis. In addition to this activity, these novel
PPAR-gamma ligands maintain many of the beneficial effects of known
PPARgamma ligands, such as antidiabetic activity.
[0084] Thus, synthetic PPAR-gamma ligands that block adipogenesis
while mimicking the beneficial effects of natural PPARgamma
hormones will be useful for the treatment of human disease,
including diabetes, obesity, dyslipidemia, metabolic syndrome,
osteoporosis, acne, cardiovascular disease, inflammation, or
cancer.
[0085] The compounds of this invention can be prepared by standard
organic chemistry as illustrated by the accompanying working
examples. The following examples are set forth to illustrate the
synthesis of some particular compounds of the present invention and
to exemplify general processes. Accordingly, the following Examples
section is in no way intended to limit the scope of the invention
contemplated herein.
EXAMPLES
[0086] As used herein the symbols and conventions used in these
processes, schemes and examples are consistent with those used in
the contemporary scientific literature, for example, the Journal of
the American Chemical Society. Unless otherwise noted, all starting
materials were obtained from commercial suppliers and used without
further purification. Specifically, the following abbreviations may
be used in the examples and throughout the specification: g
(grams); mg (milligrams); L (liters); mL (milliliters); .mu.L
(microliters); N (normal); mM (millimolar); mmol (millimoles); i.
v. (intravenous); Hz (Hertz); MHz (megahertz); mol (moles); RT or
rt (room temperature); min (minutes); h (hours); mp. (melting
point); TLC (thin layer chromatography); HPLC (high pressure liquid
chromatography); ms (mass spectrum); ES+ (electrospray); R.sub.f
(retention fraction); (t.sub.r (retention time); RP (reverse
phase); MeOH (methanol); TFA (trifluoroacetic acid); HCl
(hydrochloric acid); HCO.sub.2H (formic acid); THF
(tetrahydrofuran); CH.sub.3CN (acetonitrile); EtOH (ethanol);
CDCl.sub.3 (deuterated chloroform); DMSO (dimethylsulfoxide);
DMSO-d.sub.6 (dimethylsulfoxide-deuterated); EtOAc (ethyl acetate);
DCM or CH.sub.2Cl.sub.2 (dichloromethane) ; DMF
(dimethylformamide); Et.sub.3N (triethylamine); MgSO.sub.4
(magnesium sulfate); H.sub.2O (water); LAH (lithium aluminum
hydride; NaH (sodium hydride); Na.sub.2CO.sub.3 (sodium carbonate);
Na.sub.2SO.sub.4 (sodium sulfate); MnO.sub.2 (manganese dioxide);
NaOH (sodium hydroxide; LiOH (lithium hydroxide); DIEA
(diisopropylethylamine); Et.sub.2O (diethyl ether; diethyl
azodicaboxylate (DEAD); tert-butyloxycarbonyl (BOC); NaHCO.sub.3
(saturated aqueous sodium bicarbonate). Brine refers to a saturated
aqueous solution of NaCl. Unless otherwise indicated, all
temperatures are expressed in .degree. C. (degrees Centigrade). All
reactions conducted at room temperature unless otherwise noted.
[0087] The .sup.1H NMR spectra were recorded on a Varian VXR-300, a
Varian Unity-300, or a Varian Unity-400 instrument. Chemical shifts
are expressed in parts per million (ppm, .delta. units). Coupling
constants are in units of hertz (Hz). Splitting patterns are
designated as s, singlet; d, doublet; t, triplet; q, quartet; m,
multiplet; br, broad; hept, heptuplet.
[0088] Low-resolution mass spectra (MS) were recorded on a JOEL
JMS-AX505HA, JOEL SX-102 or a SCIEX-APliii spectrometers. All mass
spectra were taken under electrospray ionization (ES, either in the
positive ion mode or negative ion mode) or by fast atom bombardment
(FAB) methods. Infrared (IR) spectra were obtained on a Nicolet 510
FT-IR spectrometer using a 1-mm NaCl cell. All reactions were
monitored by thin-layer chromatography on 0.25 mm E. Merck silica
gel plates (60F-254), visualized with UV light, iodine staining, or
7% ethanolic phosphomolybdic acid or p-anisidehyde solutions. Flash
column chromatography was performed on silica gel (230-400 mesh,
Merck).
[0089] Analytical purity was assessed on a Hewlett Packard series
1050 or 1100 system equipped with a diode array spectrometer. The
stationary phase was either a Dynamax C8 column (25 cm.times.4.1
mm), a Dynamax 60A C18 column (25 cm.times.4.6 mm), a Vydac C18
column (5 m, 4.6 mm.times.250 mm), a Supelco C18 column (5 m, 4.6
mm.times.150 mm), or a Rainin C18 column (5 m, 4.6 mm.times.250
mm). The flow rate was 1.0 to 1.5 ml/min. (t=2.8 or 3.0 min.) and
the solvent systems were as described below. Enantiomeric purity
was assessed using either a Chiralpak AD column (25 cm.times.4.6
mm) or a Chiralpak OD column (25 cm.times.4.6 mm) on either a
Hewlet Packard series 1050 HPLC system equipped with a diode array
spectrometer or on a Supercritical Fluid (SFC) system using
CO.sub.2/methanol as the mobile phase.
Example 1
4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidine
N,N-dibenzylacetamide
[0090] 9-Fluorenylmethyl chloroformate (6.9 g) was added to
biphasic mixture of tert-butyl 4-(4-aminobutyl)benzoate (5.7 g) in
THF (100 mL) and saturated aqueous NaHCO.sub.3 (50 mL). The
resulting mixture was stirred at r.t. for 1 h and then diluted with
EtOAc and H.sub.2O. The organic layer was separated and washed with
brine, dried over MgSO.sub.4, filtered and concentrated. The crude
material was purified by flash chromatography to give 7.4 g of
tert-butyl 4-(4-(9-fluorenylmethoxycarbon-
yl)-aminobutyl)benzoate.
[0091] Trifluoroacetic acid (100 mL) was added to a solution of
tert-butyl 4-(4-(9-fluorenylmethoxycarbonyl)aminobutyl)benzoate
(6.9 g) in CH.sub.2Cl.sub.2. The resulting solution was stirred at
rt. for 3 h and then concentrated. The crude product was triturated
with hexane and the solid collected by filtration to give 5.5 g of
4-(4-(9-fluorenylmethoxyca- rbonyl)amino)butylbenzoic acid.
[0092] A mixture of
4-(4-(9-fluorenylmethoxycarbonyl)amino)butylbenzoic acid (2.8 g) in
CH.sub.2Cl.sub.2 (30 mL) was treated with
2-fluoro-1,3-dimethylpyridinium tosylate (2.1 g) followed by
N,N-diisopropylethylamine (3.7 mL). The solution was shaken for 15
min, and then Sasrin resin (4 g, 0.89 mmol/g) was added followed by
4-dimethylaminopyridine (0.1 g). The mixture was shaken for 5 h and
then drained. The resin was washed with DMF, CH.sub.2Cl.sub.2, MeOH
and Et.sub.2O. The resulting resin was treated with 0.5 M acetic
anhydride in CH.sub.2Cl.sub.2 (20 mL) and 0.5 M pyrdine in
CH.sub.2Cl.sub.2 (20 mL) and shaken for 30 min and then drained.
The resin was washed with DMF, CH.sub.2Cl.sub.2, MeOH and
Et.sub.2O. FMOC analysis showed a loading of 0.5 mmol/g.
[0093] A solution of 20% piperidine in DMF (8 mL) was added a
reaction vessel containing an aliquot of the resin (0.5 g). The
mixture was shaken for 45 min and then drained and washed with DMF,
CH.sub.2Cl.sub.2, MeOH, Et.sub.2O and THF. The resin was
transferred to a 40 mL glass scintillation vial with a Teflon lined
cap. A THF solution (15 mL) of octanal (0.75 M) and
mercaptosuccinic acid (2 M) was added followed by activated 3 .ANG.
sieves (100 mg). The mixture was heated at 70.degree. C. with
shaking for 4 h. The resin was drained and washed with DMF,
CH.sub.2C.sub.2, MeOH and Et.sub.2O. An aliquot of the resulting
resin (50 mg) was treated with a solution of pentafluorophenyl
trifluoroacetate (0.09 mL) and pyridine (0.09 mL) in DMF (0.18 mL)
and shaken for 4 h. The resin was drained and washed with DMF. A
DMF solution (0.75 mL) of N,N-dibenzylamine (0.5 M),
N,N-diisopropylethylamine (0.6 M) and 4-dimethylaminopyridine (0.01
M) was added. The mixture was shaken at rt. for 18 h. The resin was
drained and washed with DMF, CH.sub.2Cl.sub.2, MeOH and Et.sub.2O.
The resin was cleaved with 10% TFA in CH.sub.2Cl.sub.2 (0.25 mL)
for 30 min. The resulting solution was collected and the resin
washed with CH.sub.2Cl.sub.2. The combined CH.sub.2Cl.sub.2
solutions were concentrated to yield 7 mg of
4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4oxo-5-thiazolidine
N,N-dibenzylacetamide as a mixture of cis:trans (2S*,5R*:2S*,5S*)
isomers: MS (ES+) 601. The isomers could be separated by C18
reverse phase HPLC.
Example 2
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidine
N,N-dibenzylacetamide
[0094] In a seated tube tert-butyl 4-iodobenzoate (28.1 g) was
dissolved in triethylamine (26 mL). N-phthalyl-3-butene (19.5 g),
palladium (II) acetate (1.0 g) and tri-o-tolylphosphine (2.8 g)
were added. Nitrogen was bubbled through the reaction for 10 min
and then the reaction vessel was sealed. The reaction was placed in
an oil bath preheated to 110.degree. C. and stirred for 4.5 h. The
reaction was cooled to r.t. Ethyl acetate (250 mL) and 1N HCl (100
mL) were added and the reaction stirred until solids were
dissolved. The mixture was filtered through a plug of celite and
then the layers separated. The aqueous fraction was extracted with
ethyl acetate (3.times.50 mL) and the combined organic extracts
were dried over MgSO.sub.4, filtered and concentrated. The solids
were triturated with hexane and the solids filtered and collected
to afford 11.8 g of tert-butyl
4-[(E)-4-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1--
butenyl]benzoate: 1H NMR (CDCl.sub.3, 400 MHz) 7.88 (d, 2H, J=8.4
Hz), 7.82 (dd, 2H, J=3.1, 3.1 Hz), 7.77 (dd, 2H, J=3.0, 3.1 Hz),
7.32 (d, 2H, J=8.2 Hz), 6.45 (d, 1H, J=15.9 Hz), 6.28 (dt, 1H,
J=7.2, 15.9, 7.2 Hz), 3.85 (t, 2h, J=7.0 Hz), 2.65 (q, 2H, J=6.9
Hz), 1.60 (s, 9H). tert-Butyl
4-[(E)-4-(1,3-dioxo-1,3dihydro-2H-isoindol-2-yl)-1-butenyl]benzoate
(15.1 g) was dissolved in 1:1 THF/EtOH (150 mL). 10% Pd on carbon
(6.0 g) was added and the reaction placed on a Parr hydrogenator
under 50 psi of H.sub.2. The reaction was shaken for 6 h. The
reaction was filtered through a pad of celite and the celite washed
with ethanol (3.times.100 mL). The organic extracts were
concentrated to afford 15.3 g of tert-butyl
4-[4-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)butyl]benzoate: 1H NMR
(CDCl.sub.3, 400 MHz) 7.88 (d, 2H, J=8.1 Hz), 7.82 (dd, 2H, J=3.0,
3.1 Hz), 7.77 (dd, 2H, J=3.0, 3.2 Hz), 7.2 (d, 2H, J=8.1 Hz), 3.7
(t, 2h, J=6.9 Hz), 2.7 (t, 2H, J=6.9 Hz), 1.7 (m, 4H), 1.60 (s,
9H).
[0095] To tert-butyl
4-[4-(1,3dioxo-1,3-dihydro-2H-isoindol-2-yl)butyl]ben- zoate (33.2
g) in ethanol (600 mL) was added hydrazine (16.9 g). The reaction
was heated to reflux and stirred for 2 h. The reaction was then
cooled to r.t. and filtered. The solids were washed with ethanol
(4.times.200 mL). The combined organic extracts were concentrated
and then dissolved in CH.sub.2C1.sub.2 and insoluble material
removed by filtration. The organic extracts were concentrated to
afford 17.1 g of tert-butyl 4-(4-aminobutyl)benzoate: 1H NMR
(CDCl.sub.3, 400 MHz) 7.90 (d. 2H, J=8.2 Hz), 7.2 (d, 2H, J=8.0
Hz), 2.7 (m, 2H,), 1.7 (m, 4H), 1.60 (s, 9H), 1.4 (m, 2H).
[0096] tert-Butyl 4-(4-aminobutyl)benzoate (8.1 g),
mercaptosuccinic acid (10.3 g) and octanal (8.4 g) in toluene (320
mL) were heated to reflux for 18 h. The reaction was cooled to r.t.
and diluted with ethyl acetate (300 mL). The organic extracts were
washed with water (3.times.300 mL), brine (300 mL), dried over
MgSO.sub.4, filtered and concentrated to afford 28.5 g of a golden
oil. The oil was dissolved in CH.sub.2Cl.sub.2 (250 mL) and cooled
to 0.degree. C. HOBT (5.5 g) and EDC (7.8 g) were added and the
reaction stirred for 30 min. Dibenzylamine (8.1 g) was added and
the reaction stirred for 2 d. The resulting mixture was washed with
water (500 mL), brine (500 mL), dried over MgSO.sub.4, filtered and
concentrated. The crude material was purified and the cis:trans
isomers separated by flash chromatography to afford 4.3 g of
(2S*,5S*)-4-4-(4tert-butylcarboxyphenyl)butyl)-2-heptyl4-oxo-5-thiazolidi-
ne N,N-dibenzylacetamide, 4.8 g of
(2R*,5S*)-4-(4-(4-tert-butylcarboxyphen-
yl)butyl)-2heptyl-4oxo-5-thiazolidine N,N-dibenzylacetamide, and
5.86 g of the cis:trans mixture: 1H NMR (CDCl.sub.3, 400 MHz) trans
isomer 7.88 (d, 2H, J=8.0 Hz), 7.38-7.26 (m, 6H), 7.22-7.12 (m,
6H), 4.74 (d, 1 H, J=14.7 Hz), 4.52- 4.30 (m, 4H), 3.72-3.62 (m,
1H), 3.47 (dd, 1 H, J=3.0, 13.7 Hz), 3.04-2.95 (m, 1 H), 2.78-2.60
(m, 3H), 1.62-1.55 (m, 14H), 1.31-1.25 (m, 8H), 0.88 (t, 3H, J=6.8
Hz).
[0097]
(2S*,5S*)-4-(4-(4-tert-butylcarboxyphenyl)butyl)-2-heptyl-4-oxo-5-t-
hiazolidine N,N-dibenzylacetamide (13.3 g) was dissolved in a 1:1
solution of TFA/CH.sub.2Cl.sub.2 and stirred at r.t. for 2 h. The
reaction was concentrated and azetroped with CH.sub.2Cl.sub.2
(3.times.200 mL) and ether (3.times.200 mL). The residue was
dissolved in ether (15 mL) and hexane (200 mL). The solution was
stirred overnight and filtered to afford 9.2 g of
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-t-
hiazolidine N,N-dibenzylacetamide as a white solid: m.p.
94-97.degree. C.; 1H NMR (CDCl.sub.3, 400 MHz 7.96 (d, 2H, J=8.2
Hz), 7.40-7.20 (m, 1 OH), 7.14 (d, 2H, J=7.2 Hz), 4.74 (d, 1H,
J=14.7 Hz), 4.54 4.28 (m, 5H), 3.72-3.62 (m, 1H), 3.47 (dd, 1H,
J=3.0, 13.8 Hz), 3.04-2.95 (m, 1H), 2.78-2.60 (m, 3H), 1.90-1.80
(m, 1H), 1.70-1.50 (m, 8H), 1.44-1.3 m, 8H), 0.88 (t, 3H, J=6.8
Hz); Anal. (C.sub.37H.sub.46N.sub.2S.sub.1O.sub.4)C, 72.31; H,
7.49; N, 4.56; S, 5.21 Found C, 72.24; H, 7.54; N, 4.56; S,
5.16.
Example 3
(2S,5S)-4-(4-(4-carboxyphenyl)buty)-2-heptyl4-oxo-5-thiazolidine
N,N-dibenzylacetamide
[0098]
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N,N-dibenzylacetamide was a resolved by chiral HPLC using a
Daicel AD column (2.times.25 cm) with 15% isopropanol/hexane as the
mobile phase at a flow rate of 8 mL/min. Monitoring at 230 nM, peak
1 was observed at 220 min and peak 2 as observed at 260 min. Peak 1
was collected and concentrated to give
(2S,5S)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo--
5-thiazolidine N,N-dibenzylacetamide. Absolute stereochemistry was
assigned by X-ray crystallography. Peak 2 was collected and
concentrated to give
(2R,5R)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-thiazolidin-
e N,N-dibenzylacetamide.
Examples 4-36
[0099] In a similar manner to the preparation of Examples 1-3, the
following examples were prepared.
[0100] Example 4
(2S*,5S*)-4(4-(4-carboxyphenyl)butyl)-2-hexyl-4-oxo-5-thi-
azolidine N,N-dibutylacetamide
[0101] Example 5
(2S*,5S*)4-(2-(4-carboxyphenyl)ethyl)-2-octyl4-oxo-5-thia- zolidine
N,N-dibenzylacetamide
[0102] Example 6
(2R*,5S*)-4-(2-(4-carboxyphenyl)ethyl)-2-octyl4-oxo-5-thi-
azolidine N,N-dibenzylacetamide
[0103] Example 7
(2S*,5S*)-4-(2-(4-carboxyphenyl)ethyl)-2-octyl-4-oxo-5-th-
iazolidine N,N-di-(3-iodo)benzylacetamide
[0104] Example 8
(2S*,5S*)-4-(3-(4-carboxyphenyl)propyl)-2-heptyl-4-oxo-5--
thiazolidine N,N-benzylacetamide
[0105] Example 9
(2S*,5S*)-4-(4-(4carboxyphenyl)butyl)-2-heptyl-4-oxo-5-th-
iazolidine N,N-di-(3-benzylacetamide
[0106] Example 10
(2S*,5S*)-4-(2-(4-carboxyphenyl)ethyl)-2-(6-phenylhexyl)-
-4-oxo-5-thiazolidine N,N-dibenzylacetamide
[0107] Example 11
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-(6-phenylhexyl)-
-4-oxo-5-thiazolidine N,N-dibenzylacetamide
[0108] Example 12
4-(4-(4-carboxyphenyl)butyl)-2-(4-phenylbutyl)-4-oxo-5-t-
hiazolidine N,N-dibenzylacetamide
[0109] Example 13
(2S*,5S*)-4-(2-(4-ureidophenyl)ethyl)-2-octyl-4-oxo-5-th-
iazolidine N,N-dibenzylacetamide
[0110] Example 14
(2S*,5S*)-4-(2-(4-methysulfonamidophenyl)ethyl)-2-octyl--
4-oxo-5-thiazolidine N,N-dibenzylacetamide
[0111] Example 15
(2S*,5S*)-4-(2-(4-aminosulfonylphenyl)ethyl)-2-octyl-4-o-
xo-5-thiazolidine N,N-dibenzylacetamide
[0112] Example 16
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-hepyl-4-oxo-5-t-
hiazolidine N-benzyl-N-(4-trifluorobenzyl)acetamide
[0113] Example 17
(2R*,5S*)-4-(4-(4carboxyphenyl)butyl)-2-hepyl-4-oxo-5-th-
iazolidine N-benzyl-N-(4-trifluorobenzyl)acetamide
[0114] Example 18
(2S,5S*)-4-(2-(4-(3-hydantoino)phenyl)ethyl)-2-octyl4-ox-
o-5-thiazolidine N,N-dibenzylacetamide
[0115] Example 19
(2S*,5S*)-4-(2-(3,4-dioxomethylenephenyl)ethyl-2-heptyl4-
-oxo-5-thiazolidine N,N-dibenzylacetamide
[0116] Example 20
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-octyl-4-oxo-5-t-
hiazolidine N-benzyl-N-(4-hydroxybutyl)acetamide
[0117] Example 21
(2S*,5S*)-4-(2-(3,4-dioxomethylenephenyl)ethyl)-2-octyl--
4-oxo-5-thiazolidine N,N-dibenzylacetamide
[0118] Example 22
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4oxo-5-t-
hiazolidine N-benzyl-N-(4-pyridyl)acetamide
[0119] Example 23
(2S*,5S*)-4-(4-(carboxyphenyl)butyl)-2-heptyl-4-oxo-5-th-
iazolidine N-benzyl-N-(2-pyridyl)acetamide
[0120] Example 24
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5--
thiazolidine N-benzyl-N-(2-ethoxycarboxyethyl)acetamide
[0121] Example 25
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5--
thiazolidine N-benzyl-N-butylacetamide
[0122] Example 26
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5--
thiazolidine N-benzyl-N-isopropylacetamide
[0123] Example 27
(2S*,5S*)-4-(2-(4-hydroxyphenyl)ethyl)-2-heptyl-4-oxo-5--
thiazolidine N,N-dibenzylacetamide
[0124] Example 28
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5--
thiazolidine N-benzyl-N-ethoxycarboxymethylacetamide
[0125] Example 29
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5--
thiazolidine N,N-di-(4-fluorobenzyl)acetamide
[0126] Example 30
(2S*,5S*)-4-(2-(4-carboxymethoxyphenyl)ethyl)-2-heptyl-4-
-oxo-5-thiazolidine N,N-dibenzylacetamide
[0127] Example 31
(2S*,5S*)-4-(2-(4-carboxyamidomethoxyphenyl)ethyl-2-hept-
yl-4-oxo-5-thiazolidine N,N-dibenzylacetamide
[0128] Example 32
(2S*,5S*)-4-(2-(4-methoxyphenyl)ethyl)-2-heptyl-4-oxo-5--
thiazolidine N,N-dibenzylacetamide
[0129] Example 33
(2S*,5S*)-4-(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5--
thiazolidine N-benzyl-N-(2-thienylmethyl)acetamide
[0130] Example 34
(2S*,5S*)-4(4-(4-carboxyphenyl)butyl)-2-heptyl-4-oxo-5-t-
hiazolidine N-benzyl-N-(2,3-dioxomethylenebenzyl)acetamide
[0131] Example 35
(2S*,5S*)-4-(4-(4carboxyphenyl)butyl)-2-heptyl-4-oxo-5-t-
hiazolidine N-benzyl-N-(2-thiazolemethyl)acetamide
[0132] Example 36
(2S*,5S*)-4-(4-(4arboxyphenyl)butyl)-2-heptyl-4-oxo5-thi- azolidine
N-benzyl-N-(2-furfuryl)acetamide
[0133] Binding Assay
[0134] Test compounds were assayed for binding to the human
PPAR-gamma receptor ligand binding domain as described in Nichols,
J. S., Parks, D. J., Consler, T. G., and Blanchard, S. G., Anal.
Biochem., (1998), Vol. 257, pp 112-119. Each of the above Examples
1-36 had a pK.sub.i>5 in this binding assay.
[0135] Cell-Based Reporter Assay
[0136] CV-1 cells were maintained in DME High Glucose medium
(Irvine Scientific) supplemented with 10% fetal bovine serum and 2
mM Glutamine. Cells were split into D-MEM/F-12 medium (Gibco)
supplemented with 10% charcoal stripped fetal bovine serum for 3 d
before harvesting. Cells were harvested into D-MEM/F-12 medium
(Gibco) supplemented with 10% charcoal stripped fetal bovine serum
and counted. Cells were seeded at a density of 24,000 cells per
well into 96-well plates and incubated overnight at 5% CO.sub.2 and
37.degree. C. Cells were transfected for 6 to 20 hours based on the
Lipofectamine protocol (Gibco) With the following amounts of DNA
per well: 2 ng PSG5 GAL4-human PPAR-gamma, 8 ng UAS-tk-SPAP, 25 ng
beta-gal, 45 ng pBluescript. See Lehmann, J. M. et al., J. Biol.
Chem., (1995), Vol. 270, pp 12953-12956 and Brown, P. J. et al.,
Chem. Biol., (1997), Vol. 4, pp 909-918. Cells were incubated
overnight at 5% CO.sub.2 and 37.degree. C. Test compounds were
solublized to 10 mM in DMSO. Test compounds were then serially
diluted from 1e-5 M to 1e-10 M into D-MEM/F-12 (Gibco) medium
supplemented with 10% delipidated and charcoal stripped calf serum
(Sigma) heat inactivated at 60.degree. C. for 30 minutes, 2 mM
Glutamine, and Pen-Strep. This medium into which the test compounds
were diluted also contained 100 nM rosiglitazone. These test
compound dilutions were added 100 microliters/well to the
transfected cell plates after the transfection media were
aspirated. DMSO controls and 1 micromolar rosiglitazone controls
were added to each cell plate. Cells were incubated overnight at 5%
CO.sub.2 and 37.degree. C. Cells were lysed with 25 microliters
0.5% Triton X-100. Two daughter plates were made from each mother
plate. One daughter received 200 microliters/well SPAP substrate
(Sigma 104) and the other daughter received 200 microliters/well
beta-gal substrate (Sigma N-1127). Once developed, cell plates were
read at 405 nM. SPAP data were normalized to beta-gal, and %
maximum inhibition of transactivation was calculated relative to
the 1 micromolar rosiglitazone positive control. Each of the above
Examples 1-36 had >50% inhibition of transactivation by 100 nM
rosiglitazone in this PPAR-gamma cell based reporter gene
assay.
[0137] Adipocyte Differentiation Assay
[0138] C3H10T1/2 clone 8 murine fibroblasts (American Type Culture
Collection) below passage 22 were maintained in Dulbecco's modified
Eagle's medium (Life Technologies, Inc.) supplemented with 10%
fetal calf serum and 100 units/mL penicillin G and 100 microgram/mL
streptomycin. One day after passage into 96-well microtiter plates
(12.5.times.103 cells/cm.sup.2), the cells were treated with 150 nM
rosiglitazone plus 1 micromolar insulin and 1 micromolar
9-cis-retinoic acid (Sigma, St. Louis, Mo.). Vehicle or test
compounds, which had been solublized to 10 mM in DMSO and then
serially diluted from 1e-5 M to 1e-10 M into medium, were added.
After 7 days, cells were lysed in 0.01% Digitonin (Sigma, St.
Louis, Mo.) and the lipogenic activity determined by measuring
total triglycerides using a Glycerol-Triglyceride (GPO-Trinder) kit
(337-B,Sigma, St. Louis, Mo.). The mixture was incubated at
37.degree. C. for 2 h and the absorbance read at 550 nm. The %
maximum inhibition of lipogenesis was calculated relative to the
vehicle treated cells. Each of the above Examples 1-36 had >50%
inhibition of lipogenesis induced by 150 nM rosiglitazone in this
adipocyte differentiation assay.
[0139] Osteoblast Differentiation Assay (Alkaline Phosphatase
Activity):
[0140] C3H10T1/2 clone 8 murine fibroblasts (American Type Culture
Collection) below passage 22 were maintained in Dulbecco's modified
Eagle's medium (Life Technologies, Inc.) supplemented with 10%
fetal calf serum and 100 units/mL penicillin G and 100 microgram/mL
streptomycin. One day after passage into 96-well microtiter plates
(12.5.times.103 cells/cm.sup.2), the cells were treated with 625
nanomolar all-trans retinoic acid, 1 micromolar rosiglitazone and 1
micromolar insulin (Sigma, St. Louis, Mo.). Vehicle or test
compounds, which had been solublized to 10 mM in DMSO and then
serially diluted from 2e-5 M to 4e-8 M into medium, were added.
After 7 days, alkaline phosphatase activity, a surrogate measure
for osteogenesis, was determined using Sigma-Fast pNPP substrate
(N-2770) according to the manufacturer'sspecifications (Sigma, St.
Louis, Mo.) (see Paulik, M. A. and Lenhard, J. M., Cell Tissue
Res., (1997), Vol. 290, pp 7987). The substrate (50
microliters/well) was incubated with the cells at 37.degree. C. for
10 min and the absorbance read at 405 nm. The % maximum stimulation
of alkaline phosphatase activity was calculated relative to cells
treated with 10 micromolar all-trans retinoic acid.
1 Compound pEC.sub.50 % Stimulation Example 4 5.7 71
[0141] In Vivo Assay
[0142] Age and weight matched male C57BL/KsJ db/db mice (Jackson
Labs, Bar Harbor, Me.) were housed 5 animals/cage at 72.degree. F.
and 50% relative humidity with a 12 h light and dark cycle, and fed
chow diet (NIH R&M/Auto 6F-Ovals 5K67, PMI Feeds.RTM. Inc.,
Richmond, Ind.). Animals starting at 6 weeks of age were orally
gavaged once daily (8:00-9:00 AM) with TPGS/PG (25/75) or 50 mg/kg
the compound of Example 4 in TPGS/PG (25/75). After 2 weeks of
dosing, the animals were anesthetized with isofluorane, blood was
drawn by cardiac puncture, and non-fasting measurements of glucose,
insulin, and glycosylated hemoglobin (GHB) were obtained. Blood
glucose was determined using an automated chemistry analyzer
(Technicon Axon). GHB measurements were performed using a
Columnmate Analyzer (Helena Instrument). Insulin concentrations in
serum were measured by chemiluminescence using an Origen Analyzer
(Igen Inc.). Body weights were recorded at the beginning and end of
the study. All data was calculated as the mean and standard error
from experiments performed on >9 animals per treatment
group.
2 Glucose Insulin GBH Body Weight Compound (mmol/L) (pmol/L) (%)
Change (g) Vehicle 460 .+-. 40 6.1 .+-. 0.8 7.6 .+-. 0.4 0.9 .+-.
0.4 Example 4 380 .+-. 30 3.3 .+-. 0.4 6.6 .+-. 0.3 1.1 .+-.
0.5
* * * * *