U.S. patent application number 10/530757 was filed with the patent office on 2006-10-19 for novel heterocyclic analogs of diphenylethylene compounds.
This patent application is currently assigned to THERACOS, INC.. Invention is credited to Debenbdranath Dey, Arthur Lee, Satyanarayana Medicherla, Bishwajit Nag, Partha Neogi.
Application Number | 20060235062 10/530757 |
Document ID | / |
Family ID | 32092375 |
Filed Date | 2006-10-19 |
United States Patent
Application |
20060235062 |
Kind Code |
A1 |
Neogi; Partha ; et
al. |
October 19, 2006 |
Novel heterocyclic analogs of diphenylethylene compounds
Abstract
Novel diphenylethylene compounds and derivatives thereof
containing thiazolidinedione or oxazolidinedione moieties are
provided which are effective in lowering blood glucose level, serum
insulin, triglyceride and free fatty acid levels in animal models
of Type II diabetes. The compounds are disclosed as useful for a
variety of treatments including the treatment of inflammation,
inflammatory and immunological diseases, insulin resistance,
hyperlipidemia, coronary artery disease, cancer and multiple
sclerosis.
Inventors: |
Neogi; Partha; (FREMONT,
CA) ; Dey; Debenbdranath; (Fremont, CA) ;
Medicherla; Satyanarayana; (Cupertino, CA) ; Nag;
Bishwajit; (Union City, CA) ; Lee; Arthur;
(Cambridge, MA) |
Correspondence
Address: |
MAYER, BROWN, ROWE & MAW LLP
1909 K STREET, N.W.
WASHINGTON
DC
20006
US
|
Assignee: |
THERACOS, INC.
525 DEL REY AVENUE, SUITE A
SUNNYVALE
CA
94085
|
Family ID: |
32092375 |
Appl. No.: |
10/530757 |
Filed: |
October 8, 2003 |
PCT Filed: |
October 8, 2003 |
PCT NO: |
PCT/US03/31803 |
371 Date: |
May 8, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
10265902 |
Oct 8, 2002 |
|
|
|
10530757 |
May 8, 2006 |
|
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Current U.S.
Class: |
514/369 ;
514/376; 514/389; 548/186; 548/227; 548/317.1 |
Current CPC
Class: |
A61P 25/00 20180101;
A61P 35/00 20180101; A61P 3/06 20180101; C07D 277/20 20130101; A61P
9/10 20180101; C07D 277/34 20130101; A61P 3/10 20180101; C07D
277/24 20130101; A61P 37/02 20180101; A61P 29/00 20180101 |
Class at
Publication: |
514/369 ;
514/376; 514/389; 548/186; 548/227; 548/317.1 |
International
Class: |
A61K 31/426 20060101
A61K031/426; A61K 31/421 20060101 A61K031/421; A61K 31/4166
20060101 A61K031/4166 |
Claims
1. A compound represented by the following formula 1: ##STR28##
wherein Z is ##STR29## n, m, q and r independently represent
integers from zero to 4 provided that n+m.ltoreq.4 and
q+r.ltoreq.4; p and s independently represent integers from zero to
5 provided that p+s.ltoreq.5; a, b and c represent double bonds
which may be present or absent; when present, the double bonds may
be in the E or Z configuration and, when absent, the resulting
stereocenters may have the R- or S-configuration; R and R' each
independently represent a hydrogen atom; linear or branched
C.sub.1-C.sub.20 alkyl; linear or branched C.sub.2-C.sub.20
alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2; --NHR''';
--NR.sub.2'''; --OH; --OR'''; --CONR.sub.2''''; halogen atom;
optionally substituted linear or branched C.sub.1-C.sub.20alkyl;
optionally substituted linear or branched C.sub.2-C.sub.20 alkenyl;
R'' independently represents a hydrogen atom; linear or branched
C.sub.1-C.sub.20 alkyl; linear or branched C.sub.2-C.sub.20
alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2; --NHR''';
--NR.sub.2'''; --OH; --OR'''; halogen atom; optionally substituted
linear or branched C.sub.1-C.sub.20 alkyl; optionally substituted
linear or branched C.sub.2-C.sub.20 alkenyl; R''' independently
represents a linear or branched C.sub.1-C.sub.20 alkyl; linear or
branched C.sub.2-C.sub.20 alkenyl; or --(CH.sub.2).sub.x--Ar, where
x represents an integer from 1 to 6 and Ar represents aryl; R''''
independently represents a hydrogen atom; optionally substituted
C.sub.1-C.sub.20 alkyl; optionally substituted C.sub.1-C.sub.20
alkoxy; optionally substituted C.sub.2-C.sub.20 alkenyl; optionally
substituted C.sub.6-C.sub.10 aryl; or NR.sub.2'''' represents a
cyclic moiety; Z' represents a hydrogen atom or a pharmaceutically
acceptable counter-ion; A, A' and A'' each independently represent
a hydrogen atom; C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20
acyloxy; C.sub.1-C.sub.20 alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl;
C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
C.sub.1-C.sub.20 alkylcarboxylamino; carboxyl; cyano; halo; or
hydroxy; B, B' and B'' each independently represent;
C.sub.2-C.sub.20alkenoyl; aroyl; aralkanoyl; nitro; optionally
substituted, linear or branched C.sub.1-C.sub.20 alkyl; or
optionally substituted, linear or branched C.sub.2-C.sub.20
alkenyl; or A and B jointly, A' and B' jointly, or A'' and B''
jointly, independently represent a methylenedioxy or ethylenedioxy
group; and X and X' independently represent >NH, >NR''',
--O--, or --S--.
2. A compound represented by the following formula 1: ##STR30##
wherein Z is ##STR31## n, m, q and r independently represent
integers from zero to 4 provided that n+m.ltoreq.4 and
q+r.ltoreq.4; p and s independently represent integers from zero to
5 provided that p+s.ltoreq.5; a, b and c represent double bonds
which may be present or absent; when present, the double bonds may
be in the E or Z configuration and, when absent, the resulting
stereocenters may have the R- or S-configuration; R independently
represents a hydrogen atom; linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z';
--CO.sub.2R'''; --NH.sub.2; --NHR'''; --NR.sub.2'''; --OH; --OR''';
--CONR.sub.2''''; halogen atom; optionally substituted linear or
branched C.sub.1-C.sub.20 alkyl; optionally substituted linear or
branched C.sub.2-C.sub.20 alkenyl; R' independently represents a
hydrogen atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or
branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R''';
--NH.sub.2; --NHR'''; --NR.sub.2'''; --OR'''; --CONR.sub.2'''';
halogen atom; optionally substituted linear or branched
C.sub.1-C.sub.20 alkyl; optionally substituted linear or branched
C.sub.2-C.sub.20 alkenyl; R'' independently represents a hydrogen
atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2;
--NHR'''; --NR.sub.2'''; --OH; --OR'''; halogen atom; optionally
substituted linear or branched C.sub.1-C.sub.20 alkyl; optionally
substituted linear or branched C.sub.2-C.sub.20 alkenyl; R'''
independently represents a linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; or
--(CH.sub.2).sub.x--Ar, where x represents an integer from 1 to 6
and Ar represents aryl; R'''' independently represents a hydrogen
atom; optionally substituted C.sub.1-C.sub.20 alkyl; optionally
substituted C.sub.1-C.sub.20 alkoxy; optionally substituted
C.sub.2-C.sub.20 alkenyl; optionally substituted C.sub.6-C.sub.10
aryl; or NR.sub.2'''' represents a cyclic moiety; Z' represents a
hydrogen atom or a pharmaceutically acceptable counter-ion; A, A'
and A'' each independently represent a hydrogen atom;
C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20 acyloxy;
C.sub.1-C.sub.20 alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl;
C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
C.sub.1-C.sub.20 alkylcarboxylamino; carboxyl; cyano; halo; or
hydroxy; B, B' and B'' each independently represent;
C.sub.2-C.sub.20 alkenoyl; aroyl; aralkanoyl; nitro; optionally
substituted, linear or branched C.sub.1-C.sub.20 alkyl; or
optionally substituted, linear or branched C.sub.2-C.sub.20
alkenyl; or A and B jointly, A' and B' jointly, or A'' and B''
jointly, independently represent a methylenedioxy or ethylenedioxy
group; and X and X' independently represent >NH, >NR''',
--O--, or --S--.
3. A pharmaceutical composition comprising: a therapeutically
effective amount of a compound represented by the following formula
1: ##STR32## wherein Z is ##STR33## n, m, q and r independently
represent integers from zero to 4 provided that n+m.ltoreq.4 and
q+r.ltoreq.4; p and s independently represent integers from zero to
5 provided that p+s.ltoreq.5; a, b and c represent double bonds
which may be present or absent; when present, the double bonds may
be in the E or Z configuration and, when absent, the resulting
stereocenters may have the R- or S-configuration; R and R' each
independently represent a hydrogen atom; linear or branched
C.sub.1-C.sub.20 alkyl; linear or branched C.sub.2-C.sub.20
alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2; --NHR''';
--NR.sub.2'''; --OH; --OR'''; --CONR.sub.2''''; halogen atom;
optionally substituted linear or branched C.sub.1-C.sub.20 alkyl;
optionally substituted linear or branched C.sub.2-C.sub.20 alkenyl;
R'' independently represents a hydrogen atom; linear or branched
C.sub.1-C.sub.20 alkyl; linear or branched C.sub.2-C.sub.20
alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2; --NHR''';
--NR.sub.2'''; --OH; --OR'''; halogen atom; optionally substituted
linear or branched C.sub.1-C.sub.20 alkyl; optionally substituted
linear or branched C.sub.2-C.sub.20 alkenyl; R''' independently
represents a linear or branched C.sub.1-C.sub.20 alkyl; linear or
branched C.sub.2-C.sub.20 alkenyl; or --(CH.sub.2).sub.x--Ar, where
x represents an integer from 1 to 6 and Ar represents aryl; R''''
independently represents a hydrogen atom; optionally substituted
C.sub.1-C.sub.20 alkyl; optionally substituted C.sub.1-C.sub.20
alkoxy; optionally substituted C.sub.2-C.sub.20 alkenyl; optionally
substituted C.sub.6-C.sub.10 aryl; or NR.sub.2'''' represents a
cyclic moiety; Z' represents a hydrogen atom or a pharmaceutically
acceptable counter-ion; A, A' and A'' each independently represent
a hydrogen atom; C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20
acyloxy; C.sub.1-C.sub.20 alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl;
C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
C.sub.1-C.sub.20 alkylcarboxylamino; carboxyl; cyano; halo; or
hydroxy; B, B' and B'''' each independently represent;
C.sub.2-C.sub.20 alkenoyl; aroyl; aralkanoyl; nitro; optionally
substituted, linear or branched C.sub.1-C.sub.20 alkyl; or
optionally substituted, linear or branched C.sub.2-C.sub.20
alkenyl; or A and B jointly, A' and B' jointly, or A'' and B''
jointly, independently represent a methylenedioxy or ethylenedioxy
group; and X and X' independently represent >NH, >NR''',
--O--, or --S--; in a physiologically acceptable carrier.
4. A pharmaceutical composition comprising: a therapeutically
effective amount of a compound represented by the following formula
1: ##STR34## wherein Z is ##STR35## n, m, q and r independently
represent integers from zero to 4 provided that n+m.ltoreq.4 and
q+r.ltoreq.4; p and s independently represent integers from zero to
5 provided that p+s.ltoreq.5; a, b and c represent double bonds
which may be present or absent; when present, the double bonds may
be in the E or Z configuration and, when absent, the resulting
stereocenters may have the R- or S-configuration; R independently
represents a hydrogen atom; linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z';
--CO.sub.2R'''; --NH.sub.2; --NHR'''; --NR.sub.2'''; --OH; --OR''';
--CONR.sub.2''''; halogen atom; optionally substituted linear or
branched C.sub.1-C.sub.20 alkyl; optionally substituted linear or
branched C.sub.2-C.sub.20 alkenyl; R' independently represents a
hydrogen atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or
branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R''';
--NH.sub.2; --NHR'''; --NR.sub.2'''; --OR'''; --CONR.sub.2'''';
halogen atom; optionally substituted linear or branched
C.sub.1-C.sub.20 alkyl; optionally substituted linear or branched
C.sub.2-C.sub.20 alkenyl; R'' independently represents a hydrogen
atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2;
--NHR'''; --NR.sub.2'''; --OH; --OR'''; halogen atom; optionally
substituted linear or branched C.sub.1-C.sub.20 alkyl; optionally
substituted linear or branched C.sub.2-C.sub.20 alkenyl; R'''
independently represents a linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; or
--(CH.sub.2).sub.x--Ar, where x represents an integer from 1 to 6
and Ar represents aryl; R'''' independently represents a hydrogen
atom; optionally substituted C.sub.1-C.sub.20 alkyl; optionally
substituted C.sub.1-C.sub.20 alkoxy; optionally substituted
C.sub.2-C.sub.20 alkenyl; optionally substituted C.sub.6-C.sub.10
aryl; or NR.sub.2''''represents a cyclic moiety; Z' represents a
hydrogen atom or a pharmaceutically acceptable counter-ion; A, A'
and A'' each independently represent a hydrogen atom;
C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20 acyloxy;
C.sub.1-C.sub.20 alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl;
C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
C.sub.1-C.sub.20 alkylcarboxylamino; carboxyl; cyano; halo; or
hydroxy; B, B' and B'' each independently represent;
C.sub.2-C.sub.20 alkenoyl; aroyl; aralkanoyl; nitro; optionally
substituted, linear or branched C.sub.1-C.sub.20 alkyl; or
optionally substituted, linear or branched C.sub.2-C.sub.20
alkenyl; or A and B jointly, A' and B' jointly, or A'' and B''
jointly, independently represent a methylenedioxy or ethylenedioxy
group; and X and X' independently represent >NH, >NR''',
--O--, or --S--; in a physiologically acceptable carrier.
5. A method of treating diabetes comprising: administering to a
subject suffering from a diabetic condition, a therapeutically
effective amount of a compound represented by the following formula
1: ##STR36## wherein Z is ##STR37## n, m, q and r independently
represent integers from zero to 4 provided that n+m.ltoreq.4 and
q+r.ltoreq.4; p and s independently represent integers from zero to
5 provided that p+s.ltoreq.5; a, b and c represent double bonds
which may be present or absent; when present, the double bonds may
be in the E or Z configuration and, when absent, the resulting
stereocenters may have the R- or S-configuration; R and R' each
independently represent a hydrogen atom; linear or branched
C.sub.1-C.sub.20 alkyl; linear or branched C.sub.2-C.sub.20
alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2; --NHR''';
--NR.sub.2'''; --OH; --OR'''; --CONR.sub.2''''; halogen atom;
optionally substituted linear or branched C.sub.1-C.sub.20 alkyl;
optionally substituted linear or branched C.sub.2-C.sub.20 alkenyl;
R'' independently represents a hydrogen atom; linear or branched
C.sub.1-C.sub.20 alkyl; linear or branched C.sub.2-C.sub.20
alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2; --NHR''';
--NR.sub.2'''; --OH; --OR'''; halogen atom; optionally substituted
linear or branched C.sub.1-C.sub.20 alkyl; optionally substituted
linear or branched C.sub.2-C.sub.20 alkenyl; R''' independently
represents a linear or branched C.sub.1-C.sub.20 alkyl; linear or
branched C.sub.2-C.sub.20 alkenyl; or --(CH.sub.2).sub.x--Ar, where
x represents an integer from 1 to 6 and Ar represents aryl; R''''
independently represents a hydrogen atom; optionally substituted
C.sub.1-C.sub.20 alkyl; optionally substituted C.sub.1-C.sub.20
alkoxy; optionally substituted C.sub.2-C.sub.20 alkenyl; optionally
substituted C.sub.6-C.sub.10 aryl; or NR.sub.2'''' represents a
cyclic moiety; Z' represents a hydrogen atom or a pharmaceutically
acceptable counter-ion; A, A' and A'' each independently represent
a hydrogen atom; C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20
acyloxy; C.sub.1-C.sub.20 alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl;
C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
C.sub.1-C.sub.20 alkylcarboxylamino; carboxyl; cyano; halo; or
hydroxy; B, B' and B'' each independently represent;
C.sub.2-C.sub.20 alkenoyl; aroyl; aralkanoyl; nitro; optionally
substituted, linear or branched C.sub.1-C.sub.20 alkyl; or
optionally substituted, linear or branched C.sub.2-C.sub.20
alkenyl; or A and B jointly, A' and B' jointly, or A'' and B''
jointly, independently represent a methylenedioxy or ethylenedioxy
group; and X and X' independently represent >NH, >NR''',
--O--, or --S--; in a physiologically acceptable carrier.
6. A method of treating diabetes comprising: administering to a
subject suffering from a diabetic condition, a therapeutically
effective amount of a compound represented by the following formula
1: ##STR38## wherein Z is ##STR39## n, m, q and r independently
represent integers from zero to 4 provided that n+m.ltoreq.4 and
q+r.ltoreq.4; p and s independently represent integers from zero to
5 provided that p+s.ltoreq.5; a, b and c represent double bonds
which may be present or absent; when present, the double bonds may
be in the E or Z configuration and, when absent, the resulting
stereocenters may have the R- or S-configuration; R independently
represents a hydrogen atom; linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z';
--CO.sub.2R'''; --NH.sub.2; --NHR'''; --NR.sub.2'''; --OH; --OR''';
--CONR.sub.2''''; halogen atom; optionally substituted linear or
branched C.sub.1-C.sub.20 alkyl; optionally substituted linear or
branched C.sub.2-C.sub.20 alkenyl; R' independently represents a
hydrogen atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or
branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R''';
--NH.sub.2; --NHR'''; --NR.sub.2'''; --OR'''; --CONR.sub.2'''';
halogen atom; optionally substituted linear or branched
C.sub.1-C.sub.20 alkyl; optionally substituted linear or branched
C.sub.2-C.sub.20 alkenyl; R'' independently represents a hydrogen
atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2;
--NHR'''; --NR.sub.2'''; --OH; --OR'''; halogen atom; optionally
substituted linear or branched C.sub.1-C.sub.20 alkyl; optionally
substituted linear or branched C.sub.2-C.sub.20 alkenyl; R'''
independently represents a linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; or
--(CH.sub.2).sub.x--Ar, where x represents an integer from 1 to 6
and Ar represents aryl; R'''' independently represents a hydrogen
atom; optionally substituted C.sub.1-C.sub.20 alkyl; optionally
substituted C.sub.1-C.sub.20 alkoxy; optionally substituted
C.sub.2-C.sub.20 alkenyl; optionally substituted C.sub.6-C.sub.10
aryl; or NR.sub.2'''' represents a cyclic moiety; Z' represents a
hydrogen atom or a pharmaceutically acceptable counter-ion; A, A'
and A'' each independently represent a hydrogen atom;
C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20 acyloxy;
C.sub.1-C.sub.20 alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl;
C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
C.sub.1-C.sub.20 alkylcarboxylamino; carboxyl; cyano; halo; or
hydroxy; B, B' and B'' each independently represent;
C.sub.2-C.sub.20alkenoyl; aroyl; aralkanoyl; nitro; optionally
substituted, linear or branched C.sub.1-C.sub.20 alkyl; or
optionally substituted, linear or branched C.sub.2-C.sub.20
alkenyl; or A and B jointly, A' and B' jointly, or A'' and B''
jointly, independently represent a methylenedioxy or ethylenedioxy
group; and X and X' independently represent >NH, >NR''',
--O--, or --S--; in a physiologically acceptable carrier.
7. A method of treating inflammation or inflammatory disease
comprising: administering to a subject suffering from such
condition, a therapeutically effective amount of a compound
represented by the following formula 1: ##STR40## wherein Z is
##STR41## n, m, q and r independently represent integers from zero
to 4 provided that n+m.ltoreq.4 and q+r.ltoreq.4; p and s
independently represent integers from zero to 5 provided that
p+s.ltoreq.5; a, b and c represent double bonds which may be
present or absent; when present, the double bonds may be in the E
or Z configuration and, when absent, the resulting stereocenters
may have the R- or S-configuration; R and R' each independently
represent a hydrogen atom; linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z';
--CO.sub.2R'''; --NH.sub.2; --NHR'''; --NR.sub.2'''; --OH; --OR''';
--CONR.sub.2''''; halogen atom; optionally substituted linear or
branched C.sub.1-C.sub.20 alkyl; optionally substituted linear or
branched C.sub.2-C.sub.20 alkenyl; R'' independently represents a
hydrogen atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or
branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R ''';
--NH.sub.2; --NHR'''; --NR.sub.2'''; --OH; --OR'''; halogen atom;
optionally substituted linear or branched C.sub.1-C.sub.20 alkyl;
optionally substituted linear or branched C.sub.2-C.sub.20 alkenyl;
R''' independently represents a linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; or
--(CH.sub.2).sub.x--Ar, where x represents an integer from 1 to 6
and Ar represents aryl; R'''' independently represents a hydrogen
atom; optionally substituted C.sub.1-C.sub.20 alkyl; optionally
substituted C.sub.1-C.sub.20 alkoxy; optionally substituted
C.sub.2-C.sub.20 alkenyl; optionally substituted C.sub.6-C.sub.10
aryl; or NR.sub.2'''' represents a cyclic moiety; Z' represents a
hydrogen atom or a pharmaceutically acceptable counter-ion; A, A'
and A'' each independently represent a hydrogen atom;
C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20 acyloxy;
C.sub.1-C.sub.20 alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl;
C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
C.sub.1-C.sub.20 alkylcarboxylamino; carboxyl; cyano; halo; or
hydroxy; B, B' and B'' each independently represent;
C.sub.2-C.sub.20 alkenoyl; aroyl; aralkanoyl; nitro; optionally
substituted, linear or branched C.sub.1-C.sub.20 alkyl; or
optionally substituted, linear or branched C.sub.2-C.sub.20
alkenyl; or A and B jointly, A' and B' jointly, or A'' and B''
jointly, independently represent a methylenedioxy or ethylenedioxy
group; and X and X' independently represent >NH, >NR''',
--O--, or --S--; in a physiologically acceptable carrier.
8. A method of treating inflammation or inflammatory disease
comprising: administering to a subject suffering from such
condition, a therapeutically effective amount of a compound
represented by the following formula 1: ##STR42## wherein Z is
##STR43## n, m, q and r independently represent integers from zero
to 4 provided that n+m.ltoreq.4 and q+r.ltoreq.4; p and s
independently represent integers from zero to 5 provided that
p+s.ltoreq.5; a, b and c represent double bonds which may be
present or absent; when present, the double bonds may be in the E
or Z configuration and, when absent, the resulting stereocenters
may have the R- or S-configuration; R independently represents a
hydrogen atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or
branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R''';
--NH.sub.2; --NHR'''; --NR.sub.2'''; --OH; --OR''';
--CONR.sub.2''''; halogen atom; optionally substituted linear or
branched C.sub.1-C.sub.20 alkyl; optionally substituted linear or
branched C.sub.2-C.sub.20 alkenyl; R' independently represents a
hydrogen atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or
branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R''';
--NH.sub.2; --NHR'''; --NR.sub.2'''; --OR'''; --CONR.sub.2'''';
halogen atom; optionally substituted linear or branched
C.sub.1-C.sub.20 alkyl; optionally substituted linear or branched
C.sub.2-C.sub.20 alkenyl; R'' independently represents a hydrogen
atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2;
--NHR'''; --NR.sub.2'''; --OH; --OR'''; halogen atom; optionally
substituted linear or branched C.sub.1-C.sub.20 alkyl; optionally
substituted linear or branched C.sub.2-C.sub.20 alkenyl; R'''
independently represents a linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; or
--(CH.sub.2).sub.x--Ar, where x represents an integer from 1 to 6
and Ar represents aryl; R'''' independently represents a hydrogen
atom; optionally substituted C.sub.1-C.sub.20 alkyl; optionally
substituted C.sub.1-C.sub.20 alkoxy; optionally substituted
C.sub.2-C.sub.20 alkenyl; optionally substituted C.sub.6-C.sub.10
aryl; or NR.sub.2'''' represents a cyclic moiety; Z' represents a
hydrogen atom or a pharmaceutically acceptable counter-ion; A, A'
and A'' each independently represent a hydrogen atom;
C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20 acyloxy;
C.sub.1-C.sub.20 alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl;
C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
C.sub.1-C.sub.20 alkylcarboxylamino; carboxyl; cyano; halo; or
hydroxy; B, B' and B'' each independently represent;
C.sub.2-C.sub.20 alkenoyl; aroyl; aralkanoyl; nitro; optionally
substituted, linear or branched C.sub.1-C.sub.20 alkyl; or
optionally substituted, linear or branched C.sub.2-C.sub.20
alkenyl; or A and B jointly, A' and B' jointly, or A'' and B''
jointly, independently represent a methylenedioxy or ethylenedioxy
group; and X and X' independently represent >NH, >NR''',
--O--, or --S--; in a physiologically acceptable carrier.
9. A method of treating immunological disease comprising:
administering to a subject suffering from an immunological disease
a therapeutically effective amount of a compound represented by the
following formula 1: ##STR44## wherein Z is ##STR45## n, m, q and r
independently represent integers from zero to 4 provided that
n+m.ltoreq.4 and q+r.ltoreq.4; p and s independently represent
integers from zero to 5 provided that p+s.ltoreq.5; a, b and c
represent double bonds which may be present or absent; when
present, the double bonds may be in the E or Z configuration and,
when absent, the resulting stereocenters may have the R- or
S-configuration; R and R' each independently represent a hydrogen
atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2;
--NHR'''; --NR.sub.2'''; --OH; --OR'''; --CONR.sub.2''''; halogen
atom; optionally substituted linear or branched C.sub.1-C.sub.20
alkyl; optionally substituted linear or branched C.sub.2-C.sub.20
alkenyl; R'' independently represents a hydrogen atom; linear or
branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2;
--NHR'''; --NR.sub.2'''; --OH; --OR'''; halogen atom; optionally
substituted linear or branched C.sub.1-C.sub.20 alkyl; optionally
substituted linear or branched C.sub.2-C.sub.20 alkenyl; R'''
independently represents a linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; or
--(CH.sub.2).sub.x--Ar, where x represents an integer from 1 to 6
and Ar represents aryl; R'''' independently represents a hydrogen
atom; optionally substituted C.sub.1-C.sub.20 alkyl; optionally
substituted C.sub.1-C.sub.20 alkoxy; optionally substituted
C.sub.2-C.sub.20 alkenyl; optionally substituted C.sub.6-C.sub.10
aryl; or NR.sub.2'''' represents a cyclic moiety; Z' represents a
hydrogen atom or a pharmaceutically acceptable counter-ion; A, A'
and A'' each independently represent a hydrogen atom;
C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20 acyloxy;
C.sub.1-C.sub.20 alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl;
C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
C.sub.1-C.sub.20 alkylcarboxylamino; carboxyl; cyano; halo; or
hydroxy; B, B' and B'' each independently represent;
C.sub.2-C.sub.20 alkenoyl; aroyl; aralkanoyl; nitro; optionally
substituted, linear or branched C.sub.1-C.sub.20 alkyl; or
optionally substituted, linear or branched C.sub.2-C.sub.20
alkenyl; or A and B jointly, A' and B' jointly, or A'' and B''
jointly, independently represent a methylenedioxy or ethylenedioxy
group; and X and X' independently represent >NH, >NR''',
--O--, or --S--; in a physiologically acceptable carrier.
10. A method of treating immunological disease comprising:
administering to a subject suffering from an immunological disease
a therapeutically effective amount of a compound represented by the
following formula 1: ##STR46## wherein Z is ##STR47## n, m, q and r
independently represent integers from zero to 4 provided that
n+m.ltoreq.4 and q+r.ltoreq.4; p and s independently represent
integers from zero to 5 provided that p+s.ltoreq.5; a, b and c
represent double bonds which may be present or absent; when
present, the double bonds may be in the E or Z configuration and,
when absent, the resulting stereocenters may have the R- or
S-configuration; R independently represents a hydrogen atom; linear
or branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2;
--NHR'''; --NR.sub.2'''; --OH; --OR'''; --CONR.sub.2''''; halogen
atom; optionally substituted linear or branched
C.sub.1-C.sub.20alkyl; optionally substituted linear or branched
C.sub.2-C.sub.20 alkenyl; R' independently represents a hydrogen
atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2;
--NHR'''; --NR.sub.2'''; --OR'''; --CONR.sub.2''''; halogen atom;
optionally substituted linear or branched C.sub.1-C.sub.20 alkyl;
optionally substituted linear or branched C.sub.2-C.sub.20 alkenyl;
R'' independently represents a hydrogen atom; linear or branched
C.sub.1-C.sub.20 alkyl; linear or branched C.sub.2-C.sub.20
alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2; --NHR''';
--NR.sub.2'''; --OH; --OR'''; halogen atom; optionally substituted
linear or branched C.sub.1-C.sub.20 alkyl; optionally substituted
linear or branched C.sub.2-C.sub.20 alkenyl; R''' independently
represents a linear or branched C.sub.1-C.sub.20 alkyl; linear or
branched C.sub.2-C.sub.20 alkenyl; or --(CH.sub.2).sub.x--Ar, where
x represents an integer from 1 to 6 and Ar represents aryl; R''''
independently represents a hydrogen atom; optionally substituted
C.sub.1-C.sub.20 alkyl; optionally substituted C.sub.1-C.sub.20
alkoxy; optionally substituted C.sub.2-C.sub.20 alkenyl; optionally
substituted C.sub.6-C.sub.10 aryl; or NR.sub.2'''' represents a
cyclic moiety; Z' represents a hydrogen atom or a pharmaceutically
acceptable counter-ion; A, A' and A'' each independently represent
a hydrogen atom; C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20
acyloxy; C.sub.1-C.sub.20 alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl;
C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
C.sub.1-C.sub.20 alkylcarboxylamino; carboxyl; cyano; halo; or
hydroxy; B, B' and B'' each independently represent;
C.sub.2-C.sub.20 alkenoyl; aroyl; aralkanoyl; nitro; optionally
substituted, linear or branched C.sub.1-C.sub.20 alkyl; or
optionally substituted, linear or branched C.sub.2-C.sub.20
alkenyl; or A and B jointly, A' and B' jointly, or A'' and B''
jointly, independently represent a methylenedioxy or ethylenedioxy
group; and X and X' independently represent >NH, >NR''',
--O--, or --S--; in a physiologically acceptable carrier.
11. A method of inhibiting the activity of TNF-alpha, IL-1, IL-6 or
COX-2 which comprises administering to a host in need of such
inhibition an effective amount of a compound according to claim
1.
12. The method of inhibiting the undesired action of cytokine or
cyclooxygenase which comprises administering to a host in need of
such inhibition an effective amount of a compound according to
claim 1.
13. The method of treating a disease mediated by cytokines or
cyclooxygenase which comprises administering to a host in need of
such treatment a compound according to claim 1.
14. The method of treating insulin resistance which comprises
administering to a host in need of such treatment an effective
amount of a compound according to claim 1.
15. The method of treating hyperlipidemia which comprises
administering to a host in need of such treatment an effective
amount of a compound according to claim 1.
16. The method of treating coronary heart disease which comprises
administering to a host in need of such treatment an effective
amount of a compound according to claim 1.
17. The method of treating multiple sclerosis which comprises
administering to a host in need of such treatment an effective
amount of a compound according to claim 1.
18. The method of treating cancer which comprises administering to
a host in need of such treatment an effective amount of a compound
according to claim 1.
19. A compound according to claim 1 selected from the group
consisting of:
2-{4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)-phenoxy]-phenyl}-3-p--
tolylacrylic acid,
2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-phenoxy]-phenyl}-3-p-tolylacryl-
ic acid,
2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-phenoxy]-phenyl}-3-p-t-
olylacrylic acid methyl ester,
3-(3,5-dimethylphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)-phe-
noxy]-phenyl}-acrylic acid,
3-(3,5-dimethylphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-phenoxy]-
-phenyl}-acrylic acid,
3-(3,5-dimethylphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-phenoxy]-
-phenyl}-acrylic acid methyl ester,
5-(4-{4-[2-(3,5-dimethylphenyl)-1-(morpholine-4-carbonyl)-vinyl]-phenoxy}-
-benzyl)-thiazolidine-2,4-dione,
5-(4-{4-[2-(4-methoxyphenyl)-vinyl]-phenoxy}-benzyl)-thiazolidine-2,4-dio-
ne,
5-(4-{4-[2-(3,5-dimethoxyphenyl)-vinyl]-phenoxy}-benzyl)-thiazolidine-
-2,4-dione,
5-[4-(4'-methoxybiphenyl-3-yloxy)-benzylidene]-thiazolidine-2,4-dione,
5-[4-(4'-methoxybiphenyl-3-yloxy)-benzyl]-thiazolidine-2,4-dione,
5-[4-(2',4'-dimethoxybiphenyl-3-yloxy)-benzylidene]-thiazolidine-2,4-dion-
e, and
5-[4-(3',5'-dimethoxybiphenyl-3-yloxy)-benzyl]-thiazolidine-2,4-di-
one.
20. A pharmaceutical composition comprising a therapeutically
effective amount of a compound selected from the group consisting
of:
2-{4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)-phenoxy]-phenyl}-3-p-tolyl-
acrylic acid,
2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-phenoxy]-phenyl}-3-p-tolylacryl-
ic acid,
2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-phenoxy]-phenyl}-3-p-t-
olylacrylic acid methyl ester,
3-(3,5-dimethylphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)-phe-
noxy]-phenyl}-acrylic acid,
3-(3,5-dimethylphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-phenoxy]-
-phenyl}-acrylic acid,
3-(3,5-dimethylphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-phenoxy]-
-phenyl}-acrylic acid methyl ester,
5-(4-{4-[2-(3,5-dimethylphenyl)-1-(morpholine-4-carbonyl)-vinyl]-phenoxy}-
-benzyl)-thiazolidine-2,4-dione,
5-(4-{4-[2-(4-methoxyphenyl)-vinyl]-phenoxy}-benzyl)-thiazolidine-2,4-dio-
ne,
5-(4-{4-[2-(3,5-dimethoxyphenyl)-vinyl]-phenoxy}-benzyl)-thiazolidine-
-2,4-dione,
5-[4-(4'-methoxybiphenyl-3-yloxy)-benzylidene]-thiazolidine-2,4-dione,
5-[4-(4'-methoxybiphenyl-3-yloxy)-benzyl]-thiazolidine-2,4-dione,
5-[4-(2',4'-dimethoxybiphenyl-3-yloxy)-benzylidene]-thiazolidine-2,4-dion-
e, and
5-[4-(3',5'-dimethoxybiphenyl-3-yloxy)-benzyl]-thiazolidine-2,4-di-
one, together with a physiologically acceptable carrier
therefor.
21. A method for treating diabetes, comprising: co-administering an
effective amount of a compound of claim 1 and an agent selected
from the group consisting of: insulin or an insulin mimetic, a
sulfonylurea or other insulin secretagogue, a thiazolidinedione, a
fibrate or other PPAR-alpha agonist, a PPAR-delta agonist, a
biguanide, a statin or other hydroxymethylglutaryl (HMG) CoA
reductase inhibitor, an alpha-glucosidase inhibitor, a bile
acid-binding resin, apoA1, niacin, probucol, and nicotinic
acid.
22. A method for treating inflammatory or immunological disease,
comprising: co-administering an effective amount of a compound of
claim 1 and an agent selected from the group consisting of: a
nonsteroidal anti-inflammatory drug (NSAID), a cyclooxygenase-2
inhibitor, a corticosteroid or other immunosuppressive agent, a
disease-modifying antirheumatic drug (DMARD), a TNF-alpha
inhibitor, other cytokine inhibitor, other immune modulating agent,
and a narcotic agent.
23. The compound of claim 1 wherein Z is represented by:
##STR48##
24. The pharmaceutical composition of claim 3 wherein Z is
represented by: ##STR49##
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This is a continuation-in-part of U.S. patent application
Ser. No. 10/265,902, filed Oct. 8, 2002, which is a
continuation-in-part of U.S. patent application Ser. No.
09/843,167, filed Apr. 27, 2001, which is a continuation-in-part of
U.S. patent application Ser. No. 09/785,554, filed Feb. 20, 2001,
which is a continuation-in-part of U.S. patent application Ser. No.
09/591,105, filed on Jun. 9, 2000, and which further is a
continuation-in-part of U.S. patent application Ser. No.
09/287,237, filed on Apr. 6, 1999.
BACKGROUND OF THE INVENTION
[0002] The present application is directed to novel compounds
formed by chemically coupling diphenylethylene compounds and
derivatives thereof with thiazolidine or oxazolidine intermediates.
These compounds are effective for providing a variety of useful
pharmacological effects. For example, the compounds are useful in
lowering blood glucose, serum insulin and triglyceride levels in
animal models of type II diabetes.
[0003] Furthermore, these compounds are useful for treatment of
disorders associated with insulin resistance, such as polycystic
ovary syndrome, as well as hyperlipidemia, coronary artery disease
and peripheral vascular disease, and for the treatment of
inflammation and immunological diseases, particularly those
mediated by cytokines and cyclooxygenase such as TNF-alpha, IL-1,
IL-6 and/or COX-2.
[0004] The causes of type I and type II diabetes are yet unknown,
although both genetics and environment seem to be major factors.
Insulin dependent type I and non-insulin dependent type II are the
types which are known. Type I is an autoimmune disease in which the
responsible autoantigen is still unknown. Patients of type I need
to take insulin parenterally or subcutaneously to survive. However,
type II diabetes, the more common form, is a metabolic disorder
resulting from the body's inability to make a sufficient amount of
insulin or to properly use the insulin that is produced. Insulin
secretion and insulin resistance are considered the major defects,
however, the precise genetic factors involved in the mechanism
remain unknown.
[0005] Patients with diabetes usually have one or more of the
following defects: [0006] Less production of insulin by the
pancreas; [0007] Over secretion of glucose by the liver; [0008]
Decreased glucose uptake by the skeletal muscles; [0009] Defects in
glucose transporters; and [0010] Desensitization of insulin
receptors.
[0011] Other than the parenteral or subcutaneous application of
insulin, there are about 4 classes of oral hypoglycemic agents
used. TABLE-US-00001 TABLE 1 Class Approved Drugs Mode of Action
Limitations Sulfonylurea 4 (1.sup.st generation) Acts on pancreas
development of and to release more resistance 2 (2.sup.nd
generation) insulin Biguanides Metformin Reduces glucose liver
problems, production by lactic acidosis liver; improves insulin
sensitivity Alpha'- Acarbose Interferes with only useful at
glucosidase digestive process; postprandial inhibitor reduces
glucose level absorption Thiazolidinedione Troglitazone Reduce
insulin ''add-on'' with (withdrawn) resistancy insulin; not
Rosiglitazone useful for Pioglitazone people with heart and liver
disease ##STR1## ##STR2## ##STR3## ##STR4##
[0012] As is apparent from the above table, each of the current
agents available for use in treatment of diabetes has certain
disadvantages. Accordingly, there is a continuing interest in the
identification and development of new agents, particularly, water
soluble agents which can be orally administered, for use in the
treatment of diabetes.
[0013] The thiazolidinedione class listed in the above table has
gained more widespread use in recent years for treatment of type II
diabetes, exhibiting particular usefulness as insulin sensitizers
to combat "insulin resistance", a condition in which the patient
becomes less responsive to the effects of insulin. However, the
known thiazolidinediones are not effective for a significant
portion of the patient population. In addition, the first drug in
this class to be approved by the FDA, troglitazone, was withdrawn
from the market due to problems of liver toxicity. Thus, there is a
continuing need for nontoxic, more widely effective insulin
sensitizers.
[0014] Pharmaceutical compositions and methods utilizing
thiazolidinediones are described in U.S. Pat. Nos. 6,133,295;
6,133,293; 6,130,216; 6,121,295; 6,121,294; 6,117,893; 6,114,526;
6,110,951; 6,110,948; 6,107,323; 6,103,742; 6,080,765; 6,046,222;
6,046;202; 6,034,110; 6,030,973; RE36,575; 6,011,036; 6,011,031;
6,008,237; 5,990,139; 5,985,884; 5,972,973 and others.
[0015] As indicated above, the present invention is also concerned
with treatment of immunological diseases or inflammation, notably
such diseases as are mediated by cytokines or cyclooxygenase. The
principal elements of the immune system are macrophages or
antigen-presenting cells, T cells and B cells. The role of other
immune cells such as NK cells, basophils, mast cells and dendritic
cells are known, but their role in primary immunologic disorders is
uncertain. Macrophages are important mediators of both inflammation
and providing the necessary "help" for T cell stimulation and
proliferation. Most importantly macrophages make IL 1, IL 12 and
TNF-alpha, all of which are potent pro-inflammatory molecules and
also provide help for T cells. In addition, activation of
macrophages results in the induction of enzymes, such as
cyclooxygenase II (COX-2), inducible nitric oxide synthase (NOS)
and production of free radicals capable of damaging normal cells.
Many factors activate macrophages, including bacterial products,
superantigens and interferon gamma (IFN.gamma.). It is believed
that phosphotyrosine kinases (PTKs) and other undefined cellular
kinases are involved in the activation process.
[0016] Macrophages take up and break down antigens into small
fragments. These fragments then associate with the major
histocompatibility complex II (MHC II). This complex of antigen
fragments and MHC II is recognized by the T cell receptor. In
association with appropriate co-stimulatory signals this
receptor-ligand interaction leads to the activation and
proliferation of T cells. Depending on the route of administration
of antigen, their dose and the conditions under which macrophages
are activated, the immune response can result in either B cell help
and antibody production or on the development of cell mediated
response. Since macrophages are sentinel to the development of an
immune response, agents that modify their function, specifically
their cytokine secretion profile, are likely to determine the
direction and potency of the immune response.
[0017] Cytokines are molecules secreted by immune cells that are
important in mediating immune responses. Cytokine production may
lead to the secretion of other cytokines, altered cellular
function, cell division or differentiation. Inflammation is the
body's normal response to injury or infection. However, in
inflammatory diseases such as rheumatoid arthritis, pathologic
inflammatory processes can lead to morbidity and mortality. The
cytokine tumor necrosis factor-alpha (TNF-alpha) plays a central
role in the inflammatory response and has been targeted as a point
of intervention in inflammatory disease. TNF-alpha is a polypeptide
hormone released by activated macrophages and other cells. At low
concentrations, TNF-alpha participates in the protective
inflammatory response by activating leukocytes and promoting their
migration to extravascular sites of inflammation (Moser et al., J
Clin Invest, 83:444-55, 1989). At higher concentrations, TNF-alpha
can act as a potent pyrogen and induce the production of other
pro-inflammatory cytokines (Haworth et al., Eur J Immunol,
21:2575-79, 1991; Brennan et al., Lancet, 2:244-7, 1989). TNF-alpha
also stimulates the synthesis of acute-phase proteins. In
rheumatoid arthritis, a chronic and progressive inflammatory
disease affecting about 1% of the adult U.S. population, TNF-alpha
mediates the cytokine cascade that leads to joint damage and
destruction (Arend et al., Arthritis Rheum, 38:151-60, 1995).
Inhibitors of TNF-alpha, including soluble TNF receptors
(etanercept) (Goldenberg, Clin Ther, 21:75-87, 1999) and
anti-TNF-alpha antibody (infliximab) (Luong et al., Ann
Pharmacotherapy, 34:743-60, 2000), have recently been approved by
the U.S. Food and Drug Administration (FDA) as agents for the
treatment of rheumatoid arthritis.
[0018] Elevated levels of TNF-alpha have also been implicated in
many other disorders and disease conditions, including cachexia
(Fong et al., Am J Physiol, 256:8659-65, 1989), septic shock
syndrome (Tracey et al., Proc Soc Exp Biol Med, 200:233-9, 1992),
osteoarthritis (Venn et al., Arthritis Rheum, 36:819-26, 1993),
inflammatory bowel disease such as Crohn's disease and ulcerative
colitis (Murch et al., Gut, 32:913-7, 1991), Behoet's disease
(Akoglu et al., J Rheumatol, 17:1107-8, 1990), Kawasaki disease
(Matsubara et al., Clin Immunol Immunopathol, 56:29-36, 1990),
cerebral malaria (Grau et al., N Engl J Med, 320:1586-91, 1989),
adult respiratory distress syndrome (Millar et al., Lancet 2:712-4,
1989), asbestosis and silicosis (Bissonnette et al., Inflammation,
13:329-39, 1989), pulmonary sarcoidosis (Baughman et al., J Lab
Clin Med, 115:36-42, 1990), asthma (Shah et al., Clin Exp Allergy,
25:1038-44, 1995), AIDS (Dezube et al., J Acquir Immune Defic
Syndr, 5:1099-104, 1992), meningitis (Waage et al., Lancet,
1:355-7, 1987), psoriasis (Oh et al., J Am Acad Dermatol,
42:829-30, 2000), graft versus host reaction (Nestel et al., J Exp
Med, 175:405-13, 1992), multiple sclerosis (Sharief et al., N Engl
J Med, 325:467-72, 1991), systemic lupus erythematosus (Maury et
al., Int J Tissue React, 11:189-93, 1989), diabetes (Hotamisligil
et al., Science, 259:87-91, 1993) and atherosclerosis (Bruunsgaard
et al., Clin Exp Immunol, 121:255-60, 2000).
[0019] It can be seen from the references cited above that
inhibitors of TNF-alpha are potentially useful in the treatment of
a wide variety of diseases. Compounds that inhibit TNF-alpha have
been described in U.S. Pat. Nos. 6,090,817; 6,080,763; 6,080,580;
6,075,041; 6,057,369; 6,048,841; 6,046,319; 6,046,221; 6,040,329;
6,034,100; 6,028,086; 6,022,884; 6,015,558; 6,004,974; 5,990,119;
5,981,701; 5,977,122; 5,972,936; 5,968,945; 5,962,478; 5,958,953;
5,958,409; 5,955,480; 5,948,786; 5,935,978; 5,935,977; 5,929,117;
5,925,636; 5,900,430; 5,900,417; 5,891,883; 5,869,677 and
others.
[0020] Interleukin-6 (IL-6) is another pro-inflammatory cytokine
that exhibits pleiotropy and redundancy of action. IL-6
participates in the immune response, inflammation and
hematopoiesis. It is a potent inducer of the hepatic acute phase
response and is a powerful stimulator of the
hypothalamic-pituitary-adrenal axis that is under negative control
by glucocorticoids. IL-6 promotes the secretion of growth hormone
but inhibits release of thyroid stimulating hormone. Elevated
levels of IL-6 are seen in several inflammatory diseases, and
inhibition of the IL-6 cytokine subfamily has been suggested as a
strategy to improve therapy for rheumatoid arthritis (Carroll et
al., Inflamm Res, 47:1-7, 1998). In addition, IL-6 has been
implicated in the progression of atherosclerosis and the
pathogenesis of coronary heart disease (Yudkin et al.,
Atherosclerosis, 148:209-14, 1999). Overproduction of IL-6 is also
seen in steroid withdrawal syndrome, conditions related to
deregulated vasopressin secretion, and osteoporosis associated with
increased bone resorption, such as in cases of hyperparathyroidism
and sex-steroid deficiency (Papanicolaou et al., Ann Intern Med,
128:127-37, 1998).
[0021] Since excessive production of IL-6 is implicated in several
disease states, it is highly desirable to develop compounds that
inhibit IL-6 secretion. Compounds that inhibit IL-6 have been
described in U.S. Pat. Nos. 6,004,813; 5,527,546 and 5,166,137.
[0022] Cyclooxygenase is an enzyme that catalyzes a
rate-determining step in the biosynthesis of prostaglandins, which
are important mediators of inflammation- and pain. The enzyme
occurs as at least two distinct isomers, cyclooxygenase-1 (COX-1)
and cyclooxygenase-2 (COX-2). The COX-1 isomer is constitutively
expressed in the gastric mucosa, platelets and other cells and is
involved in the maintenance of homeostasis in mammals, including
protecting the integrity of the digestive tract. The COX-2 isomer,
on the other hand, is not constitutively expressed but rather is
induced by various agents, such as cytokines, mitogens, hormones
and growth factors. In particular, COX-2 is induced during the
inflammatory response (DeWitt D L, Biochim Biophys Acta,
1083:121-34, 1991; Seibert et al., Receptor, 4:17-23, 1994.).
Aspirin and other conventional non-steroid anti-inflammatory drugs
(NSAIDs) are non-selective inhibitors of both COX-1 and COX-2. They
can be effective in reducing inflammatory pain and swelling, but
since they hamper the protective action of COX-1, they produce
undesirable side effects of gastrointestinal pathology. Therefore,
agents that selectively inhibit COX-2 but not COX-1 are preferable
for treatment of inflammatory diseases. Recently, a diarylpyrazole
sulfonamide (celecoxib) that selectively inhibits COX-2 has been
approved by the FDA for use in the treatment of rheumatoid
arthritis (Luong et al., Ann Pharmacother, 34:743-60, 2000; Penning
et al., J Med Chem, 40:1347-65, 1997). COX-2 is also expressed in
many cancers and precancerous lesions, and there is mounting
evidence that selective COX-2 inhibitors may be useful for treating
and preventing colorectal, breast and other cancers (Taketo M M, J
Natl Cancer Inst, 90:1609-20, 1998; Fournier et al., J Cell Biochem
Suppl, 34:97-102, 2000; Masferrer et al., Cancer Res, 60:1306-11,
2000). In 1999 celecoxib was approved by the FDA as an adjunct to
usual care for patients with familial adenomatous polyposis, a
condition which, left untreated, generally leads to colorectal
cancer.
[0023] Compounds that selectively inhibit COX-2 have been described
in U.S. Pat. Nos. 5,344,991; 5,380,738; 5,434,178; 5,466,823;
5,474,995; 5,510,368; 5,521,207; 5,521,213; 5,536,752; 5,550,142;
5,552,422; 5,604,260; 5,639,780; 5,643,933; 5,677,318; 5,691,374;
5,698,584; 5,710,140; 5,733,909; 5,789,413; 5,811,425; 5,817,700;
5,849,943; 5,859,257; 5,861,419; 5,905,089; 5,922,742; 5,925,631;
5,932,598; 5,945,539; 5,968,958; 5,981,576; 5,994,379; 5,994,381;
6,001,843; 6,002,014; 6,004,950; 6,004,960; 6,005,000; 6,020,343;
6,034,256; 6,046,191; 6,046,217; 6,057,319; 6,071,936; 6,071,954;
6,077,850; 6,077,868; 6,077,869 and 6,083,969.
[0024] The cytokine IL-1 beta also participates in the inflammatory
response. It stimulates thymocyte proliferation, fibroblast growth
factor activity, and the release of prostaglandin from synovial
cells.
[0025] Elevated or unregulated levels of the cytokine IL-1 beta
have been associated with a number of inflammatory diseases and
other disease states, including but not limited to adult
respiratory distress syndrome (Meduri et al, Chest 107:1062-73,
1995), allergy (Hastie et al, Cytokine 8:730-8, 1996), Alzheimer's
disease (O'Barr et al, J Neuroimmunol 109:87-94, 2000), anorexia
(Laye et al, Am J Physiol Regul Integr Comp Physiol 279:893-8,
2000), asthma (Sousa et al, Thorax 52:407-10, 1997),
atherosclerosis (Dewberry et al, Arterioscler Thromb Vasc Biol
20:2394-400, 2000), brain tumors (Ilyin et al, Mol Chem Neuropathol
33:125-37, 1998), cachexia (Nakatani et al, Res Commun Mol Pathol
Pharmacol 102:241-9, 1998), carcinoma (Ikemoto et al, Anticancer
Res 20:317-21, 2000), chronic arthritis (van den Berg et al, Clin
Exp Rheumatol 17:S105-14, 1999), chronic fatigue syndrome (Cannon
et al, J Clin Immunol 17:253-61, 1997), CNS trauma (Herx et al, J
Immunol 165:2232-9, 2000), epilepsy (De Simoni et al, Eur J
Neurosci 12:2623-33, 2000), fibrotic lung diseases (Pan et al,
Pathol Int 46:91-9, 1996), fulminant hepatic failure (Sekiyama et
al, Clin Exp Immunol 98:71-7, 1994), gingivitis (Biesbrock et al,
Monogr Oral Sci 17:20-31, 2000), glomerulonephritis (Kluth et al, J
Nephrol 12:66-75, 1999), Guillain-Barre syndrome (Zhu et al, Clin
Immunol Immunopathol 84:85-94, 1997), heat hyperalgesia (Opree et
al, J Neurosci 20:6289-93, 2000), hemorrhage and endotoxemia
(Parsey et al, J Immunol 160:1007-13, 1998), inflammatory bowel
disease (Olson et al, J Pediatr Gastroenterol Nutr 16:241-6, 1993),
leukemia (Estrov et al, Leuk Lymphoma 24:379-91, 1997), leukemic
arthritis (Rudwaleit et al, Arthritis Rheum 41:1695-700, 1998),
systemic lupus erythematosus (Mao et al, Autoimmunity 24:71-9,
1996), multiple sclerosis (Martin et al, J Neuroimmunol 61:241-5,
1995), osteoarthritis (Hernvann et al, Osteoarthritis Cartilage
4:13942, 1996), osteoporosis (Zheng et al, Maturitas 26:63-71,
1997), Parkinson's disease (Bessler et al, Biomed Pharmacother
53:141-5, 1999), POEMS syndrome (Gherardi et al, Blood 83:2587-93,
1994), pre-term labor (Dudley, J Reprod Immunol 36:93-109, 1997),
psoriasis (Bonifati et al, J Biol Regul Homeost Agents 11:133-6,
1997), reperfusion injury (Clark et al, J Surg Res 58:675-81,
1995), rheumatoid arthritis (Seitz et al, J Rheumatol 23:1512-6,
1996), septic shock (van Deuren et al, Blood 90:1101-8, 1997),
systemic vasculitis (Brooks et al, Clin Exp Immunol 106:273-9,
1996), temporal mandibular joint disease (Nordahl et al, Eur J Oral
Sci 106:559-63, 1998), tuberculosis (Tsao et al, Tuber Lung Dis
79:279-85, 1999), viral rhinitis (Roseler et al, Eur Arch
Otorhinolaryngol Suppl 1:S61-3, 1995), and pain and/or inflammation
resulting from strain, sprain, trauma, surgery, infection or other
disease processes.
[0026] Since overproduction of IL-1 beta is associated with
numerous disease conditions, it is desirable to develop compounds
that inhibit the production or activity of IL-1 beta. Methods and
compositions for inhibiting IL-1 beta are described in U.S. Pat.
Nos. 6,096,728; 6,090,775; 6,083,521; 6,036,978; 6,034,107;
6,034,100; 6,027,712; 6,024,940; 5,955,480; 5,922,573; 5,919,444;
5,905,089; 5,874,592; 5,874,561; 5,874,424; 5,840,277; 5,837,719;
5,817,670; 5,817,306; 5,792,778; 5,780,513; 5,776,979; 5,776,954;
5,767,064; 5,747,444; 5,739,282; 5,731,343; 5,726,148; 5,684,017;
5,683,992; 5,668,143; 5,624,931; 5,618,804; 5,527,940; 5,521,185;
5,492,888; 5,488,032 and others.
[0027] It will be appreciated from the foregoing that, while there
have been extensive prior efforts to provide compounds for
inhibiting, for example, TNF-alpha, IL-1, IL-6, COX-2 or other
agents considered responsible for immune response, inflammation or
inflammatory diseases, e.g. arthritis, there still remains a need
for new and improved compounds for effectively treating or
inhibiting such diseases. A principal object of the invention is to
provide compounds which are effective for such treatments as well
as for the treatment of, for example, insulin resistance,
hyperlipidemia, coronary heart disease, multiple sclerosis and
cancer.
SUMMARY OF THE INVENTION
[0028] In one aspect of the invention, compounds of the following
formula 1 are provided: ##STR5## wherein Z is ##STR6## wherein n,
m, q and r are independently integers from zero to 4 provided that
n+m.ltoreq.4, and q+r.ltoreq.4; p and s are independently integers
from zero to 5 provided that p+s.ltoreq.5; a, b and c are double
bonds which may be present or absent; when present, the double
bonds may be in the E or Z configuration and, when absent, the
resulting stereocenters can have the R-- or S-configuration;
[0029] R and R' are independently H, C.sub.1-C.sub.20 linear or
branched alkyl, C.sub.2-C.sub.20 linear or branched alkenyl,
--CO.sub.2Z', wherein Z' is H, sodium, potassium, or other
pharmaceutically acceptable counter-ion such as calcium, magnesium,
ammonium, tromethamine, tetramethylammonium, and the like;
--CO.sub.2R''', --NH.sub.2, --NHR''', --NR.sub.2''', --OH, --OR''',
halo, substituted C.sub.1-C.sub.20 linear or branched alkyl or
substituted C.sub.2-C.sub.20 linear or branched alkenyl, wherein
R''' is independently C.sub.1-C.sub.20 linear or branched alkyl,
linear or branched alkenyl or aralkyl --(CH.sub.2).sub.x--Ar, where
x is 1-6; CONR.sub.2'''', where R'''' is independently H,
optionally substituted C.sub.1-C.sub.20 alkyl, optionally
substituted C.sub.1-C.sub.20 alkoxy, preferably an optionally
substituted C.sub.1-C.sub.6 alkoxy (for example methoxy, ethoxy or
propoxy), optionally substituted C.sub.2-C.sub.20 alkenyl or
optionally substituted C.sub.6-C.sub.10 aryl or where NR.sub.2''''
represents a cyclic moiety such as morpholine, piperidine,
piperazine and the like;
[0030] R'' is independently H, C.sub.1-C.sub.20 linear or branched
alkyl, C.sub.2-C.sub.20 linear or branched alkenyl, --CO.sub.2Z',
wherein Z' is H, sodium, potassium, or other pharmaceutically
acceptable counter-ion such as calcium, magnesium, ammonium,
tromethamine, tetramethylammonium, and the like; --CO.sub.2R''',
--NH.sub.2, --NHR''', --NR.sub.2''', --OH, --OR''', halo,
substituted C.sub.1-C.sub.20 linear or branched alkyl or
substituted C.sub.2-C.sub.20 linear or branched alkenyl wherein
R''' is independently C.sub.1-C.sub.20 linear or branched alkyl,
linear or branched alkenyl or aralkyl --(CH.sub.2).sub.x--Ar, where
x is 1-6;
[0031] A, A' and A'' are independently H, C.sub.1-C.sub.20
acylamino;
[0032] C.sub.1-C.sub.20 acyloxy; C.sub.1-C.sub.20 alkanoyl;
[0033] C.sub.1-C.sub.20 alkoxycarbonyl; C.sub.1-C.sub.20
alkoxy;
[0034] C.sub.1-C.sub.20 alkylamino; C.sub.1-C.sub.20
alkylcarboxylamino; carboxyl; cyano; halo; hydroxy;
[0035] B, B' and B'' are independently H;
[0036] C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20 acyloxy;
C.sub.1-C.sub.20 alkanoyl;
[0037] C.sub.1-C.sub.20 alkenoyl; C.sub.1-C.sub.20
alkoxycarbonyl;
[0038] C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
[0039] C.sub.1-C.sub.20 alkylcarboxylamino; aroyl, aralkanoyl;
carboxyl; cyano; halo; hydroxy; nitro;
[0040] optionally substituted, linear or branched C.sub.1-C.sub.20
alkyl or C.sub.2-C.sub.20 alkenyl; or A and B together, or A' and
B' together, or A'' and B'' together, may be joined to form a
methylenedioxy or ethylenedioxy group;
[0041] X, X' are independently --NH, --NR''', O or S.
[0042] In a further aspect of the invention, the following
preferred compounds of formula 1 are provided:
[0043] A) ##STR7## [0044] wherein Z is ##STR8## [0045] n, m, q and
r independently represent integers from zero to 4 provided that
n+m.ltoreq.4 and q+r.ltoreq.4; p and s independently represent
integers from zero to 5 provided that p+s.ltoreq.5; a, b and c
represent double bonds which may be present or absent; when
present, the double bonds may be in the E or Z configuration and,
when absent, the resulting stereocenters may have the R-- or
S-configuration; [0046] R and R' each independently represent a
hydrogen atom; linear or branched C.sub.1-C.sub.20 alkyl; linear or
branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R''';
--NH.sub.2; --NHR'''; --NR.sub.2'''; --OH; --OR''';
--CONR.sub.2''''; halogen atom; optionally substituted linear or
branched C.sub.1-C.sub.20 alkyl; optionally substituted linear or
branched C.sub.2-C.sub.20 alkenyl; [0047] R'' independently
represents a hydrogen atom; linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z';
--CO.sub.2R'''; --NH.sub.2; --NHR'''; --NR.sub.2'''; --OH; --OR''';
halogen atom; optionally substituted linear or branched
C.sub.1-C.sub.20 alkyl; optionally substituted linear or branched
C.sub.2-C.sub.20 alkenyl; [0048] R''' independently represents a
linear or branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; or --(CH.sub.2).sub.x--Ar, where x
represents an integer from 1 to 6 and Ar represents aryl; [0049]
R'''' independently represents a hydrogen atom; optionally
substituted C.sub.1-C.sub.20 alkyl; optionally substituted
C.sub.1-C.sub.20 alkoxy; optionally substituted C.sub.2-C.sub.20
alkenyl; optionally substituted C.sub.6-C.sub.10 aryl; or
NR.sub.2'''' represents a cyclic moiety; [0050] Z' represents a
hydrogen atom or a pharmaceutically acceptable counter-ion; [0051]
A, A' and A'' each independently represent a hydrogen atom;
C.sub.1-C.sub.20 acylamino; C.sub.1-C.sub.20 acyloxy;
C.sub.1-C.sub.20 alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl;
C.sub.1-C.sub.20 alkoxy; C.sub.1-C.sub.20 alkylamino;
C.sub.1-C.sub.20 alkylcarboxylamino; carboxyl; cyano; halo; or
hydroxy; [0052] B, B' and B'' each independently represent;
C.sub.2-C.sub.20 alkenoyl; aroyl; aralkanoyl; nitro; optionally
substituted, linear or branched C.sub.1-C.sub.20 alkyl; or
optionally substituted, linear or branched C.sub.2-C.sub.20
alkenyl; [0053] or A and B jointly, A' and B' jointly, or A'' and
B'' jointly, independently represent a methylenedioxy or
ethylenedioxy group; and [0054] X and X' independently represent
>NH, >NR''', --O--, or --S--.
[0055] B) ##STR9## [0056] wherein Z is ##STR10## [0057] n, m, q and
r independently represent integers from zero to 4 provided that
n+m.ltoreq.4 and q+r.ltoreq.4; p and s independently represent
integers from zero to 5 provided that p+s.ltoreq.5; a, b and c
represent double bonds which may be present or absent; when
present, the double bonds may be in the E or Z configuration and,
when absent, the resulting stereocenters may have the R-- or
S-configuration; [0058] R independently represents a hydrogen atom;
linear or branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2;
--NHR'''; --NR.sub.2'''; --OH; --OR'''; --CONR.sub.2''''; halogen
atom; optionally substituted linear or branched C.sub.1-C.sub.20
alkyl; optionally substituted linear or branched C.sub.2-C.sub.20
alkenyl; [0059] R' independently represents a hydrogen atom; linear
or branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2;
--NHR'''; --NR.sub.2'''; --OR'''; --CONR.sub.2''''; halogen atom;
optionally substituted linear or branched C.sub.1-C.sub.20 alkyl;
optionally substituted linear or branched C.sub.2-C.sub.20 alkenyl;
[0060] R'' independently represents a hydrogen atom; linear or
branched C.sub.1-C.sub.20 alkyl; linear or branched
C.sub.2-C.sub.20 alkenyl; --CO.sub.2Z'; --CO.sub.2R'''; --NH.sub.2;
--NHR'''; --NR.sub.2'''; --OH; --OR'''; halogen atom; optionally
substituted linear or branched C.sub.1-C.sub.20 alkyl; optionally
substituted linear or branched C.sub.2-C.sub.20 alkenyl; [0061]
R''' independently represents a linear or branched C.sub.1-C.sub.20
alkyl; linear or branched C.sub.2-C.sub.20 alkenyl; or
--(CH.sub.2).sub.x--Ar, where x represents an integer from 1 to 6
and Ar represents aryl; [0062] R'''' independently represents a
hydrogen atom; optionally substituted C.sub.1-C.sub.20 alkyl;
optionally substituted C.sub.1-C.sub.20 alkoxy; optionally
substituted C.sub.2-C.sub.20 alkenyl; optionally substituted
C.sub.6-C.sub.10 aryl; or NR.sub.2'''' represents a cyclic moiety;
[0063] Z' represents a hydrogen atom or a pharmaceutically
acceptable counter-ion; [0064] A, A' and A'' each independently
represent a hydrogen atom; C.sub.1-C.sub.20 acylamino;
C.sub.1-C.sub.20 acyloxy; C.sub.1-C.sub.20
alkanoyl;C.sub.1-C.sub.20 alkoxycarbonyl; C.sub.1-C.sub.20 alkoxy;
C.sub.1-C.sub.20 alkylamino; C.sub.1-C.sub.20 alkylcarboxylamino;
carboxyl; cyano; halo; or hydroxy; [0065] B, B' and B'' each
independently represent; C.sub.2-C.sub.20 alkenoyl; aroyl;
aralkanoyl; nitro; optionally substituted, linear or branched
C.sub.1-C.sub.20 alkyl; or optionally substituted, linear or
branched C.sub.2-C.sub.20 alkenyl; [0066] or A and B jointly, A'
and B' jointly, or A'' and B'' jointly, independently represent a
methylenedioxy or ethylenedioxy group; and [0067] X and X'
independently represent >NH, >NR''', --O--, or --S--.
[0068] These compounds are useful for treating diabetes,
hyperlipidemia and other diseases linked to insulin resistance,
such as coronary artery disease and peripheral vascular disease,
and also for treating or inhibiting inflammation or inflammatory
diseases such as inflammatory arthritides and collagen vascular
diseases, which are caused by, for example, cytokines or
cyclooxygenase such as TNF-alpha, IL-1, IL-6 and/or COX-2. The
compounds are also useful for treating or preventing other diseases
mediated by cytokines and/or cyclooxygenase, such as cancer.
[0069] Accordingly, the invention also provides a method of
treating diabetes and related diseases comprising the step of
administering to a subject suffering from a diabetic or related
condition a therapeutically effective amount of a compound of
formula 1. Additionally, the invention provides a method of
treating inflammation or inflammatory diseases or diseases mediated
by cytokines and/or cyclooxygenase by administering to a subject in
need of such treatment an effective amount of a compound according
to Formula 1. Other uses will also be evident from this
specification.
[0070] Pharmaceutical compositions containing a therapeutically
effective amount of one or more compounds according to formula 1
together with a pharmaceutically or physiologically acceptable
carrier, for use in the treatments contemplated herein, are also
provided.
BRIEF DESCRIPTION OF THE DRAWINGS
[0071] FIGS. 1A and 1B show graphs of the blood glucose levels and
body weights, respectively, of db/db (spontaneous diabetic) male
mice given a compound according to the invention of a period of 15
days.
[0072] FIGS. 2A and 2B show graphs of the blood glucose levels and
body weights of ob/ob (genetically obese and spontaneously
diabetic) male mice given a compound according to the invention
over a period of 15 days.
[0073] FIGS. 3A and 3B with graphs of blood glucose levels of db/db
mice and ob/ob mice, respectively, given a compound according to
the invention over a period of 20-25 days.
[0074] FIG. 4 shows a graph of blood glucose level in db/db male
mice over 72 hours following a dosage of the compound.
[0075] FIGS. 5A, 5B, 5C and 5D show graphs of the triglyceride
levels, free fatty acid levels, glyc-Hb levels and leptin levels in
serum of the db/db mice treated with a compound according to the
present invention.
[0076] FIGS. 6A, 6B, 6C and 6D are graphs showing the serum insulin
levels, triglyceride levels, free fatty acid levels and Glyc-Hb
levels of serum of ob/ob mice treated with a compound according to
the present invention.
[0077] FIGS. 7A and 7B show the assays of liver enzymes in mice 21
days after treatment with a compound according to the
invention.
[0078] FIG. 8 shows glucose uptake in 3T3-L1 cells for a compound
of the invention.
[0079] FIG. 9 shows a graph of the comparison of a compound of the
invention with rosiglitazone for inhibition of LPS-induced TNF
production.
[0080] FIG. 10 is a graph of the comparison of a compound of the
invention with rosiglitazone for inhibition of LPS-induced IL-6
production.
[0081] FIG. 11 is a graph of the comparison of a compound of the
invention with rosiglitazone for inhibition of LPS-induced
IL-1-beta production.
[0082] FIGS. 12A and 12B show a comparison of a compound of the
invention (FIG. 12A) with rosiglitazone (FIG. 12B) for inhibition
of LPS-induced COX-2 activity.
[0083] FIGS. 13A, 13B, 13C and 13D illustrate the suppression of
collagen-induced arthritis by using a compound according to the
invention.
[0084] FIG. 14 illustrates the suppression of experimental allergic
encephalomyelitis (EAE) by using a compound according to the
invention.
[0085] FIG. 15. NF-kB activation in stimulated RAW 264.7 cells.
[0086] FIG. 16A. Comparison of percent reduction in blood glucose
in db/db mice using compound 11 and rosiglitazone. 16B. Body
weights of the animals.
[0087] FIG. 17A. Percent reduction in blood glucose in ob/ob mice
using compound 11. 17B. Body weights of the animals.
[0088] FIG. 18. Serum profile in db/db mice after treatment of 14
days of treatment.
[0089] FIG. 19. Dose response of compound 11 on glucose levels in
ob/ob mice.
[0090] FIG. 20. A. In vitro glucose uptake measured in
differentiated 3T3-L1 adipocytes after treatment with increasing
concentrations of Compound 11 or vehicle. B. Glucose uptake in
differentiated 3T3-L1 adipocytes measured in the presence of
increasing concentrations of insulin in the presence of vehicle,
rosiglitazone or Compound 11.
[0091] FIG. 21. In Vivo Antihyperglycemic Activity of Compound 11
in Diabetic Animals.
[0092] FIG. 22. In Vivo Antihyperglycemic Activity of Compound 11
vs. Rosiglitazone in ob/ob mice.
[0093] FIG. 23. In Vitro Adipogenic Activity of Compound 11 vs.
Rosiglitazone in 3T3-L1 cells. (A) Quantitative measurement of
accumulated triglyceride. (B) Qualitative assessment of
triglyceride accumulation by Oil Red O.
[0094] FIG. 24. Induction of PPARgamma-Mediated Transactivation of
PPRE-Luc Reporter by Compound 11 and Rosiglitazone.
[0095] FIG. 25. Effect of Compound 11 on In Vitro Glycogen
Synthesis in HepG2 Cells. A. Dose-dependent stimulation of glycogen
synthesis from glucose by Compound 11 in the absence of insulin. B.
Time-dependent increase in Compound 11-stimulated glycogen
synthesis. C. Cycloheximide blocks glycogen synthesis induced by
Compound 11.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0096] A preferred compound according to formula 1 is 5-(4-(4-(1
carbomethoxy-2-(3,5-dimethoxyphenyl)-ethenyl)-phenoxy)-benzyl)-2,4-thiazo-
lidinedione, hereinafter referred to as compound 11. However, it
will be appreciated that the invention also contemplates the
provision and use of other compounds according to formula 1.
[0097] The compounds according to the present invention may be
combined with a physiologically acceptable carrier or vehicle to
provide a pharmaceutical composition, such as, lyophilized powder
in the form of tablet or capsule with various fillers and binders.
The effective dosage of a compound in the composition can be widely
varied as selected by those of ordinary skill in the art and may be
empirically determined.
[0098] As earlier indicated, the compounds of the invention are
useful for the treatment of diabetes, characterized by the presence
of elevated blood glucose levels, that is, hyperglycemic disorders
such as diabetes mellitus, including both type I and II diabetes,
as well as other hyperglycemic related disorders such as obesity,
increased cholesterol, hyperlipidemia such as hypertriglyceridemia,
kidney related disorders and the like. The compounds are also
useful for the treatment of disorders linked to insulin resistance
and/or hyperinsulinemia, which include, in addition to diabetes,
hyperandrogenic conditions such as polycystic ovary syndrome
(Ibanez et al., J. Clin Endocrinol Metab, 85:3526-30, 2000; Taylor
A. E., Obstet Gynecol Clin North Am, 27:583-95, 2000), coronary
artery disease such as atherosclerosis and vascular restenosis, and
peripheral vascular disease. Additionally, the compounds of the
present invention are also useful for the treatment of inflammation
and immunological diseases that include those mediated by signaling
pathways linked to pro-inflammatory cytokines, such as rheumatoid
arthritis and other inflammatory arthritides, multiple sclerosis,
inflammatory bowel disease, psoriasis, psoriatic arthritis,
ankylosing spondylitis and other spondylarthritides, and contact
and atopic dermatitis.
[0099] By "treatment", it is meant that the compounds of the
invention are administered in an amount which is at least
sufficient to, for example, reduce the blood glucose level in a
patient suffering from hyperglycemic disorder or to inhibit or
prevent the development of pro-inflammatory cytokine or like
responses in a patient suffering from inflammatory or immunological
disease. In the case of diabetes, the compound is usually
administered in the amount sufficient to reduce the blood glucose
level, free fatty acid level, cholesterol level, and the like to an
acceptable range, where an acceptable range means + or -10%, and
usually + or -5% of the normal average blood glucose level and like
level of the subject, or sufficient to alleviate the symptoms
and/or reduce the risk of complications associated with elevated
levels of these parameters. A variety of subjects may be treated
with the present compounds to reduce blood glucose levels such as
livestock, wild or rare animals, pets, as well as humans. The
compounds may be administered to a subject suffering from
hyperglycemic disorder using any convenient administration
technique, including intravenous, intradermal, intramuscular,
subcutaneous, oral and the like. However, oral daily dosage is
preferred. The dosage delivered to the host will necessarily depend
upon the route by which the compound is delivered, but generally
ranges from about 0.1-500 mg/kg human body weight or typically from
about 1 to 50 mg/kg human body weight. Generally similar types of
administration and dosages are also contemplated when the compounds
of the invention are used to treat inflammatory or immunological
disease.
[0100] The compounds of this invention may be used in formulations
using acceptable pharmaceutical vehicles for enteral, or
parenteral, administration, such as, for example, water, alcohol,
gelatin, gum arabic, lactose, amylase, magnesium stearate, talc,
vegetable oils, polyalkylene glycol, and the like. The compounds
can be formulated in solid form, e.g., as tablets, capsules, drages
and suppositories, or in the liquid form, e.g., solutions,
suspensions and emulsions. The preparations may also be delivered
transdermally or by topical application.
[0101] Representative compounds according to the present invention
may be synthesized by the methods disclosed below in Schemes 1
through 11, wherein Scheme 1 illustrates the preparation of
exemplary compounds 10, 11 and 14; Scheme 2 illustrates the
preparation of exemplary compounds 17 and 18; Scheme 3 illustrates
the preparation of exemplary compounds 22 and 23; Scheme 6
illustrates the synthesis of exemplary compounds 40, 41 and 42;
Scheme 7 illustrates the preparation of exemplary compounds 46, 47,
49 and 50; Scheme 8 illustrates the synthesis of compound 54;
Scheme 9 illustrates the preparation of compounds 58 and 59; and
Schemes 4, 5, 10 and 11 describe the synthesis methods more
generally. ##STR11##
[0102] .sup.aReagents and conditions: (a) acetic anhydride,
Et.sub.3N, 6 h, 130.degree. C., 47%; (b) MeOH, H.sub.2SO.sub.4, 20
h, reflux, 97%; (c) 4-fluorobenzaldehyde, NaH, DMF, 18 h,
80.degree. C., 77%; (d) 2,4-thiazolidinedione, piperidine, benzoic
acid, toluene, 5 h, reflux, 86%; (e) Pd/C(10%), HCOONH.sub.4/AcOH,
48 h, reflux, 49%; (f) NaBH.sub.4, EtOH, 1 h, 25.degree. C.,
quantitative; (g) PBr.sub.3, CH.sub.2Cl.sub.2, 25.degree. C., 1 h,
99%; (h) BuLi, 2,4-thiazolidinedione, THF, 0.degree. C., 45 min,
15%; (i) aqueous NaOH, MeOH, 15 h, 25.degree. C., 73%.
##STR12##
[0103] .sup.aReagents and conditions: (a) Pd/C (10%), H.sub.2, 18
h, 25.degree. C., quantitative; (b) 4-fluorobenzaldehyde, NaH, DMF,
18 h, 80.degree. C., 69%; (c) 2,4-thiazolidinedione, piperidine,
benzoic acid, toluene, 2 h, reflux, 81%; (d) Pd/C(10%), H.sub.2(60
psi), 34 h, 25.degree. C., 38%. ##STR13##
[0104] .sup.aReagents and conditions: (a) acetic anhydride,
Et.sub.3N, 24 h, 125.degree. C., 13%; (b) MEOH, H.sub.2SO.sub.4, 18
h, reflux, 35%; (c) 4-fluorobenzaldehyde, NaH, DMF, 18 h,
80.degree. C., 74%; (d) 2,4-thiazolidinedione, piperidine, benzoic
acid, toluene, 5 h, reflux, 91%; (e) Pd/C(10%), ammonium formate,
acetic acid, 20 h, reflux. ##STR14## ##STR15## ##STR16## ##STR17##
##STR18## ##STR19## ##STR20## ##STR21## ##STR22##
[0105] Referring to Scheme 1, the aldehyde 5 and acid 6 may be
condensed in acetic anhydride and triethylamine to form the
unsaturated acid 7. After esterification of the acid to provide
compound 8, the phenolic hydroxy group is formed into an ether 9
with p-fluorobenzaldehyde. The aldehyde 9 is then condensed with
the thiazolidinedione to provide compound 10 and the bond exo to
the heterocycle in compound 10 is reduced with hydrogen to form the
object compound 11.
[0106] The steps in Scheme 1 are generalized in Scheme 4. The
general formulas 2b, 3b, 4b, 6b, 7b and 8b correspond respectively
to formulas 5, 6, 7, 9, 10 and 11 in Scheme 1.
[0107] In Scheme 5, the general synthesis of the tricyclic products
35 and 36 is shown. The aldehyde or ketone 32 is condensed with 33
to form the bicyclic compound 34. The compound 34 is condensed with
the heterocyclic dione to form the tricyclic product 35, which can
be optionally hydrogenated to 36.
[0108] In Scheme 10, the general synthesis of compounds where
R.dbd.R'.dbd.H is shown. The aldehyde or ketone 62 is condensed
with the heterocyclic dione to form the bicyclic compound 63, which
can be optionally hydrogenated to form the product 64. Coupling of
64 with optionally substituted aldehyde yields the tricyclic
compound 65. Wittig reaction of 65 results in the formation of
stilbene derivative 66.
[0109] In Scheme 11, the general synthesis of biphenyl products 69
and 70 is shown. Coupling of optionally substituted hydroxyl
biphenyl 67 with optionally substituted aldehyde or ketone yields
68. The aldehyde or ketone 68 is condensed with the heterocyclic
dione to form the compound 69, which can be optionally hydrogenated
to form the product 70.
[0110] In Formula 1, C.sub.1-C.sub.20 linear or branched alkyl
means groups such as methyl, ethyl, propyl, isopropyl, n-butyl,
isobutyl, t-butyl, sec-butyl, isopentyl, neopentyl, etc. The
C.sub.2-C.sub.20 linear or branched alkenyl means unsaturated
groups such as ethenyl, propenyl, n-butenyl, isobutenyl, including
groups containing multiple sites of unsaturation such as
1,3-butadiene, and the like. The halo groups include chloro,
fluoro, bromo, iodo. Substituted C.sub.1-C.sub.20 linear or
branched alkyl or substituted C.sub.2-C.sub.20 linear or branched
alkenyl means that the alkyl or alkenyl groups may be substituted
with groups such as halo, hydroxy, carboxyl, cyano, amino, alkoxy,
and the like. The C.sub.1-C.sub.20 acylamino or acyloxy group means
an oxygen or amino group bonded to an acyl group (RCO) where R can
be hydrogen, C.sub.1-C.sub.20 linear or branched alkyl or
C.sub.2-C.sub.20 linear or branched alkenyl. Alkenyl groups are
--C.dbd.C--, where R can be hydrogen, C1-C.sub.20 linear or
branched alkyl or C.sub.2-C.sub.20 linear or branched alkyl.
Alkoxycarbonyl means a group ROCO-- where R can be hydrogen,
C.sub.1-C.sub.20 linear or branched alkyl or C.sub.2-C.sub.20
linear or branched alkenyl. The C.sub.1-C.sub.20 alkyl carboxyl
amino group means a group RCON(R)-- where R can be independently
hydrogen, C.sub.1-C.sub.20 linear or branched alkyl or
C.sub.2-C.sub.20 linear or branched alkenyl. Carboxyl is the group
H0.sub.2C--, and alkanoyl is the group RCO-- wherein R is a linear
or branched carbon chain. The group aroyl is Ar--CO-- wherein Ar is
an aromatic group such as phenyl, naphthyl, substituted phenyl, and
the like. Aralkanoyl is the group Ar--R--CO-- wherein Ar is a
aromatic group such as phenyl, naphthyl, substituted phenyl, etc.
and R is a linear branched alkyl chain.
[0111] As indicated earlier, the compounds of the invention where
a, b or c represents a double bond may have either the E or Z
configuration. On the other hand, when a, b or C is absent, i.e. a
single bond is present, the resulting compounds may be R-- and/or
S-stereoisomers. The invention contemplates racemic mixtures of
such stereoisomers as well as the individual, separated
stereoisomers. The individual stereoisomers may be obtained by the
use of an optically active resolving agent. Alternatively, a
desired enantiomer may be obtained by stereospecific synthesis
using an optically pure starting material of known
configuration.
[0112] The preparation of compound 11, i.e.
5-(4-(4-(1-carbomethoxy)-2-(3,5-dimethoxy
phenyl)-ethenyl)-phenoxy)-benzyl)-2,4-thiazolidinedione (also known
as
3-(3,5-dimethoxy-phenyl)-2-{4-[4-(2,4-dioxo-thiazolidin-5-ylmethyl)-pheno-
xy]-phenyl}-acrylic acid methyl ester), is described below with
reference to Scheme 1.
[0113] Perkin condensation of 3,5-dimethoxybenzaldehyde 5 with
4-hydroxyphenylacetic acid 6 yielded the alpha-phenyl substituted
cinnamic acid 7 exclusively as E-isomer. The geometry of the double
bond was confirmed by .sup.1HNMR comparison with the reported
compound (Pettit et al, J Nat Prod 51:517-27, 1998). Esterification
of 7 followed by condensation with 4-fluorobenzaldehyde yielded 9.
Knovenagel condensation of aldehyde 9 with 2,4-thiazolidinedione in
the presence of piperidinium benzoate with azeotropic removal of
water gave a good yield of 10.
[0114] A major challenge was selective hydrogenation of the double
bonds in order to produce compounds 11, 17 and 18. Reduction of 10
with magnesium/methanol was non-selective and yielded a mixture of
products. Zinc-acetic acid reduction gave a mixture of polar
product. Hydrogenation with 10% palladium on carbon as catalyst in
1,4-dioxane yielded a mixture of 11 and 18 in a ratio of 6:4.
Separation of the compounds from this mixture was only possible by
reverse phase chromatography on C-18 silica. These problems were
overcome in several ways. Hydrogenation of 10 using ammonium
formate as hydrogen donor in the presence of palladium catalyst
(Hudlicky, ACS Monograph 188:46-7, 1996; Ram and Ehrenkaufer,
Synthesis 91-5, 1988) produced minimal amounts of the over-reduced
product 18, and isolation of 11 in high purity was possible by
repeated crystallization from methanol. In a preferred variation of
this approach, platinum catalyst was substituted for palladium, and
the crude product was recrystallized from dichloromethane; with
these modifications both the amount of catalyst required and the
reaction time were significantly reduced, while the overall yield
was considerably improved. In an another attempt to make 11, the
aldehyde 9 was reduced to alcohol 12 which upon treatment with
PBr.sub.3 yielded the bromo compound 13 in high yield. The bromo
compound was condensed with 2,4-thiazolidinedione anion generated
by BuLi to yield 11 in low yield.
[0115] It was difficult to synthesize 18 in good yield from either
10 or 11 by palladium-catalyzed hydrogenation due to poisoning of
the catalyst by the 2,4-thiazolidinedione moiety in the molecule;
the resulting mixtures contained 18 as a minor product. To solve
this problem (as shown in Scheme 2), 8 was first reduced, by using
10% palladium on carbon as catalyst, to 15 quantitatively followed
by coupling with 4-fluorobenzaldehyde and 2,4-thiazolidinedione to
furnish 17 in good yield. Reduction of 17 with palladium on carbon
catalyst for a longer period of time and catalyst renewal half-way
through the reaction, followed by chromatographic purification over
C-18 reverse phase silica gel, produced 18 in moderate yield.
[0116] The synthetic strategy adopted to prepare 23, the
corresponding Z-isomer of 11, is outlined in Scheme 3. Prolonged
heating of 7 with acetic anhydride and triethylamine (Kessar et al,
Indian J Chem 20B:1-3, 1981) yielded the corresponding Z-isomer 19
in 13% yield. Interestingly, the reaction of 2,4-thiazolidinedione
with 21, in order to produce 22, showed minimal isomerization of
the cinnamic acid double bond and resulted in a mixture of E- and
Z-isomers in a ratio of 1:7 respectively. Reduction was carried out
without further purification and the final product was purified by
preparative HPLC to yield 23.
EXAMPLE 1
[0117] General Methods. Melting points were measured on a Mel-Temp
melting point apparatus and are uncorrected. The .sup.1H NMR and
.sup.13C NMR spectra were recorded on a JEOL Eclipse (400 MHz) or
Nicolet NT 36 (360 MHz) spectrometer and are reported as parts per
million (ppm) downfield from TMS. The infrared spectra were
recorded on a Nicolet Impact 410 FT-IR spectrophotometer. The mass
spectra were recorded on Fison VG Platform II of HP 1100 MDS 1964A
mass spectrophotometer. UV spectra were recorded on a Beckman DU650
spectrophotometer. TLC was done on Merck silica gel F.sub.254
precoated plates. The silica gel used for column chromatography was
`Baker` silica gel (40 .mu.m) for flash chromatography.
[0118] 3-(3,5-Dimethoxyphenyl)-2-(4-hydroxyphenyl)-acrylic acid
(7). To a mixture of 3,5-dimethoxybenzaldehyde, 5, (500 g, 3.0 mol)
and 4-hydroxyphenyl acetic acid, 6, (457 g, 3.0 mol) was added
acetic anhydride (1.0 L, 10.60 mole) and triethylamine (420 mL, 3.0
mol). After stirring at 130-140.degree. C. for 6 h, the mixture was
cooled to room temperature. Concentrated HCl (1 L) was added to the
reaction mixture slowly over 50 min while keeping the temperature
between 20-30.degree. C. The light yellow precipitate obtained was
filtered and washed with water. The solid was dissolved in 3N NaOH
(5 L) and stirred for 1 h and filtered. The filtrate was acidified
to pH 1 with concentrated HCl while maintaining a temperature at
25-30.degree. C. The precipitated product was filtered and washed
with water to give crude product that was recrystallized from
MeOH--H.sub.2O and dried at 40.degree. C. for 6 h to yield 7 (428
g, 47%): mp 225-227.degree. C. (lit. 226-228.degree. C.).sup.17;
.sup.1HNMR (360 MHz, DMSO-d.sub.6) .delta.12.48 (br s, 1H), 9.42
(s, 1H), 7.59 (s, 1H), 6.95 (d, J=8.0 Hz, 2H), 6.76 (d, J=8.0 Hz,
2H), 6.35 (t, J=2.2 Hz, 1H), 6.27 (d, J=2.2 Hz, 2H), 3.56 (s, 6H);
MS (EI) m/z 299[M].sup.-.
[0119] 3-(3,5-Dimethoxyphenyl)-2-(4-hydroxyphenyl)-acrylic acid
methyl ester (8). Methanol (3.0 L) was added to a thoroughly dried
7 (427.5 g, 1.42 mol) under argon. To this stirred suspension
concentrated sulfuric acid (100 mL) was added and heated at reflux
for 20 h under nitrogen. Methanol was evaporated under reduced
pressure at 30.degree. C. The residue was taken up in ethyl acetate
(3.0 L) and washed with water (2.times.1.0 L), saturated aqueous Na
HCO.sub.3 (2.times.1.0 L), brine (2.times.1.0 L). The organic layer
was dried on anhydrous magnesium sulfate, filtered and the solvent
was evaporated. The residue obtained was dried thoroughly under
high vacuum as white solid, (433.6 g 97%): mp 106-108.degree. C.;
.sup.1HNMR(360 MHz, CDCl.sub.3): .delta. 7.72 (s, 1H), 7.06 (d,
J=7.9 Hz, 2H), 6.77 (d, J=7.9 Hz, 2H), 6.33 (t, J=2.2 Hz, 1H), 6.26
(d, J=2.2 Hz, 2H), 5.74 (s, 1H), 3.81 (s, 3H), 3.60 (s, 6H); MS
(EI) m/z 315[M].sup.+.
[0120]
3-(3,5-Dimethoxyphenyl)-2-[4-(4-formylphenoxy)-phenyl]-acrylic acid
methyl ester (9). Under argon, 8 (433.0 g, 1.37 mol) was dissolved
in dry DMF (1.6 L) and to this sodium hydride (60.4 g, 1.51 mol)
was added. To the resulting orange solution 4-fluorobenzaldehyde
(185.0 mL, 1.71 mol) was added and heated at 80.degree. C. for 18
h. The reaction mixture was cooled to room temperature, diluted
with ethyl acetate (3.0 L) and extracted with water (3.times.1.0
L), then brine (1.times.1.0 L). The organic layer was dried over
anhydrous sodium sulfate, filtered and solvent was evaporated. The
residue was suspended in methanol (3.0 L) and stirred overnight.
Solid was filtered and dried under vacuum at 40.degree. C. to yield
9 as pale yellow solid (445 g, 77%): mp 108-110.degree. C.
.sup.1HNMR(360 MHz, CDCl.sub.3) .delta. 9.94 (s, 1H), 7.86 (d,
J=8.6 Hz, 2H), 7.80 (s, 1H), 7.28 (d, J=8.6 Hz, 2H), 7.11
(overlapped d, J=9.0 Hz, 2H), 7.08 (overlapped d, J=9.0 Hz, 2H),
6.36 (t, J=2.2 Hz, 1H), 6.25 (d, J=2.2 Hz, 2H), 3.83 (s, 3H), 3.63
(s, 6H).
[0121]
3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylidenemet-
hyl)-phenoxy]-phenyl}-acrylic acid methyl ester (10). To a stirred
suspension of 9 (352 g, 0.82 mol) in anhydrous toluene (2.5 L),
2,4-thiazolidinedione (98.6 g, 0.84 mol), benzoic acid (134 g, 1.10
mol) and piperidine (107.4 g, 1.26 mol) was added sequentially and
heated at reflux temperature with continuous removal of water with
the help of Dean-Stark apparatus for 5 h. Toluene (1.0 L) was
removed from the reaction mixture and cooled overnight at 4.degree.
C. Solid separated was filtered and mother liquor was evaporated to
dryness under reduced pressure. The residue obtained was
re-dissolved in a mixture of MeOH--diethylether (1:1, 3.0 L). On
standing overnight at 4.degree. C., the solution yielded more
solids. Solid from both the lots were combined and dried overnight
in vacuum oven at 40.degree. C. to give 10 as yellow solid (362.5
g, 86%): mp 106-108.degree. C.; mp 225-226.degree. C.; .sup.1H NMR
(360 MHz, DMSO-d.sub.6): 12.53 (br s, 1H), 7.78 (s,1H), 7.73
(s,1H), 7.63 (d, J=9.2 Hz, 2H), 7.25 (d, J=9.2 Hz, 2H), 7.13
(overlapped d, J=8.3 Hz, 2H), 7.11 (overlapped d, J=8.6 Hz, 2H),
6.42 (t, J=2.2 Hz, 1H), 6.27 (d, J=2.2 Hz, 2H), 3.73 (s, 3H), and
3.59 (s,6H); MS (EI) m/z 518[M].sup.+.
[0122]
5-(4-(4-(1-carbomethoxy-2-(3,5-dimethoxyphenyl)-ethenyl)-phenoxy)--
benzyl)-2,4-thiazolidinedione, also called
3-(3,5-dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-phenoxy-
]-phenyl}-acrylic acid methyl ester (compound 11). Compound 10 (30
g, 58 mmol) was dissolved in warm dioxane (900 mL), transferred to
a 2 L hydrogenation bottle and 10% Pd--C (.about.50% water, 15 g)
was added to this and hydrogenated in a Parr hydrogenator at 60 psi
for 24 h. Following this period, an additional 15 g Pd--C was added
and hydrogenation was allowed to continue for another 24 h.
Catalyst was filtered through a bed of Celite and solvent was
evaporated. The residue was taken up in acetonitrile (500 mL) and
adsorbed on C-18 silica (50 g). The adsorbed material was placed on
the top of a column containing C-18 reverse phase silica gel (400).
Column was eluted with CH.sub.3CN in H.sub.2O (45%, 2 L),
CH.sub.3CN in H.sub.2O (50%, 2 L), CH.sub.3CN in H.sub.2O (55%, 2
L) to elute the undesired fractions. Fractions were collected with
the start of 60% CH.sub.3CN in H.sub.2O elution for the desired
compound. Fractions were mixed on the basis of their HPLC purity.
Acetonitrile was evaporated under reduced pressure. Water was
removed by lyophilization. Yield: 12 g (40%). White solid; m.p.
126-128.degree. C. .sup.1H NMR (DMSO-d.sub.6) .delta. 12.01 (br, 1
H), 7.73 (s, 1 H), 7.28 (d, J=8.6 Hz, 2H), 7.19(d, J=8.6 Hz, 2H),
7.02 (d, J=8.6 Hz, 2H), 6.96 (d, J=8.6 Hz, 2H), 6.40 (t, J=2.2 Hz,
1 H), 6.27 (d, J=2.2 Hz, 2H), 4.92 (dd, J=9.2 and 4.4 Hz, 1 H),
3.73 (s, 3H), 3.57 (s, 6H), 3.37 (dd, J=14.8 and 4.3 Hz, 1 H) and
3.12 (dd, J=14.8 and 9.4 Hz, 1 H); IR (KBr) .nu..sub.max 3200,
2950, 2850, 1700, 1600, 1500, 1350, 1150, and 850 cm.sup.-1;
EIMS:m/z, 518, [M-H].sup.-265, 249, and 113.
[0123]
3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)--
phenoxy]-phenyl}-acrylic acid methyl ester (11). To a solution of
10 (599 g, 1.16 mol) in glacial acetic acid (11.5 L), ammonium
formate (4.0 kg, 62.9 mol) was added and stirred for 30 min. A
slurry of Pd on carbon (10%, dry, 300 g) in glacial acetic acid
(500 mL) was added to the flask (caution: in a large scale reaction
exothermicity may be a problem; rigorous exclusion of oxygen is
desirable) and heated at 120.degree. C. for 24 h followed by
stirring at room temperature for 48 h. The resulting mixture was
filtered through a bed of Celite.RTM.. The filtrate was poured
slowly into vigorously stirred water (12 L) and the separated solid
was filtered and dried. Resulting solid was purified by slurring
twice in hot methanol followed by once from ethanol to yield pure
11 as white solid (296 g, 49.2%): mp 126-128.degree. C.; .sup.1H
NMR (400 MHz, DMSO-d.sub.6) 12.01 (br s, 1H), 7.73 (s, 1H), 7.28
(d, J=8.6 Hz, 2H), 7.19 (d, J=8.6Hz, 2H), 7.02 (d, J=8.6 Hz, 2H),
6.96 (d, J=8.6 Hz, 2H), 6.40 (t, J=2.2 Hz, 1H), 6.27 (d, J=2.2 Hz,
2H), 4.92(dd, J=9.2 and 4.4 Hz, 1H), 3.73 (s, 3H), 3.57 (s, 6H),
3.37 (dd, J=14.8 and 4.3 Hz, 1H) and 3.12 (dd, J=14.8 and 9.4 Hz,
1H); IR (KBr) .nu..sub.max 3200, 2950, 2850, 1700, 1600, 1500,
1350, 1150, and 850 cm.sup.-1; MS (EI) m/z 518[M-H].sup.-, 265,
249, and 113.
[0124]
3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)--
phenoxy]-phenyl}-acrylic acid methyl ester (11). Compound 10 (20 g,
38.6 mmol), ammonium formate (150 g, 2.38 mol), 10% Pt/C (dry, 4 g)
and acetic acid (660 mL) were combined into a round bottom flask
equipped with reflux condenser, thermometer and mechanical stirrer.
The reactor was evacuated and purged three times with nitrogen
then, heated to a steady reflux (ca. 124.degree. C). Reaction was
completed within 15 h and allowed to cool with stirring to ambient
room temperature. After cooling, the mixture was filtered though a
pad of Celite.RTM. (5 g) and the filter pad washed with fresh
acetic acid (2.times.100 mL). The mother liquor and washes were
combined and concentrated. The residue was then diluted with
dichloromethane (400 mL), and the combined organics were extracted
twice with water (400 mL) and 5% bicarbonate (400 mL). The organic
portion was then dried and poured through silica gel (30 g) and
washed with dichloromethane (2.times.100 mL). The washes were
combined and concentrated. The residue was diluted with ethanol,
allowed to cool to 60.degree. C. and seed crystals were added. This
slurry was stirred at 50.degree. C. for about 30 min then allowed
to cool to ambient room temperature to yield compound 11 (12.85 g,
64%) with an HPLC assay of 98.1%.
[0125]
3-(3,5-Dimethoxyphenyl)-2[4-(4-hydroxymethylphenoxy)-phenyl]-acryl-
ic acid methyl ester (12). Compound 9 (5.0 g, 11.9 mmol) was
suspended in anhydrous ethanol (60 mL) at room temperature and
sodium borohydride (0.23 g, 6.1 mmol) was added with efficient
stirring. Reaction was complete in 1 h, solvent was evaporated and
the residue was dissolved in ethyl acetate. The organic layer was
extracted with water (50 mL), brine (25 mL), dried on anhydrous
magnesium sulfate, filtered and solvent was evaporated to yield the
title compound 12 as white solid (5.1 g, 100%): mp 93-95.degree.
C.; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.72 (s, 1H), 7.35
(d, J=8.8 Hz, 2H), 7.19 (d, J=8.8 Hz, 2H), 7.02 (d, J=8.4 Hz, 2H),
7.00 (d, J=8.4 Hz, 2H), 6.41 (t, J=2.4 Hz, 1H), 6.29 (d, J=2.0 Hz,
2H), 5.18 (t, J=6.4 Hz, 1H), 4.49 (d, J=4.8 Hz, 2H), 3.73 (s, 3H),
3.57 (s, 6H); MS (EI) m/z 315[M].sup.+.
[0126]
2-[4-(4-Bromomethylphenoxy)-phenyl]-3-(3,5-dimethoxyphenyl)-acryli-
c acid methyl ester (13). A solution of PBr.sub.3 (4.8 mL of 1.0 M
in CH.sub.2Cl.sub.2) was added dropwise to 12 (5.0 g, 11.9 mmol)
dissolved in CH.sub.2Cl.sub.2 (20 mL) at temperature with good
stirring. After 1 h, the solution was extracted with water
(2.times.60 mL) and brine (20 mL). The organic phase was dried over
anhydrous magnesium sulfate, filtered through a small bed of silica
gel (20 g) and solvent was evaporated. The resulting tacky syrup
was dried under high vacuum for 48 h at room temperature to yield
the title compound (5.7 g, 99%): mp 79-81.degree. C.; .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.73 (s, 1H), 7.49 (d, J=8.4 Hz,
2H), 7.22 (d, J=8.4 Hz, 2H), 7.07 (d, J=8.4 Hz, 2H), 7.00 (d, J=8.4
Hz, 2H), 6.42 (t, J=2.4 Hz, 1H), 6.28 (d, J=2.0 Hz, 2H), 4.73 (d,
J=4.8 Hz, 2H), 3.68 (s, 3H), 3.58 (s, 6H); Anal.
(C.sub.25H.sub.23BrO.sub.5) C: calculated, 61.12; found, 62.26; H:
calculated 4.80; found 4.88.
[0127]
3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)--
phenoxy]-phenyl}-acrylic acid methyl ester (11).
2,4-Thiazolidinedione (2.83 g, 24.2 mmol) was dissolved in dry THF
(170 mL) and cooled to 0.degree. C. under argon. Butyllithium (1.6
M in hexanes, 30 mL, 48.0 mmol) was added dropwise. Stirring
continued for 0.5 h at 0.degree. C. Under argon, 13 (5.7 g, 11.8
mmol) was dissolved in dry THF (30 mL) and was added rapidly via
syringe to the above suspension with rapid stirring. The
temperature was maintained at 0.degree. C. for 45 min before
quenching with aqueous HCl (5%, 40 mL). Additional H.sub.2O (40 mL)
was added and extracted with ethyl acetate (3.times.30 mL). Organic
layers were combined, washed with brine, dried over anhydrous
magnesium sulfate, filtered and the solvent was evaporated. Flash
chromatography over silica gel using hexanes-ethyl acetate (3:2) as
eluting solvent yielded the title compound, 11, (0.93 g, 15%). The
melting point and .sup.1H NMR of compound 11 made by this method
were identical to those for compound 11 produced by the synthetic
route starting from compound 10 described above.
[0128]
3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)--
phenoxy]-phenyl acrylic acid (14). To a stirred, cooled below
10.degree. C., suspension of 11 (10 g, 19.27 mmol) in methanol (50
mL), aqueous sodium hydroxide (2N, 33.7 mL, 67.4 mmol) was added
and stirred for 15 h at room temperature. The resulting pale yellow
solution was cooled to 10.degree. C. and acidified with aqueous HCl
(5%, 115 mL). Solid separated was filtered and washed with water
(3.times.30 mL), recrystallized from ethanol to give 14 as white
solid (7.14 g, 73%): mp 138-140.degree. C.; .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.69 (s, 1H), 7.28 (d, J=8.8 Hz, 2H), 7.19
(d, J=8.8 Hz, 2H), 7.02 (d, J=8.8 Hz, 2H), 6.97 (d, J=8.8 Hz, 2H),
6.41 (t, J=2.4 Hz, 1H), 6.28 (d, J=2.4 Hz, 2H), 4.92 (dd, J=9.2 and
4.4 Hz, 1H), 3.58 (s, 6H), 3.38 (dd, J=14.0 and 4.0 Hz, 1H) and
3.13 (dd, J=14.4 and 9.2 Hz, 1H); MS (EI) m/z 506[M].sup.+.
[0129] 3-(3,5-Dimethoxyphenyl)-2-(4-hydroxyphenyl)-propionic acid
methyl ester (15). To a suspension of 8 (6.28 g, 20.0 mmol) in
ethanol (200 mL) palladium on carbon (10%, wet, 0.63 g) was added
and stirred under H.sub.2 at atmospheric pressure at room
temperature for 18 h. Catalyst was filtered through a bed of
Celite.RTM. and solvent was evaporated under reduced pressure to
yield 15 as white solid (6.32 g, 100%): mp 63-65.degree. C.;
.sup.1HNMR (400 MHz, DMSO-d.sub.6): .delta. 7.15 (d, J=8.7 Hz, 2H),
6.74 (d, J=8.7 Hz, 2H), 6.29 (t, J=2.4 Hz, 1H), 6.25 (d, J=2.4 Hz,
2H), 3.78(t, J=8.7 Hz, 1H), 3.72(s, 6H), 3.62(s, 3H), 3.31(dd,
J=13.5 and 8.4 Hz, 1H), 2.93(dd, J=13.5 and 6.9 Hz, 1H); MS (EI)
m/z 317[M].sup.+.
[0130]
3-(3,5-Dimethoxyphenyl)-2-[4-(4-formylphenoxy)-phenyl]-propionic
acid methyl ester (16). To a suspension of sodium hydride (60% in
oil, 0.25 g, 6.3 mmol) in DMF (2 mL) under argon, 15 (2.0 g, 6.3
mmol) in dry DMF (3 mL) was added. To the resulting yellow
solution, 4-fluorobenzaldehyde (0.68 mL, 6.3 mmol) was added and
heated at 80.degree. C. for 18 h. The reaction mixture was cooled
to room temperature, water (20 mL) was added and extracted with
ethyl acetate (3.times.50 mL). The organic layer was dried over
anhydrous sodium sulfate, filtered and solvent was evaporated. An
ethyl acetate solution of crude product was filtered through a
small bed of silica gel to yield 16 (1.83 g, 69%) as oil:
.sup.1HNMR(400 MHz, DMSO-d.sub.6): .delta. 9.91 (s, 1H), 7.84 (d,
J=8.7 Hz, 2H), 7.33 (d, J=8.7 Hz, 2H), 7.04 (d, J=5.4 Hz, 2H), 7.01
(d, J=5.4 Hz, 2H), 6.30 (t, J=2.1 Hz, 1H), 6.25 (d, J=2.1 Hz, 2H),
3.86 (t, J=7.8 Hz, 1 Hz), 3.76 (s, 6H), 3.66 (s, 3H), 3.36 (dd,
J=12.6 and 8.1 Hz, 1H), 2.97 (dd, J=13.5 and 7.5 Hz, 1H); MS (EI)
m/z 421 [M].sup.+.
[0131]
3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylidenemet-
hyl)-phenoxy]-phenyl}-propionic acid methyl ester (17). To a
stirred suspension of 16 (1.81 g, 4.3 mmol) in anhydrous toluene
(25 mL), 2,4-thiazolidinedione (0.56 g, 4.74 mmol), benzoic acid
(0.68 g, 5.60 mmol) and piperidine (0.60 mL, 6.03 mmol) was added
sequentially and heated at reflux temperature with continuous
removal of water using a Dean-Stark apparatus for 2 h. Solvent was
evaporated to dryness under reduced pressure. The residue obtained
was purified by silica gel chromatography, eluted with hexane-ethyl
acetate (1:1) to yield 17 (1.82 g, 81%): mp 104-106.degree. C.;
.sup.1H NMR (400 MHz, DMSO-d.sub.6) 12.53 (br s, 1H), 7.76 (s,1H),
7.60 (d, J=8.7 Hz, 2H), 7.35 (d, J=8.7 Hz, 2H), 7.07 (d, J=4.8 Hz,
2H), 7.03 (d, J=4.8 Hz, 2H), 6.33-6.28 (m, 3H), 4.01 (t, J=7.5 Hz,
1 Hz), 3.66 (s, 6H), 3.56 (s, 3H), 3.22 (dd, J=13.8 and 8.4 Hz,
1H), 2.90 (dd, J=13.5 and 7.2 Hz, 1H); MS (EI) m/z 520[M].sup.+;
Anal. (C.sub.28H.sub.25NO.sub.7S) C: calculated, 64.73; found,
65.89; H: calculated, 4.85; found, 5.08, N: calculated, 2.70;
found, 2.56.
[0132]
3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)--
phenoxy]-phenyl}-propionic acid methyl ester (18). 17 (1.6 g, 3.08
mmol) was dissolved in dioxane (45 mL), transferred in a
hydrogenation bottle and Pd on carbon (10%, 1.0 g) was added.
Hydrogenation was done at 65 psi for 34 h. Following this period,
additional Pd on carbon (10%, 0.6 g) was added and hydrogenation
was allowed to continue for another 18 h. Catalyst was filtered
through a bed of Celite.RTM. and solvent was evaporated. The
residue was purified by column chromatography on reverse phase
silica gel (C-18) using acetonitrile-water (1:1) mixture to elute
18 as white solid (0.60 g, 38%): mp 125-128.degree. C.; .sup.1H NMR
(400 MHz, DMSO-d.sub.6) 12.04 (br s, 1H), 7.31 (d, J=8.4 Hz, 2H),
7.25 (d, J=8.4 Hz, 2H), 6.92 (d, J=8.6 Hz, 4H), 6.30 (d, J=2.0 Hz,
2H), 6.29 (t, J=2.0 Hz, 1H), 4.90 (dd, J=9.2 and 4.4 Hz, 1H), 3.98
(t, J=8.0 Hz, 1H), 3.67 (s, 6H), 3.56 (s, 3H), 3.37 (dd, J=13.6 and
4.0 Hz, 1H), 3.21 (dd, J=14.0 and 8.8 Hz, 1H); 3.11 (dd, J=14.0 and
9.2 Hz, 1H) and 2.90 (dd, J=13.6 and 7.6 Hz, 1H); MS (EI) m/z
522[M].sup.+.
[0133] Z-3-(3,5-Dimethoxyphenyl)-2-(4-hydroxyphenyl)-acrylic acid
(19). E-3-(3,5-dimethoxyphenyl)-2-(4-hydroxyphenyl)-acrylic acid, 7
(10.0 g, 33.3 mmol) was dissolved in a mixture of acetic anhydride
(40 mL, 0.42 mole) and triethylamine (40 mL, 0.29 mole) and heated
at 125.degree. C. for 24 h. The mixture was cooled to room
temperature. Ethyl acetate (150 mL) was added, further cooled to
5.degree. C., acidified with concentrated HCl (30 mL) and stirred
at this temperature for 90 min. The organic layer was separated and
the aqueous layer was extracted with ethyl acetate (100 mL). The
combined organic layers were washed with water (2.times.50 mL) and
extracted with aqueous NaOH (5M, 3.times.70 mL). The aqueous
alkaline layer was acidified with glacial acetic acid (65 mL) to pH
5.2 and stirred at 0.degree. C. for 30 min. Solid that separated
was filtered and mother liquor was acidified with concentrated HCl
(90 mL) and stirred at 5.degree. C. for 1 h. Solid that separated
was filtered, washed with cold water (2.times.50 mL) and dried at
45.degree. C. for 6 h to yield, 19, (1.3 g, 13%): mp
135-137.degree. C.; .sup.1HNMR (400 MHz, DMSO-d.sub.6) .delta.
13.28 (br, 1H), 9.70 (br, 1H), 7.32 (d, J=10.4 Hz, 2H), 6.81(s, 1H,
overlapped), 6.79 (d, J=9.7 Hz, 2H), 6.67 (d, J=2.5 Hz, 2H), 6.64
(t, J=2.5 Hz, 1H), and 3.73 (s, 6H); MS (EI) m/z 299[M].sup.-.
[0134] Z-3-(3,5-Dimethoxyphenyl)-2-(4-hydroxyphenyl)-acrylic acid
methyl ester (20). Concentrated sulfuric acid (10 drops) was added
to a stirred methanol suspension of thoroughly dried 19 (0.60 g,
2.0 mmol) under argon and heated at reflux for 18 h. Methanol was
evaporated under reduced pressure, residue was taken up in ethyl
acetate (20 mL) and washed with water (20 mL), saturated aqueous Na
HCO.sub.3 (10 mL) and brine (10 mL). The organic layer was dried on
anhydrous magnesium sulfate, filtered and the solvent was
evaporated. The crude product obtained was purified by
chromatography over silica gel and eluted with hexane-ethyl acetate
(7:3) to yield 20 as white solid (0.24 g, 35%): .sup.1HNMR (400
MHz, CDCl.sub.3) .delta. 7.26 (d, J=8.4 Hz, 2H), 6.82 (s, 1H), 6.76
(d, J=8.4 Hz, 2H), 6.45 (d, J=2.0 Hz, 2H), 6.34 (t, J=2.0 Hz, 1H),
4.97 (s, 1H), 3.73(s, 3H), 3.72(s, 6H).
[0135]
Z-3-(3,5-Dimethoxyphenyl)-2-[4-(4-formylphenoxy)-phenyl]-acrylic
acid methyl ester (21). Under argon, 20 (0.60 g, 1.9 mmol) was
dissolved in dry DMF (4 mL) and to this sodium hydride (60% in oil,
0.09 g, 2.28 mmol) was added. To the resulting orange solution,
4-fluorobenzaldehyde (0.25 mL, 2.28 mmol) was added and heated at
80.degree. C. for 18 h. The reaction mixture was cooled to room
temperature, water (10 mL) was added and the mixture was extracted
with ethyl acetate (3.times.20 mL). The crude product obtained
after evaporation was purified by chromatography over silica gel
and elution with a mixture of hexane-ethyl acetate (4:1) to yield
21 as white solid (0.59 g, 74%): .sup.1HNMR(400 MHz, CDCl.sub.3)
.delta. 9.93 (s, 1H), 7.86 (d, J=8.8 Hz, 2H), 7.49 (d, J=8.8 Hz,
2H), 7.10 (overlapped d, J=8.8 Hz, 2H), 7.08 (overlapped d, J=8.8
Hz, 2H), 6.96 (s, 1H), 6.53 (dd, J=2.8 Hz, 2H), 6.43 (t, J=2.0 Hz,
1H), 3.80(s, 3H), 3.79 (s, 6H).
[0136]
Z-3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylidenem-
ethyl)-phenoxy]-phenyl}-acrylic acid methyl ester (22). To a
stirred suspension of 21 (0.53 g, 1.3 mmol) in anhydrous toluene
(10 mL), 2,4-thiazolidinedione (0.15 g, 1.30 mmol), benzoic acid
(0.21 g, 1.69 mmol) and piperidine (0.19 g, 1.95 mmol) were added
sequentially and the mixture was heated at reflux temperature with
continuous removal of water using a Dean-Stark apparatus for 5 h.
Toluene was evaporated and the residue was chromatographed over
silica gel and eluted with hexane-ethyl acetate (1:1) to yield a
mixture of 22 and 10 (0.60 g, 91%) in a ratio of 7:1 on the basis
of proton NMR analysis: .sup.1H NMR (400 MHz, DMSO-d.sub.6): 12.53
(br s, 1H), 7.79 (s,1H), 7.65 (d, J=8.8 Hz, 2H), 7.54 (d, J=8.8 Hz,
2H), 7.18 (overlapped d, J=8.8 Hz, 2H), 7.16 (overlapped d, J=8.8
Hz, 2H), 7.17 (overlapped s,1H), 6.57 (d, J=2.0 Hz, 2H), 6.50 (t,
J=2.0 Hz, 1H), 3.79 (s, 3H), and 3.75 (s, 6H).
[0137]
Z-3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl-
)-phenoxy]-phenyl}-acrylic acid methyl ester (23). To a solution of
22 (0.60 g, 1.6 mmol) in acetic acid (15 mL) was added Pd on carbon
(10%, 300 mg) and ammonium formate (4.3 g, 55.8 mmol) (caution: in
a large scale reaction exothermicity may be a problem; rigorous
exclusion of oxygen is desirable) and heated at 120.degree. C. for
20 h. Catalyst was filtered through a bed of Celite.RTM. and acetic
acid was evaporated under reduced pressure. Water (50 mL) was added
to the residue and solid separated was filtered. Pure Z-isomer was
isolated by preparative HPLC using Intersil ODS-3 preparative
column (250.times.4.6 mm, 5 .mu.m) running at a rate of 15 mL per
minute using methanol:acetonitrile:water (3:3:2) containing formic
acid (0.05%): mp 65-66.degree. C.; .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 12.05 (br s, 1H), 7.48 (d, J=9.2 Hz, 2H),
7.29 (d, J=8.4 Hz, 2H), 7.13(s, 1H), 7.03 (overlapped d, J=8.8 Hz,
2H), 7.01 (overlapped d, J=8.4 Hz, 2H), 6.56 (d, J=2.0 Hz, 2H),
6.49 (t, J=2.0 Hz, 1H), 4.90(dd, J=9.2 and 4.4 Hz, 1H), 3.77 (s,
3H), 3.75 (s, 6H), 3.38 (dd, J=14.8 and 4.8 Hz, 1H) and 3.13 (dd,
J=14.4 and 9.2 Hz, 1H); MS (I) m/z 300[M].sup.+.
[0138] Referring to the drawings, compound 11 was administered in a
single oral dose (50 mg/kg body weight) for 15 days to db/db male
mice as shown in FIG. 1A. A substantial reduction in blood glucose
level was observed. There was no increase in body weight in the
treatment group as compared to the control treated with the vehicle
without the active ingredient, FIG. 1B.
[0139] The compound was orally administered to ob/ob mice with a
single oral dose (50 mg/kg body weight). As shown in FIG. 2A, there
was a 62% drop in blood glucose level and, similar to db/db mice,
there was no significant increase in body weight between the
control and the treatment groups as shown in FIG. 2B. This is in
contrast to treatment of diabetic animals by thiazolidinedione type
compounds which are known to be associated with increase in body
weight. See Okuno et al., J. Clin. Invest., 101, 1354-1361 (1998)
and Yoshioka et al., Metabolism, 42, 75-80 (1993). By stopping
treatment after day 15 in both models, there was shown an increase
in glucose level as depicted in FIGS. 3A and 3B. The time course of
the drug effect is shown in FIG. 4. Oral administration of a single
dose of the compound in db/db mice was effective for 24 hours and
beyond.
[0140] The triglyceride levels were also measured. Triglycerides,
which are esters of fatty acids and glycerol, do not freely
circulate in plasma but are bound to proteins and transported as
macromolecular complexes called lipoproteins. The triglycerides
were measured by the enzymatic method described by McGowan et al.,
Clin Chem 29:538-42, 1983, with a modification to determine the
triglyceride levels in db/db and ob/ob mice. There was shown a 24%
drop in triglyceride levels in db/db mice (FIG. 5A) after 15 days
of treatment with the compound and in ob/ob mice, a 65% decrease in
triglyceride as compared to the control (FIG. 6B) after treatment
for 10 days.
[0141] The free fatty acids (FFA) were enzymatically measured using
coenzyme A in the presence of acyl CoA synthase (Wako Chemicals
USA). The free fatty acid levels in db/db and ob/ob mice treated
with the compound were significantly lower compared to the control
animals. A 34% drop in FFA levels in db/db mice (FIG. 5B) was shown
after 15 days of treatment with the compound. In ob/ob mice, after
10 days of treatment, a lowering of 33% of FFA was shown compared
to the control (FIG. 6C).
[0142] The percentage of glycohemoglobin (GHb) in blood reflects
the average blood glucose concentration. It is a measure of overall
diabetic control and can be use to monitor the average blood
glucose levels. The glycosylation of hemoglobin occurs continuously
in the red blood cells. But since the reaction is non-enzymatic and
irreversible, the concentration of glycohemoglobin in a cell
reflects the average blood glucose levels seen by the cell during
its life. An assay was conducted using affinity chromatography with
boronate as described by Abraham et al., J. Lab. Clin. Med., 102,
187 (1983). There is a 0.7% drop in the GHb level in db/db mice
(FIG. 5C) after 15 days of treatment with the compound and in ob/ob
mice after 14 days of treatment, there is 1.3% decrease (FIG. 6D)
in the GHb level compared to the control. The blood insulin level
was measured by ELISA following a standard protocol. A 58% drop of
serum insulin in ob/ob mice (FIG. 6A) was shown after 10 days of
treatment with the compound, thus, demonstrating its ability to act
as an insulin sensitizer.
[0143] Obesity is considered a significant risk factor for various
adult diseases such as diabetes and cardiac disease. Leptin, an
obese gene product, has been identified from the investigation of
ob/ob mice, where the leptin is lacking because of a mutation in
that gene (Zhiang et al., Nature, 372, 425 (1994). Leptin is a
protein of about 16 kDa, which is expressed in adipose tissue, and
which promotes weight loss by suppressing appetite and stimulating
metabolism. It is currently believed that leptin plays a key role
in the obesity syndrome. In the db/db mice according to the
experiment, the leptin level was measured by an ELISA, following a
standard protocol. After 15 days of treatment with the compound,
there is a 23.degree./a increase (FIG. 5D) in the serum leptin
level compared to the control group.
[0144] The liver enzymes glutamic oxalacetic transaminase/aspartate
aminotransferase (AST/GOT) and glutamic pyruvic
transaminase/alanine aminotransferase (ALT/GPT) were assayed in the
sera of ob/ob mice after 21 days of treatment (orally, 50 mg/kg) of
the test compound. The test was also conducted using troglitazone.
These enzyme levels are found to elevate in several kinds of
hepatic disorders or liver necrosis. In FIG. 7A, the AST level in
the mice was not elevated compared to untreated mice or to mice
treated with troglitazone. Similarly, FIG. 7B shows that the ALT
level did not elevate compared to untreated mice or mice treated
with troglitazone.
[0145] Referring to FIG. 8 glucose uptake in 3T3-L1 differentiated
adipocytes was measured after treatment with the test compound. The
assay was conducted according to the method of Tafuri,
Endocrinology, 137, 4706-4712 (1996). The serum-starved cells were
treated with the test compound for 48 hours at different
concentrations, then washed and incubated in glucose-free media for
30 minutes at 37.degree. C. Then .sup.14C-deoxyglucose was added
and uptake was monitored for 30 minutes under incubation. After
washing, the cells were lysed (0.1% SIDS) and counted. As shown in
FIG. 8, there is a 3.5 to 4-fold increase in glucose uptake at the
indicated concentrations of the test compound with respect to basal
levels.
[0146] Referring to FIG. 9, RAW cells were preincubated with either
compound 11 or rosiglitazone (0.1, 1, 10, 50 or 100 pM) for 1 hour
at 37.degree. C. in RPMI-1640 containing 10% FBS. After 1 hour,
LIPS (0.1 pg/ml) was added and cells were incubated an additional 6
hours. Cell supernatant was then collected, aliquoted and frozen at
-70.degree. C., and an aliquot used to determine TNF-alpha
concentration by ELISA. Compound 11 was a better inhibitor of
TNF-alpha than rosiglitazone.
[0147] Referring to FIG. 10, RAW cells were preincubated with
either compound 11 or rosiglitazone (0.1, 1, 10, 50 or 100 pM) for
1 hour at 37.degree. C. in RPMI-1640 containing 10% FBS. After 1
hour LIPS (0.1 pg/ml) was added and cells were incubated an
additional 6 hours. Cell supernatant was then collected, aliquoted
and frozen at -70.degree. C., and an aliquot used to determine the
concentration of IL-6 by ELISA. Compound 11 was a better inhibitor
of IL-6 than rosiglitazone.
[0148] Referring to FIG. 11, RAW cells were preincubated with
either compound 11 or rosiglitazone (0.1, 1, 10, 50 or 100 pM) for
1 hour at 37.degree. C. in RPM 1-1640 containing 10% FIBS. After 1
hour LPS (0.1 pg/mL) was added and cells were incubated an
additional 6 hours. Cell supernatant was then collected, aliquoted
and frozen at -70.degree. C., and an aliquot used to determine the
concentration of IL-1-beta by ELISA. Compound 11 inhibited
IL-1-beta better than rosiglitazone.
[0149] Referring to FIGS. 12A AND 12B, RAW cells were preincubated
with either compound 11 (12A) or rosiglitazone (12B) (0.1, 1.0, 10,
50 or 100 .mu.M) for 1 hour at 37.degree. C. in RPMI-1640
containing 10% FIBS. After 1 hour LPS (0.1 pg/mL) was added and
cells were incubated an additional 6 hours. Cell supernatant was
then collected, aliquoted and frozen at -70.degree. C., and
aliquots used to determine COX-2 and COX-1 activity. Compound 11,
but not rosiglitazone, inhibited the activity of COX-2 (as measured
by PGE2 production in a 50 .mu.l sample). Neither compound
inhibited COX-1 activity.
[0150] The transcription factor NF-kappaB coordinates the
activation of many genes involved in the response to
pro-inflammatory cytokines, and, therefore, plays a key role in the
development of inflammatory diseases. NF-kappaB is activated by
phosphorylation of the inhibitory protein IkappaB. To examine the
effect of compound 11 on the LPS-stimulated phosphorylation of
IkappaB, RAW 264.7 cells were preincubated with vehicle only,
15-deoxy-.DELTA..sup.12,14-prostaglandin J2 (15dPGJ.sub.2) (3 pM)
as a positive control, compound 11 (3, 10 or 30 pM), or
rosiglitazone (3, 10 or 30 .mu.M) for 1 hr. at 37.degree. C. Then,
cells were treated with or without LPS (10 .mu.g/mL) plus IFN-gamma
(10 U/mL) for 5 min. or 15 min. at 37.degree. C. Cells were then
lysed, and the cell lysates (27 .mu.g/lane) were separated by
electrophoresis on a 4-20% polyacrylamnide gel, blotted onto a
nitrocellulose membrane and probed with anti-phospho-IkappaB
antibody. The results revealed that compound 11, but not
rosiglitazone, exhibited dose-dependent inhibition of the
phosphorylation of IkappaB.
[0151] To further confirm the ability of Compound 11 to inhibit the
activation of NF-kB, the production of free p65 (activated) NF-kB
in LPS-stimulated cells was measured. RAW cells were seeded at
5.times.10.sup.5/well in 6-well plates at 37.degree. C. overnight
in 10% FBS complete medium. Cells were washed 2.times. with 0.5%
FBS medium and then pretreated with 10 .mu.M of Compound 11,
rosiglitazone, or 15dPGJ.sub.2 at 37.degree. C. for 1 hr. After
pretreatment, cells were incubated with 0.5% FBS medium or
stimulated with 1 .mu.g/ml LPS at 37.degree. C. for 15 min. After
being washed 3.times. with cold PBS, cells were lysed to generate
whole cell lysates. The protein concentrations were determined and
5 .mu.g protein of whole cell lysate was used to determine the
NF-kB p65 activity for each sample using a commercial ELISA kit
(Active Motif, Carlsbad, Calif.). In this ELISA, micro-well plates
are coated with an oligonucleotide containing the consensus binding
site for NF-kB. After addition of the cell lysate, DNA-bound
transcription factor is detected using anti-p65 antibodies. Two
wells of each sample were assayed and the mean of two ELISA
readings for each sample was determined. The specificity of the
binding was checked by subsequent addition of 20 pmol of wild-type
consensus oligonucleotide to each well.
[0152] As shown in FIG. 15, both Compound 11 and 15dPGJ.sub.2
inhibited the activation of NF-kB. By contrast, rosiglitazone, a
strong agonist of PPAR-gamma, did not inhibit the production of p65
transcription factor.
[0153] FIGS. 13A-D illustrate the suppression of collagen-induced
arthritis by treatment with Compound 11. Arthritis was induced by
intradermal administration of collagen (100 .mu.g/mouse) in
complete adjuvant in make DBA/1 Lac mice of 7 weeks. The booster
(100 .mu.g/mouse) immunization in incomplete adjuvant was given
subcutaneously on Day-21. Two days later when arthritic scores were
around 1, the animals were divided into two groups. One group
received 50 mg/kg dose of Compound 11 orally for 17 days daily. The
second group received 10% PEG in water and was used as a vehicle
treated group. Body weight (FIG. 13A), Clinical score (FIG. 13B),
Joints affected (FIG. 13C) and Paw thickness (FIG. 13D) were
monitored 24 hours after the drug administration at different time
intervals. As shown in the Figures, the mice treated with Compound
11 showed significantly lower clinical scores, joints affected and
paw thickness when compared to the vehicle treated group. There was
no change in body weight between the vehicle and the treatment
groups.
[0154] Experimental allergic encephalomyelitis (EAE) is an
autoimmune demyelinating inflammatory disease of the central
nervous system. EAE exhibits many of the clinical and pathological
manifestations of human multiple sclerosis (MS), and it serves as
an animal model to test potential therapeutic agents for MS
(Scolding et al, Prog Neurobiol, 43:143-73, 2000). FIG. 14
illustrates the suppression of EAE by Compound 11. Active EAE was
induced in SJL-/J mice essentially according to the method of Owens
and Sriram (Neurol Clin, 13:51-73, 1995). Naive mice were immunized
subcutaneously with 400 pg each of mouse spinal cord homogenate in
complete Freund's adjuvant on day 0 and day 7. Mice were then
treated once daily by subcutaneous injection with 50 .mu.g or 200
.mu.g of compound 11 or with vehicle only. Paralysis was graded
according to the numeric scale indicated. As evidenced by the
dramatic reduction in clinical score shown in FIG. 14, treatment
with the 200 .mu.g dose of compound 11 was highly effective in
ameliorating EAE. It will be evident from the above that the
compounds according to the present invention, as represented by
compound 11, not only lower blood glucose level, triglyceride
level, free fatty acid level, glycohemoglobin and serum insulin,
but also raise the leptin level while showing no significant
increase in body weight or liver toxicity. The compounds also
inhibit TNF-alpha, IL-6, IL-1-beta production and COX-2 activity in
vitro and, as shown by FIGS. 13A-13D and 14, the compounds can be
used to suppress arthritis and potentially to treat multiple
sclerosis, respectively. The properties demonstrated above indicate
that the compounds of the invention should be useful in the
treatment of disorders associated with insulin resistance,
hyperlipidemia, coronary artery disease and peripheral vascular
disease and for the treatment of inflammation, inflammatory
diseases, immunological diseases and cancer, especially those
mediated by cytokines and cyclooxygenase.
[0155] While the invention has been exemplified above by reference
to the preparation and use of compound 11, it will be understood
that the invention is of broader application consistent with the
scope of compounds represented by formula 1. This includes, for
example, compound 10, which is not only useful as an intermediate
for preparing compound 11 as shown but also demonstrates useful
biological activity of its own consistent with the activities of
compound 11.
[0156] The synthesis of other compounds representative of the scope
of the invention is illustrated by the examples which follow.
EXAMPLE 2
3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-phenoxy]-
-phenyl}-acrylamide (24)
[0157] Compound 24, which may be represented by the formula:
##STR23## was prepared as follows from compound 14. A clean dry
flask with stirbar was charged with compound 14 (0.423 g, 0.837
mmol) and dry DMF (10 mL). Then with stirring carbonyldiimidazole
(0.271 g, 1.67 mmol) was added and the reaction was heated to
60.degree. C. for 1 h. while vented through an oil-bubbler. The
reaction mixture was then cooled to 0.degree. C. and 2M ammonia in
methanol (2.1 mL, 4.2 mmol) was added. The reaction was worked up
by partitioning the mixture with 10% citric acid (10 mL), ethyl
acetate (50 mL), and water (40 mL). The organic phase was then
rinsed sequentially with water (2.times.30 mL), brine (1.times.20
mL) and dried with anhydrous MgSO.sub.4. Concentration of the
organics afforded crude product. The crude product was purified by
silica gel chromatography using ethyl acetate-hexanes (1:1)
containing 1% acetic acid to ethyl acetate-hexanes (3:2) containing
1% acetic acid gradient elution. Concentration of the appropriate
fractions yielded 200 mg (47%) of the white-light yellow primary
amide as a solid. Analysis: .sup.1H NMR, 400 MHz (DMSO-d.sub.6):
.delta.12.06 (br, 1 H), 7.40 (s, 1 H), 7.34 (br, 1 H), 7.27 (d,
J=8.4 Hz, 2H), 7.18 (d, J=8.8 Hz, 2H), 7.05 (d, J=9.2 Hz, 2H),
6.98(d, J=8.4 Hz, 2H), 6.93 (br, 1 H), 6.36(m, 1 H), 6.20 (s, 1 H),
6.19 (s, 1 H), 4.91(dd, J=4.0 Hz, 1 H), 3.57 (s, 6H), 3.12 (dd,
J=9.2 Hz, 1 H).
EXAMPLE 3
3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5ylmethyl)-phenoxy]--
phenyl}-N,N-dimethylacrylamide (25)
[0158] Compound 25, represented by the formula: ##STR24## was
prepared as follows: A clean dry flask with stirbar was charged
with compound 14 (0.422 g, 0.835 mmol) and dry DMF (1 mL). Then
with stirring carbonyldiimidazole (0.271 g, 1.67 mmol) was added
and the reaction was heated to 60.degree. C. for 1 h. while vented
through an oil-bubbler. The reaction mixture was then cooled to
0.degree. C. and a 2M dimethylamine in THF (2.1 mL, 4.2 mmol)
solution was added. The reaction was worked up by partitioning the
mixture with 10% citric acid (10 mL), ethyl acetate (50 mL), water
(40 mL). The organic phase was then rinsed sequentially with water
(2.times.30 mL), brine (1.times.20 mL) and dried with anhydrous
MgSO.sub.4. Concentration of the organics afforded crude product.
The crude product was purified by silica gel chromatography using
ethyl acetate-hexanes (3:2) containing 1% acetic acid elution.
Concentration of the fractions yielded 381 mg (86%) of the
off-white tertiary dimethylamide as a solid. Analysis: .sup.1H NMR,
400 MHz (DMSO-d.sub.6): .delta.11.97 (br, 1H), 7.29 (d, J=8.8 Hz,
2H), 7.27 (d, J=8 Hz, 2H), 6.99 (d, J=8.8 Hz), 6.95(d, J=8.8 Hz,
2H), 6.57 (s, 1 H), 6.35 (m, 1 H), 6.29 (s, 1 H), 6.28 (s, 1 H),
4.91(dd, J=4.4 Hz, 1 H), 3.58 (s, 6H), 3.12 (dd, J=9.2 Hz, 1 H),
3.05 (br, 3H), 2.91 (s, 3H).
EXAMPLE 4
3-(3,5-Dimethoxyphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-phenoxy]-
-phenyl}-N-methoxy,-N-methylacrylamide (compound 26)
[0159] Compound 26 may be structurally shown as follows: ##STR25##
was prepared as follows. A clean dry flask with stirbar was charged
with compound 14 (0.450 g, 0.890 mmol) and dry DMF (1 mL). Then,
with stirring, carbonyldiimidazole (0.29 g, 1.78 mmol) was added
and the reaction was heated to 60.degree. C. for 1 h. while vented
through an oil-bubbler. The reaction mixture was then cooled to
0.degree. C. and N-methyl-N-methoxyhydroxylamine hydrochloride
(0.434 g, 4.45 mmol) in water (1 mL) and triethylamine (0.62 mL)
was added and stirred overnight. The reaction was worked up by
partitioning the mixture with 10% citric acid (10 mL), ethyl
acetate (50 mL), and water (40 mL). The organic phase was then
rinsed sequentially with water (2.times.30 mL), brine (1.times.20
mL) and dried with anhydrous magnesium sulfate. Concentration of
the organics afforded crude product. The crude product was purified
by silica gel chromatography using ethyl acetate-chloroform (1:5)
elution. Concentration of the appropriate fractions yielded 400 mg
(82%) of the off-white tertiary N-methyl-N-methoxyamide as a solid.
Analysis: .sup.1H NMR, 400 MHz (DMSO-d6): .delta.12.06 (br, 1 H),
7.27 (d, J=9.2 Hz, 2H), 7.26 (d, J=8.8 Hz, 2H), 7.00 (d, J=8.8 Hz),
6.95 (d, J=8.4 Hz, 2H), 6.57 (s, 1 H), 6.35 (m, 1 H), 6.29 (s, 1
H), 6.28 (s, 1 H), 4.91(dd, J=4.4 Hz, 1 H), 3.58 (s, 6H), 3.12 (dd,
J=9.2 Hz, 1 H), 3.05 (br, 3H), 2.91 (s, 3H).
EXAMPLE 5
[0160] The syntheses shown in this example are illustrated in
Scheme 6.
[0161] 2-(4-Acetoxyphenyl)-3-p-tolylacrylic acid (37). To a mixture
of (4-hydroxyphenyl)-acetic acid (18.3 g, 120.3 mmol) and
4-methylbenzaldehyde (12.0 g, 100 mmol) in 250 mL acetic anhydride
was added potassium carbonate (11.9 g, 121.2 mmol). The reaction
mixture was stirred at 80.degree. C. for 16 h before it was cooled
to room temperature. To the mixture was added 100 mL H.sub.2O, 5%
HCl in water to pH 1 and 200 mL ethyl acetate. The mixture was then
heated to 80.degree. C. until all ethyl acetate was evaporated. The
precipitate was filtered and washed with water and hexane. The
filter cake was recrystallized out of toluene, filtered, washed
with hexane and dried under vacuum to yield a pale yellow powder
(20.16 g, 68.1%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.62
(br s, 1H), 7.74 (s, 1H), 7.19 (d, J=8.8 Hz, 2H), 7.13 (d, J=8.8
Hz, 2H), 7.01 (d, J=8.0 Hz, 2H), 6.94 (d, J=8.4 Hz, 2H), 2.29 (s,
3H), 2.23 (s, 3H).
[0162] 2-(4-Hydroxyphenyl)-3-p-tolylacrylic acid (38). To a
solution of compound 37 (20.16 g, 68.1 mmol) in 100 mL THF was
added a solution of lithium hydroxide (5.7 g, 237.5 mmol) in 100 mL
water. The reaction was allowed to stir at room temperature for 16
h after which 5% HCl in water was added to pH=1. The yellow solid
was filtered and recrystallized out of toluene, washed with hexane
and dried under vacuum to yield a white solid (14.27 g, 82.5%).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.46 (br s, 1H), 9.47
(br s, 1H), 7.63 (s, 1H), 7.02 (d, J=8.0 Hz, 2H), 6.97 (d, J=8.4
Hz, 2H), 6.93 (d, J=8.4 Hz, 2H), 6.74 (d, J=8.8 Hz, 2H), 2.22 (s,
3H).
[0163] 2-[4-(4-Formylphenoxy)-phenyl]-3-p-tolylacrylic acid (39).
To a solution of 38 (7.27 g, 28.6 mmol) and potassium carbonate
(8.68 g, 62.9 mmol) in 200 mL N,N-dimethylacetamide was added
4-fluorobenzaldehyde (3.9 mL, 36.4 mmol). The reaction mixture was
heated to 190.degree. C. for 1.5 h under argon then cooled to room
temperature. The addition of 5% HCl in water to pH 1 resulted in
the product separating out as oil. Approximately 50 mL ethyl
acetate was added to the mixture which is allowed to stir 16 h. The
solid was collected and recrystallized out of toluene, rinsed with
hexane and dried under vacuum to yield a white powder (8.1 g,
79.0%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.71 (s, 1H),
9.94 (s, 1H), 7.97 (d, J=8.8 Hz, 2H), 7.76 (s, 1H), 7.25 (d, J=8.4
Hz, 2H), 7.18 (d, J=6.8 Hz, 2H), 7.16 (d, J=6.4 Hz, 2H), 7.07 (d,
J=8.0 Hz, 2H), 6.98 (d, J=8.4 Hz, 2H), 2.25 (s, 3H).
[0164]
2-{4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-phenoxy]-phenyl}-3--
p-tolylacrylic acid (40). To a solution of 39 (4.0 g, 11.2 mmol),
thiazolidine-2,4-dione (1.31 g, 11.2 mmol), and benzoic acid (1.64
g, 13.4 mmol) in 100 mL toluene was added piperidine (1.66 mL, 16.8
mmol). The mixture was vigorously refluxed with Dean Stark
apparatus for 1.5 h under argon then cooled to room temperature. 5%
HCl was added to pH 1. The solid was filtered, recrystallized out
of toluene, filtered, and washed with hexane before drying under
vacuum to a yellow solid (quantitative). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 12.70 (br s, 1H), 12.59 (br s, 1H), 7.80 (s,
1H), 7.75 (s, 1H), 7.67 (d, J=8.8 Hz, 2H), 7.22 (d, J=9.2 Hz, 2H),
7.18 (d, J=7.6 Hz, 2H), 7.12 (d, J=9.2 Hz, 2H), 7.07 (d, J=8.0 Hz,
2H), 6.99 (d, J=8.0 Hz, 2H), 2.25 (s, 3H).
[0165]
2-{4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)-phenoxy]-phenyl}-3-p-tol-
ylacrylic acid (41). To a solution of 40 (1.0 g, 2.2 mmol) and
ammonium formate (8.32 g, 132 mmol) in 25 mL glacial acetic acid
was added 5% Pd/C (1.0 g). The mixture was refluxed for 7.5 h,
cooled to room temperature and filtered over Celite. The mixture
was concentrated in vacuum then added to 200 mL water. The product
was filtered and washed with hexanes. The solid was recrystallized
out of toluene, cooled to room temperature and sonicated until the
solid was observed. The mixture was then stirred at room
temperature for 16 h. The precipitate was collected and washed with
hexanes to yield a white solid (0.609 g, 59.1 %). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 12.65 (s, 1H), 12.05 (br s, 1H), 7.72
(s, 1H), 7.30 (d, J=8.8 Hz, 2H), 7.16 (d, J=8.4 Hz, 2H), 7.05 (d,
J=8.4 Hz, 2H), 7.01 (d, J=8.4 Hz, 2H), 6.99 (d, J=9.2 Hz, 2H), 6.98
(d, J=8.0 Hz, 2H), 4.92 (dd, J=4.4 and 9.6 Hz, 1H), 3.38 (dd, J=4.4
and 14.0 Hz, 1H), 3.21 (dd, J=9.2 and 14.0 Hz, 1H), 2.24 (s,
H).
[0166]
2-{4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)-phenoxy]-phenyl}-p-tolyl-
acrylic acid methyl ester (42). To a mixture of 41 (0.1 g, 0.218
mmol) and BOP [Castro's Reagent,
Benzotriazole-1-yl-oxy-tris-(dimethylamino)-phosphonium
hexafluorophosphate] (0.144 g, 0.326 mmol) in 5 mL dichloromethane
was added triethylamine (0.067 mL, 0.477 mmol). The mixture was
stirred for 1 h, and then sodium methoxide (0.5 M solution in
methanol, 0.07 mL, 0.035 mmol) was added with 5 mL MeOH. The
reaction was allowed to stir at room temperature for 16 h. 5% HCl
was added to pH 0 and the mixture was extracted with 25 mL
dichloromethane three times. The combined organic layers were
washed with brine, dried over MgSO.sub.4, filtered and
concentrated. The residue was loaded onto silica gel column as a
solution in dichloromethane. The product was eluted with
hexanes-ethyl acetate (3:2). Fractions were concentrated in vacuum
to a white solid (0.037 g, 36.4%). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 12.05 (br s, 1H), 7.75 (s, 1H), 7.30 (d,
J=8.8 Hz, 2H), 7.18 (d, J=8.8 Hz, 2H), 7.06 (d, J=8.0 Hz, 2H), 7.02
(d, J=8.8 Hz, 2H), 6.99 (d, J=8.8 Hz, 2H), 6.98 (d, J=8.0 Hz, 2H),
4.91 (dd, J=4.8 and 9.6 Hz, 1H), 3.72 (s, 3H), 3.39 (dd, J=4.0 and
13.6 Hz, 1H), 3.13 (dd, J=9.2 and 14.0 Hz, 1H), 2.25 (s, 3H).
EXAMPLE 6
[0167] The syntheses shown in this example are illustrated in
Scheme 7.
[0168] 3,5-dimethylbenzaldehyde (43). To a mixture of
3,5-dimethylbenzoic acid (7.51 g, 50 mmol) and triethylamine (21
mL, 150 mmol) in dichloromethane (200 mL) was added BOP reagent
(22.11 g, 50 mmol). The solution was stirred at room temperature
for 20 min and then N,O-dimethylhydroxylamine hydrochloride (5.0 g,
50 mmol) was added. After an additional 10 min, triethylamine (7
mL, 50 mmol) was added and the mixture was stirred for another 0.5
h. The solvent was removed in vacuo and the mixture was redissolved
in ethyl acetate (300 mL), washed with 1N HCl (200 mL), 1N NaOH
(200 mL), water, brine then dried (MgSO.sub.4), filtered and
concentrated in vacuo to yield a colorless syrup (6.25 g). This
material was dissolved in THF (250 mL) and cooled to 0.degree. C.
under argon atmosphere. A solution of DIBAL [diisobutylaluminum
hydride] (1M in THF, 50 mL) was added to the stirring solution over
5 min. After 20 min of stirring, additional DIBAL (20 mL) was
added. After an additional 15 min, the reaction was quenched with
careful addition of 1N HCl (300 mL) and the product was extracted
into ethyl acetate (300 mL), washed with water (2.times.200 mL),
brine (200 mL), dried (MgSO.sub.4), filtered and concentrated in
vacuo to yield 43 (4.97 g, 74% overall).
[0169] 3-(3,5-Dimethylphenyl)-2-(4-hydroxyphenyl)-acrylic acid
(44). To a mixture of 43 (3.23 g, 24 mmol), 4-hydroxyphenylacetic
acid (3.66 g, 24 mmol) and potassium acetate (2.83 g, 28 mmol) was
added acetic anhydride (100 mL). The mixture was heated to reflux
for 4 h, cooled to room temperature, and then poured over water
(400 mL). After stirring for 1.5 h, a solid gum settled to the
bottom and the supernatant was decanted. To the residue was added
THF (100 mL) and 1N NaOH (150 mL) and the mixture was stirred for
30 min. The mixture was acidified with 1N HCl (200 mL) and the
product was extracted into ethyl acetate (300 mL), washed with
water (300 mL), brine (300 mL), dried (MgSO.sub.4), filtered and
concentrated in vacuo. The crude solid was crystallized in toluene
to yield 2.65 g (42%) of a pale yellow solid 44.
[0170]
3-(3,5-Dimethylphenyl)-2-[4-(4-formylphenoxy)-phenyl]-acrylic acid
(45). To a solution of 44 (2.65 g, 10 mmol) in DMF (20 mL) was
added sodium hydride (60% dispersion in mineral oil, 0.88 g, 22
mmol). After gas evolution ceased 4-fluorobenzaldehyde (1.60 g, 15
mmol) was added and the reaction was stirred for 16 h. The mixture
was poured over 10% citric acid (100 mL), after which a bright
yellow solid formed. The solid was washed with water and then the
wet solid was azeotroped and recrystallized from toluene to yield
2.97 g (80%) of a yellow solid 45.
[0171]
3-(3,5-Dimethylphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylidenemeth-
yl)-phenoxy]-phenyl}-acrylic acid (46). In a 100 mL round-bottomed
flask equipped with a Dean-Stark apparatus, a mixture of 45 (1.55
g, 4.2 mmol), 2,4-thiazolidinedione (0.5 g, 4.2 mmol), benzoic acid
(0.62 g, 5.0 mmol) and piperidine (0.62 mL, 6.3 mmol) was
azeotroped in toluene (60 mL) for 45 min under vigorous reflux. The
reaction mixture was cooled then poured over 10% citric acid (40
mL) and stirred until a bright yellow solid formed. The solid was
filtered and washed with water and the wet solid was azeotroped and
recrystallized from toluene to yield 1.83 g (93%) of 46. .sup.1H
NMR (400 MHz, DMSO-d.sub.6): .delta. 12.72 (br s, 1H), 12.57 (br s,
1H), 7.77 (s, 1H), 7.70 (s, 1H), 7.63 (d, J=8.8 Hz, 2H), 7.22 (d,
J=8.4 Hz, 2H), 7.14 (d, J=8.4 Hz, 2H), 7.12 (d, J=8.8 Hz, 2H), 6.92
(s, 1H), 6.69 (s, 2H), 2.16 (s, 6H).
[0172]
3-(3,5-Dimethylphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-p-
henoxy]-phenyl}-acrylic acid (47). A mixture of 46 (1.83 g, 3.9
mmol), ammonium formate (4.90 g, 78 mmol) and 10% Pd on alumina
(2.0 g) was refluxed for 15 h. The reaction mixture was cooled to
room temperature and the catalyst was filtered off. Product was
separated out with addition of water and the solid was filtered.
The wet solid was azeotroped and recrystallized from toluene to
yield 0.81 g (44%) 5. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
12.68 (br s, 1H), 12.05 (br s, 1H), 7.68 (s, 1H), 7.27 (d, J=8.8
Hz, 2H), 7.16 (d, J=8.4 Hz, 2H), 7.02 (d, J=8.4 Hz, 2H), 6.96 (d,
J=8.8 Hz, 2H), 6.90 (s, 1H), 6.62 (s, 2H), 4.92 (dd, J=8.8 and 4.4
Hz, 1H), 3.37 (dd, J=14.0 and 4.4 Hz, 1H), 3.12 (dd, J=14.0 and 8.8
Hz, 1H), 2.12 (s, 6H).
[0173]
3-(3,5-Dimethylphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-p-
henoxy]-phenyl}-acrylic acid benztriazol-1-yl ester (48). To a
mixture of 47 (0.81 g, 1.7 mmol) and N,N-diisopropylethylamine
(0.33 mL, 1.9 mmol) in dichloromethane (20 mL) was added BOP
reagent (0.76 g, 1.7 mmol). After 45 min of stirring at room
temperature, the mixture was diluted with ethyl acetate (150 mL)
then washed with 1N HCl (100 mL), water (100 mL), brine (100 mL),
dried (MgSO.sub.4), filtered and concentrated in vacuo. The crude
product was purified by flash chromatography using hexanes:ethyl
acetate (3:2). The solid obtained after concentration was further
triturated with hexanes:ethyl acetate (4:1) to yield 0.75 g (76%)
of 48.
[0174]
3-(3,5-Dimethylphenyl)-2-{4-[4-(2,4-dioxothiazolidin-5-ylmethyl)-p-
henoxy]-phenyl}-acrylic acid methyl ester (49). To a solution of 48
(240 mg, 0.4 mmol) in methanol (10 mL) was added sodium methoxide
(0.5 N in methanol, 2 mL). After 10 min the mixture was diluted
with 1N HCl (2 mL) and the product was extracted into ethyl acetate
(50 mL), washed with water (50 mL), brine (50 mL), dried
(MgSO.sub.4), filtered and concentrated in vacuo. The crude
material was purified by flash chromatography using hexanes:ethyl
acetate (3:2) to yield 49 (122 mg, 62%) as a white foam. .sup.1H
NMR (400 MHz, DMSO-d.sub.6): .delta. 12.10 (br s, 1H), 7.71 (s,
1H), 7.28 (d, J=8.8 Hz, 2H), 7.18 (d, J=8.8 Hz, 2H), 7.03 (d, J=8.8
Hz, 2H), 6.97 (d, J=8.8 Hz, 2H), 6.92 (s, 1H), 6.68 (s, 2H), 4.91
(dd, J=8.8 and 4.4 Hz, 1H), 3.73 (s, 3H), 3.37 (dd, J=14.0 and 4.4
Hz, 1H), 3.12 (dd, J=14.0 and 8.8 Hz, 1H), 2.12 (s, 6H).
[0175]
5-(4-{4-[2-(3,5-Dimethylphenyl)-1-(morpholine-4-carbonyl)-vinyl]-p-
henoxy}-benzyl)-thiazolidine-2,4-dione (50). To a suspension of 48
(116 mg, 0.2 mmol) in dichloromethane (5 mL) at room temperature
was added morpholine (87 .mu.L, 1 mmol). Solution became clear.
After 10 min the mixture was treated with 10% citric acid (4 mL).
The dichloromethane layer was dried (MgSO.sub.4) and directly
loaded onto a column which was eluted with hexanes:ethyl acetate
(2:3) to yield 50 (104 mg, 86%) as a white foam. .sup.1H NMR (400
MHz, DMSO-d.sub.6): .delta. 12.10 (br s, 1H), 7.27 (d, J=8.8 Hz,
2H), 7.26 (d, J=8.8 Hz, 2H), 6.98 (d, J=8.8 Hz, 2H), 6.96 (d, J=8.8
Hz, 2H), 6.85 (s, 1H), 6.72 (s, 2H), 6.61 (s, 1H), 4.91 (dd, J=8.8
and 4.4 Hz, 1H), 3.59 (bs, 8H), 3.37 (dd, J=14.0 and 4.4 Hz, 1H),
3.12 (dd, J=14.0 and 8.8 Hz, 1H), 2.13 (s, 6H).
EXAMPLE 7
Synthesis of
5-(4-{4-[2-(4-methoxyphenyl)-vinyl]-phenoxy}-benzyl)-thiazolidine-2,4-dio-
ne (54) (see Scheme 8)
[0176] 5-(4-Hydroxybenzylidene)-thiazolidine-2,4-dione (51). To a
mixture of 4-hydroxybenzaldehyde (3.67 g, 30 mmol),
2,4-thiazolidinedione (3.51 g, 30 mmol) and benzoic acid (4.40 g,
36 mmol) in toluene (100 mL) was added piperidine (4.5 mL, 45 mmol)
and the mixture was equipped with a Dean Stark apparatus and
brought to a vigorous reflux. After 45 min the mixture was cooled
in an ice bath and the supernatant was decanted. The bright yellow
solid was made into a suspension by the addition of glacial acetic
acid (100 mL) and filtered through a Buchner funnel to yield a pale
yellow solid (6.00g, 90%). .sup.1H NMR: (400 MHz, DMSO-d.sub.6):
.delta. 12.45 (bs, 1H), 10.30 (s, 1H), 7.70 (s, 1H), 7.45 (d, J=8.8
Hz, 2H), 6.91 (d, J=8.8 Hz, 2H).
[0177] 5-(4-Hydroxybenzyl)-thiazolidine-2,4-dione (52). To a
suspension of 51 (6.00 g, 27 mmol) in glacial acetic acid (100 mL)
was added ammonium formate (6.27 g, 100 mmol) and 10% Pd on carbon
(5.80 g) and the mixture was heated to vigorous reflux for 16 h.
The mixture was cooled to room temperature then filtered through
Celite. Most of the acetic acid was removed in vacuo then the crude
product was dissolved in ethyl acetate (250 mL), washed with water
(2.times.250 mL) then brine (250 mL). The organic layer was dried
(MgSO.sub.4), filtered and concentrated in vacuo to yield a beige
solid (5.35 g, 85%). .sup.1H NMR: (400 MHz, DMSO-d.sub.6): .delta.
11.98 (bs, 1H), 9.32 (s, 1H), 7.02 (d, J=8.4 Hz, 2H), 6.68 (d,
J=8.4 Hz, 2H), 4.82 (dd, J=8.4 and 4.0 Hz, 1H), 3.25 (dd, J=14.4
and 4.4 Hz, 1H), 2.99 (dd, J=14.0 and 9.2 Hz, 1H).
[0178] 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)-phenoxy]-benzaldehyde
(53). To a solution of 52 (5.35 g, 24 mmol) in DMF (150 mL) was
added 4-fluorobenzaldehyde (3.00 g, 24 mmol) and Cs.sub.2CO.sub.3
(20 g, 62 mmol) and the mixture was heated to 100.degree. C. for 2
h. The mixture was poured over vigorously string 10% citric acid
(200 mL) and ethyl acetate (200 mL). The organic layer was washed
with water (300 mL), brine (300 mL), dried (MgSO.sub.4), filtered
and concentrated in vacuo. The crude product was triturated in
hexanes-ethyl acetate (2:1) to yield a white solid (5.15 g, 65%).
.sup.1H NMR: (400 MHz, DMSO-d.sub.6): .delta. 12.07 (bs, 1H), 9.92
(s, 1H), 7.92 (d, J=8.8 Hz, 2H), 7.35 (d, J=8.4 Hz, 2H), 7.12 (d,
J=8.4 Hz, 2H), 7.09 (d, J=8.4 Hz, 2H) 4.94 (dd, J=8.4 and 4.4 Hz,
1H), 3.41 (dd, J=14.4 and 4.4 Hz, 1H), 3.17 (dd, J=14.4 and 9.2 Hz,
1H).
[0179]
5-(4-{4-[2-(4-Methoxyphenyl)-vinyl]-phenoxy}-benzyl)-thiazolidine--
2,4-dione (54). To a suspension of
4-methoxybenzyltriphenylphosphonium-chloride (419 mg, 1.0 mmol) in
THF (10 mL) at 0.degree. C. was added solid potassium tert-butoxide
(224 mg, 2.0 mmol). The resultant orange-red solution was stirred
for 15 min at 0.degree. C. then cooled to -45.degree. C. Solid
compound 53 (327 mg, 1.0 mmol) was added and the reaction mixture
stirred for 30 min at this temperature. To this pale yellow
solution glacial acetic acid (60 .mu.L, 1 mmol) was added and the
solvent was removed in vacuo. The crude product was suspended in
dichloromethane, adsorbed onto silica gel and purified by flash
chromatography using hexanes-ethyl acetate (7:3) to yield a white
solid (143 mg, 33%) after drying under high vacuum. .sup.1H NMR:
(400 MHz, DMSO-d.sub.6): .delta. 12.04 (bs, 1H), 7.27 (d, J=8.0 Hz,
2H), 7.24 (d, J=8.4 Hz, 2H), 7.18 (d, J=8.4 Hz, 2H), 6.97 (d, J=8.4
Hz, 2H), 6.88 (d, J=8.8 Hz, 2H), 6.84 (d, J=8.8 Hz, 2H), 6.53 (d,
J=12.4 Hz, 2H), 6.48 (d, J=12.0 Hz, 2H), 4.90 (dd, J=9.2 and 4.4
Hz, 1H), 3.36 (dd, J=14.0 and 4.4Hz, 1H), 3.11 (dd, J=14.0 and 8.8
Hz, 1H).
EXAMPLE 8
[0180] ##STR26##
[0181]
5-(4-{4-[2-(3,5-Dimethoxyphenyl)-vinyl]-phenoxy}-benzyl)-thiazolid-
ine-2,4-dione (55). To a suspension of
3,5-dimethoxybenzyltriphenylphosphonium bromide (0.82 g, 2.0 mmol)
in THF (10 mL) at 0.degree. C. was added solid potassium
tert-butoxide (224 mg, 2.0 mmol). The resultant red solution was
stirred for 15 min at 0.degree. C. then cooled to -.sub.78.degree.
C. Solid 53 (0.3 mg, 0.90 mmol) was added and the reaction was
allowed to warm to room temperature. After 30 min 10% citric acid
(50 mL) was added and the mixture was partitioned between water (50
mL) and ethyl acetate (75 mL). The organic layer was washed with
water (50 mL) and brine (50 mL) then dried (MgSO.sub.4), filtered
and concentrated in vacuo. The crude product was purified by flash
chromatography using hexanes-ethyl acetate (7:3) to yield 55 as a
slightly opaque film (25 mg, 6%) after concentration and drying
under high vacuum. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
12.04 (b s, 1H), 7.26 (d, J=8.8 Hz, 2H), 7.25 (d, J=8.8 Hz, 2H),
6.93 (d, J=8.4 Hz, 2H), 6.91 (d, J=8.4 Hz, 2H), 6.61 (d, J=12.4 Hz,
1H), 6.53 (d, J=12.0 Hz, 1H), 6.39 (d, J=2.4 Hz, 1H), 6.36 (t,
J=2.4 Hz, 1H), 4.90 (dd, J=9.2 and 4.4 Hz, 1H), 3.62 (s, 6H), 3.36
(dd, J=14.0 and 4.4 Hz, 1H), 3.11 (dd, J=14.4, 8.8 Hz, 1H).
EXAMPLE 9
[0182] The syntheses shown in this example are illustrated in
Scheme 9.
[0183] 4'-Methoxybiphenyl-3-ol (56). To a solution of
3-hydroxyphenylboronic acid (1.00 g, 7.3 mmol) in 2M aqueous
K.sub.2CO.sub.3 (4 mL) was added a solution of 4-iodoanisole (1.70
g, 7.3 mmol) in acetone (4 mL). A homogeneous mixture was obtained
by sequential addition of water (60 mL) and acetone (30 mL).
Catalytic Pd(OAc).sub.2 was added (160 mg, 0.73 mmol) and the
mixture was stirred for 10 min at room temperature. The acetone was
removed from the dark brown solution in vacuo and the resultant
aqueous mixture was acidified with 1N HCl (20 mL) and extracted
with ethyl acetate (75 mL). The organic layer was washed with water
(50 mL), brine (50 mL), dried (MgSO.sub.4), filtered and
concentrated in vacuo. The crude product was suspended in
dichloromethane and adsorbed onto silica gel and purified by flash
chromatography using hexanes-ethyl acetate (3:1) to yield 1.20 g
(82%) 56 as a white solid after solvent removal.
[0184] 4-(4'-Methoxybiphenyl-3-yloxy)-benzaldehyde (57). To a
solution of 56 (1.20 g, 6.0 mmol) and 4-fluorobenzaldehyde (745 mg,
6.0 mmol) in DMF (25 mL) was added cesium carbonate (3.90 g, 12.0
mmol). After stirring for 1 h at 100.degree. C., the product was
partitioned between ethyl acetate (100 mL) and water (100 mL). The
organic layer was washed with water (100 mL), brine (100 mL), dried
(MgSO.sub.4), filtered and concentrated in vacuo. The crude product
was dissolved in dichloromethane and adsorbed onto silica gel and
purified by flash chromatography using hexanes-ethyl acetate (6:1)
to yield 57 (1.23 g, 67%) as a white solid after solvent
removal.
[0185]
5-[4-(4'-Methoxybiphenyl-3-yloxy)-benzylidene]-thiazolidine-2,4-di-
one (58). A mixture of 57 (1.23 g, 4.0 mmol), 2,4-thiazolidinedione
(0.48 g, 4.0 mmol), benzoic acid (0.60 g, 4.8 mmol) and piperidine
(0.61 mL, 6.0 mmol) in toluene (30 mL) was heated to a vigorous
reflux until most of the solvent had evaporated. A yellow
suspension was achieved by addition of acetic acid (25 mL) followed
by sonication. Filtration of the suspension followed by washing
with acetic acid (10 mL) yielded 58 (1.25 g, 75%) after filtration
and oven drying. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 12.57
(br s, 1H), 7.78 (s, 1H), 7.63 (d, J=8.8 Hz, 2H), 7.62 (d, J=8.8
Hz, 2H), 7.49 (d, J=5.2 Hz, 2H), 7.36 (t, J=1.6 Hz, 1H), 7.16 (d,
J=8.8 Hz, 2H), 7.03 (m, 1H), 7.01 (d, J=8.8 Hz, 2H), 3.7 (s,
3H).
[0186]
5-[4-(4'-Methoxybiphenyl-3-yloxy)-benzyl]-thiazolidine-2,4-dione
(59). To a suspension of 58 (1.25 g, 3.0 mmol) in acetic acid (30
mL) was added ammonium formate (1.50 g, 24 mmol) and 10% Pd on
carbon (1.30 g). After vigorous refluxing for 16 h, the mixture was
filtered through Celite, which was subsequently washed with ethyl
acetate (100 mL). The mixture was washed with water (2.times.75
mL), 1N NaHCO.sub.3 (50 mL), brine (75 mL) then dried (MgSO.sub.4),
filtered and concentrated in vacuo. The crude product was dissolved
in dichloromethane, adsorbed onto silica gel and purified by flash
chromatography using hexanes:ethyl acetate (3:7) to yield a white
solid (0.48 g, 38%) after concentration then trituration and
filtration from hexanes-ethyl acetate (4:1). .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 12.04 (br s, 1H), 7.58 (d, J=8.8 Hz, 2H),
7.43 (t, J=7.6 Hz, 1H), 7.39 (dt, J=8.0, 1.6 Hz, 1H), 7.27 (d,
J=8.8 Hz, 2H), 7.22 (t, J=2.0 Hz, 1H), 7.01 (d, J=8.8 Hz, 2H), 7.00
(d, J=9.2 Hz, 2H), 4.91 (dd, J=8.8 and 4.4 Hz, 1H), 3.79 (s, 3H),
3.37 ( dd, J=14.4 and 4.4 Hz, 1H), 3.13 (dd, J=14.4 and 8.8 Hz,
1H).
EXAMPLE 10
[0187] ##STR27##
[0188]
5-[4-(3',5'-Diethoxybiphenyl-3-yloxy)-benzyl]-thiazolidine-2,4-dio-
ne (61). First, 5-[4-(2',440
-dimethoxybiphenyl-3-yloxy)-benzylidene]-thiazolidine-2,4-dione
(60), was synthesized using a scheme analogous to the synthesis of
58 depicted in Scheme 9. To a suspension of 60 (0.28 g, 0.65 mmol)
in acetonitrile (20 mL) was added triethylamine (180 .mu.L, 1.3
mmol), ammonium formate (0.41 g, 6.5 mmol) and 10% Pd on alumina
(0.5 g). After refluxing for 2.5 h, the mixture was filtered
through Celite, which was subsequently washed with ethyl acetate
(60 mL). The mixture was acidified with 10% citric acid (50 mL)
then washed with water (50 mL), brine (50 mL) then dried
(MgSO.sub.4), filtered and concentrated in vacuo. The crude product
was purified by flash chromatography using hexanes:ethyl acetate
(3:7) to yield a light film (59 mg, 21%) after concentration and
drying under high vacuum. .sup.1H NMR (400 MHz, DMSO-d.sub.6):
.delta. 12.04 (br s, 1H), 7.38 (t, J=8.4 Hz, 1H), 7.27 (d, J=9.2
Hz, 2H), 7.22 (d, J=8.4, 1H), 7.20 (dt, J=8.4 and 0.8 Hz, 1H), 7.06
(t, J=2.0 Hz, 1H), 7.00 (d, J=8.8 Hz, 2H), 6.90 (ddd, J=8.0 2.4 and
0.8 Hz, 1H), 6.64 (d, J=2.0 Hz, 1H), 6.60 (dd, J=8.4 and 2.4 Hz,
1H), 4.91 (dd, J=8.8 and 4.4 Hz, 1H), 3.79 (s, 6H), 3.37 ( dd,
J=14.4 and 4.4 Hz, 1H), 3.13 (dd, J=14.4 and 8.8 Hz, 1H).
EXAMPLE 11
Prevention of Cancellous Bone Loss in Adjuvant Induced Arthritic
(AIA) rats
[0189] Inflammatory arthritis results in significant peri-articular
bone loss due to activation of cytokines that activate osteoclast
activity. Thiazolidinediones (TZD) are insulin sensitizing agents
that may inhibit TNF-alpha production, an important factor leading
to bone loss in inflammatory arthritis The purpose of this
investigation was to demonstrate that the TZD compound 11 and
etanercept (p55 TNF soluble receptor) could prevent bone loss in
AIA rats. Arthritis was induced in male Lewis rats (Harlan, weight
150 g) by immunizing them with Freund's Complete Adjuvant
containing Mycobacterium butyricum (100 .mu.g) on the tail base.
When the arthritic symptoms began to appear (by day 12-14), the
animals were randomized to one of three treatment groups: Group I:
vehicle (20% PEG-400 in water, per oral gavage). Group II: compound
11 (50 mg/kg, per oral gavage once daily) Group III: etanercept
(1.67 mg/kg, intra-peritoneal once daily). Each limb was
individually scored from 0 (no change or normal) to 4 (marked
arthritis with swelling, erythema, nodules, deformation, rigidity)
by an observer blinded to the treatment groups. The body weights,
number of limbs affected and hind paw volume were also noted. On
day 11 AIA rats were sacrificed and the right femur and tibia
harvested, dissected and fixed in 70% EtOH. Cancellous bone volume
and microstructure of the right proximal tibia and distal femur
were accessed by MicroCT (.mu.CT-20, Scanco Medical, Bassersdorf,
Switzerland). Results of right proximal tibia are shown in the
following table. TABLE-US-00002 TABLE 2 BV/TV Tb.N Conn. Dens C.Th
SMI Groups (%) (l/mm) (l/mm3) (.mu.m) (0-3) Sham 21.4 .+-. 2.6 4.5
.+-. 0.4 70.5 .+-. 14.9 250 .+-. 24 2.3 .+-. 0.9 (n = 7) Vehicle
6.2 .+-. 4.0 2.0 .+-. 0.4 10.3 .+-. 15.3 167 .+-. 27 3.5 .+-. 0.3
(n = 12) Comp. 11 13.7 .+-. 4.7 2.6 .+-. 0.5 38.3 .+-. 19.7 186
.+-. 25 2.8 .+-. 0.3 (n = 9) Etanercept 12.9 .+-. 5.2 2.8 .+-. 0.5
31.9 .+-. 19.7 198 .+-. 25 2.8 .+-. 0.4 (n = 9)
[0190] In summary, significant cancellous bone loss and
microarchitecture changes occurred in AIA rats. However, in this
AIA model, nearly 50% less cancellous bone was lost in animals
treated with either compound 11 or etanercept. Therefore, compound
11 may be effective in reducing bone loss in inflammatory arthritis
and similar rapid bone loss states.
EXAMPLE 12
Glucose Uptake
[0191] Basal glucose uptake was measured in differentiated 3T3-L1
adipocytes following the protocol of Tafuri (6) with modifications.
Briefly, 3T3-L1 fibroblasts, obtained from ATCC (Manassas, Va.),
were differentiated to adipocytes by treating cells with porcine
insulin (1 .mu.g/ml for 4 days), dexamethasone (0.25 .mu.M for
first 2 days) and isobutyl methyl xanthine (IBMX, 0.5 mM for first
2 days) (all from Sigma Chemicals, St Louis, Mo.) following the
protocol of Frost and Lane (7). Differentiated adipocytes were
incubated in Dulbecco's Modified Eagle Medium (DMEM) containing 10%
fetal bovine serum (GibcoBRL, Gaithersburg, Md.) with various
concentrations of Compound 11 or vehicle (0.1% DMSO) for 48 h in
24-well plates, in triplicate. Cells were washed with phosphate
buffered saline (PBS, 150 mM NaCl, 1 mM KH.sub.2PO.sub.4, 3 mM
Na.sub.2HPO.sub.4; pH 7.4) and incubated in glucose-free DMEM for 2
h at 37.degree. C. The cells were washed 3 times with Krebs Ringer
Phosphate Buffer (KRP). Glucose uptake was initiated by addition of
0.25 .mu.Ci 2-.sup.14C(U)-deoxy-D-glucose (300 .mu.Ci/mmol,
American Radiolabeled Chemicals Inc., St Louis, Mo.) per well and
the cells incubated for 10 min at room temperature in the presence
of 0.1 mmol cold 2-deoxy-D-glucose. Finally, the cells were washed
three times with ice-cold PBS containing 10 mM cold glucose, lysed
with 0.5% SDS, and counted in a scintillation counter (Beckman
LS6500).
Transfection and Luciferase Activity Assay
[0192] Human PPAR-.gamma.2 expression vector was constructed by
inserting PPAR-.gamma.2 encoding region into pcDNA3.1+ vector
(Invitrogen, Carlsbad, Calif.). Luciferase reporter vector was
constructed by ligating PPRE response element adjacent to the
upstream of Firefly luciferase coding region. Control vector,
pRL-SV40 expressing Renilla luciferase was purchased from Promega
(Madison, Wis.).
[0193] About 2.7.times.10.sup.4 293 cells (ATCC, Manassas, Va.)
were plated into a 35 mm culture well and maintained in Eagle's
Minimal Essential Medium (ATCC, Manassas, Va.) supplemented with
10% heat inactivated horse serum (ATCC, Manassas, Va.) for 24
hours. Expression, reporter and control vectors (2.5 ng control and
100 ng others per culture well) were transfected by LIPOFECTAMIN
PLUS.TM. Reagent (Invitrogen, Carlsbad, Calif.). Transfection
reagent and DNA were prepared according to manufacture's
recommendations and incubated with cells for 3 hours followed by
adding equal volume EMEM supplemented with 20% horse serum.
Twenty-four hours after transfection, cells were treated with
vehicle or compounds at indicated final concentration for 24 hours.
Final concentration of vehicle was 0.001% DMSO (Sigma, ST Louis,
Mo.) in medium. Vehicle and compound treatment were all conducted
in triplicate. Each culture well was then assayed for a response
characterized by increased Firefly luciferase activity normalized
with Renilla luciferase activity.
[0194] Assays for Firefly luciferase activity and Renilla
luciferase activity followed the standard protocol of
Dual-luciferase Reporter.RTM. Assay System (Promega, Madison,
Wis.). Briefly, 400 .mu.l Passive Lysis Buffer was added into each
culture well and all wells were placed on a shaker for 15 minutes.
Five .mu.l cell lysate of each well was added to a reaction tube.
Luciferase reagent II and Stop & Glo.RTM. were injected into
the reaction tube sequentially by Sirius Luminometer (Berthold
Detection Systems, Pforzheim, Germany). Final reporter activity was
calculated as the ratio of Firefly luciferase activity over by
Renilla luciferase activity.
Results
[0195] The three possible methyl ester analogs with double bond(s)
at different positions (10, 11, 17), the methyl ester without any
double bond 18, the free acid analog 14 of compound 11, and the
Z-isomer 23 of 11 were made and tested on in vitro glucose uptake
in 3T3-L1 cells (Tafuri et al, Endocrinology, 137:4706-12, 1996;
Frost and Lane, J Biol Chem 260:2646-52, 1985) at 0.1 and 1 .mu.M
concentrations (Table 3). At a concentration of 1 .mu.M compounds
10, 11, and 14 increased glucose uptake to a level comparable to
rosiglitazone. Compounds 17 and 18 showed modest activity at 1
.mu.M concentration and compound 23 was essentially inactive. At a
concentration of 0.1 .mu.M, compounds 10 and 11 retained activity,
but less than that of rosiglitazone. Compound 10, containing two
double bonds, showed lower increased glucose uptake compared to 11.
Compound 17 with only one double bond joined to the TZD ring and
the doubly reduced product 18 were devoid of activity at 0.1 .mu.M.
We infer from this that the absence of the double bond joined to
the TZD ring and the presence of the cinnamic acid double bond is
important for increased glucose uptake in this system. The lack of
activity of 23, the corresponding Z-isomer of 11, indicates that
the geometry of the double bond is critical to activity. The lower
activity of the free acid 14 compared to the methyl ester 11 may be
due to difference in lipophilicity of the compounds.
[0196] Antidiabetic compounds of the TZD class increase peripheral
tissue sensitivity to insulin via PPAR-gamma activation. It was our
goal to introduce into diphenylethylene compounds, by chemical
modification, this additional mechanism of action. Agonist activity
on PPAR-gamma was explored in an in vitro system using cells
transfected with human PPAR-gamma2 ligated to firefly luciferase as
the reporter element. The results from this in vitro
transactivation assay for the tested compounds are summarized in
Table 4. The most potent compound in this series was found to be 11
(EC.sub.50 of 0.28 .mu.M) which had approximately one-thirtieth the
activity of rosiglitazone (EC.sub.50 of 0.009 .mu.M) in the same
assay. Compound 10 (two double bonds) and compound 14 (the free
acid of 11) also showed reasonable potency although less than that
of compound 11. Compounds 17 and 18, in which the cinnamic acid
double bond was reduced, were essentially inactive.
[0197] The Z-isomer 23 showed less than one-tenth the potency of
compound 11 in this assay. Based on these in vitro results,
compound 11 was evaluated in two widely-used mouse models of
NIDDM.
[0198] Initially, in vivo glucose-lowering efficacy of 11 was
explored in the genetically hyperinsulinemic, diabetic mouse
(db/db) model (8 weeks old male mice, 5 animals per group)
following a single oral dose of 50 mg/kg (in 0.5% CMC and 10% PEG).
Blood glucose levels were monitored at time intervals of 0 min, 1,
4, 6, 24, 48 and 72 h. Compound 11 showed a time dependent glucose
lowering effect which was maximal at 24 hrs (23% of 0 min, data not
shown). Side by side comparison of rosiglitazone and compound 11
(each at 50 mg/kg/day, orally, for 14 days) showed dramatic,
comparable blood glucose lowering effects which increased with
duration of dosing (FIG. 16A). Body weights (FIG. 16B) were not
affected by either compound in this short term experiment.
[0199] Compound 11 was also evaluated in the genetically obese
(ob/ob) male diabetic mouse (7 weeks old male mice, 5 animals per
group) with treatment continued for 14 days at a dose of 50 mg/kg
body weight. In 11 treated mice blood glucose levels were
significantly decreased within 48 hours and normalized by three
days (*p value<0.05; paired T test) (FIG. 17A). Blood glucose
levels in these mice rebounded slowly after stopping treatment
(FIG. 17A). Interestingly, we did not see any increase in body
weight of drug treated animals compared to vehicle treated group
(FIG. 17B). This may be an advantageous situation compared to
strong PPAR-gamma agonists which showed a large body weight gain in
different animal models.
[0200] After a treatment of fourteen days blood samples were
collected from both the groups for the determination of
glycosylated hemoglobin, serum insulin, triglycerides, FFA levels
(FIG. 18). Glycosylated hemoglobin, which is being used as marker
of diabetes management, was reduced from 5.2% to 3.8% (p=0.15). 11
reduced serum insulin by 58%, triglycerides by 65% and free fatty
acid (FFA) levels by 33%, all p<0.05 compared to vehicle treated
animals.
[0201] To establish the effective dose of 11, a further experiment
was carried out in ob/ob mice (n=8) with three different doses. As
shown in FIG. 19 there was a dose dependent glucose lowering
effect. Animals treated with a dose of 3.12 mg/kg showed a similar
extent of glucose reduction compared to 12.5 mg/kg after twelve
days of treatment. Thus, the effect of 11 is comparable with
rosiglitazone in both db/db and ob/ob mice.
[0202] It has been proposed earlier that the compounds which show
high degree of activation of PPAR-gamma generally have superior
glucose lowering activities in animal experiments (Wilson et al, J
Med Chem 39:665-8, 1996; Foreman et al, Cell 83:803-12, 1995).
However, findings reported here differ from this interpretation. We
have found a less potent PPAR-gamma agonist 11 which showed a
glucose lowering activity in animals comparable to that of
rosiglitazone. Attempts are being made to elucidate further the
mechanism behind this potent glucose lowering activity of 11.
Glycogen synthesis in the liver, but particularly in the muscle is
major site for the disposal of postprandial glucose. One possible
explanation for strong glucose lowering activity in spite of modest
PPAR-gamma agonist activity is our recent observation of higher
glycogen synthesis induced by 11 compared to rosiglitazone (see
below). TABLE-US-00003 TABLE 3 In vitro Glucose Uptake in 3T3-L1
Adipocytes.sup.a % Glucose uptake Compound no. 0.1 .mu.m 1.0 .mu.m
1 (rosiglitazone) 204.2 .+-. 8.5 214.7 .+-. 36.5 10 142.6 .+-. 7.4
209.3 .+-. 22.6 11 161.6 .+-. 11.4 218.1 .+-. 10.0 14 108.6 .+-.
19.9 199.5 .+-. 11.6 17 101.9 .+-. 6.8 130.0 .+-. 27.0 18 104.4
.+-. 5.7 137.0 .+-. 10.1 23 89.5 .+-. 5.6 118.0 .+-. 3.6 Insulin
335.2 .+-. 21.7 345.6 .+-. 8.3 .sup.aBasal glucose uptake is
expressed as % basal of the non treated cells (each point is the
average of quadruplicate determinations .+-. SD values).
[0203] TABLE-US-00004 TABLE 4 Induction of PPAR-.gamma. Mediated
Luciferase Activity by Thiazolidinediones.sup.a Compound no.
EC.sub.50 (.mu.M) 1 (rosiglitazone) 0.009 .+-. 0.007 10 1.136 11
0.284 .+-. 0.036 (n = 5).sup.b 14 0.690 .+-. 0.038 (n = 2) 17 23.9
18 57.7 23 3.686 .+-. 1.454 (n = 2) .sup.aResults are based on
several independent experiments. Each experiment contains at least
8 different concentration of drug treatment (between 0.1 to 30
.mu.M), with each concentration in triplicate. EC.sub.50 values
were calculated by non-linear regression analysis using GraphPad
Prism software. .sup.bn = independent experiments.
Conclusion
[0204] TZD's based on the alpha-phenyl cinnamic acid motif have
glucose lowering activity in genetic models of diabetic mice.
Compound 11 showed strong glucose lowering activity even though it
is a weak PPAR-gamma agonist. Strong glucose lowering activity of
11 in spite of being a weaker PPAR-gamma agonist may be due to its
higher glycogen synthesis compared to rosiglitazone. Further
investigation is required to fully elucidate the mechanism(s) of
action of this new family of TZDs.
EXAMPLE 13
Glycogen Synthesis
[0205] Glycogen synthesis was measured as net conversion of
.sup.14C-D-glucose to cellular glycogen in HepG2 cells as described
by Ciaraldi et al, Diabetes 41:975-81, 1992. Briefly, HepG2 cells
(ATCC, Manassas, Va.) in 6-well plates were treated with Compound
11 or other compounds for 48 h. They were washed with 10 mM HEPES
buffer (150 mM NaCl, 5 mM KCl, 1.2 mM MgSO.sub.4, 1.2 mM
CaCl.sub.2, 2.5 mM CaCl.sub.2, 10 mM HEPES; pH 7.4) containing 1%
BSA. Cells were incubated in the same buffer for 30 min prior to
addition of 0.2 .mu.Ci/well of .sup.14C-D-glucose (5 mM final
concentration, 10 .mu.Ci/mmol, American Radiolabeled Chemicals
Inc., St Louis, Mo.). After incubation for 2 h at 37.degree. C. the
cells were washed with ice-cold PBS and solubilized with 1 M KOH at
55.degree. C. Converted glycogen was precipitated by ethanol after
addition of 10 mM carrier glycogen. The pellet was washed and
resuspended in water, and an aliquot was counted in a scintillation
counter (Beckman LS6500). Total protein was assayed and results
were reported as cpm/mg of protein.
Adipogenesis
[0206] Adipogenesis in 3T3-L1 fibroblasts was carried out as
described by Wu et al, J Clin Invest 101:22-32, 1998. After two
days of growth in 6-well plates, cells were treated either with
vehicle (0.1% DMSO) or with compounds for ten days. Fresh medium
with compounds or vehicle was replenished every 48 h. Cells were
washed with PBS twice and fixed in 10% formalin (Sigma) in PBS.
After washing in PBS, cells were stained with freshly diluted Oil
Red O in isopropanol for 1 h at room temperature. The cells were
washed five times with PBS and visualized under an Olympus BH2
microscope. Quantitative accumulation of triglyceride was also
measured under similar experimental conditions, except in this case
cells were plated in 100-mm tissue culture dishes. Triglyceride was
extracted with methanol:chloroform (2:1) mixture. To monitor the
efficiency of recovery, .sup.3H-cholesterol oleate (50,000
cpm/well, American Radiolabeled Chemicals Inc., St Louis, Mo.) was
added in each tube as tracer before extraction following the
protocol of Brown et al, J Clin Invest 55:783-93, 1975. Extracted
triglyceride was measured by a colorimetric assay (GPO-Trinder,
Sigma) according to manufacturer instructions.
Transfection and Transactivation Assays
[0207] Human PPAR.gamma.2 expression vector was constructed by
inserting the PPAR.gamma.2 cDNA coding region into pcDNA3.1+ vector
(Invitrogen, Carlsbad, Calif.). The PPRE-luciferase reporter gene
was the kind gift of Dr. Kenneth Feingold. The control vector,
pRL-SV40 containing the Renilla luciferase cDNA was purchased from
Promega (Madison, Wis.). About 2.7.times.10.sup.4 HEK293 human
embryonal kidney cells (ATCC) were plated into a 35-mm tissue
culture dish and maintained in Eagle Modified Essential Medium
(EMEM, ATCC) containing 10% heat-inactivated horse serum for 24 h.
Expression, reporter (100 ng/dish) and control (2.5 ng/dish)
vectors were transfected using LIPOFECTAMIN PLUS.TM. Reagent
(GibcoBRL) according to manufacturer's recommendation. At 24 h
after transfection, cells were treated with vehicle (0.001% DMSO in
medium) or compounds at the indicated concentration and incubated
for 24 h. Each treatment was conducted in triplicate. Each culture
dish was assayed for firefly luciferase activity normalized by
Renilla luciferase activity to account for differences in
transfection efficiency. Luciferase activity was measured using the
Dual-luciferase Reporter.RTM. Assay System (Promega) and a Sirius
luminometer (Berthold Detection System, Pforzheim, Germany).
In vivo Studies
[0208] All procedures performed were in compliance with the Animal
Welfare Act and U.S. Department of Agriculture regulations and were
approved by the Calyx Therapeutics Institutional Animal Care and
Use Committee. Animals were housed at 22.degree. C. and 50%
relative humidity, with a 12-h light and dark cycle, and received a
regular rodent diet (Harlan Teklad, Madison, Wis.) ad libitum with
free access to water. Male C57BL/KsJ-db/db and C57BL/6J-ob/ob mice
were obtained from Jackson Laboratories (Bar Harbor, Maine) when
their age was 5 weeks. Seven-week-old animals were dosed with
Compound 11, rosiglitazone maleate (recrystallized from
commercially available tablets) or vehicle (0.5% carboxymethyl
cellulose (Sigma, St. Louis, Mo.) in water) orally once daily by
gavage. Blood glucose measurements were made with a One Touch
Glucose Meter (Life Scan, Inc., Milpitas, Calif.) and/or a glucose
oxidase assay (Glucose Trinder, Sigma, St. Louis, Mo.) prior to
administering the next dose and in the fed state. Body weights were
monitored throughout the study. Eight-week-old male Zucker diabetic
fatty (ZDF-fa/fa) rats (Genetic Models, Indianapolis, Ind.) were
kept on 6.5% fat Formulab Diet 5008 (PMI Feeds, Richmond, Ind.) for
two weeks prior to dosing as described above.
Statistical Analysis
[0209] Data are presented as the mean.+-.standard error (SE) and
statistical comparisons were made by t test or ANOVA with
Tukey/Kramer post hoc testing where appropriate using StatView 5
software (SAS Institute).
Results
Compound 11 Stimulates Glucose Uptake in vitro
[0210] Differentiating 3T3-L1 adipocytes represent an
insulin-sensitive cell-culture model for studying glucose uptake
and is often used to characterize potential antidiabetic compounds.
Although TZDs increase glucose uptake in these cells, both in the
absence and presence of insulin, the majority of this effect
appears to be the result of non-insulin-mediated glucose disposal.
As shown in FIG. 20A, glucose uptake was increased to a maximum of
1.33.+-.0.02 (mean.+-.SE) fold over basal levels in response to
increasing concentrations of Compound 11(0.01, 0.1, 1.0 and 10
.mu.M). We also examined the effect of Compound 11 and
rosiglitazone on insulin-stimulated glucose uptake in 3T3-L1
adipocytes (FIG. 20B).
[0211] There was no difference in dose response curves of glucose
uptake in response to insulin in the presence (5 .mu.M) or absence
of the TZDs. Differences in the maximal responses can be accounted
for by the increased amount of basal glucose uptake in the absence
of insulin and either Compound 11 or rosiglitazone, indicating an
additive, not synergistic, effect on glucose uptake. These results
suggest this enhancement of glucose uptake is mediated through a
non-insulin-dependent mechanism, such as an increase in GLUT-1
transporters.
In vivo Antihyperglycemic Effect of Compound 11
[0212] The antihyperglycemic activity of Compound 11 was examined
in several models of type 2 diabetes mellitus. FIG. 21 summarizes
the effect of Compound 11 given as single daily oral doses of 50
mg/kg (96.2 .mu.mol/kg) over 8 to 9 days. At the end of each study
the drug led to marked decreases in blood glucose levels in ob/ob
mice (59% vs. baseline), db/db mice (32.% vs. baseline) and ZDF
rats (50% vs baseline).
[0213] Weight gain in treated and control animals was similar
except for ZDF rats, where Compound 11 treated animals gained 13%
more weight than control animals. In a subsequent study, the in
vivo potency of Compound 11 was compared to rosiglitazone in ob/ob
mice (FIG. 22). Compound 11 and rosiglitazone treatment (both at 10
mg/kg/day; 19.2 and 28.0 .mu.mol/kg/day for Compound 11 and
rosiglitazone, respectively) demonstrated similar antidiabetic
potency over the 8-day treatment period.
[0214] Weight gain was similar in vehicle and drug-treated groups.
Both compounds significantly lower serum insulin, free fatty acids
and triglycerides in this model (data not shown).
Compound 11 is Less Adipogenic than Rosiglitazone
[0215] Because TZDs are ligands for PPAR-gamma and induce adipocyte
differentiation (13-15), we sought to determine the adipogenic
potential of Compound 11 using the 3T3-L1 preadipocyte model. In
these studies, 3T3-L1 fibroblasts were incubated with various
concentrations (0.1, 1, 10 .mu.M) of Compound 11, rosiglitazone or
vehicle in the absence of dexamethasone, insulin and IBMX. After 14
days cells were stained with Oil Red O, counterstained with
methylene blue for visual assessment (FIG. 23B) and assayed for
triglyceride accumulation. As shown in FIG. 23A there was a
dose-dependent increase in triglyceride accumulation in response to
Compound 11 and rosiglitazone. The dose response of triglyceride
accumulation in response to Compound 11 is right-shifted in
comparison to rosiglitazone. Moreover, the maximal amount of
triglyceride accumulated in response to Compound 11 was
significantly less than that seen in response to rosiglitazone
(3.96 vs. 9.22 fold increase over control, respectively;
P<0.0001, ANOVA). Compound 11 at concentrations of 100 .mu.M and
higher were cytotoxic in this system (data not shown).
Compound 11 is a Partial Agonist of PPAR.gamma..sup.2
[0216] We examined the ability of rosiglitazone and Compound 11 to
transactivate a PPRE-Luc reporter gene in HEK293 cells
cotransfected with a human PPAR-gamma2 expression vector. Compound
11 was a substantially less potent activator of PPAR-gamma than
rosiglitazone (FIG. 24). The EC.sub.50 for transactivation in this
system was 0.009.+-.0.0007 .mu.M (SE) for rosiglitazone, and
0.284.+-.0.036 .mu.M for Compound 11 (n=5). Similar dose-response
curves were obtained using a reporter-gene assay in cells
transfected with heterologous cDNA constructs of GAL4-DNA binding
domain/PPAR-gamma ligand binding domain and 5.times. upstream
activator sequence (UAS)-luciferase constructs (data not
shown).
Compound 11 Increases Glycogen Synthesis in HepG2 Hepatocytes
[0217] The ability of Compound 11 to increase glycogen synthesis
was examined in HepG2 hepatocytes. FIG. 25A shows that there is a
dose-dependent increase in .sup.14C-glucose incorporated into
glycogen in response to Compound 11 in the absence of insulin; this
response was maximal (3.1-fold increase over baseline) at 48 to 72
h (FIG. 25B). In contrast, rosiglitazone did not increase glycogen
synthesis (0.9-fold decrease at 10 .mu.M, FIG. 25A).
[0218] In separate experiments, concentrations of rosiglitazone
higher than 30 .mu.M produced only minimal increases in glycogen
synthesis (1.4.+-.0.06, SD fold increase over baseline, data not
shown). The increase in glycogen synthesis induced by Compound 11
was dependent upon new protein synthesis as it was blocked by
cotreatment with cycloheximide (FIG. 25C).
Discussion
[0219] Our results indicate that Compound 11 is effective in
lowering blood glucose in several animal models of type 2 diabetes.
It has a robust antihyperglycemic effect in ZDF rats, where it
normalizes glucose levels. This drug also has potent
glucose-lowering activity in ob/ob mice, where it appears to be
equipotent to rosiglitazone on a mass basis. In actuality Compound
11 appears to be 46% more potent than rosiglitazone in vivo, on a
mole-per-mole basis; the molecular weight of rosiglitazone is
substantially less than Compound 11 (357 vs. 520 g/mol), yet the
two drugs produced the same degree of glucose-lowering in ob/ob
mice. Our results also indicate that Compound 11 is substantially
less adipogenic than rosiglitazone. This effect likely reflects the
lower affinity of Compound 11 for PPAR-gamma in comparison to
rosiglitazone. We did detect a small increase in weight gain in ZDF
rats treated with Compound 11, and we were unable to detect
differences in weight gain between rosiglitazone and Compound 11
treated animals in this short-term study. These data are difficult
to interpret because the studies were conducted in genetically
obese animals, which may have obscured a differential effect of
these drugs on weight gain. Several studies in normal animals, up
to one month in duration, have failed to demonstrate clinically
meaningful weight gain (data not shown).
[0220] It is widely held that the weight gain associated with TZDs
is partly due to their adipogenic potential, and there has been
much effort directed at finding compounds which are potent
PPAR-gamma activators but do not cause weight gain. Because
Compound 11 has less adipogenic activity, but maintains
antihyperglycemic activity, it appears that it may be possible to
develop pharmacologic PPAR-gamma activators that produce less
weight gain than current commercially available PPAR-gamma
activators. In addition to the contribution of adipogenesis, edema
is an important factor in the weight gain associated with
PPAR-gamma agonists. This toxicity is believed to be directly
related to the PPAR-gamma activation potency of the molecule.
Because Compound 11 is a weak agonist of PPAR-gamma it may be
associated with less edema. Clinical studies underway will help
define the effect of Compound 11 on body weight in humans.
[0221] The affinity (Ki) of PPAR-gamma for Compound 11 was 6.5-fold
less than its affinity for rosiglitazone in preliminary competition
binding assays (data not shown), and the transactivation potency
was as much as 30-fold less than that of rosiglitazone. In general,
there is a relatively strong correlation between PPAR-gamma
affinity and glucose-lowering activity; however, recent data
indicate that this relationship may not be true for all ligands of
this receptor. Recently a non-TZD PPAR-gamma activator,
FMOC-L-Leucine, has been shown to have a similar profile to
Compound 11. FMOC-L-Leucine has approximately 400-fold less
affinity for PPAR-gamma, is only weakly adipogenic, but has potent
in vivo antihyperglycemic activity (Rocchi et al, Mol Cell
8:737-47, 2001). Differences in in vivo metabolism may explain part
of the apparent discrepancy between PPAR-gamma affinity and in vivo
antidiabetic potency for Compound 11 and other ligands. Studies by
Reginato et al (J Biol Chem 273:32679-84, 1998), Mukherjee et al
(Mol Endocrinol 14:1425-33, 2000) and Rocchi et al (Mol Cell
8:737-47, 2001) indicate that ligand-mediated recruitment of the
coactivator SRC-1 to PPAR-gamma is an important determinant for
differential activities of ligands. At present, we can only
speculate on the way in which Compound 11 influences coactivator
recruitment to the PPARgamma-RXR complex. It may be that Compound
11 is less conducive for SRC-1 or PGC-1 recruitment and, as a
result, transcriptional activation. A recent study (Nugent et al,
Mol Endocrinol 15:1729-38, 2001) reported that in vitro glucose
uptake into adipocytes is partially independent of
PPAR.quadrature.. Whether non-PPARgamma-mediated activities or
coactivator recruitment explains the unique properties of Compound
11 will require further investigation.
[0222] Presently, the mechanism by which PPAR-gamma ligands,
including TZDs, produce their antihyperglycemic effects is not
known. The prevailing wisdom suggests that the glucose-lowering
effect of these drugs is mediated through the PPAR-gamma receptor,
which enhances insulin sensitivity. Recent data suggest that the
relationship between PPAR-gamma, its ligands and insulin
sensitivity is more complex. For example, heterozygous PPAR-gamma
null mice actually demonstrate increased insulin sensitivity, and
the insulin sensitizing effect of synthetic ligands may result from
a balance between transcriptional activation and repression (Miles
et al, J Clin Invest 105:287-92, 2000). Additionally, a
non-receptor mediated mechanism of action for Compound 11 and other
PPAR-gamma agonists cannot be excluded. Indeed, abrogating
endogenous PPAR-gamma does not result in the elimination of TZD
activity (Nugent et al, Mol Endocrinol 15:1729-38, 2001; Chawla et
al, Nat Med 7:48-52, 2001). Our data indicate that Compound 11, in
contrast to rosiglitazone, increases glycogen synthesis in liver
cells, possibly providing an added mechanism for lowering glucose
levels in diabetic animals. Recent data suggest that some non-TZD
PPAR-gamma agonists may upregulate genes involved in glycogen
synthesis (Way et al, Endocrinology 142:1269-77, 2001). It is
becoming apparent that ligands for this receptor will have a
spectrum of affinities, transcriptional activities, and in vivo
pharmacodynamic profiles. Therefore, there is substantial clinical
value in generating compounds with selective PPAR-gamma modulating
activities. The acronym SPRM, for "selective PPAR modulator"
(Mukherjee et al, Mol Endocrinol 14:1425-33, 2000), may best
describe these molecules. SPRMs may ultimately prove to have both
specific and tailored activities, including the potential avoidance
of weight gain associated with currently marketed TZDs. Such agents
would have the potential to be of great benefit in treating
patients with type 2 diabetes.
[0223] FIG. 20 shows glucose uptake in 3T3-L1 Cells. A. In vitro
glucose uptake was measured in differentiated 3T3-L1 adipocytes
after 48-h treatment with increasing concentrations of Compound 11
(black bars) or 0.1% DMSO as vehicle (hatched bars).
*P.ltoreq.0.001. B. Glucose uptake in differentiated 3T3-L1
adipocytes was measured in the presence of increasing
concentrations of insulin in the presence of vehicle (circles),
rosiglitazone (5 .mu.M) (triangles) or Compound 11 (5 .mu.M)
(squares). *P.ltoreq.0.05, #P=0.02 vs. vehicle.
[0224] FIG. 21 shows in vivo Antihyperglycemic Activity of Compound
11 in Diabetic Animals. Diabetic ob/ob mice (A) and db/db mice (B)
and diabetic ZDF rats (C) were treated with single daily doses of
Compound 11 (50 mg/kg=96.2 .mu.mol/kg) (squares) or vehicle (0.5%
carboxymethyl cellulose) (circles) by oral gavage for 8 or 9 days.
Blood glucose measurements were made in the fed state.
*P.ltoreq.0.05, #*P.ltoreq.0.01, .dagger.P.ltoreq.0.001.
[0225] FIG. 22 shows in vivo Antihyperglycemic Activity of Compound
11 vs. Rosiglitazone in ob/ob mice. Diabetic ob/ob mice were
treated with single daily doses of Compound 11 (squares) and
rosiglitazone (triangles) at 10 mg/kg (19.2 and 28.0 .mu.mol/kg,
respectively) or vehicle (0.5% carboxymethyl cellulose) (circles)
by oral gavage. Blood glucose (A) and body weight (B) were measured
during the 8-day treatment period.
[0226] FIG. 23 shows in vitro Adipogenic Activity of Compound 11.
3T3-L1 cells were cultured with vehicle, Compound 11 or
rosiglitazone for 10 days. Total accumulated triglyceride was
measured. (A) Quantitative measurement of accumulated triglyceride
after 10 days of treatment with increasing concentrations of
Compound 11 (black bars) or rosiglitazone (hatched bars) or vehicle
(white bar). (B) Qualitative assessment of triglyceride
accumulation by Oil Red O after 10-day treatment with 1 .mu.M
Compound 11, rosiglitazone or vehicle.
[0227] FIG. 24 shows induction of PPARgamma-Mediated
Transactivation of PPRE-Luc Reporter by Compound 11 and
Rosiglitazone. HEK293 cells were transiently cotransfected with a
PPAR-gamma expression vector and a PPRE-Luc reporter construct.
Cells were also transfected with a cDNA construct containing
Renilla luciferase, which was used to control for transfection
efficiency. Transfected cells were treated with increasing
concentrations of Compound 11 and rosiglitazone. Luciferase
activity is expressed as enhancement over basal levels (no drug)
and is corrected for transfection efficiency. The figure shows a
representative result of five experiments.
[0228] FIG. 25 shows the effect of Compound 11 on In Vitro Glycogen
Synthesis in HepG2 Cells. A. Dose-dependent stimulation of glycogen
synthesis from glucose by Compound 11 in the absence of insulin at
48 h. Stimulation of glycogen synthesis is expressed as a
percentage of basal (vehicle), which is defined as 100%. B.
Time-dependent increase in Compound 11-stimulated glycogen
synthesis. The maximal effect occurs at 48 to 72 h of treatment
with Compound 11. C. Cycloheximide blocks glycogen synthesis
induced by Compound 11 (30 .mu.M, 48 h). Vehicle=white bars,
Compound 11=black bars, rosiglitazone=hatched bars,
CHX=cycloheximide, checked bars, rosi=rosiglitazone.
Co-Administration
[0229] The compounds according to the present invention may be
combined with a physiologically acceptable carrier or vehicle to
provide a pharmaceutical composition, such as, lyophilized powder
in the form of tablet or capsule with various fillers and binders.
Similarly, the compounds may be co-administered with other agents.
Co-administration shall mean the administration of at least two
agents to a subject so as to provide the beneficial effects of the
combination of both agents. For example, the agents may be
administered simultaneously or sequentially over a period of time.
The effective dosage of a compound in the composition can be widely
varied as selected by those of ordinary skill in the art and may be
empirically determined. Moreover, the compounds of the present
invention can be used alone or in combination with one or more
additional agents depending on the indication and the desired
therapeutic effect. For example, in the case of diabetes, insulin
resistance and associated conditions or complications, including
obesity and hyperlipidemia, such additional agent(s) may be
selected from the group consisting of: insulin or an insulin
mimetic, a sulfonylurea (such as acetohexamide, chlorpropamide,
glimepiride, glipizide, glyburide, tolbutamide and the like) or
other insulin secretagogue (such as nateglinide, repaglinide and
the like), a thiazolidinedione (such as pioglitazone, rosiglitazone
and the like) or other peroxisome proliferator-activated receptor
(PPAR)-gamma agonist, a fibrate (such as bezafibrate, clofibrate,
fenofibrate, gemfibrozol and the like) or other PPAR-alpha agonist,
a PPAR-delta agonist, a biguanide (such as metformin), a statin
(such as fluvastatin, lovastatin, pravastatin, simvastatin and the
like) or other hydroxymethylglutaryl (HMG) CoA reductase inhibitor,
an alpha-glucosidase inhibitor (such as acarbose, miglitol,
voglibose and the like), a bile acid-binding resin (such as
cholestyramine, celestipol and the like), a high density
lipoprotein (HDL)-lowering agent such as apolipoprotein A-I
(apoA1), niacin and the like, probucol and nicotinic acid. In the
case of inflammation, inflammatory diseases, autoimmune disease and
other such cytokine mediated disorders, the additional agent(s) may
be selected from the group consisting of: a nonsteroidal
anti-inflammatory drug (NSAID) (such as diclofenac, diflunisal,
ibuprofen, naproxen and the like), a cyclooxygenase-2 inhibitor
(such as celecoxib, rofecoxib and the like), a corticosteroid (such
as prednisone, methylprednisone and the like) or other
immunosuppressive agent (such as methotrexate, leflunomide,
cyclophosphamide, azathioprine and the like), a disease-modifying
antirheumatic drug (DMARD) (such as injectable gold, penicillamine,
hydroxychloroquine, sulfasalazine and the like), a TNF-alpha
inhibitor (such as etanercept, infliximab and the like), other
cytokine inhibitor (such as soluble cytokine receptor,
anti-cytokine antibody and the like), other immune modulating agent
(such as cyclosporin, tacrolimus, rapamycin and the like) and a
narcotic agent (such as hydrocodone, morphine, codeine, tramadol
and the like). The combination therapy contemplated by the
invention includes, for example, administration of the inventive
compound and additional agent(s) in a single pharmaceutical
formulation as well as administration of the inventive compound and
additional agent(s) in separate pharmaceutical formulations.
[0230] It will be appreciated that various modifications may be
made in the invention as described above and as defined in the
following claims.
* * * * *