U.S. patent application number 12/627895 was filed with the patent office on 2010-06-03 for small molecule modulators of cytokine activity.
This patent application is currently assigned to ANGION BIOMEDICA CORP.. Invention is credited to Rama K. Mishra, Jasbir Singh, David E. Zembower.
Application Number | 20100137285 12/627895 |
Document ID | / |
Family ID | 35840190 |
Filed Date | 2010-06-03 |
United States Patent
Application |
20100137285 |
Kind Code |
A1 |
Zembower; David E. ; et
al. |
June 3, 2010 |
SMALL MOLECULE MODULATORS OF CYTOKINE ACTIVITY
Abstract
The present invention provides compounds having formula (I) or
(II): ##STR00001## and pharmaceutically acceptable derivatives
thereof, wherein m, p, R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are as
described generally and in classes and subclasses herein, and
additionally provides pharmaceutical compositions thereof, and
methods for the use thereof for the treatment of any of a number of
diseases, disorders or conditions associated with HGF/SF or other
cytokine activity.
Inventors: |
Zembower; David E.; (La
Grange, IL) ; Singh; Jasbir; (Naperville, IL)
; Mishra; Rama K.; (Chicago, IL) |
Correspondence
Address: |
CHOATE, HALL & STEWART LLP
TWO INTERNATIONAL PLACE
BOSTON
MA
02110
US
|
Assignee: |
ANGION BIOMEDICA CORP.
Great Neck
NY
|
Family ID: |
35840190 |
Appl. No.: |
12/627895 |
Filed: |
November 30, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11238285 |
Sep 28, 2005 |
7648978 |
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12627895 |
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60675241 |
Apr 27, 2005 |
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60613740 |
Sep 28, 2004 |
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Current U.S.
Class: |
514/217.05 ;
514/248; 540/599; 544/237 |
Current CPC
Class: |
A61P 43/00 20180101;
C07D 403/14 20130101; A61P 25/28 20180101; A61P 3/06 20180101; A61P
25/00 20180101; A61P 35/00 20180101; A61P 17/02 20180101; C07D
403/10 20130101; A61P 25/32 20180101; A61P 25/02 20180101; A61P
11/00 20180101; C07D 401/10 20130101; A61P 9/10 20180101; A61P
13/12 20180101; C07D 401/14 20130101; A61P 9/00 20180101; A61P 3/10
20180101; A61P 1/16 20180101; C07D 237/32 20130101 |
Class at
Publication: |
514/217.05 ;
540/599; 544/237; 514/248 |
International
Class: |
A61K 31/501 20060101
A61K031/501; C07D 403/10 20060101 C07D403/10; C07D 401/10 20060101
C07D401/10; A61K 31/55 20060101 A61K031/55; A61P 9/00 20060101
A61P009/00 |
Goverment Interests
GOVERNMENT SUPPORT
[0002] This work was supported in part by the U.S. Government,
grant 1R43CA096077 from the Public Health Service, National
Institutes of Health. The U.S. Government may have certain rights
in this invention.
Claims
1-71. (canceled)
72-108. (canceled)
109. An isolated compound having the structure: ##STR00061## or
pharmaceutically acceptable salt, ester, or salt of such an ester
thereof; wherein Cy is an N-linked 5- to 10-membered heterocyclic
or heteroaryl group other than an optionally substituted
homopiperidinyl group, wherein Cy is optionally substituted with p
occurrences of R.sup.4; p is an integer from 1 to 6; each
occurrence of R.sup.4 is independently hydrogen, halogen, hydroxyl,
--NO.sub.2, --NH.sub.2, --CN, --CONH.sub.2, --SO.sub.2OH, an
optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a,
--OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic, or acyl
moiety; m is an integer from 1 to 4; each occurrence of R.sup.4 is
independently hydrogen, halogen, hydroxyl, --NO.sub.2, --NH.sub.2,
--CN, --CONH.sub.2, --SO.sub.2OH, an optionally substituted
aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic moiety, --OR.sup.R, --S(.dbd.O).sub.nR.sup.d,
--NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, heteroaromatic or acyl moiety; R.sup.a, for each
occurrence, is hydrogen or an optionally substituted aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aromatic, or
heteroaromatic moiety; R.sup.b and R.sup.c, for each occurrence,
are independently hydrogen, hydroxy, SO.sub.2R.sup.d, or an
optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic, heteroaromatic or acyl moiety; R.sup.d, for
each occurrence, is independently hydrogen, --N(R.sup.e).sub.2, or
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety; and R.sup.e, for
each occurrence, is independently hydrogen or aliphatic; with the
proviso that the compound does not have one of the following
structures: ##STR00062## ##STR00063## wherein p and R.sup.4 are as
defined above, ##STR00064## wherein R.sup.4A is hydrogen, methyl,
methoxy, chloro or --NO.sub.2,
110. The compound of claim 109, wherein Cy is one of: ##STR00065##
wherein: q is 1, 2 or 4; and R.sup.4A is hydrogen, hydroxyl,
SO.sub.2R.sup.d, or an alkyl, heteroalkyl, cycloalkyl,
heterocyclic, aryl, heteroaryl, or acyl moiety.
111. The compound of claim 109, wherein Cy is one of: ##STR00066##
wherein: q is 1, 2 or 4; and R.sup.4A is hydrogen, hydroxyl,
SO.sub.2R.sup.d, or an alkyl, heteroalkyl, cycloalkyl,
heterocyclic, aryl, heteroaryl, or acyl moiety.
112. The compound of claim 109, wherein: each occurrence of R.sup.4
is independently hydrogen, halogen, hydroxyl, --NO.sub.2,
--NH.sub.2, --CN, --CONH.sub.2, --SO.sub.2OH, an optionally
substituted alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl or
heteroaryl moiety, --OR.sup.R, --S(.dbd.O).sub.nR.sup.d,
--NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl,
heteroaryl or acyl moiety; each occurrence of R.sup.1 is
independently hydrogen, halogen, hydroxyl, --NO.sub.2, --NH.sub.2,
--CN, --CONH.sub.2, --SO.sub.2OH, an optionally substituted alkyl,
heteroalkyl, cycloalkyl, heterocyclic, aryl or heteroaryl moiety,
--OR.sup.R, --S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c,
--C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a;
wherein n is 0-2, R.sup.R is an optionally substituted alkyl,
heteroalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl or acyl
moiety; wherein R.sup.a, for each occurrence, is independently
hydrogen or an optionally substituted alkyl, heteroalkyl,
cycloalkyl, heterocyclic, aryl or heteroaryl moiety; R.sup.b and
R.sup.c, for each occurrence, are independently hydrogen, hydroxy,
SO.sub.2R.sup.d, or an alkyl, heteroalkyl, cycloalkyl,
heterocyclic, aryl, heteroaryl or acyl moiety; R.sup.d, for each
occurrence, is independently hydrogen, --N(R.sup.e).sub.2, alkyl,
heteroalkyl, cycloalkyl, heterocyclic, aryl or heteroaryl; and
R.sup.e, for each occurrence, is independently hydrogen or
alkyl.
113. The compound of claim 109, having the structure: ##STR00067##
wherein q is 1, 2, or 4.
114. The compound of claim 109, having one of the following
structures: ##STR00068## ##STR00069## ##STR00070##
115. The compound of claim 109, wherein at least one occurrence of
--R.sup.1 is --NO.sub.2.
116. The compound of claim 109, wherein at least one occurrence of
R.sup.1 is --NH.sub.2.
117. The compound of claim 109, wherein at least one occurrence of
R.sup.1 is --COOH, --C(.dbd.O)OCH.sub.3, --COCH.sub.3,
--CONH.sub.2, --SO.sub.2OH, --SO.sub.2CH.sub.3, --SO.sub.2CF.sub.3,
--OPO.sub.2OH, --NHC(.dbd.O)CH.sub.3, --NHC(.dbd.O)CF.sub.3,
--NHSO.sub.2CH.sub.3 or --NHSO.sub.2CF.sub.3.
118. The compound of claim 109, wherein at least one occurrence of
R.sup.1 is halogen.
119. The compound of claim 109, wherein at least one occurrence of
R.sup.1 is an optionally substituted N-linked heterocyclic
group.
120. The compound of claim 119, wherein the N-linked heterocyclic
group is an optionally substituted N-pyrrolyl.
121. The compound of claim 109, wherein at least one occurrence of
R.sup.1 is an aliphatic moiety.
122. The compound of claim 109, wherein at least one occurrence of
R.sup.1 is a lower alkyl moiety.
123. The compound of claim 109, wherein at least one occurrence of
R.sup.4 is an aliphatic group.
124. The compound of claim 109, wherein each occurrence of R.sup.4
is independently an aliphatic group.
125. The compound of claim 124, wherein the aliphatic group is an
optionally substituted cyclic or acyclic C.sub.6-12 alkyl,
C.sub.6-12 alkenyl, or C.sub.6-12 alkynyl group.
126. The compound of claim 124, wherein the aliphatic group is an
optionally substituted -(alkyl)aryl group.
127. The compound of claim 109, wherein at least one occurrence of
R.sup.4 is --NR.sup.bR.sup.c; wherein R.sup.b and R.sup.c are
independently hydrogen, hydroxy, SO.sub.2R.sup.d, or an alkyl,
heteroalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl or acyl
moiety; R.sup.d is hydrogen, --N(R.sup.e).sub.2, alkyl,
heteroalkyl, cycloalkyl, heterocyclic, aryl or heteroaryl; and
R.sup.e is hydrogen or alkyl.
128. The compound of claim 109, wherein at least one occurrence of
R.sup.4 is --NH.sub.2.
129. The compound of claim 109, wherein at least one occurrence of
R.sup.4 is --C(.dbd.O)OR.sup.a; wherein R.sup.a is hydrogen or an
optionally substituted alkyl, heteroalkyl, cycloalkyl,
heterocyclic, aryl or heteroaryl moiety.
130. The compound of claim 109, wherein at least one occurrence of
R.sup.4 is --CO.sub.2H.
131. The compound of claim 109, wherein m is 0 or 1.
132. The compound of claim 109, wherein p is 0 or 1.
133. A pharmaceutical composition comprising: a pharmaceutically
acceptable carrier, adjuvant or vehicle; and a compound having the
structure: ##STR00071## or pharmaceutically acceptable salt, ester,
or salt of such an ester thereof; wherein Cy is an N-linked 5- to
10-membered heterocyclic or heteroaryl group other than an
optionally substituted homopiperidinyl group, wherein Cy is
optionally substituted with p occurrences of R.sup.4; p is an
integer from 1 to 6; each occurrence of R.sup.4 is independently
hydrogen, halogen, hydroxyl, --NO.sub.2, --NH.sub.2, --CN,
--CONH.sub.2, --SO.sub.2OH, an optionally substituted aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic moiety, --OR.sup.R, --S(.dbd.O).sub.nR.sup.d,
--NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, heteroaromatic, or acyl moiety; m is an integer from 1 to
4; each occurrence of R.sup.1 is independently hydrogen, halogen,
hydroxyl, --NO.sub.2, --NH.sub.2, --CN, --CONH.sub.2, --SO.sub.2OH,
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a,
--OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic or acyl
moiety; R.sup.a, for each occurrence, is hydrogen or an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, or heteroaromatic moiety; R.sup.b and R.sup.c, for each
occurrence, are independently hydrogen, hydroxy, SO.sub.2R.sup.d,
or an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic, heteroaromatic or acyl moiety; R.sup.d, for
each occurrence, is independently hydrogen, --N(R.sup.e).sub.2, or
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety; and R.sup.e, for
each occurrence, is independently hydrogen or aliphatic; with the
proviso that the compound does not have one of the following
structures: ##STR00072## wherein R.sup.4A is hydrogen, methyl,
wherein p and R.sup.4 are as defined above methoxy, chloro or
--NO.sub.2
134. A method for: agonizing HGF/SF activity; agonizing c-met;
stimulating cell proliferation; promoting angiogenic activity,
promoting the formation of new blood vessels; inducing
proliferation of epithelial cells, neuronal cells, Schwann cells or
oligodendrocyte cells; promoting axonal growth; inducing myelin
production; and/or reducing fibrosis; in a subject comprising:
administering to a subject in need thereof, optionally with a
pharmaceutically acceptable carrier, adjuvant or vehicle, a
therapeutically effective amount of a compound having the
structure: ##STR00073## or pharmaceutically acceptable salt, ester,
or salt of such an ester thereof; wherein Cy is an N-linked 5- to
10-membered heterocyclic or heteroaryl group other than an
optionally substituted homopiperidinyl group, wherein Cy is
optionally substituted with p occurrences of R.sup.4; p is an
integer from 1 to 6; each occurrence of R.sup.4 is independently
hydrogen, halogen, hydroxyl, --NO.sub.2, --NH.sub.2, --CN,
--CONH.sub.2, --SO.sub.2OH, an optionally substituted aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic moiety, --OR.sup.R, --S(.dbd.O).sub.nR.sup.d,
--NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, heteroaromatic, or acyl moiety; m is an integer from 1 to
4; each occurrence of R.sup.1 is independently hydrogen, halogen,
hydroxyl, --NO.sub.2, --NH.sub.2, --CN, --CONH.sub.2, --SO.sub.2OH,
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a,
--OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic or acyl
moiety; R.sup.a, for each occurrence, is hydrogen or an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, or heteroaromatic moiety; R.sup.b and R.sup.c, for each
occurrence, are independently hydrogen, hydroxy, SO.sub.2R.sup.d,
or an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic, heteroaromatic or acyl moiety; R.sup.d, for
each occurrence, is independently hydrogen, --N(R.sup.e).sub.2, or
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety; and R.sup.e, for
each occurrence, is independently hydrogen or aliphatic.
135. A method for protecting from cell apoptosis in a subject
comprising: administering to a subject in need thereof, optionally
with a pharmaceutically acceptable carrier, adjuvant or vehicle, a
therapeutically effective amount of a compound having the
structure: ##STR00074## or pharmaceutically acceptable salt, ester,
or salt of such an ester thereof; wherein Cy is an N-linked 5- to
10-membered heterocyclic or heteroaryl group other than an
optionally substituted homopiperidinyl group, wherein Cy is
optionally substituted with p occurrences of R.sup.4; p is an
integer from 1 to 6; each occurrence of R.sup.4 is independently
hydrogen, halogen, hydroxyl, --NO.sub.2, --NH.sub.2, --CN,
--CONH.sub.2, --SO.sub.2OH, an optionally substituted aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic moiety, --OR.sup.R, --S(.dbd.O).sub.nR.sup.d,
--NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, heteroaromatic, or acyl moiety; m is an integer from 1 to
4; each occurrence of R.sup.1 is independently hydrogen, halogen,
hydroxyl, --NO.sub.2, --NH.sub.2, --CN, --CONH.sub.2, --SO.sub.2OH,
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a,
--OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic or acyl
moiety; R.sup.a, for each occurrence, is hydrogen or an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, or heteroaromatic moiety; R.sup.b and R.sup.c, for each
occurrence, are independently hydrogen, hydroxy, SO.sub.2R.sup.d,
or an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic, heteroaromatic or acyl moiety; R.sup.d, for
each occurrence, is independently hydrogen, --N(R.sup.e).sub.2, or
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety; and R.sup.e, for
each occurrence, is independently hydrogen or aliphatic; with the
proviso that the compound does not have the following structure:
##STR00075##
136. A method for: agonizing HGF/SF activity; agonizing c-met;
stimulating cell proliferation; promoting angiogenic activity,
promoting the formation of new blood vessels; inducing
proliferation of epithelial cells, neuronal cells, Schwann cells or
oligodendrocyte cells; promoting axonal growth; inducing myelin
production; and/or reducing fibrosis; in a subject comprising:
administering to a subject in need thereof, optionally with a
pharmaceutically acceptable carrier, adjuvant or vehicle, a
therapeutically effective amount of a compound having the
structure: ##STR00076## or pharmaceutically acceptable salt, ester,
or salt of such an ester thereof; wherein m is an integer from 1 to
4; each occurrence of R.sup.1 is independently hydrogen, halogen,
hydroxyl, --NO.sub.2, --NH.sub.2, --CN, --CONH.sub.2, --SO.sub.2OH,
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a,
--OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic or acyl
moiety; R.sup.2 and R.sup.3 are independently hydrogen, hydroxyl,
--NH.sub.2, an optionally substituted aliphatic, heteroaliphatic,
alicyclic, heterocyclic, aromatic or heteroaromatic moiety,
--OR.sup.R, --S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c,
--C(.dbd.O)R.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, heteroaliphatic,
alicyclic, heterocyclic, aromatic or heteroaromatic or acyl moiety;
R.sup.a, for each occurrence, is independently selected from the
group consisting of hydrogen and an optionally substituted
aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, or
heteroaromatic moiety; R.sup.b and R.sup.c, for each occurrence,
are independently selected from the group consisting of hydrogen;
hydroxy; SO.sub.2R.sup.d; and aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic or acyl
moiety; R.sup.d, for each occurrence, is independently selected
from the group consisting of hydrogen; --N(R.sup.e).sub.2;
aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic; and R.sup.e, for each occurrence, is independently
hydrogen or aliphatic.
137. A method for protecting from cell apoptosis in a subject
comprising: administering to a subject in need thereof a
therapeutically effective amount of a compound having the
structure: ##STR00077## or pharmaceutically acceptable salt, ester,
or salt of such an ester thereof; wherein m is an integer from 1 to
4; each occurrence of R.sup.1 is independently hydrogen, halogen,
hydroxyl, --NO.sub.2, --NH.sub.2, --CN, --CONH.sub.2, --SO.sub.2OH,
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a,
--OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic or acyl
moiety; R.sup.2 and R.sup.3 are independently hydrogen, hydroxyl,
--NH.sub.2, an optionally substituted aliphatic, heteroaliphatic,
alicyclic, heterocyclic, aromatic or heteroaromatic moiety,
--OR.sup.R, --S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.e,
--C(.dbd.O)R.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, heteroaliphatic,
alicyclic, heterocyclic, aromatic or heteroaromatic or acyl moiety;
R.sup.a, for each occurrence, is independently selected from the
group consisting of hydrogen and an optionally substituted
aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, or
heteroaromatic moiety; R.sup.b and R.sup.c, for each occurrence,
are independently selected from the group consisting of hydrogen;
hydroxy; SO.sub.2R.sup.d; and aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic or acyl
moiety; R.sup.d, for each occurrence, is independently selected
from the group consisting of hydrogen; --N(R.sup.e).sub.2;
aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic; and R.sup.e, for each occurrence, is independently
hydrogen or aliphatic; with the proviso that the compound does not
have one of the following structures: ##STR00078##
138. A method for treating or lessening the severity of fibrotic
liver disease, hepatic ischemia-reperfusion injury, cerebral
infarction, ischemic heart disease, renal disease or lung
(pulmonary) fibrosis, multiple sclerosis or a neurodegenerative
disease in a subject comprising administering to a subject in need
thereof, optionally with a pharmaceutically acceptable carrier,
adjuvant or vehicle, a therapeutically effective amount of a
compound having the structure: ##STR00079## or pharmaceutically
acceptable salt, ester, or salt of such an ester thereof; wherein m
is an integer from 1 to 4; each occurrence of R.sup.1 is
independently hydrogen, halogen, hydroxyl, --NO.sub.2, --NH.sub.2,
--CN, --CONH.sub.2, --SO.sub.2OH, an optionally substituted
aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic moiety, --OR.sup.R, --S(.dbd.O).sub.nR.sup.d,
--NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, heteroaromatic or acyl moiety; R.sup.2 and R.sup.3 are
independently hydrogen, hydroxyl, --NH.sub.2, an optionally
substituted aliphatic, heteroaliphatic, alicyclic, heterocyclic,
aromatic or heteroaromatic moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted aliphatic, heteroaliphatic, alicyclic, heterocyclic,
aromatic or heteroaromatic or acyl moiety; or R.sup.2 and R.sup.3
taken together with the nitrogen to which they are attached form an
optionally substituted heteroaromatic or heterocyclic group other
than an optionally substituted homopiperidinyl group comprising
4-10 ring members and 0-3 additional heteroatoms selected from the
group consisting of O, N and S; the heteroaromatic or heterocyclic
group optionally further substituted with one or more optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, heteroaromatic or acyl groups; R.sup.b and R.sup.c, for
each occurrence, are independently selected from the group
consisting of hydrogen; hydroxy; SO.sub.2R.sup.d; and aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic
or acyl moiety; R.sup.d, for each occurrence, is independently
selected from the group consisting of hydrogen; --N(R.sup.e).sub.2;
aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic; and R.sup.e, for each occurrence, is independently
hydrogen or aliphatic; with the proviso that the compound does not
have one of the following structures: ##STR00080##
139. The method of claim 138, wherein the neurodegenerative disease
is metachromatic leukodystrophy, Refsum's disease,
adrenoleukodystrophy, Krabbe's disease, phenylketonuria, Canavan
disease, Pelizaeus-Merzbacher disease or Alexander's disease.
140. The method of claim 138, wherein the method is for treating or
lessening the severity of a disease or condition selected from
liver fibrosis associated with hepatitis C, hepatitis B, delta
hepatitis, chronic alcoholism, non-alcoholic steatohepatitis,
extrahepatic obstructions (stones in the bile duct),
cholangiopathies (primary biliary cirrhosis and sclerosing
cholangitis), autoimmune liver disease, and inherited metabolic
disorders (Wilson's disease, hemochromatosis, and alpha-1
antitrypsin deficiency); damaged and/or ischemic organs,
transplants or grafts; ischemia/reperfusion injury; stroke,
traumatic head injury, spinal cord injury, and other
cerebrovascular diseases; myocardial ischemia; atherosclerosis;
peripheral vascular disease; cardiovascular diseases; diabetes;
renal failure; renal fibrosis, lung fibrosis or idiopathic
pulmonary fibrosis; and multiple sclerosis.
141. The method of claim 138, wherein the method is for the
treatment of wounds for acceleration of healing; promoting
vascularization of a damaged and/or ischemic organ, transplant or
graft; amelioration of ischemia/reperfusion injury in the brain,
heart, liver, kidney, or other tissues or organs; normalization of
myocardial perfusion as a consequence of chronic cardiac ischemia
or myocardial infarction; development or augmentation of collateral
vessel development after vascular occlusion or to ischemic tissues
or organs; fibrotic diseases; hepatic disease including fibrosis
and cirrhosis; lung fibrosis; radiocontrast nephropathy; fibrosis
secondary to renal obstruction; renal trauma and transplantation;
renal failure secondary to chronic diabetes and/or hypertension;
and/or diabetes mellitus.
142. The method of claim 138, wherein the compound has the
following structure: ##STR00081## wherein Cy is an N-linked 5- to
10-membered heterocyclic or heteroaryl group other than an
optionally substituted homopiperidinyl group, wherein Cy is
optionally substituted with p occurrences of R.sup.4.
143. A method for treating or lessening the severity of liver
disease, lung disease, fibrotic disease, renal fibrosis, renal
fibrosis secondary to renal obstruction, renal trauma, liver
fibrosis, lung fibrosis, idiopathic pulmonary fibrosis,
radiocontrast nephropathy, promotion of vascularization of organs,
peripheral vascular disease, liver fibrosis associated with
hepatitis C, hepatitis B, delta hepatitis, chronic alcoholism,
non-alcoholic steatohepatitis, extrahepatic obstructions (stones in
the bile duct), cholangiopathies (primary biliary cirrhosis and
sclerosing cholangitis), autoimmune liver disease, or inherited
metabolic disorders (Wilson's disease, hemochromatosis, and alpha-1
antitrypsin deficiency), comprising administering to a subject in
need thereof a therapeutically effective amount of a compound
having the structure: ##STR00082##
144. An isolated compound having the structure: ##STR00083## or
pharmaceutically acceptable salt, ester, or salt of such an ester
thereof; wherein Cy is an N-linked pyrrolyl group optionally
substituted with p occurrences of R.sup.4; p is 1, 3, or 4; each
occurrence of R.sup.4 is independently hydrogen, halogen, hydroxyl,
--NO.sub.2, --NH.sub.2, --CN, --CONH.sub.2, --SO.sub.2OH, an
optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a,
--OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic or acyl
moiety; m is an integer from 1 to 4; each occurrence of R.sup.1 is
independently hydrogen, halogen, hydroxyl, --NO.sub.2, --NH.sub.2,
--CN, --CONH.sub.2, --SO.sub.2OH, an optionally substituted
aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic moiety, --OR.sup.R, --S(.dbd.O).sub.nR.sup.d,
--NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, heteroaromatic or acyl moiety; R.sup.a, for each
occurrence, is hydrogen or an optionally substituted aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aromatic, or
heteroaromatic moiety; R.sup.b and R.sup.c, for each occurrence,
are independently hydrogen, hydroxy, SO.sub.2R.sup.d, or an
optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic, heteroaromatic or acyl moiety; R.sup.d, for
each occurrence, is independently hydrogen, --N(R.sup.e).sub.2, or
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety; and R.sup.e, for
each occurrence, is independently hydrogen or aliphatic.
145. An isolated compound having the structure: ##STR00084## or
pharmaceutically acceptable salt, ester, or salt of such an ester
thereof; wherein Cy is an N-linked pyrrolyl group optionally
substituted with p occurrences of R.sup.4; p is 2; each occurrence
of R.sup.4 is independently hydrogen, halogen, hydroxyl,
--NO.sub.2, --NH.sub.2, --CN, --CONH.sub.2, --SO.sub.2OH, an
optionally substituted C.sub.2-20 aliphatic moiety, an optionally
substituted alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic moiety, --OR.sup.R, --S(.dbd.O).sub.nR.sup.d,
--NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, heteroaromatic or acyl moiety; m is an integer from 1 to
4; each occurrence of R.sup.1 is independently hydrogen, halogen,
hydroxyl, --NO.sub.2, --NH.sub.2, --CN, --CONH.sub.2, --SO.sub.2OH,
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a,
--OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic or acyl
moiety; R.sup.a, for each occurrence, is hydrogen or an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, or heteroaromatic moiety; R.sup.b and R.sup.c, for each
occurrence, are independently hydrogen, hydroxy, SO.sub.2R.sup.d,
or an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic, heteroaromatic or acyl moiety; R.sup.d, for
each occurrence, is independently hydrogen, --N(R.sup.e).sub.2, or
an optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety; and R.sup.e, for
each occurrence, is independently hydrogen or aliphatic.
Description
PRIORITY
[0001] This application claims priority to provisional applications
Ser. No. 60/613,740, filed Sep. 28, 2004, and Ser. No. 60/675,241,
filed Apr. 27, 2005, both of which are incorporated by reference
herein in their entireties.
BACKGROUND OF THE INVENTION
[0003] Scatter factor (SF; also known as hepatocyte growth factor
[HGF], and hereinafter referred to and abbreviated as HGF/SF) is a
pleiotropic' growth factor that stimulates cell growth, cell
motility, morphogenesis and angiogenesis. HGF/SF is produced as an
inactive monomer (.about.100 kDa) which is proteolytically
converted to its active form. Active HGF/SF is a heparin binding
heterodimeric protein composed of a 62 kDa a chain and a 34 kDa
.beta. chain. HGF/SF is a potent mitogen for parenchymal liver,
epithelial and endothelial cells (Matsumoto, K, and Nakamura, T.,
1997, Hepatocyte growth factor (HGF) as a tissue organizer for
organogenesis and regeneration. Biochem. Biophys. Res. Commun. 239,
639-44; Boros, P. and Miller, C. M., 1995, Hepatocyte growth
factor: a multifunctional cytokine. Lancet 345, 293-5). It
stimulates the growth of endothelial cells and also acts as a
survival factor against endothelial cell death (Morishita, R,
Nakamura, S, Nakamura, Y, Aoki, M, Moriguchi, A, Kida, I, Yo, Y,
Matsumoto, K, Nakamura, T, Higaki, J, Ogihara, T, 1997, Potential
role of an endothelium-specific growth factor, hepatocyte growth
factor, on endothelial damage in diabetes. Diabetes 46:138-42).
HGF/SF synthesized and secreted by vascular smooth muscle cells
stimulates endothelial cells to proliferate, migrate and
differentiate into capillary like tubes in vitro (Grant, D. S,
Kleinman, H. K., Goldberg, I. D., Bhargava, M. M., Nickoloff, B.
J., Kinsella, J. L., Polyerini, P., Rosen, E. M., 1993, Scatter
factor induces blood vessel formation in vivo. Proc. Natl. Acad.
Sci. USA 90:1937-41; Morishita, R., Nakamura, S., Hayashi, S.,
Taniyama, Y., Moriguchi, A., Nagano, T., Taiji, M., Noguchi, H.,
Takeshita, S., Matsumoto, K., Nakamura, T., Higaki, J., Ogihara,
T., 1999, Therapeutic angiogenesis induced by human recombinant
hepatocyte growth factor in rabbit hind limb ischemia model as
cytokine supplement therapy. Hypertension 33:1379-84). HGF/SF
containing implants in mouse subcutaneous tissue and rat cornea
induce growth of new blood vessels from surrounding tissue. HGF/SF
protein is expressed at sites of neovascularization including in
tumors (Jeffers, M., Rong, S., Woude, G. F., 1996, Hepatocyte
growth factor/scatter factor-Met signaling in tumorigenicity and
invasion/metastasis. J. Mol. Med. 74:505-13; Moriyama, T., Kataoka,
H., Koono, M., Wakisaka, S., 1999, Expression of hepatocyte growth
factor/scatter factor and its receptor c-met in brain tumors:
evidence for a role in progression of astrocytic tumors Int. J.
Mol. Med. 3:531-6). These findings suggest that HGF/SF plays a
significant role in the formation and repair of blood vessels under
physiologic and pathologic conditions. Further discussion of
angiogenic proteins may be found in U.S. Pat. Nos. 6,011,009 and
5,997,868, both of which are incorporated herein by reference in
their entireties.
[0004] In certain embodiments, the present invention is directed
toward the identification of small organic molecules that exhibit
HGF/SF activity and are thus useful in the treatment or prevention
of conditions or diseases in which HGF/SF activity is
desirable.
[0005] All citations in the present application are incorporated
herein by reference in their entireties. The citation of any
reference herein should not be construed as an admission that such
reference is available as "Prior Art" to the instant
application.
SUMMARY OF THE INVENTION
[0006] As discussed above, there remains a need for the development
of novel therapeutics that mimic or modulate HGF/SF activity. The
present invention is directed to novel therapeutics capable of
mimicking or modulating the activities of various cytokines,
including but not limited to hepatocyte growth factor (HGF; also
known as scatter factor (SF)), epidermal growth factor (EGF),
vascular endothelial growth factor (VEGF) and nerve growth factor
(NGF), by way of non-limiting examples, or at least providing one
or more of the same biological activities as the foregoing
exemplary but non-limiting cytokines. Moreover, the compounds of
the invention are capable of activating, agonizing or inducing
phosphorylation of, and/or directly activating, the signaling
pathways of various receptor tyrosine kinases, including but not
limited to the HGF/SF receptor (c-met), the EGF receptor, the VEGF
receptor and the NGF receptor.
[0007] In general, certain novel inventive compounds have the
structure shown in Formula (I) below:
##STR00002##
[0008] wherein m, p, R.sup.1 and R.sup.4 are as described generally
and in classes and subclasses herein, tautomers thereof,
pharmaceutical compositions thereof, which compounds are useful as
modulators of HGF/SF activity as well as that of other receptor
tyrosine kinases.
[0009] In certain other embodiments, the present invention provides
compounds of Formula (II.sup.A):
##STR00003##
[0010] wherein m, p, R.sup.1 and R.sup.4 are as described generally
and in classes and subclasses herein, tautomers thereof,
pharmaceutical compositions thereof, which compounds are useful as
modulators of HGF/SF activity as well as that of other receptor
tyrosine kinases.
[0011] In another aspect, the invention is directed to compositions
comprising of any of the compounds disclosed herein.
[0012] In another aspect, the invention provides methods for the
use of any of the compounds disclosed herein for modulating HGF/SF
activity in a patient or a biological sample, in particular
providing anti-fibrotic and anti-apoptotic activities. The
compounds and pharmaceutical compositions of the invention have
properties of HGF/SF and are useful in the treatment of any
disease, disorder or condition in which prophylactic or therapeutic
administration of HGF/SF would be useful.
[0013] In another aspect, the invention provides methods for the
use of any of the compounds disclosed herein for treating or
lessening the severity of a disease, disorder or condition
associated with HGF/SF or other cytokine activity.
[0014] In yet other aspects, the invention provides methods for the
prophylaxis or treatment of conditions and diseases in which
promoting or mimicking the activity of cytokines is desired, or
biological activities resulting from activating, agonizing or
inducing phosphorylation of c-met or other receptor tyrosine
kinases. In a preferred embodiment, the activity is inducing
endothelial cell proliferation or angiogenesis. In another
embodiment, the activity is to induce proliferation of other cells,
such as epithelial cells, neuronal cells, Schwann cells, or
oligodendrocyte cells. In a further embodiment, the activity is to
induce growth of neuronal axons. In yet another embodiment, the
activity is induction of myelin production. In yet another
embodiment, the activity is protection against apoptosis. In yet
another embodiment, the activity is anti-fibrotic. The compounds
described herein are useful in the treatment of conditions and
diseases where inducing endothelial cell proliferation or
therapeutic angiogenesis is beneficial, where inducing
proliferation of cells such as epithelial cells, neuronal cells,
Schwann cells, and oligodendrocyte cells is beneficial, where
inducing axonal growth is beneficial, where induction of myelin
production is beneficial, where protection against apoptosis is
beneficial, where anti-fibrosis is beneficial, or where all or some
of the foregoing activities are beneficial, including but not
limited to fibrotic liver disease, hepatic ischemia-reperfusion
injury, cerebral infarction, ischemic heart disease, renal disease
or lung (pulmonary) fibrosis, multiple sclerosis or various
neurodegenerative diseases. In certain embodiments, the method is
useful for treating a disease or condition, or lessening the
severity of a disease or condition selected from liver fibrosis
associated with hepatitis C, hepatitis B, delta hepatitis, chronic
alcoholism, non-alcoholic steatohepatitis, extrahepatic
obstructions (stones in the bile duct), cholangiopathies (primary
biliary cirrhosis and sclerosing cholangitis), autoimmune liver
disease, and inherited metabolic disorders (Wilson's disease,
hemochromatosis, and alpha-1 antitrypsin deficiency); damaged
and/or ischemic organs, transplants or grafts; ischemia/reperfusion
injury; stroke, traumatic head injury, spinal cord injury, and
other cerebrovascular diseases; myocardial ischemia;
atherosclerosis; peripheral vascular disease; other cardiovascular
diseases; diabetes; renal failure; renal fibrosis, lung fibrosis or
idiopathic pulmonary fibrosis; multiple sclerosis; and
neurodegenerative diseases such as but not limited to metachromatic
leukodystrophy, Refsum's disease, adrenoleukodystrophy, Krabbe's
disease, phenylketonuria, Canavan disease, Pelizaeus-Merzbacher
disease and Alexander's disease. In certain exemplary embodiments,
the method is for the treatment of wounds for acceleration of
healing; promoting vascularization of a damaged and/or ischemic
organ, transplant or graft; amelioration of ischemia/reperfusion
injury in the brain, heart, liver, kidney, or other tissues or
organs; normalization of myocardial perfusion as a consequence of
chronic cardiac ischemia or myocardial infarction; development or
augmentation of collateral vessel development after vascular
occlusion or to ischemic tissues or organs; fibrotic diseases;
hepatic disease including fibrosis and cirrhosis; lung fibrosis;
radiocontrast nephropathy; fibrosis secondary to renal obstruction;
renal trauma and transplantation; renal failure secondary to
chronic diabetes and/or hypertension; and/or diabetes mellitus. Use
of the compound is also provided for prophylaxis or preventing the
occurrence of the diseases in subjects, and in particular subjects
susceptible to of exhibiting risk factors for, the aforementioned
diseases and conditions. Common among the foregoing conditions is
benefit therein by promoting endothelial cell growth, angiogenesis
or formation of new blood vessels. Moreover, the compounds of the
invention are beneficial in providing biological activities
resulting from activating, agonizing, phosphorylating, or in any
other way activating the signaling pathway of the HGF/SF receptor,
c-met, or other receptor tyrosine kinases.
DEFINITIONS
[0015] It is understood that the compounds, as described herein,
may be substituted with any number of substituents or functional
moieties. In general, the term "substituted" whether preceded by
the term "optionally" or not, and substituents contained in
formulas of this invention, refer to the replacement of hydrogen
radicals in a given structure with the radical of a specified
substituent. When more than one position in any given structure may
be substituted with more than one substituent selected from a
specified group, the substituent may be either the same or
different at every position. As used herein, the term "substituted"
is contemplated to include all permissible substituents of organic
compounds. In a broad aspect, the permissible substituents include
acyclic and cyclic, branched and unbranched, carbocyclic and
heterocyclic, aromatic and non-aromatic, carbon and heteroatom
substituents of organic compounds. For purposes of this invention,
heteroatoms such as nitrogen may have hydrogen substituents and/or
any permissible substituents of organic compounds described herein
which satisfy the valencies of the heteroatoms. Furthermore, this
invention is not intended to be limited in any manner by the
permissible substituents of organic compounds. Combinations of
substituents and variables envisioned by this invention are
preferably those that result in the formation of stable compounds
useful in the treatment and prevention, for example of disorders,
as described generally above. Examples of substituents include, but
are not limited to aliphatic; heteroaliphatic; alicyclic;
heterocyclic; aromatic, heteroaromatic; aryl; heteroaryl;
alkylaryl; alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy;
heteroaryloxy; alkylthio; arylthio; heteroalkylthio;
heteroarylthio; F; Cl; Br; I; --NO.sub.2; --CN; --CF.sub.3;
--CH.sub.2CF.sub.3; --CHCl.sub.2; --CH.sub.2OH;
--CH.sub.2CH.sub.2OH; --CH.sub.2NH.sub.2;
--CH.sub.2SO.sub.2CH.sub.3; or -GR.sup.G1 wherein G is --O--,
--S--, --NR.sup.G2--, --C(.dbd.O)--, --S(.dbd.O)--, --SO.sub.2--,
--C(.dbd.O)O--, --C(.dbd.O)NR.sup.G2--, --OC(.dbd.O)--,
--NR.sup.G2C(.dbd.O)--, --OC(.dbd.O)O--, --OC(.dbd.O)NR.sup.G2--,
--NR.sup.G2C(.dbd.O)O--, --NR.sup.G2C(.dbd.O)NR.sup.G2--,
--C(.dbd.S)--, --C(.dbd.S)S--, --SC(.dbd.S)--, --SC(.dbd.S)S--,
--C(.dbd.NR.sup.G2)--, --C(.dbd.NR.sup.G2)O--,
--C(.dbd.NR.sup.G2)NR.sup.G3--, --OC(.dbd.NR.sup.G2)--, NR.sup.G2C
(.dbd.NR.sup.G3)--, --NR.sup.G2SO.sub.2--,
--NR.sup.G2SO.sub.2NR.sup.G3--, or --SO.sub.2NR.sup.G2--, wherein
each occurrence of R.sup.G1, R.sup.G2 and R.sup.G3 independently
includes, but is not limited to, hydrogen, halogen, or an
optionally substituted aliphatic, heteroaliphatic, alicyclic,
heterocyclic, aromatic, heteroaromatic, aryl, heteroaryl,
alkylaryl, or alkylheteroaryl moiety. Additional examples of
generally applicable substituents are illustrated by the specific
embodiments shown in the Examples that are described herein.
[0016] The term "stable", as used herein, preferably refers to
compounds which possess stability sufficient to allow manufacture
and which maintain the integrity of the compound for a sufficient
period of time to be detected and preferably for a sufficient
period of time to be useful for the purposes detailed herein.
[0017] The term "aliphatic", as used herein, includes both
saturated and unsaturated, straight chain (i.e., unbranched) or
branched aliphatic hydrocarbons as defined by IUPAC, which are
optionally substituted with one or more functional groups. As
defined herein, "aliphatic" is intended to include optionally
substituted alkyl, alkenyl and alkynyl moieties. Thus, as used
herein, the term "alkyl" includes straight and branched alkyl
groups. An analogous convention applies to other generic terms such
as "alkenyl", "alkynyl" and the like. Furthermore, as used herein,
the terms "alkyl", "alkenyl", "alkynyl" and the like encompass both
substituted and unsubstituted groups. In certain embodiments, as
used herein, "lower alkyl" is used to indicate those alkyl groups
(substituted, unsubstituted, branched or unbranched) having about
1-6 carbon atoms.
[0018] In certain embodiments, the alkyl, alkenyl and alkynyl
groups employed in the invention contain 1-20; 2-20; 3-20; 4-20;
5-20; 6-20; 7-20 or 8-20 aliphatic carbon atoms. In certain other
embodiments, the alkyl, alkenyl, and alkynyl groups employed in the
invention contain 1-10; 2-10; 3-10; 4-10; 5-10; 6-10; 7-10 or 8-10
aliphatic carbon atoms. In yet other embodiments, the alkyl,
alkenyl, and alkynyl groups employed in the invention contain 1-8;
2-8; 3-8; 4-8; 5-8; 6-20 or 7-8 aliphatic carbon atoms. In still
other embodiments, the alkyl, alkenyl, and alkynyl groups employed
in the invention contain 1-6; 2-6; 3-6; 4-6 or 5-6 aliphatic carbon
atoms. In yet other embodiments, the alkyl, alkenyl, and alkynyl
groups employed in the invention contain 1-4; 2-4 or 3-4 carbon
atoms. Illustrative aliphatic groups thus include, but are not
limited to, for example, methyl, ethyl, n-propyl, isopropyl, allyl,
n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, sec-pentyl,
isopentyl, tert-pentyl, n-hexyl, sec-hexyl, moieties and the like,
which again, may bear one or more substituents. Alkenyl groups
include, but are not limited to, for example, ethenyl, propenyl,
butenyl, 1-methyl-2-buten-1-yl, and the like. Representative
alkynyl groups include, but are not limited to, ethynyl, 2-propynyl
(propargyl), 1-propynyl and the like.
[0019] The term "alicyclic", as used herein, refers to compounds
which combine the properties of aliphatic and cyclic compounds and
include but are not limited to cyclic, or polycyclic aliphatic
hydrocarbons and bridged cycloalkyl compounds, which are optionally
substituted with one or more functional groups. As will be
appreciated by one of ordinary skill in the art, "alicyclic" is
intended herein to include, but is not limited to, cycloalkyl,
cycloalkenyl, and cycloalkynyl moieties, which are optionally
substituted with one or more functional groups. Illustrative
alicyclic groups thus include, but are not limited to, for example,
cyclopropyl, --CH.sub.2-cyclopropyl, cyclobutyl,
--CH.sub.2-cyclobutyl, cyclopentyl, --CH.sub.2-cyclopentyl-n,
cyclohexyl, --CH.sub.2-cyclohexyl, cyclohexenylethyl,
cyclohexanylethyl, norborbyl moieties and the like, which again,
may bear one or more substituents.
[0020] The term "cycloalkyl", as used herein, refers to cyclcic
alkyl groups, specifically to groups having three to seven,
preferably three to ten carbon atoms. Suitable cycloalkyls include,
but are not limited to cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl and the like, which, as in the case of
aliphatic, heteroaliphatic or heterocyclic moieties, may optionally
be substituted. An analogous convention applies to other generic
terms such as "cycloalkenyl", "cycloalkynyl" and the like.
Additional examples of generally applicable substituents are
illustrated by the specific embodiments shown in the Examples that
are described herein.
[0021] The term "heteroaliphatic", as used herein, refers to
aliphatic moieties in which one or more carbon atoms in the main
chain have been replaced with a heteroatom. Thus, a heteroaliphatic
group refers to an aliphatic chain which contains one or more
oxygen, sulfur, nitrogen, phosphorus or silicon atoms in place of
carbon atoms in the aliphatic main chain. Heteroaliphatic moieties
may be branched or linear unbranched. In certain embodiments,
heteroaliphatic moieties are substituted by independent replacement
of one or more of the hydrogen atoms thereon with one or more
moieties including, but not limited to aliphatic; heteroaliphatic;
alicyclic; heterocyclic; aromatic, heteroaromatic; aryl;
heteroaryl; alkylaryl; alkylheteroaryl; alkoxy; aryloxy;
heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio;
heteroarylthio; F; Cl; Br; I; --NO.sub.2; --CN; --CF.sub.3;
--CH.sub.2CF.sub.3; --CHCl.sub.2; --CH.sub.2OH;
--CH.sub.2CH.sub.2OH; --CH.sub.2NH.sub.2;
--CH.sub.2SO.sub.2CH.sub.3; or -GR.sup.G1 wherein G is --O--,
--S--, --NR.sup.G2--, --C(.dbd.O)--, --S(.dbd.O)--, --SO.sub.2--,
--C(.dbd.O)O--, --C(.dbd.O)NR.sup.G2--, --OC(.dbd.O)--,
--NR.sup.G2C(.dbd.O)--, --OC(.dbd.O)O--, --OC(.dbd.O)NR.sup.G2--,
--NR.sup.G2C(.dbd.O)O--, --NR.sup.G2C(.dbd.O)NR.sup.G2--,
--C(.dbd.S)--, --C(.dbd.S)S--, --SC(.dbd.S)--, --SC(.dbd.S)S--,
--C(.dbd.NR.sup.G2)--, --C(.dbd.NR.sup.G2)O--,
--C(.dbd.NR.sup.G2)NR.sup.G3--, --OC(.dbd.NR.sup.G2)--,
NR.sup.G2C(.dbd.NR.sup.G3)--, --NR.sup.G2SO.sub.2NR.sup.G3--, or
--SO.sub.2NR.sup.G2--, wherein each occurrence of R.sup.G1,
R.sup.G2 and R.sup.G3 independently includes, but is not limited
to, hydrogen, halogen, or an optionally substituted aliphatic,
heteroaliphatic, alicyclic, heterocyclic, aromatic, heteroaromatic,
aryl, heteroaryl, alkylaryl, or alkylheteroaryl moiety. Additional
examples of generally applicable substituents are illustrated by
the specific embodiments shown in the Examples that are described
herein.
[0022] The term "heteroalicyclic", "heterocycloalkyl" or
"heterocyclic", as used herein, refers to compounds which combine
the properties of heteroaliphatic and cyclic compounds and include
but are not limited to saturated and unsaturated mono- or
polycyclic ring systems having 5-16 atoms wherein at least one ring
atom is a heteroatom selected from O, S and N (wherein the nitrogen
and sulfur heteroatoms may be optionally be oxidized), wherein the
ring systems are optionally substituted with one or more functional
groups, as defined herein. In certain embodiments, the term
"heterocyclic" refers to a non-aromatic 5-, 6- or 7-membered ring
or a polycyclic group, including, but not limited to a bi- or
tri-cyclic group comprising fused six-membered rings having between
one and three heteroatoms independently selected from oxygen,
sulfur and nitrogen, wherein (i) each 5-membered ring has 0 to 2
double bonds and each 6-membered ring has 0 to 2 double bonds, (ii)
the nitrogen and sulfur heteroatoms may optionally be oxidized,
(iii) the nitrogen heteroatom may optionally be quaternized, and
(iv) any of the above heterocyclic rings may be fused to an aryl or
heteroaryl ring. Representative heterocycles include, but are not
limited to, pyrrolidinyl, pyrazolinyl, pyrazolidinyl imidazolinyl,
imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl,
isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, and
tetrahydrofuryl. In certain embodiments, a "substituted
heterocycloalkyl or heterocycle" group is utilized and as used
herein, refers to a heterocycloalkyl or heterocycle group, as
defined above, substituted by the independent replacement of one or
more hydrogen atoms thereon with aliphatic; heteroaliphatic;
alicyclic; heterocyclic; aromatic, heteroaromatic; aryl;
heteroaryl; alkylaryl; alkylheteroaryl; alkoxy; aryloxy;
heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio;
heteroarylthio; F; Cl; Br; I; --NO.sub.2; --CN; --CF.sub.3;
--CH.sub.2CF.sub.3; --CHCl.sub.2; --CH.sub.2OH;
--CH.sub.2CH.sub.2OH; --CH.sub.2NH.sub.2;
--CH.sub.2SO.sub.2CH.sub.3; or -GR.sup.G1 wherein G is --O--,
--S--, --NR.sup.G2--, --C(.dbd.O)--, --S(.dbd.O)--, --SO.sub.2--,
--C(.dbd.O)O--, --C(.dbd.O)NR.sup.G2--, --OC(.dbd.O)--,
--NR.sup.G2C(.dbd.O)--, --OC(.dbd.O)O--, --OC(.dbd.O)NR.sup.G2--,
--NR.sup.G2C(.dbd.O)O--, --NR.sup.G2C(.dbd.O)NR.sup.G2--,
--C(.dbd.S)--, --C(.dbd.S)S--, --SC(.dbd.S)--, --SC(.dbd.S)S--,
--C(.dbd.NR.sup.G2)--, --C(.dbd.NR.sup.G2)O--,
--C(.dbd.NR.sup.G2)NR.sup.G3--, --OC(.dbd.NR.sup.G2)--,
--NR.sup.G2C(.dbd.NR.sup.G3)--, --NR.sup.G2SO.sub.2--,
--NR.sup.G2SO.sub.2NR.sup.G3--, or --SO.sub.2NR.sup.G2--, wherein
each occurrence of R.sup.G1, R.sup.G2 and R.sup.G3 independently
includes, but is not limited to, hydrogen, halogen, or an
optionally substituted aliphatic, heteroaliphatic, alicyclic,
heterocyclic, aromatic, heteroaromatic, aryl, heteroaryl,
alkylaryl, or alkylheteroaryl moiety. Additional examples or
generally applicable substituents are illustrated by the specific
embodiments shown in the Examples, which are described herein.
[0023] Additionally, it will be appreciated that any of the
alicyclic or heterocyclic moieties described above and herein may
comprise an aryl or heteroaryl moiety fused thereto. Additional
examples of generally applicable substituents are illustrated by
the specific embodiments shown in the Examples that are described
herein.
[0024] In general, the term "aromatic moiety", as used herein,
refers to a stable mono- or polycyclic, unsaturated moiety having
preferably 3-14 carbon atoms, each of which may be substituted or
unsubstituted. In certain embodiments, the term "aromatic moiety"
refers to a planar ring having p-orbitals perpendicular to the
plane of the ring at each ring atom and satisfying the Huckel rule
where the number of pi electrons in the ring is (4n+2) wherein n is
an integer. A mono- or polycyclic, unsaturated moiety that does not
satisfy one or all of these criteria for aromaticity is defined
herein as "non-aromatic", and is encompassed by the term
"alicyclic". Examples of aromatic moieties include, but are not
limited to, phenyl, indanyl, indenyl, naphthyl, phenanthryl and
anthracyl.
[0025] In general, the term "heteroaromatic moiety", as used
herein, refers to stable substituted or unsubstituted unsaturated
mono-heterocyclic or polyheterocyclic moieties having preferably
3-14 carbon atoms, comprising at least one ring having p-orbitals
perpendicular to the plane of the ring at each ring atom, and
satisfying the Huckel rule where the number of pi electrons in the
ring is (4n+2) wherein n is an integer. Examples of heteroaromatic
moieties include, but are not limited to, pyridyl, quinolinyl,
dihydroquinolinyl, isoquinolinyl, quinazolinyl, dihydroquinazolyl,
and tetrahydroquinazolyl.
[0026] It will also be appreciated that aromatic and heteroaromatic
moieties, as defined herein, may be attached via an aliphatic
(e.g., alkyl) or heteroaliphatic (e.g., heteroalkyl) moiety and
thus also include moieties such as -(aliphatic)aromatic,
-(heteroaliphatic)aromatic, -(aliphatic)heteroaromatic,
-(heteroaliphatic)heteroaromatic, -(alkyl)aromatic,
-(heteroalkyl)aromatic, -(alkyl)heteroaromatic, and
-(heteroalkyl)heteroaromatic moieties. Thus, as used herein, the
phrases "aromatic or heteroaromatic moieties" and "aromatic,
heteroaromatic, -(alkyl)aromatic, -(heteroalkyl)aromatic,
-(heteroalkyl)heteroaromatic, and -(heteroalkyl)heteroaromatic" are
interchangeable. Substituents include, but are not limited to, any
of the previously mentioned substituents, i.e., the substituents
recited for aliphatic moieties, or for other moieties as disclosed
herein, resulting in the formation of a stable compound.
[0027] In general, the term "aryl" refers to aromatic moieties, as
described above, excluding those attached via an aliphatic (e.g.,
alkyl) or heteroaliphatic (e.g., heteroalkyl) moiety. In certain
embodiments of the present invention, "aryl" refers to a mono- or
bicyclic carbocyclic ring system having one or two rings satisfying
the Huckel rule for aromaticity, including, but not limited to,
phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl and the
like.
[0028] Similarly, the term "heteroaryl" refers to heteroaromatic
moieties, as described above, excluding those attached via an
aliphatic (e.g., alkyl) or heteroaliphatic (e.g., heteroalkyl)
moiety. In certain embodiments of the present invention, the term
"heteroaryl", as used herein, refers to a cyclic unsaturated
radical having from about five to about ten ring atoms of which one
ring atom is selected from S, O and N; zero, one or two ring atoms
are additional heteroatoms independently selected from S, O and N;
and the remaining ring atoms are carbon, the radical being joined
to the rest of the molecule via any of the ring atoms, such as, for
example, pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl,
imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl,
oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, and
the like.
[0029] As defined herein, "aryl" and "heteroaryl" groups (including
bicyclic aryl groups) can be unsubstituted or substituted, wherein
substitution includes replacement of one or more of the hydrogen
atoms thereon independently with any of the previously mentioned
substitutents, i.e., the substituents recited for aliphatic
moieties, or for other moieties as disclosed herein, resulting in
the formation of a stable compound. For example, aryl and
heteroaryl groups (including bicyclic aryl groups) can be
unsubstituted or substituted, wherein substitution includes
replacement of one or more of the hydrogen atoms thereon
independently with any one or more of the following moieties
including, but not limited to: aliphatic; heteroaliphatic;
alicyclic; heterocyclic; aromatic, heteroaromatic; aryl;
heteroaryl; alkylaryl; alkylheteroaryl; alkoxy; aryloxy;
heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio;
heteroarylthio; F; Cl; Br; I; --NO.sub.2; --CN; --CF.sub.3;
--CH.sub.2CF.sub.3; --CHCl.sub.2; --CH.sub.2OH;
--CH.sub.2CH.sub.2OH; --CH.sub.2NH.sub.2;
--CH.sub.2SO.sub.2CH.sub.3; or -GR.sup.G1 wherein G is --O--,
--S--, --NR.sup.G2--, --C(.dbd.O)--, --S(O)--, --SO.sub.2--,
--C(.dbd.O)O--, --C(.dbd.O)NR.sup.G2--, --OC(.dbd.O)--,
--NR.sup.G2C(.dbd.O)--, --OC(.dbd.O)O--, --OC(.dbd.O)NR.sup.G2--,
--NR.sup.G2C(.dbd.O)O--, --NR.sup.G2C(.dbd.O)NR.sup.G2--,
--C(.dbd.S)--, --C(.dbd.S)S--, --SC(.dbd.S)--, --SC(.dbd.S)S--,
--C(.dbd.NR.sup.G2)--, --C(.dbd.NR.sup.G2)O--,
--C(.dbd.NR.sup.G2)NR.sup.G3--, --OC(.dbd.NR.sup.G2)--,
--NR.sup.G2C(NR.sup.G3)--, --NR.sup.G2SO.sub.2--,
--NR.sup.G2SO.sub.2NR.sup.G3--, or --SO.sub.2NR.sup.G2--, wherein
each occurrence of R.sup.G1, R.sup.G2 and R.sup.G3 independently
includes, but is not limited to, hydrogen, halogen, or an
optionally substituted aliphatic, heteroaliphatic, alicyclic,
heterocyclic, aromatic, heteroaromatic, aryl, heteroaryl,
alkylaryl, or alkylheteroaryl moiety. Additionally, it will be
appreciated, that any two adjacent groups taken together may
represent a 4, 5, 6, or 7-membered substituted or unsubstituted
alicyclic or heterocyclic moiety. Additional examples of generally
applicable substituents are illustrated by the specific embodiments
shown in the Examples that are described herein.
[0030] The term "alkoxy" or "alkyloxy", as used herein refers to a
saturated (i.e., O-alkyl) or unsaturated (i.e., O-alkenyl and
O-alkynyl) group attached to the parent molecular moiety through an
oxygen atom. In certain embodiments, the alkyl group contains 1-20;
2-20; 3-20; 4-20; 5-20; 6-20; 7-20 or 8-20 aliphatic carbon atoms.
In certain other embodiments, the alkyl group contains 1-10; 2-10;
3-10; 4-10; 5-10; 6-10; 7-10 or 8-10 aliphatic carbon atoms. In yet
other embodiments, the alkyl, alkenyl, and alkynyl groups employed
in the invention contain 1-8; 2-8; 3-8; 4-8; 5-8; 6-20 or 7-8
aliphatic carbon atoms. In still other embodiments, the alkyl group
contains 1-6; 2-6; 3-6; 4-6 or 5-6 aliphatic carbon atoms. In yet
other embodiments, the alkyl group contains 1-4; 2-4 or 3-4
aliphatic carbon atoms. Examples of alkoxy, include but are not
limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy,
i-butoxy, sec-butoxy, tert-butoxy, neopentoxy, n-hexoxy and the
like.
[0031] The term "thioalkyl" as used herein refers to a saturated
(i.e., S-alkyl) or unsaturated (i.e., S-alkenyl and S-alkynyl)
group attached to the parent molecular moiety through a sulfur
atom. In certain embodiments, the alkyl group contains 1-20
aliphatic carbon atoms. In certain other embodiments, the alkyl
group contains 1-10 aliphatic carbon atoms. In yet other
embodiments, the alkyl, alkenyl, and alkynyl groups employed in the
invention contain 1-8 aliphatic carbon atoms. In still other
embodiments, the alkyl group contains 1-6 aliphatic carbon atoms.
In yet other embodiments, the alkyl group contains 1-4 aliphatic
carbon atoms. Examples of thioalkyl include, but are not limited
to, methylthio, ethylthio, propylthio, isopropylthio, n-butylthio,
and the like.
[0032] The term "alkylamino" refers to a group having the structure
--NHR' wherein R' is aliphatic or alicyclic, as defined herein. The
term "aminoalkyl" refers to a group having the structure
NH.sub.2R'--, wherein R' is aliphatic or alicyclic, as defined
herein. In certain embodiments, the aliphatic or alicyclic group
contains 1-20 aliphatic carbon atoms. In certain other embodiments,
the aliphatic or alicyclic group contains 1-10 aliphatic carbon
atoms. In still other embodiments, the aliphatic or alicyclic group
contains 1-6 aliphatic carbon atoms. In yet other embodiments, the
aliphatic or alicyclic group contains 1-4 aliphatic carbon atoms.
In yet other embodiments, R' is an alkyl, alkenyl, or alkynyl group
containing 1-8 aliphatic carbon atoms. Examples of alkylamino
include, but are not limited to, methylamino, ethylamino,
iso-propylamino and the like.
[0033] Some examples of substituents of the above-described
aliphatic (and other) moieties of compounds of the invention
include, but are not limited to aliphatic; alicyclic;
heteroaliphatic; heterocyclic; aromatic; heteroaromatic; aryl;
heteroaryl; alkylaryl; heteroalkylaryl; alkylheteroaryl;
heteroalkylheteroaryl; alkoxy; aryloxy; heteroalkoxy;
heteroaryloxy; alkylthio; arylthio; heteroalkylthio;
heteroarylthio; F; Cl; Br; I; --OH; --NO.sub.2; --CN; --CF.sub.3;
--CH.sub.2CF.sub.3; --CHCl.sub.2; --CH.sub.2OH;
--CH.sub.2CH.sub.2OH; --CH.sub.2NH.sub.2;
--CH.sub.2SO.sub.2CH.sub.3; --C(.dbd.O)R.sub.x;
--CO.sub.2(R.sub.x); --C(.dbd.O)N(R.sub.x).sub.2;
--OC(.dbd.O)R.sub.x; --OCO.sub.2R.sub.x;
--OC(.dbd.O)N(R.sub.x).sub.2; --N(R.sub.x).sub.2; --OR.sub.x;
--SR.sub.x; --S(O)R.sub.x; --S(O).sub.2R.sub.x;
--NR.sub.x(CO)R.sub.x; --N(R.sub.x)CO.sub.2R.sub.x;
--N(R.sub.x)S(O).sub.2R.sub.x;
--N(R.sub.x)C(.dbd.O)N(R.sub.x).sub.2;
--S(O).sub.2N(R.sub.x).sub.2; wherein each occurrence of R.sub.x
independently includes, but is not limited to, aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl,
alkylaryl, alkylheteroaryl, heteroalkylaryl or
heteroalkylheteroaryl, wherein any of the aliphatic, alicyclic,
heteroaliphatic, heterocyclic, alkylaryl, or alkylheteroaryl
substituents described above and herein may be substituted or
unsubstituted, branched or unbranched, saturated or unsaturated,
and wherein any of the aryl or heteroaryl substituents described
above and herein may be substituted or unsubstituted. Additional
examples of generally applicable substituents are illustrated by
the specific embodiments shown in the Examples that are described
herein.
[0034] The terms "halo" and "halogen" as used herein refer to an
atom selected from fluorine, chlorine, bromine and iodine.
[0035] The term "haloalkyl" denotes an alkyl group, as defined
above, having one, two, or three halogen atoms attached thereto and
is exemplified by such groups as chloromethyl, bromoethyl,
trifluoromethyl, and the like.
[0036] The term "amino", as used herein, refers to a primary
(--NH.sub.2), secondary (--NHR.sub.x), tertiary (--NR.sub.xR.sub.y)
or quaternary (--N.sup.+R.sub.xR.sub.yR.sub.z) amine, where
R.sub.x, R.sub.y and R.sub.z, are independently an aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic moiety, as defined herein. Examples of amino groups
include, but are not limited to, methylamino, dimethylamino,
ethylamino, diethylamino, diethylaminocarbonyl, methylethylamino,
iso-propylamino, piperidino, trimethylamino, and propylamino.
[0037] The term "acyl", as used herein, refers to a group having
the general formula --C(.dbd.O)R, where R is an aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic moiety, as defined herein.
[0038] The term "C.sub.2-6alkenylene", as used herein, refers to a
substituted or unsubstituted, linear or branched unsaturated
divalent radical consisting solely of carbon and hydrogen atoms,
having from two to six carbon atoms, having a free valence "-" at
both ends of the radical, and wherein the unsaturation is present
only as double bonds and wherein a double bond can exist between
the first carbon of the chain and the rest of the molecule.
[0039] As used herein, the terms "aliphatic", "heteroaliphatic",
"alkyl", "alkenyl", "alkynyl", "heteroalkyl", "heteroalkenyl",
"heteroalkynyl", and the like encompass substituted and
unsubstituted, saturated and unsaturated, and linear and branched
groups. Similarly, the terms "alicyclic", "heterocyclic",
"heterocycloalkyl", "heterocycle" and the like encompass
substituted and unsubstituted, and saturated and unsaturated
groups. Additionally, the terms "cycloalkyl", "cycloalkenyl",
"cycloalkynyl", "heterocycloalkyl", "heterocycloalkenyl",
"heterocycloalkynyl", "aromatic", "heteroaromatic", "aryl",
"heteroaryl" and the like encompass both substituted and
unsubstituted groups.
[0040] The phrase, "pharmaceutically acceptable derivative", as
used herein, denotes any pharmaceutically acceptable salt, ester,
or salt of such ester, of such compound, or any other adduct or
derivative which, upon administration to a patient, is capable of
providing (directly or indirectly) a compound as otherwise
described herein, or a metabolite or residue thereof.
Pharmaceutically acceptable derivatives thus include among others
pro-drugs. A pro-drug is a derivative of a compound, usually with
significantly reduced pharmacological activity, which contains an
additional moiety, which is susceptible to removal in vivo yielding
the parent molecule as the pharmacologically active species. An
example of a pro-drug is an ester, which is cleaved in vivo to
yield a compound of interest. Another example is an N-methyl
derivative of a compound, which is susceptible to oxidative
metabolism resulting in N-demethylation. Pro-drugs of a variety of
compounds, and materials and methods for derivatizing the parent
compounds to create the pro-drugs, are known and may be adapted to
the present invention. Certain exemplary pharmaceutical
compositions and pharmaceutically acceptable derivatives will be
discussed in more detail herein below.
[0041] The term "tautomerization" refers to the phenomenon wherein
a proton of one atom of a molecule shifts to another atom. See,
Jerry March, Advanced Organic Chemistry: Reactions, Mechanisms and
Structures, Fourth Edition, John Wiley & Sons, pages 69-74
(1992). The term "tautomer" as used herein, refers to the compounds
produced by the proton shift.
[0042] By the term "protecting group", as used herein, it is meant
that a particular functional moiety, e.g., O, S, or N, is
temporarily blocked so that a reaction can be carried out
selectively at another reactive site in a multifunctional compound.
In preferred embodiments, a protecting group reacts selectively in
good yield to give a protected substrate that is stable to the
projected reactions; the protecting group must be selectively
removed in good yield by readily available, preferably nontoxic
reagents that do not attack the other functional groups; the
protecting group forms an easily separable derivative (more
preferably without the generation of new stereogenic centers); and
the protecting group has a minimum of additional functionality to
avoid further sites of reaction. As detailed herein, oxygen,
sulfur, nitrogen and carbon protecting groups may be utilized. For
example, in certain embodiments, as detailed herein, certain
exemplary oxygen protecting groups are utilized. These oxygen
protecting groups include, but are not limited to methyl ethers,
substituted methyl ethers (e.g., MOM (methoxymethyl ether), MTM
(methylthiomethyl ether), BOM (benzyloxymethyl ether), PMBM or MPM
(p-methoxybenzyloxymethyl ether), to name a few), substituted ethyl
ethers, substituted benzyl ethers, silyl ethers (e.g., TMS
(trimethylsilyl ether), TES (triethylsilylether), TIPS
(triisopropylsilyl ether), TBDMS (t-butyldimethylsilyl ether),
tribenzyl silyl ether, TBDPS (t-butyldiphenyl silyl ether), to name
a few), esters (e.g., formate, acetate, benzoate (Bz),
trifluoroacetate, dichloroacetate, to name a few), carbonates,
cyclic acetals and ketals. In certain other exemplary embodiments,
nitrogen protecting groups are utilized. These nitrogen protecting
groups include, but are not limited to, carbamates (including
methyl, ethyl and substituted ethyl carbamates (e.g., Troc), to
name a few) amides, cyclic imide derivatives, N-alkyl and N-aryl
amines, imine derivatives, and enamine derivatives, to name a few.
Certain other exemplary protecting groups are detailed herein,
however, it will be appreciated that the present invention is not
intended to be limited to these protecting groups; rather, a
variety of additional equivalent protecting groups can be readily
identified using the above criteria and utilized in the present
invention. Additionally, a variety of protecting groups are
described in "Protective Groups in Organic Synthesis" Third Ed.
Greene, T. W. and Wuts, P. G., Eds., John Wiley & Sons, New
York: 1999, the entire contents of which are hereby incorporated by
reference.
[0043] As used herein, the term "isolated" when applied to the
compounds of the present invention, refers to such compounds that
are (i) separated from at least some components with which they are
associated in nature or when they are made and/or (ii) produced,
prepared or manufactured by the hand of man.
[0044] As used herein the term "biological sample" includes,
without limitation, cell cultures or extracts thereof; biopsied
material obtained from an animal (e.g., mammal) or extracts
thereof; and blood, saliva, urine, feces, semen, tears, or other
body fluids or extracts thereof; or purified versions thereof. For
example, the term "biological sample" refers to any solid or fluid
sample obtained from, excreted by or secreted by any living
organism, including single-celled microorganisms (such as bacteria
and yeasts) and multicellular organisms (such as plants and
animals, for instance a vertebrate or a mammal, and in particular a
healthy or apparently healthy human subject or a human patient
affected by a condition or disease to be diagnosed or
investigated). The biological sample can be in any form, including
a solid material such as a tissue, cells, a cell pellet, a cell
extract, cell homogenates, or cell fractions; or a biopsy, or a
biological fluid. The biological fluid may be obtained from any
site (e.g. blood, saliva (or a mouth wash containing buccal cells),
tears, plasma, serum, urine, bile, seminal fluid, cerebrospinal
fluid, amniotic fluid, peritoneal fluid, and pleural fluid, or
cells therefrom, aqueous or vitreous humor, or any bodily
secretion), a transudate, an exudate (e.g. fluid obtained from an
abscess or any other site of infection or inflammation), or fluid
obtained from a joint (e.g. a normal joint or a joint affected by
disease such as rheumatoid arthritis, osteoarthritis, gout or
septic arthritis). The biological sample can be obtained from any
organ or tissue (including a biopsy or autopsy specimen) or may
comprise cells (whether primary cells or cultured cells) or medium
conditioned by any cell, tissue or organ. Biological samples may
also include sections of tissues such as frozen sections taken for
histological purposes. Biological samples also include mixtures of
biological molecules including proteins, lipids, carbohydrates and
nucleic acids generated by partial or complete fractionation of
cell or tissue homogenates. Although the sample is preferably taken
from a human subject, biological samples may be from any animal,
plant, bacteria, virus, yeast, etc. The term animal, as used
herein, refers to humans as well as non-human animals, at any stage
of development, including, for example, mammals, birds, reptiles,
amphibians, fish, worms and single cells. Cell cultures and live
tissue samples are considered to be pluralities of animals. In
certain exemplary embodiments, the non-human animal is a mammal
(e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat,
a sheep, cattle, a primate, or a pig). An animal may be a
transgenic animal or a human clone. If desired, the biological
sample may be subjected to preliminary processing, including
preliminary separation techniques.
BRIEF DESCRIPTION OF THE DRAWING
[0045] FIG. 1A-D show the increase in HUVEC proliferation by
compounds of the invention (A) compared to control and HGF, and
dose-response curves using three compounds of the invention
(B-D).
[0046] FIG. 2 A-C show that a compound of the invention protects
HUVEC from apoptosis induced by serum starvation (A), hydrogen
peroxide (B) and CRP (C), respectively.
[0047] FIG. 3 shows that a compound of the invention increases
phosphorylation of the HGF/SF receptor, c-met, in Schwann
cells.
[0048] FIG. 4 A-C show the a compound of the invention stimulates
proliferation of Schwann cells (A) and PC12 neuronal cells (B), and
stimulates myelin production by Schwann cells (C).
[0049] FIG. 5 shows oligodendrocyte proliferation is stimulated by
two compounds of the invention.
[0050] FIG. 6 A-D show the effect of two compounds of the invention
on axonal growth in vitro.
[0051] FIG. 7 shows that a compound of the invention increases
affected limb blood flow in rats in a hindlimb ischemia model.
[0052] FIG. 8 shows that a compound of the invention administered
to rats in which a stroke has been induced, reduces the infarct
size.
[0053] FIG. 9 A-B show the reduction in plaque area in the arteries
of ApoE knockout mice by a compound of the invention.
[0054] FIG. 10 shows that a compound of the invention improves
cardiac function after ischemia-reperfusion in vitro.
[0055] FIG. 11 shows the reduction in alpha-SMA RNA expression in
TGFbetal-induced renal fibrosis treated with a compound of the
invention.
[0056] FIG. 12 A-D show the effect of a compound of the invention
on bleomycin-induced pulmonary fibrosis in mice, through survival
(A), lung collagen-1 gene expression (B), lung collagen content
measured by hydroxyproline (C), and fibrotic score (D).
[0057] FIG. 13 A-C shows the effects of a compound of the invention
on renal fibrosis in a doxorubicin induction model, with regard to
BUN (A), serum creatinine (B) and prefibrotic score (C).
[0058] FIG. 14 A-F show the effect of a compound of the invention
on liver fibrosis induced by TAA. FIGS. 14 A-C: treatment initiated
at the time of TAA induction, showing hydroxyproline content (A),
alpha-SMA level (B) and collagen-1 gene expression (C). Delayed
oral treatment, FIGS. 14 D-F: showing hydroxyproline (D), portal
pressure (E) and fibrotic score (F).
[0059] FIG. 15 shows that a compound of the invention reduces
hyperglycemia in streptozotocin-treated (diabetic) mice.
[0060] FIG. 16 shows that a compound of the invention improves the
locomotor deficit induced in mice in the EAE multiple sclerosis
model.
[0061] FIG. 17 shows that a compound of the invention decreases
migration of monocytes.
DETAILED DESCRIPTION OF CERTAIN PREFERRED EMBODIMENTS OF THE
INVENTION
[0062] The compounds of the invention are capable of mimicking or
modulating the activities of cytokines, such as hepatocyte growth
factor (HGF; also known as scatter factor (SF)), epidermal growth
factor (EGF), vascular endothelial growth factor (VEGF) and nerve
growth factor (NGF), or provide one or more of the same biological
activities as the foregoing exemplary but non-limiting cytokines.
Moreover, the compounds of the invention are capable of activating,
agonizing or inducing phosphorylation of, and/or directly
activating the signaling pathways of various receptor tyrosine
kinases, including but not limited to the HGF/SF receptor (c-met),
EGF receptor, VEGF receptor or NGF receptor. The compounds of the
invention also induce the phosphorylation of c-met or agonize,
activate or phosphorylate other receptor tyrosine kinases, such as
but not limited to those mentioned above. In preferred embodiments,
inventive compounds are small molecule HGF/SF mimics or agonists.
Without wishing to be bound to any particular theory, in certain
other embodiments, small-molecule compounds of the invention
modulate the activity of the HGF/SF receptor, c-met. In further
embodiments, compounds of the invention bind to c-met.
[0063] Having such biological activities, the compounds of the
invention, optionally provided in a pharmaceutical composition,
find use in the prophylaxis or treatment of conditions and diseases
in which promoting or mimicking the activity of the aforementioned
cytokines, among others, is desired, or exhibiting biological
activities resulting from activating, agonizing or inducing
phosphorylation of c-met or other receptor tyrosine kinases. In a
preferred embodiment, the activity is inducing endothelial cell
proliferation or angiogenesis. In another embodiment, the activity
is to induce proliferation of other cells, such as epithelial
cells, neuronal cells, Schwann cells, or oligodendrocyte cells. In
a further embodiment, the activity is to induce growth of neuronal
axons. In yet another embodiment, the activity is induction of
myelin production. In yet another embodiment, the activity is
protection against apoptosis. In yet another embodiment, the
activity is anti-fibrotic.
[0064] The compounds of the invention are useful in the treatment
of conditions and diseases where inducing endothelial cell
proliferation or therapeutic angiogenesis is beneficial, where
inducing proliferation of cells such as epithelial cells, neuronal
cells, Schwann cells, or oligodendrocyte cells is beneficial, where
inducing axonal growth is beneficial, where induction of myelin
production is beneficial, where protection against apoptosis is
beneficial, where anti-fibrosis is beneficial, or where all or some
of these activities are beneficial, including but not limited to
fibrotic liver disease, hepatic ischemia-reperfusion injury,
cerebral infarction, ischemic heart disease, renal disease or lung
(pulmonary) fibrosis. In certain embodiments, the method is useful
for treating a disease or condition, or lessening the severity of a
disease or condition selected from liver fibrosis associated with
hepatitis C, hepatitis B, delta hepatitis, chronic alcoholism,
non-alcoholic steatohepatitis, extrahepatic obstructions (stones in
the bile duct), cholangiopathies (primary biliary cirrhosis and
sclerosing cholangitis), autoimmune liver disease, and inherited
metabolic disorders (Wilson's disease, hemochromatosis, and alpha-1
antitrypsin deficiency); damaged and/or ischemic organs,
transplants or grafts; ischemia/reperfusion injury; stroke,
traumatic head injury, spinal cord injury, and other
cerebrovascular diseases; diabetes; myocardial ischemia;
atherosclerosis; peripheral vascular disease; other cardiovascular
diseases; renal failure; renal fibrosis; and idiopathic pulmonary
fibrosis. In certain exemplary embodiments, the method is for the
treatment of wounds for acceleration of healing; promoting
vascularization of a damaged and/or ischemic organ, transplant or
graft; amelioration of ischemia/reperfusion injury in the brain,
heart, liver, kidney, and other tissues and organs; normalization
of myocardial perfusion as a consequence of chronic cardiac
ischemia or myocardial infarction; development or augmentation of
collateral vessel development after vascular occlusion or to
ischemic tissues or organs; fibrotic diseases; hepatic disease
including fibrosis and cirrhosis; lung fibrosis; radiocontrast
nephropathy; fibrosis secondary to renal obstruction; renal trauma
and transplantation; renal failure secondary to chronic diabetes
and/or hypertension; and/or diabetes mellitus.
[0065] Another disease amenable to treatment by the compounds and
compositions of the invention is multiple sclerosis (MS). MS
usually manifests itself between the 20th and 50th years of life.
Current estimates are that approximately 2.5 million people
worldwide have MS, with between 250,000 and 350,000 cases in the
United States, 50,000 cases in Canada, 130,000 cases in Germany,
85,000 cases in the United Kingdom, 75,000 cases in France, 50,000
cases in Italy, and 11,000 cases in Switzerland. MS attacks the
white matter of the central nervous system (CNS). In its classic
manifestation (90% of all cases), it is characterized by
alternating relapsing/remitting phases with periods of remission
growing shorter over time. Its symptoms include any combination of
spastic paraparesis, unsteady gait, diplopia, and incontinence.
[0066] Another category of diseases also amenable to treatment
herein are the hereditary neurodegenerative disorders. This
category includes the eight identified leukodystrophies:
metachromatic leukodystrophy, Refsum's disease,
adrenoleukodystrophy, Krabbe's disease, phenylketonuria, Canavan
disease, Pelizaeus-Merzbacher disease and Alexander's disease. The
first six are storage disorders. The lack or the malfunctioning of
an enzyme causes a toxic buildup of chemical substances. In
Pelizaeus-Merzbacher disease myelin is never formed
(dysmyelination) because of a mutation in the gene that produces a
basic protein of CNS myelin. The etiology of Alexander's disease
remains largely unknown. The clinical course of hereditary
demyelinating disorders, which usually tend to manifest themselves
in infancy or early childhood, is tragic. Previously normal
children are deprived, in rapid progression, of sight, hearing,
speech, and ambulation. Equally tragic is their prognosis: death
within a few years.
[0067] Use of the compound for prophylaxis or preventing the
occurrence of the diseases in subjects, and in particular subjects
susceptible to of exhibiting risk factors for, the aforementioned
diseases and conditions. Common among the foregoing conditions is
benefit therein by promoting endothelial cell growth, angiogenesis
or formation of new blood vessels. These are merely exemplary of
the biological activities of the present compounds.
[0068] Compounds of this invention include those generally set
forth above and described specifically herein, and are illustrated
in part by the various classes, subgenera and species disclosed
herein.
[0069] Additionally, the present invention provides
pharmaceutically acceptable derivatives of the inventive compounds,
and methods of treating a subject using these compounds,
pharmaceutical compositions thereof, or either of these in
combination with one or more additional therapeutic agents.
[0070] 1) General Description of Compounds of the Invention
[0071] In certain embodiments, compounds of the invention include
compounds of the general Formula (I) as further defined below:
##STR00004##
[0072] or pharmaceutically acceptable derivative thereof;
[0073] wherein m is an integer from 1 to 4;
[0074] p is an integer from 1 to 6;
[0075] each occurrence of R.sup.1 and R.sup.4 is independently
hydrogen, halogen, hydroxyl, --NO.sub.2, --NH.sub.2, --CN, an
optionally substituted aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic or heteroaromatic moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a,
--OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic or acyl
moiety;
[0076] R.sup.a, for each occurrence, is independently selected from
the group consisting of hydrogen and an optionally substituted
aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, or
heteroaromatic moiety;
[0077] R.sup.b and R.sup.c, for each occurrence, are independently
selected from the group consisting of hydrogen; hydroxy;
SO.sub.2R.sup.d; and aliphatic, alicyclic, heteroaliphatic,
heterocyclic, aromatic, heteroaromatic or acyl moiety;
[0078] R.sup.d, for each occurrence, is independently selected from
the group consisting of hydrogen; --N(R.sup.e).sub.2; aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic; and
[0079] R.sup.e, for each occurrence, is independently hydrogen or
aliphatic.
[0080] In certain other embodiments, compounds of formula (I) are
defined as follows:
[0081] m is an integer from 1 to 4;
[0082] p is an integer from 1 to 6;
[0083] each occurrence of R.sup.1 and R.sup.4 is independently
hydrogen, halogen, hydroxyl, --NO.sub.2, --NH.sub.2, --CN, an
optionally substituted alkyl, heteroalkyl, cycloalkyl,
heterocyclic, aryl or heteroaryl moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a,
--OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted alkyl, heteroalkyl,
cycloalkyl, heterocyclic, aryl, heteroaryl or acyl moiety;
[0084] R.sup.a, for each occurrence, is independently hydrogen or
an optionally substituted alkyl, heteroalkyl, cycloalkyl,
heterocyclic, aryl or heteroaryl moiety;
[0085] R.sup.b and R.sup.c, for each occurrence, are independently
hydrogen, hydroxy, SO.sub.2R.sup.d, or an alkyl, heteroalkyl,
cycloalkyl, heterocyclic, aryl, heteroaryl or acyl moiety;
[0086] R.sup.d, for each occurrence, is independently hydrogen,
--N(R.sup.e).sub.2, alkyl, heteroalkyl, cycloalkyl, heterocyclic,
aryl or heteroaryl; and
R.sup.e, for each occurrence, is independently hydrogen or
alkyl.
[0087] In another aspect, the invention provides compounds of
formula (II):
##STR00005##
[0088] or pharmaceutically acceptable derivatives thereof;
[0089] m is an integer from 1 to 4;
[0090] each occurrence of R.sup.1 is independently hydrogen,
halogen, hydroxyl, --NO.sub.2, --NH.sub.2, --CN, an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic or heteroaromatic moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a,
--OPO.sub.2OR.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, alicyclic,
heteroaliphatic, heterocyclic, aromatic, heteroaromatic or acyl
moiety;
[0091] R.sup.2 and R.sup.3 are independently hydrogen, hydroxyl,
--NH.sub.2, an optionally substituted aliphatic, heteroaliphatic,
alicyclic, heterocyclic, aromatic or heteroaromatic moiety,
--OR.sup.R, --S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c,
--C(.dbd.O)R.sup.a or --C(.dbd.O)OR.sup.a; wherein n is 0-2,
R.sup.R is an optionally substituted aliphatic, heteroaliphatic,
alicyclic, heterocyclic, aromatic or heteroaromatic or acyl moiety;
or R.sup.2 and R.sup.3 taken together with the nitrogen to which
they are attached form an optionally substituted heteroaromatic or
heterocyclic group other than an optionally substituted
homopiperidinyl group comprising 4-10 ring members and 0-3
additional heteroatoms selected from the group consisting of O, N
and S; the heteroaromatic or heterocyclic group optionally further
substituted with one or more optionally substituted aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic
or acyl groups;
[0092] R.sup.a, for each occurrence, is hydrogen or an optionally
substituted aliphatic, alicyclic, heteroaliphatic, heterocyclic,
aromatic, or heteroaromatic moiety;
[0093] R.sup.b and R.sup.c, for each occurrence, are independently
hydrogen, hydroxy, SO.sub.2R.sup.d, or an optionally substituted
aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic,
heteroaromatic or acyl moiety;
[0094] R.sup.d, for each occurrence, is independently hydrogen,
--N(R.sup.e).sub.2, or an optionally substituted aliphatic,
alicyclic, heteroaliphatic, heterocyclic, aromatic or
heteroaromatic moiety; and
[0095] R.sup.e, for each occurrence, is independently hydrogen or
aliphatic.
[0096] In certain other embodiments, compounds of formula (II) are
defined as follows:
[0097] m is an integer from 1 to 4;
[0098] each occurrence of R.sup.1 is independently hydrogen,
halogen, hydroxyl, --NO.sub.2, --NH.sub.2, --CN, an optionally
substituted alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl or
heteroaryl moiety, --OR.sup.R, --S(.dbd.O)R.sup.d,
--NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl,
heteroaryl or acyl moiety;
[0099] R.sup.2 and R.sup.3 are independently hydrogen, hydroxyl,
--NH.sub.2, an optionally substituted alkyl, heteroalkyl,
cycloalkyl, heterocyclic, aryl or heteroaryl moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl,
heteroaryl or acyl moiety; or R.sup.2 and R.sup.3 taken together
with the nitrogen to which they are attached form a optionally
substituted heteroaryl or heterocyclic group other than an
optionally substituted homopiperidinyl group comprising 4-10 ring
members and 0-3 additional heteroatoms selected from the group
consisting of O, N and S; the heteroaryl or heterocyclic group
optionally further substituted with one or more optionally
substituted alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl,
heteroaryl or acyl groups;
[0100] wherein R.sup.a, for each occurrence, is independently
hydrogen or an optionally substituted alkyl, heteroalkyl,
cycloalkyl, heterocyclic, aryl or heteroaryl moiety;
[0101] R.sup.b and R.sup.c, for each occurrence, are independently
hydrogen, hydroxy, SO.sub.2R.sup.d, or an alkyl, heteroalkyl,
cycloalkyl, heterocyclic, aryl, heteroaryl or acyl moiety;
[0102] R.sup.d, for each occurrence, is independently hydrogen,
--N(R.sup.e).sub.2, alkyl, heteroalkyl, cycloalkyl, heterocyclic,
aryl or heteroaryl; and
[0103] R.sup.e, for each occurrence, is independently hydrogen or
alkyl.
[0104] In certain embodiments, the present invention defines
certain classes of compounds which are of special interest. For
example, one class of compounds of special interest includes those
compounds having the structure of formula (II.sup.A) in which the
compound has the structure:
##STR00006##
[0105] wherein m, p, R.sup.1 and R.sup.4 are as defined in classes
and subclasses herein; and Cy is an optionally substituted N-linked
5- to 10-membered heterocyclic group other than an optionally
substituted homopiperidinyl group.
[0106] Another class of compounds of special interest includes
those compounds having the structure of formula (II.sup.B) in which
the compound has the structure:
##STR00007##
[0107] wherein m, p, R.sup.1 and R.sup.4 are as defined in classes
and subclasses herein; and q is an integer selected from 1, 2 or
4.
[0108] In certain embodiments, for compounds of formula (II.sup.A)
having the structure:
##STR00008##
[0109] --Cy--(R.sup.4).sub.p is not one of the following
structures:
##STR00009##
[0110] In certain embodiments, for compounds of formula (II.sup.A),
the following groups do not occur simultaneously as defined:
[0111] m is 1; R.sup.1 is H and --Cy--(R.sup.4).sub.p is one of the
following structures:
##STR00010##
[0112] wherein R.sup.4A is hydrogen, methyl, methoxy, chloro or
--NO.sub.2 and p and R.sup.4 are as defined above.
[0113] A number of important subclasses of each of the foregoing
classes of compounds of formulae (I) and (II) deserve separate
mention; these subclasses include subclasses of the foregoing
classes in which:
[0114] i) each occurrence of R.sup.1 is independently hydrogen,
halogen, hydroxyl, --NO.sub.2, --NH.sub.2, --CN, an optionally
substituted alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl or
heteroaryl moiety, --OR.sup.R, --S(.dbd.O).sub.nR.sup.d,
--NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl,
heteroaryl or acyl moiety; wherein R.sub.a is as defined in subset
lxvi) below;
[0115] ii) at least one occurrence of R.sup.1 is hydrogen;
[0116] iii) at least one occurrence of R.sup.1 is NO.sub.2;
[0117] iv) at least one occurrence of R.sup.1 is NH.sub.2;
[0118] v) at least one occurrence of R.sup.1 is --COOH,
--C(.dbd.O)OCH.sub.3, --COCH.sub.3, --CONH.sub.2, --SO.sub.2OH,
--SO.sub.2CH.sub.3, --SO.sub.2CF.sub.3, --OPO.sub.2OH,
--NHC(.dbd.O)CH.sub.3, --NHC(.dbd.O)CF.sub.3,
--NHC(.dbd.O)CH.sub.3, --NHC(.dbd.O)CF.sub.3, --NHSO.sub.2CH.sub.3
or --NHSO.sub.2CF.sub.3.
[0119] vi) at least one occurrence of R.sup.1 is halogen;
[0120] vii) at least one occurrence of R.sup.1 is an optionally
substituted N-linked heterocyclic group; viii) at least one
occurrence of R.sup.1 is an optionally substituted N-pyrrolyl
group;
[0121] ix) at least one occurrence of R.sup.1 is an aliphatic
moiety;
[0122] x) at least one occurrence of R.sup.1 is an alkyl
moiety;
[0123] xi) at least one occurrence of R.sup.1 is a lower alkyl
moiety;
[0124] xii) m is 1 and at least one occurrence of R.sup.1 is ortho
to the bond to the phthalazinone ring;
[0125] xiii) m is 1 and at least one occurrence of R.sup.1 is meta
to the bond to the phthalazinone ring;
[0126] xiv) each occurrence of R.sup.1 is independently hydrogen,
--NO.sub.2, --NH.sub.2, --COOH, --C(.dbd.O)OCH.sub.3, --COCH.sub.3,
--CONH.sub.2, --SO.sub.2OH, --SO.sub.2CH.sub.3, --SO.sub.2CF.sub.3,
--OPO.sub.2OH, --NHC(.dbd.O)CH.sub.3, --NHC(.dbd.O)CF.sub.3,
--NHSO.sub.2CH.sub.3, --NHSO.sub.2CF.sub.3, halogen, an optionally
substituted N-linked heterocyclic group or an aliphatic moiety;
[0127] xv) each occurrence of R.sup.1 is independently hydrogen,
--NO.sub.2, --NH.sub.2, --COOH, --C(.dbd.O)OCH.sub.3, --COCH.sub.3,
--CONH.sub.2, --SO.sub.2OH, --SO.sub.2CH.sub.3, --SO.sub.2CF.sub.3,
--OPO.sub.2OH, --NHC(.dbd.O)CH.sub.3, --NHC(.dbd.O)CF.sub.3,
--NHSO.sub.2CH.sub.3, --NHSO.sub.2CF.sub.3, halogen, an optionally
substituted N-pyrrolyl group or a lower alkyl moiety;
[0128] xvi) R.sup.2 and R.sup.3 are independently hydrogen,
hydroxyl, --NH.sub.2, an optionally substituted alkyl, heteroalkyl,
cycloalkyl, heterocyclic, aryl or heteroaryl moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl,
heteroaryl or acyl moiety; or R.sup.2 and R.sup.3 taken together
with the nitrogen to which they are attached form a optionally
substituted heteroaryl or heterocyclic group other than a
homopiperidinyl group comprising 4-10 ring members and 0-3
additional heteroatoms selected from the group consisting of O, N
and S; the heteroaryl or heterocyclic group optionally further
substituted with one or more optionally substituted alkyl,
cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl or acyl
groups; wherein R.sub.a is as defined in subset lxvi) below;
[0129] xvii) R.sup.2 and R.sup.3 are independently hydrogen,
hydroxyl, --NH.sub.2, an optionally substituted alkyl, heteroalkyl,
cycloalkyl, heterocyclic, aryl or heteroaryl moiety, --OR.sup.R,
--S(.dbd.O).sub.nR.sup.d, --NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl,
heteroaryl or acyl moiety; wherein R.sub.a is as defined in subset
lxvi) below;
[0130] xviii) R.sup.2 and R.sup.3 are independently hydrogen, lower
alkyl or aryl;
[0131] xix) R.sup.2 and R.sup.3 are independently hydrogen or lower
alkyl;
[0132] xx) R.sup.2 and R.sup.3 are independently a hydrophobic
group;
[0133] xxi) R.sup.2 and R.sup.3 are independently an aliphatic
group;
[0134] xxii) R.sup.2 and R.sup.3 are independently an unsubstituted
aliphatic group;
[0135] xxiii) R.sup.2 and R.sup.3 are independently a cyclic or
acyclic C.sub.6-12alkyl, C.sub.6-12alkenyl, or C.sub.6-12alkynyl
group;
[0136] xxiv) R.sup.2 and R.sup.3 are independently an unsubstituted
cyclic or acyclic C.sub.6-12alkyl, C.sub.6-12alkenyl, or
C.sub.6-12alkynyl group;
[0137] xxv) R.sup.2 and R.sup.3 are independently is an
-(alkyl)aryl group;
[0138] xxvi) R.sup.2 and R.sup.3 are independently a unsubstituted
-(alkyl)aryl group;
[0139] xxvii) R.sup.2 and R.sup.3 taken together with the nitrogen
to which they are attached form a optionally substituted heteroaryl
or heterocyclic group other than a homopiperidinyl group comprising
4-10 ring members and 0-3 additional heteroatoms selected from the
group consisting of O, N and S; the heteroaryl or heterocyclic
group optionally further substituted with one or more optionally
substituted alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl,
heteroaryl or acyl groups;
[0140] xxviii) R.sup.2 and R.sup.3 taken together with the nitrogen
atom to which they are attached form an optionally substituted
pyrrolyl, pyrrolidinyl, imidazolyl, imidazolidinyl, pyrazolyl,
pyrazolidinyl, 1,2,3-triazolyl, piperidinyl, morpholinyl,
thiomorpholinyl, piperazinyl, indolyl, isoindolyl, indolinyl,
indazolyl, benzimidazolyl or purinyl moiety;
[0141] xxix) R.sup.2 and R.sup.3 taken together with the nitrogen
to which they are attached form an optionally substituted
6-membered heterocyclic group comprising 0-3 additional heteroatoms
selected from the group consisting of O, N and S;
[0142] xxx) R.sup.2 and R.sup.3, taken together, represent the
hydrophobic portion of an optionally substituted N-linked ring;
[0143] xxxi) R.sup.2 and R.sup.3, taken together, represent the
hydrophobic portion of an N-linked ring substituted with
hydrophobic groups, such as one or more aliphatic groups;
[0144] xxxii) R.sup.2 and R.sup.3, taken together, represent the
hydrophobic portion of an optionally substituted piperidinyl
ring;
[0145] xxxiii) R.sup.2 and R.sup.3, taken together, represent the
hydrophobic portion of a piperidinyl ring substituted with
hydrophobic groups, such as one or more aliphatic groups;
[0146] xxxiv) each occurrence of R.sup.4 is independently hydrogen,
halogen, hydroxyl, --NO.sub.2, --NH.sub.2, --CN, an optionally
substituted alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl or
heteroaryl moiety, --OR.sup.R, --S(.dbd.O).sub.nR.sup.d,
--NR.sup.bR.sup.c, --C(.dbd.O)R.sup.a, --OPO.sub.2OR.sup.a or
--C(.dbd.O)OR.sup.a; wherein n is 0-2, R.sup.R is an optionally
substituted alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl,
heteroaryl or acyl moiety; wherein R.sub.a, R.sub.b, R.sub.e and
R.sub.d are as defined in subsets lxvi), lxvii) and lxviii)
below;
[0147] xxxv) at least one occurrence of R.sup.4 is hydrogen;
[0148] xxxvi) at least one occurrence of R.sup.4 is a hydrophobic
group;
[0149] xxxvii) at least one occurrence of R.sup.4 is an optionally
substituted aliphatic group;
[0150] xxxviii) at least one occurrence of R.sup.4 is an
unsubstituted aliphatic group;
[0151] xxxix) at least one occurrence of R.sup.4 is an optionally
substituted cyclic or acyclic C.sub.6-12alkyl, C.sub.6-12alkenyl,
or C.sub.6-12alkynyl group;
[0152] xl) at least one occurrence of R.sup.4 is an unsubstituted
cyclic or acyclic C.sub.6-12alkyl, C.sub.6-12alkenyl, or
C.sub.6-12alkynyl group;
[0153] xli) at least one occurrence of R.sup.4 is an optionally
substituted (alkyl)aryl group;
[0154] xlii) at least one occurrence of R.sup.4 is a unsubstituted
(alkyl)aryl group;
[0155] xliii) at least one occurrence of R.sup.4 is
--NR.sup.bR.sup.c;
[0156] xliv) at least one occurrence of R.sup.4 is NH.sub.2;
[0157] xlv) at least one occurrence of R.sup.4 is
--C(.dbd.O)OR.sup.a; wherein R.sub.a is as defined in subset lxvi)
below;
[0158] xlvi) at least one occurrence of R.sup.4 is CO.sub.2H;
[0159] xlvii) p is .gtoreq.3 and each occurrence of R.sup.4 is
independently a cyclic or acyclic C.sub.1-6alkyl, C.sub.2-6alkenyl,
C.sub.2-6alkynyl or (C.sub.1-6alkyl)aryl group;
[0160] xlviii) p is .gtoreq.3 and each occurrence of R.sup.4 is
independently methyl, ethyl, propyl, butyl, pentyl, hexyl, i-propyl
or benzyl;
[0161] xlixi) each occurrence of R.sup.4 is independently hydrogen,
halogen, an optionally substituted aliphatic group,
--NR.sup.bR.sup.c, or --C(.dbd.O)OR.sup.a, wherein R.sub.a, R.sub.b
and R.sub.e are as defined in subsets lxvi) and lxvii) below;
[0162] l) each occurrence of R.sup.4 is independently hydrogen,
halogen, an optionally substituted cyclic or acyclic
C.sub.6-12alkyl, C.sub.6-12alkenyl, or C.sub.6-12alkynyl group, an
optionally substituted -(alkyl)aryl group, --NH.sub.2 or
--CO.sub.2H;
[0163] li) m is 0;
[0164] lii) m is 1;
[0165] liii) m is 2;
[0166] liv) m is 3;
[0167] lv) m is 4;
[0168] lvi) p is 0;
[0169] lvii) p is 1;
[0170] lviii) p is 2;
[0171] lix) p is 3;
[0172] lx) p is 4;
[0173] lxi) p is 5;
[0174] lxii) p is 6;
[0175] lxiii) q is 1;
[0176] lxiv) q is 2;
[0177] xlv) q is 4;
[0178] lxvi) R.sup.a, for each occurrence, is independently
hydrogen or an optionally substituted alkyl, heteroalkyl,
cycloalkyl, heterocyclic, aryl or heteroaryl moiety;
[0179] lxvii) R.sup.b and R.sup.c, for each occurrence, are
independently hydrogen, hydroxy, SO.sub.2R.sup.d, or an alkyl,
heteroalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl or acyl
moiety;
[0180] lxviii) R.sup.d, for each occurrence, is independently
hydrogen, --N(R.sup.e).sub.2, alkyl, heteroalkyl, cycloalkyl,
heterocyclic, aryl or heteroaryl;
[0181] lxix) R.sup.e, for each occurrence, is independently
hydrogen or alkyl; and/or
[0182] lx) Cy is one of:
##STR00011##
[0183] wherein q is 1, 2 or 4 and p and R.sup.4 are as defined in
classes and subclasses herein, and R.sup.4A is hydrogen, hydroxy,
SO.sub.2R.sup.d, or an alkyl, heteroalkyl, cycloalkyl,
heterocyclic, aryl, heteroaryl or acyl moiety; wherein R.sup.d is
as defined in classes and subclasses herein.
[0184] It will be appreciated that for each of the classes and
subclasses described above and herein, any one or more occurrences
of groups such as aliphatic, heteroaliphatic, alkyl, heteroalkyl
may independently be substituted or unsubstituted, linear or
branched, saturated or unsaturated; and any one or more occurrences
of alicyclic, heterocyclic, cycloalkyl, aryl, heteroaryl,
cycloaliphatic, cycloheteroaliphatic may be substituted or
unsubstituted.
[0185] The reader will also appreciate that all possible
combinations of the variables described in i)-through lx) above
(e.g., R.sup.1-R.sup.4, m, p and q, among others) are considered
part of the invention. Thus, the invention encompasses any and all
compounds of formula I, and subclasses thereof, generated by taking
any possible permutation of variables R.sup.1-R.sup.4, m, p and q,
and other variables/substituents (e.g., R.sub.a-e, etc.) as further
defined for R.sup.1-R.sup.4, described in i)-through lx) above,
leading to a stable compound.
[0186] As the reader will appreciate, compounds of particular
interest include, among others, those which share the attributes of
one or more of the foregoing subclasses. Some of those subclasses
are illustrated by the following sorts of compounds:
[0187] I) Compounds of the Formula (and Pharmaceutically Acceptable
Derivatives Thereof):
##STR00012##
[0188] wherein p, R.sup.1 and R.sup.4 are as defined in classes and
subclasses herein. In certain embodiments, p is 1-4 and each
occurrence of R.sup.4 is independently hydrogen or lower alkyl. In
certain embodiments, at least one occurrence of R.sup.4 is a
hydrophilic group.
[0189] II) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00013##
[0190] wherein p and R.sup.4 are as defined in classes and
subclasses herein. In certain embodiments, p is 1-4 and each
occurrence of R.sup.4 is independently hydrogen or lower alkyl. In
certain embodiments, at least one occurrence of R.sup.4 is a
hydrophilic group.
[0191] III) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00014##
[0192] wherein p and R.sup.4 are as defined in classes and
subclasses herein. In certain embodiments,
[0193] p is 1-4 and each occurrence of R.sup.4 is independently
hydrogen or lower alkyl. In certain embodiments, at least one
occurrence of R.sup.4 is a hydrophilic group.
[0194] IV) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00015##
[0195] wherein p and R.sup.4 are as defined in classes and
subclasses herein. In certain embodiments, p is 1-4 and each
occurrence of R.sup.4 is independently hydrogen or lower alkyl. In
certain embodiments, at least one occurrence of R.sup.4 is a
hydrophilic group.
[0196] V) Compounds of the Formula (and Pharmaceutically Acceptable
Derivatives Thereof):
##STR00016##
[0197] wherein m, R.sup.1 and R.sub.a are as defined in classes and
subclasses herein. In certain embodiments, R.sub.a is hydrogen. In
certain embodiments, R.sub.a is lower alkyl. In certain
embodiments, R.sub.a is a hydrophilic group. In certain
embodiments, R.sub.a is an optionally substituted cyclic or acyclic
C.sub.6-12alkyl, C.sub.6-12alkenyl, or C.sub.6-12alkynyl group. In
certain embodiments, R.sub.a is an optionally substituted
(alkyl)aryl group.
[0198] VI) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00017##
[0199] wherein m, R.sup.1, R.sub.b and R.sub.c are as defined in
classes and subclasses herein. In certain embodiments, R.sub.b and
R.sub.c are independently hydrogen or lower alkyl. In certain
embodiments, R.sub.b and R.sub.c are independently a hydrophilic
group. In certain embodiments, R.sub.b and R.sub.c are
independently an optionally substituted cyclic or acyclic
C.sub.6-12alkyl, C.sub.6-12alkenyl, or C.sub.6-12alkynyl group. In
certain embodiments, R.sub.b and R.sub.e are independently an
optionally substituted -(alkyl)aryl group.
[0200] VII) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00018##
[0201] wherein Cy, p, R.sup.1 and R.sup.4 are as defined in classes
and subclasses herein. In certain embodiments, p is 1-4 and each
occurrence of R.sup.4 is independently hydrogen or lower alkyl. In
certain embodiments, at least one occurrence of R.sup.4 is a
hydrophilic group.
[0202] VIII) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof
##STR00019##
[0203] wherein Cy, p and R.sup.4 are as defined in classes and
subclasses herein; with the proviso that --Cy--(R.sup.4).sub.p is
not one of the following structures:
##STR00020##
[0204] In certain embodiments, p is 1-4 and each occurrence of
R.sup.4 is independently hydrogen or lower alkyl. In certain
embodiments, at least one occurrence of R.sup.4 is a hydrophilic
group.
[0205] IX) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof:
##STR00021##
[0206] wherein Cy, p and R.sup.4 are as defined in classes and
subclasses herein. In certain embodiments, p is 1-4 and each
occurrence of R.sup.4 is independently hydrogen or lower alkyl. In
certain embodiments, at least one occurrence of R.sup.4 is a
hydrophilic group.
[0207] X) Compounds of the Formula (and Pharmaceutically Acceptable
Derivatives Thereof):
##STR00022##
[0208] wherein Cy, p and R.sup.4 are as defined in classes and
subclasses herein, with the proviso that --Cy--(R.sup.4).sub.p is
not one of the following structures:
##STR00023##
[0209] wherein R.sup.4A is hydrogen, methyl, methoxy, chloro or
--NO.sub.2 and p and R.sup.4 are as defined in classes and
subclasses herein.
[0210] In certain embodiments, p is 1-4 and each occurrence of
R.sup.4 is independently hydrogen or lower alkyl. In certain
embodiments, at least one occurrence of R.sup.4 is a hydrophilic
group.
[0211] XI) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives thereof):
##STR00024##
[0212] wherein Cy, m, R.sup.1 and R.sub.a are as defined in classes
and subclasses herein. In certain embodiments, R.sub.a is hydrogen.
In certain embodiments, R.sub.a is lower alkyl.
[0213] XII) Compounds of the Formula (and Pharmaceutically
Acceptable Derivatives Thereof):
##STR00025##
[0214] wherein Cy, m, R.sup.1, R.sub.b and R.sub.c are as defined
in classes and subclasses herein. In certain embodiments, R.sub.b
and R.sub.c are independently hydrogen or lower alkyl.
[0215] In certain embodiments, for compounds of classes I-XII
above, at least one occurrence of R.sup.4 is a hydrophobic group.
In certain embodiments, each occurrence of R.sup.4 is independently
a hydrophobic group. In certain embodiments, the hydrophobic group
is an aliphatic group. In certain embodiments, the hydrophobic
group is an unsubstituted aliphatic group. In certain embodiments,
the hydrophobic group is a cyclic or acyclic C.sub.6-12alkyl,
C.sub.6-12alkenyl, or C.sub.6-12alkynyl group. In certain
embodiments, the hydrophobic group is an unsubstituted cyclic or
acyclic C.sub.6-12alkyl, C.sub.6-12alkenyl, or C.sub.6-12alkynyl
group. In certain embodiments, the hydrophobic group is a
(alkyl)aryl group. In certain embodiments, the hydrophobic group is
an unsubstituted (alkyl)aryl group.
[0216] In certain embodiments, for compounds of classes V and VI
above, m is 0-2. In certain embodiments, m is 0. In certain
embodiments, m is 1.
[0217] In certain embodiments, for compounds of classes I-XII
above, p is 0-2. In certain embodiments, p is 0. In certain
embodiments, p is 1.
[0218] Non-limiting examples of compounds of the invention in
Formula (I) include:
##STR00026## ##STR00027## ##STR00028## ##STR00029##
##STR00030##
[0219] Examples of compounds of Formula (II) where R.sup.2 and
R.sup.3 do not form a ring include:
##STR00031##
[0220] In certain embodiments, compounds of Formula (II) do not
have the following structure:
##STR00032##
[0221] Non-limiting examples of compounds of Formula (II) wherein
the --NR.sup.2R.sup.3 moiety forms a ring, optionally further
substituted, include the following compounds:
##STR00033## ##STR00034## ##STR00035##
[0222] In certain embodiments, compounds of Formula (II) exclude
the following compounds:
##STR00036##
wherein R.sup.4A is hydrogen, methyl, wherein p and R.sup.4 are as
defined in classes methoxy, chloro or --NO.sub.2 and subclasses
herein.
[0223] In certain embodiments, compounds of Formula (II) exclude
the following compounds, whose compositions and uses are embraced
in the present invention:
##STR00037##
[0224] Some of the foregoing compounds can comprise one or more
asymmetric centers, and thus can exist in various isomeric forms,
e.g., stereoisomers and/or diastereomers. Thus, inventive compounds
and pharmaceutical compositions thereof may be in the form of an
individual enantiomer, diastereomer or geometric isomer, or may be
in the form of a mixture of stereoisomers. In certain embodiments,
the compounds of the invention are enantiopure compounds. In
certain other embodiments, mixtures of stereoisomers or
diastereomers are provided.
[0225] Furthermore, certain compounds, as described herein may have
one or more double bonds that can exist as either the Z or E
isomer, unless otherwise indicated. The invention additionally
encompasses the compounds as individual isomers substantially free
of other isomers and alternatively, as mixtures of various isomers,
e.g., racemic mixtures of stereoisomers. In addition to the
above-mentioned compounds per se, this invention also encompasses
pharmaceutically acceptable derivatives of these compounds and
compositions comprising one or more compounds of the invention and
one or more pharmaceutically acceptable excipients or
additives.
[0226] Compounds of the invention may be prepared by
crystallization of compound of formula (I) under different
conditions and may exist as one or a combination of polymorphs of
compound of general formula (I) forming part of this invention. For
example, different polymorphs may be identified and/or prepared
using different solvents, or different mixtures of solvents for
recrystallization; by performing crystallizations at different
temperatures; or by using various modes of cooling, ranging from
very fast to very slow cooling during crystallizations. Polymorphs
may also be obtained by heating or melting the compound followed by
gradual or fast cooling. The presence of polymorphs may be
determined by solid probe NMR spectroscopy, IR spectroscopy,
differential scanning calorimetry, powder X-ray diffractogram
and/or other techniques. Thus, the present invention encompasses
inventive compounds, their derivatives, their tautomeric forms,
their stereoisomers, their polymorphs, their pharmaceutically
acceptable salts their pharmaceutically acceptable solvates and
pharmaceutically acceptable compositions containing them.
[0227] As discussed above, this invention provides novel compounds
with a range of biological properties. Preferred compounds of this
invention have biological activities relevant for the treatment of
diseases, conditions or disorders where increase of HGF/SF activity
would be beneficial.
[0228] Compounds of this invention include those specifically set
forth above and described herein, and are illustrated in part by
the various classes, subgenera and species disclosed elsewhere
herein.
[0229] Additionally, the present invention provides
pharmaceutically acceptable derivatives of the inventive compounds,
and methods of treating a subject using these compounds,
pharmaceutical compositions thereof, or either of these in
combination with one or more additional therapeutic agents. Certain
compounds of the present invention are described in more detail
below. For purposes of this invention, the chemical elements are
identified in accordance with the Periodic Table of the Elements,
CAS version, Handbook of Chemistry and Physics, 75.sup.th Ed.,
inside cover, and specific functional groups are generally defined
as described therein. Additionally, general principles of organic
chemistry, as well as specific functional moieties and reactivity,
are described in "Organic Chemistry", Thomas Sorrell, University
Science Books, Sausalito: 1999, the entire contents of which are
incorporated herein by reference. Furthermore, it will be
appreciated by one of ordinary skill in the art that the synthetic
methods, as described herein, utilize a variety of protecting
groups. It will be appreciated that the compounds, as described
herein, may be substituted with any number of substituents or
functional moieties.
[0230] It will also be appreciated that certain of the compounds of
present invention can exist in free form for treatment, or where
appropriate, as a pharmaceutically acceptable derivative thereof.
According to the present invention, a pharmaceutically acceptable
derivative includes, but is not limited to, pharmaceutically
acceptable salts, esters, salts of such esters, or a prodrug or
other adduct or derivative of a compound of this invention which
upon administration to a patient in need is capable of providing,
directly or indirectly, a compound as otherwise described herein,
or a metabolite or residue thereof.
[0231] 2) Pharmaceutical Compositions
[0232] As discussed above this invention provides compounds that
have biological properties useful for the treatment of any of a
number of conditions or diseases in which cytokines such as but not
limited to HGF/SF, EGF, VEGF or NGF, or the activities thereof,
have a therapeutically useful role.
[0233] Accordingly, in another aspect of the present invention,
pharmaceutical compositions are provided, which comprise any one or
more of the compounds described herein (or a prodrug,
pharmaceutically acceptable salt or other pharmaceutically
acceptable derivative thereof), and optionally comprise a
pharmaceutically acceptable carrier. In certain embodiments, these
compositions optionally further comprise one or more additional
therapeutic agents. Alternatively, a compound of this invention may
be administered to a patient in need thereof in combination with
the administration of one or more other therapeutic agents. For
example, additional therapeutic agents for conjoint administration
or inclusion in a pharmaceutical composition with a compound of
this invention may be an approved agent to treat the same or
related indication, or it may be any one of a number of agents
undergoing approval in the Food and Drug Administration that
ultimately obtain approval for the treatment of any disorder
related to HGF/SF activity. It will also be appreciated that
certain of the compounds of present invention can exist in free
form for treatment, or where appropriate, as a pharmaceutically
acceptable derivative thereof. According to the present invention,
a pharmaceutically acceptable derivative includes, but is not
limited to, pharmaceutically acceptable salts, esters, salts of
such esters, or a pro-drug or other adduct or derivative of a
compound of this invention which upon administration to a patient
in need is capable of providing, directly or indirectly, a compound
as otherwise described herein, or a metabolite or residue
thereof.
[0234] In one aspect, the invention is directed to compositions
including pharmaceutical compositions comprising at least one
compound of Formula (I).
[0235] In yet another embodiment, the invention is directed to
compositions including pharmaceutical compositions comprising
compounds of Formula (II). In certain embodiments, compositions
comprising the following compounds are excluded:
##STR00038##
[0236] In yet another embodiment, the invention is directed to
compositions including pharmaceutical compositions comprising
compounds of Formula (II.sup.A). In certain embodiments,
compositions comprising the following compounds are excluded:
##STR00039##
wherein R.sup.4A s hydrogen, methyl, wherein p and R.sup.4 are as
defined in classes methoxy, chloro or --NO.sub.2 and subclasses
herein.
[0237] In yet another embodiment, the invention is directed to
compositions including pharmaceutical compositions comprising
compounds of Formula (II.sup.B). In certain embodiments,
compositions comprising the following compound are excluded:
##STR00040##
[0238] As used herein, the term "pharmaceutically acceptable salt"
refers to those salts which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of humans
and lower animals without undue toxicity, irritation, allergic
response and the like, and are commensurate with a reasonable
benefit/risk ratio. Pharmaceutically acceptable salts of amines,
carboxylic acids, and other types of compounds, are well known in
the art. For example, S. M. Berge, et al. describe pharmaceutically
acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19
(1977), incorporated herein by reference. The salts can be prepared
in situ during the final isolation and purification of the
compounds of the invention, or separately by reacting a free base
or free acid function with a suitable reagent, as described
generally below. For example, a free base function can be reacted
with a suitable acid. Furthermore, where the compounds of the
invention carry an acidic moiety, suitable pharmaceutically
acceptable salts thereof may, include metal salts such as alkali
metal salts, e.g. sodium or potassium salts; and alkaline earth
metal salts, e.g. calcium or magnesium salts. Examples of
pharmaceutically acceptable, nontoxic acid addition salts are salts
of an amino group formed with inorganic acids such as hydrochloric
acid, hydrobromic acid, phosphoric acid, sulfuric acid and
perchloric acid or with organic acids such as acetic acid, oxalic
acid, maleic acid, tartaric acid, citric acid, succinic acid or
malonic acid or by using other methods used in the art such as ion
exchange. Other pharmaceutically acceptable salts include adipate,
alginate, ascorbate, aspartate, benzenesulfonate, benzoate,
bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, formate, fumarate, glucoheptonate,
glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate,
hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate,
laurate, lauryl sulfate, malate, maleate, malonate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,
oleate, oxalate, palmitate, pamoate, pectinate, persulfate,
3-phenylpropionate, phosphate, picrate, pivalate, propionate,
stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like.
Representative alkali or alkaline earth metal salts include sodium,
lithium, potassium, calcium, magnesium, and the like. Further
pharmaceutically acceptable salts include, when appropriate,
nontoxic ammonium, quaternary ammonium, and amine cations formed
using counterions such as halide, hydroxide, carboxylate, sulfate,
phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.
[0239] Additionally, as used herein, the term "pharmaceutically
acceptable ester" refers to esters that hydrolyze in vivo and
include those that break down readily in the human body to leave
the parent compound or a salt thereof. Suitable ester groups
include, for example, those derived from pharmaceutically
acceptable aliphatic carboxylic acids, particularly alkanoic,
alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl
or alkenyl moiety advantageously has not more than 6 carbon atoms.
Examples of particular esters include formates, acetates,
propionates, butyrates, acrylates and ethylsuccinates.
[0240] Furthermore, the term "pharmaceutically acceptable prodrugs"
as used herein refers to those prodrugs of the compounds of the
present invention which are, within the scope of sound medical
judgment, suitable for use in contact with the issues of humans and
lower animals with undue toxicity, irritation, allergic response,
and the like, commensurate with a reasonable benefit/risk ratio,
and effective for their intended use, as well as the zwitterionic
forms, where possible, of the compounds of the invention. The term
"prodrug" refers to compounds that are rapidly transformed in vivo
to yield the parent compound of the above formula, for example by
hydrolysis in blood, or N-demethylation of a compound of the
invention. A thorough discussion is provided in T. Higuchi and V.
Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S.
Symposium Series, and in Edward B. Roche, ed., Bioreversible
Carriers in Drug Design, American Pharmaceutical Association and
Pergamon Press, 1987, both of which are incorporated herein by
reference.
[0241] As described above, the pharmaceutical compositions of the
present invention additionally comprise a pharmaceutically
acceptable carrier, which, as used herein, includes any and all
solvents, diluents, or other liquid vehicle, dispersion or
suspension aids, surface active agents, isotonic agents, thickening
or emulsifying agents, preservatives, solid binders, lubricants and
the like, as suited to the particular dosage form desired.
Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W.
Martin (Mack Publishing Co., Easton, Pa., 1980) discloses various
carriers used in formulating pharmaceutical compositions and known
techniques for the preparation thereof. Except insofar as any
conventional carrier medium is incompatible with the compounds of
the invention, such as by producing any undesirable biological
effect or otherwise interacting in a deleterious manner with any
other component(s) of the pharmaceutical composition, its use is
contemplated to be within the scope of this invention. Some
examples of materials which can serve as pharmaceutically
acceptable carriers include, but are not limited to, sugars such as
lactose, glucose and sucrose; starches such as corn starch and
potato starch; cellulose and its derivatives such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatine; talc; excipients such as cocoa
butter and suppository waxes; oils such as peanut oil, cottonseed
oil; safflower oil, sesame oil; olive oil; corn oil and soybean
oil; glycols; such as propylene glycol; esters such as ethyl oleate
and ethyl laurate; agar; buffering agents such as magnesium
hydroxide and aluminum hydroxide; alginic acid; pyrogenfree water;
isotonic saline; Ringer's solution; ethyl alcohol, and phosphate
buffer solutions, as well as other non-toxic compatible lubricants
such as sodium lauryl sulfate and magnesium stearate, as well as
coloring agents, releasing agents, coating agents, sweetening,
flavoring and perfuming agents, preservatives and antioxidants can
also be present in the composition, according to the judgment of
the formulator.
[0242] Liquid dosage forms for oral administration include, but are
not limited to, pharmaceutically acceptable emulsions,
microemulsions, solutions, suspensions, syrups and elixirs. In
addition to the active compounds, the liquid dosage forms may
contain inert diluents commonly used in the art such as, for
example, water or other solvents, solubilizing agents and
emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in
particular, cottonseed, groundnut (peanut), corn, germ, olive,
castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol,
polyethylene glycols and fatty acid esters of sorbitan, and
mixtures thereof. Besides inert diluents, the oral compositions can
also include adjuvants such as wetting agents, emulsifying and
suspending agents, sweetening, flavoring, and perfuming agents.
[0243] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution, suspension or emulsion in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, U.S.P.
and isotonic sodium chloride solution. In addition, sterile, fixed
oils are conventionally employed as a solvent or suspending medium.
For this purpose any bland fixed oil can be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid are used in the preparation of injectables.
[0244] The injectable formulations can be sterilized, for example,
by filtration through a bacterial-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use.
[0245] In order to prolong the effect of a drug, it is often
desirable to slow the absorption of the drug from subcutaneous or
intramuscular injection. This may be accomplished by the use of a
liquid suspension or crystalline or amorphous material with poor
water solubility. The rate of absorption of the drug then depends
upon its rate of dissolution that, in turn, may depend upon crystal
size and crystalline form. Alternatively, delayed absorption of a
parenterally administered drug form is accomplished by dissolving
or suspending the drug in an oil vehicle. Injectable depot forms
are made by forming microencapsule matrices of the drug in
biodegradable polymers such as polylactide-polyglycolide. Depending
upon the ratio of drug to polymer and the nature of the particular
polymer employed, the rate of drug release can be controlled.
Examples of other biodegradable polymers include poly(orthoesters)
and poly(anhydrides). Depot injectable formulations are also
prepared by entrapping the drug in liposomes or microemulsions
which are compatible with body tissues.
[0246] Compositions for rectal or vaginal administration are
preferably suppositories which can be prepared by mixing the
compounds of this invention with suitable non-irritating excipients
or carriers such as cocoa butter, polyethylene glycol or a
suppository wax which are solid at ambient temperature but liquid
at body temperature and therefore melt in the rectum or vaginal
cavity and release the active compound.
[0247] Solid dosage forms for oral administration include capsules,
tablets, pills, powders, and granules. In such solid dosage forms,
the active compound is mixed with at least one inert,
pharmaceutically acceptable excipient or carrier such as sodium
citrate or dicalcium phosphate and/or a) fillers or extenders such
as starches, lactose, sucrose, glucose, mannitol, and silicic acid,
b) binders such as, for example, carboxymethylcellulose, alginates,
gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants
such as glycerol, d) disintegrating agents such as agar-agar,
calcium carbonate, potato or tapioca starch, alginic acid, certain
silicates, and sodium carbonate, e) solution retarding agents such
as paraffin, f) absorption accelerators such as quaternary ammonium
compounds, g) wetting agents such as, for example, cetyl alcohol
and glycerol monostearate, h) absorbents such as kaolin and
bentonite clay, and i) lubricants such as talc, calcium stearate,
magnesium stearate, solid polyethylene glycols, sodium lauryl
sulfate, and mixtures thereof. In the case of capsules, tablets and
pills, the dosage form may also comprise buffering agents.
[0248] Solid compositions of a similar type may also be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugar as well as high molecular
weight polyethylene glycols and the like. The solid dosage forms of
tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings and other
coatings well known in the pharmaceutical formulating art. They may
optionally contain opacifying agents and can also be of a
composition that they release the active ingredient(s) only, or
preferentially, in a certain part of the intestinal tract,
optionally, in a delayed manner. Examples of embedding compositions
that can be used include polymeric substances and waxes. Solid
compositions of a similar type may also be employed as fillers in
soft and hard-filled gelatin capsules using such excipients as
lactose or milk sugar as well as high molecular weight polethylene
glycols and the like.
[0249] The active compounds can also be in micro-encapsulated form
with one or more excipients as noted above. The solid dosage forms
of tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings, release
controlling coatings and other coatings well known in the
pharmaceutical formulating art. In such solid dosage forms the
active compound may be admixed with at least one inert diluent such
as sucrose, lactose and starch. Such dosage forms may also
comprise, as in normal practice, additional substances other than
inert diluents, e.g., tableting lubricants and other tableting aids
such as magnesium stearate and microcrystalline cellulose. In the
case of capsules, tablets and pills, the dosage forms may also
comprise buffering agents. They may optionally contain opacifying
agents and can also be of a composition that they release the
active ingredient(s) only, or preferentially, in a certain part of
the intestinal tract, optionally, in a delayed manner. Examples of
embedding compositions which can be used include polymeric
substances and waxes.
[0250] The present invention encompasses pharmaceutically
acceptable topical formulations of inventive compounds. The term
"pharmaceutically acceptable topical formulation", as used herein,
means any formulation which is pharmaceutically acceptable for
intradermal administration of a compound of the invention by
application of the formulation to the epidermis. In certain
embodiments of the invention, the topical formulation comprises a
carrier system. Pharmaceutically effective carriers include, but
are not limited to, solvents (e.g., alcohols, poly alcohols,
water), creams, lotions, ointments, oils, plasters, liposomes,
powders, emulsions, microemulsions, and buffered solutions (e.g.,
hypotonic or buffered saline) or any other carrier known in the art
for topically administering pharmaceuticals. A more complete
listing of art-known carriers is provided by reference texts that
are standard in the art, for example, Remington's Pharmaceutical
Sciences, 16th Edition, 1980 and 17th Edition, 1985, both published
by Mack Publishing Company, Easton, Pa., the disclosures of which
are incorporated herein by reference in their entireties. In
certain other embodiments, the topical formulations of the
invention may comprise excipients. Any pharmaceutically acceptable
excipient known in the art may be used to prepare the inventive
pharmaceutically acceptable topical formulations. Examples of
excipients that can be included in the topical formulations of the
invention include, but are not limited to, preservatives,
antioxidants, moisturizers, emollients, buffering agents,
solubilizing agents, other penetration agents, skin protectants,
surfactants, and propellants, and/or additional therapeutic agents
used in combination to the inventive compound. Suitable
preservatives include, but are not limited to, alcohols, quaternary
amines, organic acids, parabens, and phenols. Suitable antioxidants
include, but are not limited to, ascorbic acid and its esters,
sodium bisulfite, butylated hydroxytoluene, butylated
hydroxyanisole, tocopherols, and chelating agents like EDTA and
citric acid. Suitable moisturizers include, but are not limited to,
glycerine, sorbitol, polyethylene glycols, urea, and propylene
glycol. Suitable buffering agents for use with the invention
include, but are not limited to, citric, hydrochloric, and lactic
acid buffers. Suitable solubilizing agents include, but are not
limited to, quaternary ammonium chlorides, cyclodextrins, benzyl
benzoate, lecithin, and polysorbates. Suitable skin protectants
that can be used in the topical formulations of the invention
include, but are not limited to, vitamin E oil, allatoin,
dimethicone, glycerin, petrolatum, and zinc oxide.
[0251] In certain embodiments, the pharmaceutically acceptable
topical formulations of the invention comprise at least a compound
of the invention and a penetration enhancing agent. The choice of
topical formulation will depend or several factors, including the
condition to be treated, the physicochemical characteristics of the
inventive compound and other excipients present, their stability in
the formulation, available manufacturing equipment, and costs
constraints. As used herein the term "penetration enhancing agent"
means an agent capable of transporting a pharmacologically active
compound through the stratum corneum and into the epidermis or
dermis, preferably, with little or no systemic absorption. A wide
variety of compounds have been evaluated as to their effectiveness
in enhancing the rate of penetration of drugs through the skin.
See, for example, Percutaneous Penetration Enhancers, Maibach H. I.
and Smith H. E. (eds.), CRC Press, Inc., Boca Raton, Fla. (1995),
which surveys the use and testing of various skin penetration
enhancers, and Buyuktimkin et al., Chemical Means of Transdermal
Drug Permeation Enhancement in Transdermal and Topical Drug
Delivery Systems, Gosh T. K., Pfister W. R., Yum S. I. (Eds.),
Interpharm Press Inc., Buffalo Grove, Ill. (1997). In certain
exemplary embodiments, penetration agents for use with the
invention include, but are not limited to, triglycerides (e.g.,
soybean oil), aloe compositions (e.g., aloe-vera gel), ethyl
alcohol, isopropyl alcohol, octolyphenylpolyethylene glycol, oleic
acid, polyethylene glycol 400, propylene glycol,
N-decylmethylsulfoxide, fatty acid esters (e.g., isopropyl
myristate, methyl laurate, glycerol monooleate, and propylene
glycol monooleate) and N-methylpyrrolidone.
[0252] In certain embodiments, the compositions may be in the form
of ointments, pastes, creams, lotions, gels, powders, solutions,
sprays, inhalants or patches. In certain exemplary embodiments,
formulations of the compositions according to the invention are
creams, which may further contain saturated or unsaturated fatty
acids such as stearic acid, palmitic acid, oleic acid,
palmito-oleic acid, cetyl or oleyl alcohols, stearic acid being
particularly preferred. Creams of the invention may also contain a
non-ionic surfactant, for example, polyoxy-40-stearate. In certain
embodiments, the active component is admixed under sterile
conditions with a pharmaceutically acceptable carrier and any
needed preservatives or buffers as may be required. Ophthalmic
formulation, eardrops, and eye drops are also contemplated as being
within the scope of this invention. Formulations for intraocular
administration are also included. Additionally, the present
invention contemplates the use of transdermal patches, which have
the added advantage of providing controlled delivery of a compound
to the body. Such dosage forms are made by dissolving or dispensing
the compound in the proper medium. As discussed above, penetration
enhancing agents can also be used to increase the flux of the
compound across the skin. The rate can be controlled by either
providing a rate controlling membrane or by dispersing the compound
in a polymer matrix or gel.
[0253] It will also be appreciated that the compounds and
pharmaceutical compositions of the present invention can be
formulated and employed in combination therapies, that is, the
compounds and pharmaceutical compositions can be formulated with or
administered concurrently with, prior to, or subsequent to, one or
more other desired therapeutics or medical procedures. The
particular combination of therapies (therapeutics or procedures) to
employ in a combination regimen will take into account
compatibility of the desired therapeutics and/or procedures and the
desired therapeutic effect to be achieved. It will also be
appreciated that the therapies employed may achieve a desired
effect for the same disorder (for example, an inventive compound
may be administered concurrently with an anti-inflammatory agent),
or they may achieve different effects (e.g., control of any adverse
effects). In non-limiting examples, one or more compounds of the
invention may be formulated with at least one cytokine, growth
factor or other biological, such as an interferon, e.g., alpha
interferon, or with at least another small molecule compound.
Non-limiting examples of pharmaceutical agents that may be combined
therapeutically with compounds of the invention include: antivirals
and antifibrotics such as interferon alpha, combination of
interferon alpha and ribavirin, Lamivudine, Adefovir dipivoxil and
interferon gamma; anticoagulants such as heparin and warfarin;
antiplatelets e.g., aspirin, ticlopidine and clopidogrel; other
growth factors involved in regeneration, e.g., VEGF and FGF and
mimetics of these growth factors; antiapoptotic agents; and
motility and morphogenic agents.
[0254] In certain embodiments, the pharmaceutical compositions of
the present invention further comprise one or more additional
therapeutically active ingredients (e.g., anti-inflammatory and/or
palliative). For purposes of the invention, the term "Palliative"
refers to treatment that is focused on the relief of symptoms of a
disease and/or side effects of a therapeutic regimen, but is not
curative. For example, palliative treatment encompasses
painkillers, antinausea medications and anti-sickness drugs.
[0255] 3) Research Uses, Clinical Uses, Pharmaceutical Uses and
Methods of Treatment
[0256] Research Uses
[0257] According to the present invention, the inventive compounds
may be assayed in any of the available assays known in the art for
identifying compounds having the ability to (1) induce endothelial
cell growth and angiogenesis, (2) induce proliferation of other
cells such as epithelial cells, neuronal cells, Schwann cells, and
oligodendrocyte cells, (3) induce axonal growth, (4) induce myelin
production, (5) inhibit apoptosis, (6) reduce fibrosis, (7)
activate HGF signaling pathways, or (8) exhibit some or all of
these activities. For example, the assay may be cellular or
non-cellular, in vivo or in vitro, high- or low-throughput format,
etc.
[0258] Thus, in one aspect, compounds of this invention which are
of particular interest include those with HGF/SF-like activity,
which: exhibit HGF/SF activity; exhibit the ability to mimic or
agonize HGF/SF activities or the HGF/SF receptor c-met; stimulate
cell proliferation, and in particular endothelial cell
proliferation; exhibit angiogenic activity, the promotion of
formation of new blood vessels; induce oligodendrocyte cell
proliferation and axonal growth; induce myelin production; protect
against apoptosis; and reduce fibrosis. In another aspect,
compounds of this invention which are of particular interest
include those with activities that mimic other cytokines, including
but not limited to EGF, VEGF, and NGF, activate their receptors,
activate the signaling pathways of their receptors, and exhibit
various biological activities.
[0259] Clinical Uses of the Compounds of the Invention
[0260] In yet other aspects, the invention provides methods for
using a compound of Formula (I) or a composition comprising a
compound of Formula (I) for the prophylaxis or treatment of
conditions and diseases in which promoting or mimicking the
activity of cytokines is desired, or biological activities
resulting from activating, agonizing or inducing phosphorylation of
c-met or other receptor tyrosine kinases. In a preferred
embodiment, the activity is inducing endothelial cell proliferation
or angiogenesis. In another embodiment, the activity is to induce
proliferation of other cells, such as epithelial cells, neuronal
cells, Schwann cells, or oligodendrocyte cells. In a further
embodiment, the activity is to induce growth of neuronal axons. In
yet another embodiment, the activity is induction of myelin
production. In yet another embodiment, the activity is protection
against apoptosis. In yet another embodiment, the activity is
anti-fibrotic. The compounds described herein are useful in the
treatment of conditions and diseases where inducing endothelial
cell proliferation or therapeutic angiogenesis is beneficial, where
inducing proliferation of cells such as epithelial cells, neuronal
cells, Schwann cells, and oligodendrocyte cells is beneficial,
where inducing axonal growth is beneficial, where induction of
myelin production is beneficial, where protection against apoptosis
is beneficial, where anti-fibrosis is beneficial, or where all or
some of the foregoing activities are beneficial, including but not
limited to fibrotic liver disease, hepatic ischemia-reperfusion
injury, cerebral infarction, ischemic heart disease, renal disease
or lung (pulmonary) fibrosis, multiple sclerosis or various
neurodegenerative diseases. In certain embodiments, the method is
useful for treating a disease or condition, or lessening the
severity of a disease or condition selected from liver fibrosis
associated with hepatitis C, hepatitis B, delta hepatitis, chronic
alcoholism, non-alcoholic steatohepatitis, extrahepatic
obstructions (stones in the bile duct), cholangiopathies (primary
biliary cirrhosis and sclerosing cholangitis), autoimmune liver
disease, and inherited metabolic disorders (Wilson's disease,
hemochromatosis, and alpha-1 antitrypsin deficiency); damaged
and/or ischemic organs, transplants or grafts; ischemia/reperfusion
injury; stroke, traumatic head injury, spinal cord injury, and
other cerebrovascular diseases; myocardial ischemia;
atherosclerosis; peripheral vascular disease; other cardiovascular
diseases; diabetes; renal failure; renal fibrosis or idiopathic
pulmonary fibrosis; multiple sclerosis; and neurodegenerative
diseases such as but not limited to metachromatic leukodystrophy,
Refsum's disease, adrenoleukodystrophy, Krabbe's disease,
phenylketonuria, Canavan disease, Pelizaeus-Merzbacher disease and
Alexander's disease. In certain exemplary embodiments, the method
is for the treatment of wounds for acceleration of healing;
promoting vascularization of a damaged and/or ischemic organ,
transplant or graft; amelioration of ischemia/reperfusion injury in
the brain, heart, liver, kidney, or other tissues or organs;
normalization of myocardial perfusion as a consequence of chronic
cardiac ischemia or myocardial infarction; development or
augmentation of collateral vessel development after vascular
occlusion or to ischemic tissues or organs; fibrotic diseases;
hepatic disease including fibrosis and cirrhosis; lung fibrosis;
radiocontrast nephropathy; fibrosis secondary to renal obstruction;
renal trauma and transplantation; renal failure secondary to
chronic diabetes and/or hypertension; and/or diabetes mellitus. Use
of the compound is also provided for prophylaxis or preventing the
occurrence of the diseases in subjects, and in particular subjects
susceptible to of exhibiting risk factors for, the aforementioned
diseases and conditions. Common among the foregoing conditions is
benefit therein by promoting endothelial cell growth, angiogenesis
or formation of new blood vessels. Moreover, the compounds of the
invention are beneficial in providing biological activities
resulting from activating, agonizing, phosphorylating, or in any
other way activating the signaling pathway of the HGF/SF receptor,
c-met, or other receptor tyrosine kinases.
[0261] In yet other aspects, the invention provides methods for
using a compound of Formula (II) or a composition comprising a
compound of Formula (II) for the prophylaxis or treatment of
conditions and diseases in which promoting or mimicking the
activity of cytokines is desired, or biological activities
resulting from activating, agonizing or inducing phosphorylation of
c-met or other receptor tyrosine kinases. In a preferred
embodiment, the activity is inducing endothelial cell proliferation
or angiogenesis. In another embodiment, the activity is to induce
proliferation of other cells, such as epithelial cells, neuronal
cells, Schwann cells, or oligodendrocyte cells. In a further
embodiment, the activity is to induce growth of neuronal axons. In
yet another embodiment, the activity is induction of myelin
production. In yet another embodiment, the activity is protection
against apoptosis. In yet another embodiment, the activity is
anti-fibrotic. The compounds described herein are useful in the
treatment of conditions and diseases where inducing endothelial
cell proliferation or therapeutic angiogenesis is beneficial, where
inducing proliferation of cells such as epithelial cells, neuronal
cells, Schwann cells, and oligodendrocyte cells is beneficial,
where inducing axonal growth is beneficial, where induction of
myelin production is beneficial, where protection against apoptosis
is beneficial, where anti-fibrosis is beneficial, or where all or
some of the foregoing activities are beneficial, including but not
limited to fibrotic liver disease, hepatic ischemia-reperfusion
injury, cerebral infarction, ischemic heart disease, renal disease
or lung (pulmonary) fibrosis, multiple sclerosis or various
neurodegenerative diseases. In certain embodiments, the method is
useful for treating a disease or condition, or lessening the
severity of a disease or condition selected from liver fibrosis
associated with hepatitis C, hepatitis B, delta hepatitis, chronic
alcoholism, non-alcoholic steatohepatitis, extrahepatic
obstructions (stones in the bile duct), cholangiopathies (primary
biliary cirrhosis and sclerosing cholangitis), autoimmune liver
disease, and inherited metabolic disorders (Wilson's disease,
hemochromatosis, and alpha-1 antitrypsin deficiency); damaged
and/or ischemic organs, transplants or grafts; ischemia/reperfusion
injury; stroke, traumatic head injury, spinal cord injury, and
other cerebrovascular diseases; myocardial ischemia;
atherosclerosis; peripheral vascular disease; other cardiovascular
diseases; diabetes; renal failure; renal fibrosis or idiopathic
pulmonary fibrosis; multiple sclerosis; and neurodegenerative
diseases such as but not limited to metachromatic leukodystrophy,
Refsum's disease, adrenoleukodystrophy, Krabbe's disease,
phenylketonuria, Canavan disease, Pelizaeus-Merzbacher disease and
Alexander's disease. In certain exemplary embodiments, the method
is for the treatment of wounds for acceleration of healing;
promoting vascularization of a damaged and/or ischemic organ,
transplant or graft; amelioration of ischemia/reperfusion injury in
the brain, heart, liver, kidney, or other tissues or organs;
normalization of myocardial perfusion as a consequence of chronic
cardiac ischemia or myocardial infarction; development or
augmentation of collateral vessel development after vascular
occlusion or to ischemic tissues or organs; fibrotic diseases;
hepatic disease including fibrosis and cirrhosis; lung fibrosis;
radiocontrast nephropathy; fibrosis secondary to renal obstruction;
renal trauma and transplantation; renal failure secondary to
chronic diabetes and/or hypertension; and/or diabetes mellitus. Use
of the compound is also provided for prophylaxis or preventing the
occurrence of the diseases in subjects, and in particular subjects
susceptible to of exhibiting risk factors for, the aforementioned
diseases and conditions. Common among the foregoing conditions is
benefit therein by promoting endothelial cell growth, angiogenesis
or formation of new blood vessels. Moreover, the compounds of the
invention are beneficial in providing biological activities
resulting from activating, agonizing, phosphorylating, or in any
other way activating the signaling pathway of the HGF/SF receptor,
c-met, or other receptor tyrosine kinases.
[0262] 1. Fibrotic Liver Disease: Liver fibrosis is the scarring
response of the liver to chronic liver injury; when fibrosis
progresses to cirrhosis, morbid complications can develop. In fact,
end-stage liver fibrosis or cirrhosis is the seventh leading cause
of death in the United States, and afflicts hundreds of millions of
people worldwide; deaths from end-stage liver disease in the United
States are expected to triple over the next 10-15 years, mainly due
to the hepatitis C Epidemic 1. In addition to the hepatitis C
virus, many other forms of chronic liver injury also lead to
end-stage liver disease and cirrhosis, including other viruses such
as hepatitis B and delta hepatitis, chronic alcoholism,
non-alcoholic steatohepatitis, extrahepatic obstructions (stones in
the bile duct), cholangiopathies (primary biliary cirrhosis and
sclerosing cholangitis), autoimmune liver disease, and inherited
metabolic disorders (Wilson's disease, hemochromatosis, and alpha-1
antitrypsin deficiency).
[0263] Treatment of liver fibrosis has focused to date on
eliminating the primary injury. For extrahepatic obstructions,
biliary decompression is the recommended mode of treatment whereas
patients with Wilson's disease are treated with zinc acetate. In
chronic hepatitis C infection, interferon has been used as
antiviral therapies with limited response: .about.20% when used
alone or 50% response when used in combination with ribavirin. In
addition to the low-level of response, treatment with interferon
with or without ribavirin is associated with numerous severe side
effects including neutropenia, thrombocytopenia, anemia,
depression, generalized fatigue and flu-like symptoms, which are
sufficiently significant to necessitate cessation of therapy.
Treatments for other chronic liver diseases such as hepatitis B,
autoimmune hepatitis and Wilson's disease are also associated with
many side effects, while primary biliary cirrhosis, primary
sclerosing cholangitis and non-alcoholic fatty liver disease have
no effective treatment other than liver transplantation.
[0264] The advantage of treating fibrosis rather than only the
underlying etiology, is that antifibrotic therapies should be
broadly applicable across the full spectrum of chronic liver
diseases. While transplantation is currently the most effective
cure for liver fibrosis, mounting evidence indicates that not only
fibrosis, but even cirrhosis is reversible. Unfortunately patients
often present with advanced stages of fibrosis and cirrhosis, when
many therapies such as antivirals can no longer be safely used due
to their side effect profile. Such patients would benefit
enormously from effective antifibrotic therapy, because attenuating
or reversing fibrosis may prevent many late stage complications
such as infection, ascites, and loss of liver function and preclude
the need for liver transplantation. The compounds of the invention
are beneficial for the treatment of the foregoing conditions, and
generally are angiogenic and stimulate endothelial cell
proliferation in this and other organ or tissues.
[0265] 2. Hepatic Ischemia-Reperfusion Injury: Currently,
transplantation is the most effective therapeutic strategy for
liver fibrosis. However, in spite of the significant improvement in
clinical outcome during the last decade, liver dysfunction or
failure is still a significant clinical problem after
transplantation surgery. Ischemia-reperfusion (IR) injury to the
liver is a major alloantigen-independent component affecting
transplantation outcome, causing up to 10% of early organ failure,
and leading to the higher incidence of both acute and chronic
rejection. Furthermore, given the dramatic organ shortage for
transplantation, surgeons are forced to consider cadaveric or
steatotic grafts or other marginal livers, which have a higher
susceptibility to reperfusion injury. In addition to
transplantation surgery, liver IR injury is manifested in clinical
situations such as tissue resections (Pringle maneuver), and
hemorrhagic shock.
[0266] The damage to the postischemic liver represents a continuum
of processes that culminate in hepatocellular injury. Ischemia
activates Kupffer cells, which are the main sources of vascular
reactive oxygen species (ROS) formation during the initial
reperfusion period. In addition to Kupffer cell-induced oxidant
stress, with increasing length of the ischemic episode,
intracellular generation of ROS by xanthine oxidase and in
particular mitochondria may also contribute to liver dysfunction
and cell injury during reperfusion. Endogenous antioxidant
compounds, such as superoxide dismutase, catalase, glutathione,
alphatocopherol, and beta-carotene, may all limit the effects of
oxidant injury but these systems can quickly become overwhelmed by
large quantities of ROS. Work by Lemasters and colleagues, has
indicated that in addition to formation of ROS, intracellular
calcium dyshomeostasis is a key contributor to liver IR injury.
Cell death of hepatocytes and endothelial cells in this setting is
characterized by swelling of cells and their organelles, release of
cell contents, eosinophilia, karyolysis, and induction of
inflammation, characteristic of oncotic necrosis. More recent
reports indicate that liver cells also die by apoptosis, which is
morphologically characterized by cell shrinkage, formation of
apoptotic bodies with intact cell organelles and absence of an
inflammatory response.
[0267] Indeed, minimizing the adverse effects of IR injury could
significantly increase the number of patients that may successfully
undergo liver transplantation. Pharmacologic interventions that
reduce cell death and/or enhance organ regeneration represent a
therapeutic approach to improve clinical outcome in liver
transplantation, liver surgery with vascular exclusion and trauma
and can therefore reduce recipient/patient morbidity and mortality.
The compounds of the invention are beneficial for the treatment of
the foregoing conditions.
[0268] 3. Cerebral Infarction. Stroke and cerebrovascular disease
are a leading cause of morbidity and mortality in the US: at least
600,000 Americans develop strokes each year, and about 160,000 of
these are fatal. Research on the pathophysiological basis of stroke
has produced new paradigms for prevention and treatment, but
translation of these approaches into improved clinical outcomes has
proved to be painfully slow. Preventive strategies focus primarily
on reducing or controlling risk factors such as diabetes,
hypertension, cardiovascular disease, and lifestyle; in patients
with severe stenosis, carotid endarterectomy may be indicated.
Cerebral angioplasty is used investigationally, but the high
restenosis rates observed following coronary angioplasty suggest
this approach may pose unacceptable risk for many patients.
Therapeutic strategies focus primarily on acute treatment to reduce
injury in the ischemic penumbra, the region of reversibly damaged
tissue surrounding an infarct. Thrombolytic therapy has been shown
to improve perfusion to the ischemic penumbra, but it must be
administered within three hours of the onset of infarction. Several
neuroprotective agents that block specific tissue responses to
ischemia are promising, but none have yet been approved for
clinical use. While these therapeutic approaches limit damage in
the ischemic penumbra, they do not address the underlying problem
of inadequate blood supply due to occluded arteries. An alternative
strategy is to induce formation of collateral blood vessels in the
ischemic region; this occurs naturally in chronic ischemic
conditions, but stimulation of vascularization via therapeutic
angiogenesis has potential therapeutic benefit.
[0269] Recent advances in imaging have confirmed the
pathophysiological basis of the clinical observations of evolving
stroke. Analysis of impaired cerebral blood flow (CBF) in the
region of an arterial occlusion supports the hypothesis that a
central region of very low CBF, the ischemic core, is irreversibly
damaged, but damage in surrounding or intermixed zones where CBF is
of less severely reduced, the ischemic penumbra, can be limited by
timely reperfusion. Plate recently reviewed the evidence suggesting
that therapeutic angiogenesis may be useful for treatment or
prevention of stroke. First, analysis of cerebral vasculature in
stroke patients showed a strong correlation between blood vessel
density and survival and a higher density of microvessels in the
ischemic hemisphere compared to the contralateral region. Second,
studies in experimental models of cerebral ischemia indicate
expression of angiogenic growth factors such as vascular
endothelial growth factor (VEGF) or HGF/SF is induced rapidly in
ischemic brain tissue. Third, administration of VEGF or HGF/SF can
reduce neuronal damage and infarct volume in animal models. Similar
evidence provided the rationale for developing therapeutic
angiogenesis for treating peripheral and myocardial ischemia, which
has been shown to produce clinical improvements in early studies in
humans. The compounds of the invention are beneficial for the
treatment of the foregoing conditions.
[0270] 4. Ischemic heart disease is a leading cause of morbidity
and mortality in the US, afflicting millions of Americans each year
at a cost expected to exceed $300 billion/year. Numerous
pharmacological and interventional approaches are being developed
to improve treatment of ischemic heart disease including reduction
of modifiable risk factors, improved revascularization procedures,
and therapies to halt progression and/or induce regression of
atherosclerosis. One of the most exciting areas of research for the
treatment of myocardial ischemia is therapeutic angiogenesis.
Recent studies support the concept that administration of
angiogenic growth factors, either by gene transfer or as a
recombinant protein, augments nutrient perfusion through
neovascularization. The newly developed, supplemental collateral
blood vessels constitute endogenous bypass conduits around occluded
native arteries, improving perfusion to ischemic tissue. Some of
the best-studied cytokines with angiogenic activity are vascular
endothelial growth factor (VEGF), basic fibroblast growth factor
(bFGF) and hepatocyte growth factor/scatter factor (HGF/SF). The
compounds of the invention are beneficial for the treatment of the
foregoing conditions.
[0271] Furthermore, advantage may be taken of the antifibrotic
activities of the compounds of the invention in the area of heart
disease by incorporating compounds in or on indwelling devices such
as stents inserted into coronary arteries to maintain patency as
part of an angioplasty procedure. Such devices can be coated with a
controlled release formulation of one or more compounds of the
invention, optionally including other agents, to prevent or impede
fibrosis of the device and restenosis of the artery. The
anti-fibrotic activity of the inventive compounds may likewise be
used analogously in or on devices used elsewhere in the body.
[0272] 5. Renal Disease. Chronic renal dysfunction is a
progressive, degenerative disorder that ultimately results in acute
renal failure and requires dialysis as an intervention, and renal
transplantation as the only potential cure. Initiating conditions
of renal dysfunction include ischemia, diabetes, underlying
cardiovascular disease, or renal toxicity associated with certain
chemotherapeutics, antibiotics, and radiocontrast agents. Most
end-stage pathological changes include extensive fibrinogenesis,
epithelial atrophy, and inflammatory cell infiltration into the
kidneys.
[0273] Acute renal failure is often a complication of diseases
including diabetes or renal ischemia, procedures such as
heminephrectomy, or as a side effect of therapeutics administered
to treat disease. The widely prescribed anti-tumor drug
cis-diamminedichloroplatinum (cisplatin), for example, has side
effects that include a high incidence of nephrotoxicity and renal
dysfunction, mainly in the form of renal tubular damage that leads
to impaired glomerular filtration. Administration of gentamicin, an
aminoglycoside antibiotic, or cyclosporin A, a potent
immunosuppressive compound, causes similar nephrotoxicity. The
serious side effects of these effective drugs restrict their use.
The development of agents that protect renal function and enhance
renal regeneration after administration of nephrotoxic drugs will
be of substantial benefit to numerous patients, especially those
with malignant tumors, and may allow the maximal therapeutic
potentials of these drugs to be realized. The compounds of the
invention are beneficial for the treatment of the renal diseases
mentioned above.
[0274] 6. Lung (Pulmonary) Fibrosis. Idiopathic pulmonary fibrosis
(IPF) accounts for a majority of chronic interstitial lung
diseases, and has an estimated incidence rate of 10.7 cases for
100,000 per year, with an estimated mortality of 50-70%. IPF is
characterized by an abnormal deposition of collagen in the lung
with an unknown etiology. Although the precise sequence of the
pathogenic sequelae is unknown, disease progression involves
epithelial injury and activation, formation of distinctive
subepithelial fibroblast/myofibroblast foci, and excessive
extracellular matrix accumulation. The development of this
pathological process is preceded by an inflammatory response, often
dominated by macrophages and lymphocytes, which is mediated by the
local release of chemoattractant factors and upregulation of
cell-surface adhesion molecules. Lung injury leads to
vasodilatation and leakage of plasma proteins into interstitial and
alveolar spaces, as well as activation of the coagulation cascade
and deposition of fibrin. Fibroblasts migrate into this provisional
fibrin matrix where they synthesize extracellular matrix molecules.
In non-pathogenic conditions, excess fibrin is usually degraded by
plasmin, a proteinase that also has a role in the activation of
matrix metalloproteinases (MMPs). Activated MMPs degrade
extracellular matrix and participate in fibrin removal, resulting
in the clearance of the alveolar spaces and the ultimate
restoration of injured tissues. In pathological conditions,
however, these processes can lead to progressive and irreversible
changes in lung architecture, resulting in progressive respiratory
insufficiency and an almost universally terminal outcome in a
relatively short period of time. Fibrosis is the final common
pathway of a variety of lung disorders, and in this context, the
diagnosis of pulmonary fibrosis implies the recognition of an
advanced stage in the evolution of a complex process of abnormal
repair. While many studies have focused on inflammatory mechanisms
for initiating the fibrotic response, the synthesis and degradation
the extracellular matrix represent the central event of the
disease. It is this process that presents a very attractive site of
therapeutic intervention.
[0275] The course of IPF is characterized by progressive
respiratory insufficiency, leading to death within 3 to 8 years
from the onset of symptoms. Management of interstitial lung disease
in general, and in particular idiopathic pulmonary fibrosis, is
difficult, unpredictable and unsatisfactory. Attempts have been
made to use antiinflammatory therapy to reverse inflammation,
relief, stop disease progression and prolong survival.
Corticosteroids are the most frequently used antiinflammatory
agents and have been the mainstay of therapy for IPF for more than
four decades, but the efficacy of this approach is unproven, and
toxicities are substantial. No studies have compared differing
dosages or duration of corticosteroid treatment in matched
patients. Interpretation of therapy efficacy is obscured by several
factors including heterogeneous patient populations, inclusion of
patients with histologic entities other than usual interstitial
pneumonia, lack of objective, validated endpoints, and different
criteria for "response." Cytotoxic drugs such as azathioprine and
cyclophosphamide have also being used in combination with low dose
oral corticosteroids. The results of such treatments vary from no
improvement to significant prolongation of survival. Overall,
currently available treatments for lung fibrosis are sub-optimal.
Potential new therapies have emerged from the use of animal models
of pulmonary fibrosis and recent advances in the cellular and
molecular biology of inflammatory reactions. Such therapies involve
the use of cytokines, oxidants and growth factors that are
elaborated during the fibrotic reaction. Despite the use of newer
strategies for treatment, the overall prognosis for patients with
interstitial lung disease has had little quantifiable change, and
the population survival remains unchanged for the last 30 years.
Interferon gamma (IFN) may be effective in the treatment of IPF in
some patients but its role is controversial. Literature indicated
that IFN-gamma may be involved in small airway disease in silicotic
lung. Others showed that IFN-gamma mediates bleomycin-induced
pulmonary inflammation and fibrosis. Recently, hepatocyte growth
factor (HGF), also known as scatter factor (SF) has emerged as an
attractive target for the development of antifibrotic agents. The
compounds of the invention are beneficial for the treatment of the
foregoing condition, among other fibrotic diseases.
[0276] 7. Spinal Cord Injury. It is estimated that the annual
incidence of spinal cord injury (SCI), not including those who die
at the scene of the accident, is approximately 11,000 new cases
each year. The number of people in the United States who are alive
in December 2003 who have SCI has been estimated to be
approximately 243,000 persons. After initial injury, about half of
those affected will remain completely paralyzed below the level of
their spinal lesion. In the other half, the lesion is "incomplete"
and some movement and/or sensation is preserved. Only 17% of those
injured recover enough function to walk again. The higher up spinal
cord lesion, the greater the involvement of paralysis and the
greater mortality. Half of the surviving spinal cord injures are
quadriplegic (paralysis of all four limbs) and half are paraplegic
(paralysis of both legs). Compounds of the invention hold promise
as a new approach to the clinical management of SCI, because they
reduces neuronal cell death, promotes neuronal cell proliferation,
scattering, axonal growth, and functional recovery after SCI.
[0277] 8. Promotion of angiogenesis. Underlying the successful
treatment of the aforementioned diseases is the induction of
endothelial cell proliferation and formation of new blood vessels
to restore vasculature in ischemic and fibrotic tissues. As noted
above, recent studies support the concept that administration of
angiogenic growth factors, either by gene transfer or as a
recombinant protein, augments nutrient perfusion through
neovascularization. The newly developed, supplemental collateral
blood vessels constitute endogenous bypass conduits around occluded
native arteries, improving perfusion to ischemic tissue. Some of
the best-studied cytokines with angiogenic activity are vascular
endothelial growth factor (VEGF), basic fibroblast growth factor
(bFGF) and hepatocyte growth factor/scatter factor (HGF/SF). The
compounds of the invention are beneficial for the treatment of the
foregoing conditions.
[0278] 9. Multiple Sclerosis. The most common of these is multiple
sclerosis (MS), which usually manifests itself between the 20th and
50th years of life. Current estimates are that approximately 2.5
million people worldwide have MS, with between 250,000 and 350,000
cases in the United States, 50,000 cases in Canada, 130,000 cases
in Germany, 85,000 cases in the United Kingdom, 75,000 cases in
France, 50,000 cases in Italy, and 11,000 cases in Switzerland.
[0279] MS attacks the white matter of the central nervous system
(CNS). In its classic manifestation (90% of all cases), it is
characterized by alternating relapsing/remitting phases with
periods of remission growing shorter over time. Its symptoms
include any combination of spastic paraparesis, unsteady gait,
diplopia, and incontinence. The compounds of the invention have
utility in the treatment of MS.
[0280] 10. Hereditary Neurodegenerative Disorders. This category
includes the eight identified leukodystrophies: metachromatic
leukodystrophy, Refsum's disease, adrenoleukodystrophy, Krabbe's
disease, phenylketonuria, Canavan disease, Pelizaeus-Merzbacher
disease and Alexander's disease. The first six are storage
disorders. The lack or the malfunctioning of an enzyme causes a
toxic buildup of chemical substances. In Pelizaeus-Merzbacher
disease myelin is never formed (dysmyelination) because of a
mutation in the gene that produces a basic protein of CNS myelin.
The etiology of Alexander's disease remains largely unknown.
[0281] The clinical course of hereditary demyelinating disorders,
which usually tend to manifest themselves in infancy or early
childhood, is tragic. Previously normal children are deprived, in
rapid progression, of sight, hearing, speech, and ambulation.
Equally tragic is their prognosis: death within a few years.
[0282] 11. Peripheral vascular disease. Peripheral vascular disease
(PVD) is a nearly pandemic condition that has the potential to
cause loss of limb, or even loss of life. PVD manifests as
insufficient tissue perfusion caused by existing atherosclerosis
that may be acutely compounded by either emboli or thrombi. Many
people live daily with PVD; however, in settings such as acute limb
ischemia, this pandemic disease can be life threatening and can
require emergency intervention to minimize morbidity and
mortality.
[0283] PVD, also known as arteriosclerosis obliterans, is primarily
the result of atherosclerosis. The atheroma consists of a core of
cholesterol joined to proteins with a fibrous intravascular
covering. The atherosclerotic process gradually may progress to
complete occlusion of medium and large arteries. The disease
typically is segmental, with significant variation from patient to
patient. Vascular disease may manifest acutely when thrombi,
emboli, or acute trauma compromises perfusion. Thromboses are often
of an atheromatous nature and occur in the lower extremities more
frequently than in the upper extremities. Multiple factors
predispose patients for thrombosis. These factors include sepsis,
hypotension, low cardiac output, aneurysms, aortic dissection,
bypass grafts, and underlying atherosclerotic narrowing of the
arterial lumen.
[0284] The compounds of the invention have utility in treating
PVD.
[0285] Exemplary Assays
[0286] Efficacy of the compounds of the invention on the
aforementioned disorders and diseases or the potential to be of
benefit for the prophylaxis or treatment thereof may be
demonstrated in various studies, ranging from biochemical effects
evaluated in vitro and effects on cells in culture, to in-vivo
models of disease, wherein direct clinical manifestations of the
disease can be observed and measured, or wherein early structural
and/or functional events occur that are established to be involved
in the initiation or progression of the disease. The positive
effects of the compounds of the invention have been demonstrated in
a variety of such assays and models, for a number of diseases and
disorders. One skilled in the art can readily determine following
the guidance described herein that a compound of the invention is a
cytokine mimic and is useful therapeutically in the same manner as
a cytokine.
[0287] 1. In Vitro Stimulation of Cell Proliferation, Myelin
Production and Axonal Growth
[0288] a. Stimulation of Cellular proliferation. The compounds of
invention induce proliferation of human umbilical vein endothelial
cells (HUVEC), monkey bronchial epithelial cells, neuronal cells,
Schwann cells and oligodendrocytes as measured, for example, using
the method of [.sup.311]-thymidine incorporation.
[0289] b. Stimulation of Axonal growth. Human cortical neuronal
cells (HCN-2 from ATCC) are seeded in a flask in full medium (with
10% serum) and incubated for 24 hr. The medium is changed to one
containing 1% serum and compounds are added. The cells are
incubated with the compounds for 72 hr followed by observation of
axonal growth.
[0290] c. Stimulation of Myelin production. Compounds of the
invention induce myelin production by Schwann cells in vitro.
Myelin production is assessed by staining with fluoromyelin.
[0291] 2. Cellular Signaling
[0292] a. Phosphorylation of receptors and signaling proteins. In
human umbilical vein endothelial cells (HUVECs), monkey bronchial
epithelial cells, MDCK cells, and Schwann cells, the compounds of
the invention induce phosphorylation of c-met and other receptors.
The assay is performed by Western blot analysis using antibodies
specific to target proteins.
[0293] b. Intracellular signaling induced by compounds of the
invention. In cells the compounds induce phosphorylation of
extracellular receptor kinase (ERK), as determined by Western blot
analysis.
[0294] 3. Gene Expression
[0295] a. Reduced Expression of Fibrotic markers. Compounds inhibit
expression of alpha SMA in rat kidney fibroblasts.
[0296] 4. Apoptosis.
[0297] a. Apoptosis in endothelial cells and other cells is induced
by serum starvation, hydrogen peroxide, adriamycin, or ethanol when
the cells are treated with the compounds. The extent of apoptosis
and the protective effect by the compounds are measured by annexin
V staining.
[0298] 5. Angiogenesis
[0299] a. Aortic ring assay. Thoracic artery rings from rats are
embedded in Matrigel and grown for 5 days in the presence or
absence of compounds of the invention. Treatment with compounds of
the invention causes an increased outgrowth from the rings.
[0300] b. In vivo Matrigel assay. Matrigel mixed with a compound of
the invention or vehicle is injected into the abdominal
subcutaneous tissue of C57BL/6 mice. When harvested 10 days later,
the compound is found to induce blood vessel formation into the
Matrigel plugs, demonstrating that the compound can exert its
angiogenic effects in vivo.
[0301] 6. Effects on Cellular Migration
[0302] Assays demonstrate the following effects of the compounds of
the invention on cellular migration:
[0303] a. Induction of Endothelial Cell Migration.
[0304] b. Increase Schwann Cell Migrationn
[0305] c. Decrease Monocyte Migration
[0306] 7. Hepatic Disease Models
[0307] a. Antifibrotic Activity in Hepatic Stellate Cells. Serum
starved (activated) LX2 cells (an immortalized human hepatic
stellate cell line) that are treated with HGF/SF or a compound of
the invention show a decrease in collagen I mRNA expression, as
well as expression of other fibrotic marker genes, related to
significant antifibrotic activity.
[0308] b. Liver Disease endpoints. The rat model of thioacetamide
(TAA)-induced liver fibrosis and the rat bile duct ligation model
of fibrosis showed improvements by the compounds of the invention,
in a panel of functional and histological tests: gross morphology,
mass, portal pressure, presence of ascites, enzymes (AST, ALT),
collagen content, interstitial fibrosis and expression of fibrotic
marker genes such as collagen 1, alpha-smooth muscle actin and
MMP-2.
[0309] 8. Protection Against Renal Dysfunction
[0310] a. Clinical model: arterial occlusion. In a mouse model of
transient unilateral renal artery occlusion, compounds of the
invention are shown to restore function to injured kidneys.
[0311] b. Protection against HgCl.sub.2-induced renal injury. In
this model, mice are injected with a high dose of HgCl.sub.2 and
divided into treatment groups. Serum creatinine, BUN, and
development of tubular necrosis are measured to indicate positive
clinical activity.
[0312] c. Protection against ureteral obstruction. The effects of
the compounds of invention on renal injury secondary to ureteral
obstruction are examined in a mouse model of transient unilateral
renal artery occlusion. Immunohistochemical staining is performed
for fibronectin, proliferating cell nuclear antigen, and TUNEL (for
an assessment of apoptosis). Trichrome staining is also performed
to assess the extent of collagen formation as an indication of
interstitial fibrosis.
[0313] d. Protection against Doxorubicin-induced Renal Fibrosis in
Rats. Compounds of the invention attenuate renal dysfunction and
reduce interstitial collagen accumulation in this model.
[0314] 9. Cerebral infarction/Stroke Model
[0315] a. Neuroprotective Effects in Brain Tissue. Cerebral
infarction was induced in rats by middle cerebral artery occlusion
(MCAO) for 24 hr. Test compound or vehicle was administered i.p.
Sections of the brain were then examined for cell death by staining
with a tetrazolium compound. Normal rat brains exhibit a red
staining due to TTC reduction whereas areas containing dead cells
are white. The effect of inducing new vessel formation was
determined by measuring blood flow using a laser Doppler
imager.
[0316] 10. Cardiovascular Disease Models
[0317] a. Atherosclerosis in Apo E knock-out mice. Reduction in the
extent of plaque and lipid content of vessels was observed, as well
as a reduction in hair loss and skin lesions in this model.
[0318] b. Ischemia/reperfusion. Compounds of the invention have
been shown effective in ischemia/reperfusion models, such as the
isolated perfused heart model.
[0319] 11. Transplantation and Organ Preservation
[0320] The viability of organs and tissues harvested and
transported for transplant is currently optimally maintained by
bathing and transport in storage solutions such as the University
of Wisconsin (UW) cold storage solution (100 mM KH.sub.2PO.sub.4, 5
mM MgSO.sub.4 100 mM potassium lactobionate, 1 mM allopurinol, 3 mM
glutathione, 5 mM adenosine, 30 mM raffinose, 50 g/liter of
hydroxyethyl starch, 40 units/liter of insulin, 16 mg/liter of
dexamethasone, 200,000 units/liter of penicillin, pH 7.4; 320-330
mOsM) (Ploeg R J, Goossens D, Vreugdenhil P, McAnulty J F, Southard
J H, Belzer F O. Successful 72-hour cold storage kidney
preservation with UW solution. Transplant Proc. 1988 February; 20(1
Suppl 1):935-8.). To further enhance the viability of transplanted
organs and tissues, inhibit apoptosis and promote vascularization
thereof, one or more compounds of the invention can be included in
this or any other storage solution, as well as perfused into the
donor or donor organ prior to harvesting, and administered to the
recipient systemically and/or locally into the transplanted organ
or transplant site.
[0321] 12. Lung Fibrosis Model
[0322] a. Bleomycin-induced lung injury. The effects of inventive
compounds on pulmonary fibrosis can be assessed using a
well-established mouse model of bleomycin-induced lung injury. The
Ashcroft scale is used to obtain a numerical fibrotic score with
each specimen being scored independently by two histopathologists,
and the mean of their individual scores considered as the fibrotic
score. In addition, reduction in lung hydroxyproline content was
used to also assess efficacy of the compounds in reducing pulmonary
fibrosis.
[0323] 13. Diabetes Mellitus
[0324] a. Hyperglycemia. The effect of the compounds of the
invention on glycemia in streptozotocin induced diabetes was
evaluated. Compounds of the invention reduced blood glucose
levels.
[0325] 14. Multiple Sclerosis and Neurodegenerative Diseases
[0326] a. Effect on Schwann Cells. As noted above in section (1)
above, compounds of the invention promote axonal growth. As will be
shown in the examples below, compounds of the invention also
increase myelin production thereby.
[0327] b. As noted above, compounds of the invention induce robust
c-Met phosphorylation in Schwann cells.
[0328] c. Mouse model of multiple sclerosis. As is shown in the
examples below, experimental autoimmune encephalomyelitis (EAE) was
induced by immunization of female mice with MOG 35-55 emulsified in
CFA containing 1 mg/ml M. tuberculosis. Drug administration started
immediately after second injection of peptide. Disease severity was
monitored in the blind scoring by two scientists and according to a
standard scale. The result shows significant recovery effect from
developing MS in this animal model.
[0329] 15. Angiogenesis/Peripheral Ischemia Models
[0330] a. Mouse and Rat hindlimb ischemia model. In a mouse
hindlimb ischemia model treatment with a compound of the invention
produces greater recovery of hindlimb blow flow (as measured by
laser Doppler imaging). Improved flux is associated with an
increased number of capillaries in the ischemic muscle. Similar
findings are seen in a rat model.
[0331] b. Hindlimb ischemia in non-obese diabetic (NOD) mice. In
female NOD mice subjected to hindlimb ischemia, hindlimb blood flow
(measured using a Laser Doppler imager) demonstrates recovery by
administration of a compound of the invention.
[0332] c. Angiogenesis in full-thickness cutaneous wounds. In full
thickness cutaneous wounds in pigs significant increases are
observed in capillary numbers after treatment with a compound of
the invention.
[0333] As detailed in the exemplification herein, in assays to
determine the ability of compounds to stimulate cell growth, myelin
production, and axonal growth, induce angiogenesis, protect against
apoptosis, and reduce fibrosis, certain inventive compounds
exhibited ED.sub.50 values .ltoreq.50 .mu.M. In certain other
embodiments, inventive compounds exhibit ED.sub.50 values
.ltoreq.40 .mu.M. In certain other embodiments, inventive compounds
exhibit ED.sub.50 values .ltoreq.30 .mu.M. In certain other
embodiments, inventive compounds exhibit ED.sub.50 values
.ltoreq.20 .mu.M. In certain other embodiments, inventive compounds
exhibit ED.sub.50 values .ltoreq.10 .mu.M. In certain other
embodiments, inventive compounds exhibit ED.sub.50 values
.ltoreq.7.5 .mu.M. In certain embodiments, inventive compounds
exhibit ED.sub.50 values .ltoreq.5 .mu.M. In certain other
embodiments, inventive compounds exhibit ED.sub.50 values
.ltoreq.2.5 .mu.M. In certain embodiments, inventive compounds
exhibit ED.sub.50 values .ltoreq.1 .mu.M. In certain other
embodiments, inventive compounds exhibit ED.sub.50 values
.ltoreq.750 nM. In certain other embodiments, inventive compounds
exhibit ED.sub.50 values .ltoreq.500 nM. In certain other
embodiments, inventive compounds exhibit ED.sub.50 values
.ltoreq.250 nM. In certain other embodiments, inventive compounds
exhibit ED.sub.50 values .ltoreq.100 nM. In other embodiments,
exemplary compounds exhibited ED.sub.50 values .ltoreq.75 nM. In
other embodiments, exemplary compounds exhibited ED.sub.50 values
.ltoreq.50 nM. In other embodiments, exemplary compounds exhibited
ED.sub.50 values .ltoreq.40 nM. In other embodiments, exemplary
compounds exhibited ED.sub.50 values .ltoreq.30 nM. In other
embodiments, exemplary compounds exhibited ED.sub.50 values
.ltoreq.20 nM. In other embodiments, exemplary compounds exhibited
ED.sub.50 values .ltoreq.10 nM. In other embodiments, exemplary
compounds exhibited ED.sub.50 values .ltoreq.5 nM.
[0334] Pharmaceutical Uses and Methods of Treatment
[0335] As discussed above, certain of the compounds as described
herein induce endothelial cell proliferation or therapeutic
angiogenesis, induce proliferation of cells such as epithelial
cells, neuronal cells, Schwann cells, and oligodendrocyte cells,
induce axonal growth, induce myelin production, protect cells
against apoptosis, exhibit anti-fibrotic activity, or exhibit all
or some of these activities. Thus, compounds of the invention are
useful for the treatment of any condition, disease or disorder in
which these beneficial activities would have a beneficial role.
Accordingly, in another aspect of the invention, methods for the
treatment of HGF/SF activity or other cytokine activity related
disorders are provided comprising administering a therapeutically
effective amount of a compound of the invention as described
herein, to a subject in need thereof. In certain embodiments, a
method for the treatment disorders related to these activities is
provided comprising administering a therapeutically effective
amount of an inventive compound, or a pharmaceutical composition
comprising an inventive compound to a subject in need thereof, in
such amounts and for such time as is necessary to achieve the
desired result.
[0336] In certain embodiments, the method involves the
administration of a therapeutically effective amount of the
compound or a pharmaceutically acceptable derivative thereof to a
subject (including, but not limited to a human or animal) in need
of it. Subjects for which the benefits of the compounds of the
invention are intended for administration include, in addition to
humans, livestock, domesticated, zoo and companion animals.
[0337] As discussed above this invention provides novel compounds
that have the beneficial activities In certain embodiments, the
inventive compounds are useful for the treatment of wounds for
acceleration of healing (wound healing may be accelerated by
promoting cellular proliferation, particularly of vascular cells),
normalization of myocardial perfusion as a consequence of chronic
cardiac ischemia or myocardial infarction, development or
augmentation of collateral vessel development after vascular
occlusion or to ischemic tissues or organs, fibrotic diseases,
hepatic disease including fibrosis and cirrhosis, lung fibrosis,
renal failure, renal fibrosis, cerebral infarction (stroke),
diabetes mellitus, and vascularization of grafted or transplanted
tissues or organs. Renal conditions for which compounds of the
invention may prove useful include: radiocontrast nephropathy;
fibrosis secondary to renal obstruction; indication for renal
trauma and transplantation; renal failure secondary to chronic
diabetes and/or hypertension.
[0338] It will be appreciated that the compounds and compositions,
according to the method of the present invention, may be
administered using any amount and any route of administration
effective for the treatment of the conditions or diseases in which
cytokines such as but not limited to HGF/SF, VEGF, EGF, or NGF, or
the activities thereof have a therapeutically useful role. Thus,
the expression "effective amount" as used herein, refers to a
sufficient amount of agent to exhibit these activities modulate
cytokine activity (e.g., mimic cytokine activity), and to exhibit a
therapeutic effect. The exact amount required will vary from
subject to subject, depending on the species, age, and general
condition of the subject, the severity of the infection, the
particular therapeutic agent, its mode and/or route of
administration, and the like. The compounds of the invention are
preferably formulated in dosage unit form for ease of
administration and uniformity of dosage. The expression "dosage
unit form" as used herein refers to a physically discrete unit of
therapeutic agent appropriate for the patient to be treated. It
will be understood, however, that the total daily usage of the
compounds and compositions of the present invention will be decided
by the attending physician within the scope of sound medical
judgment. The specific therapeutically effective dose level for any
particular patient or organism will depend upon a variety of
factors including the disorder being treated and the severity of
the disorder; the activity of the specific compound employed; the
specific composition employed; the age, body weight, general
health, sex and diet of the patient; the time of administration,
route of administration, and rate of excretion of the specific
compound employed; the duration of the treatment; drugs used in
combination or coincidental with the specific compound employed;
and like factors well known in the medical arts. Furthermore, after
formulation with an appropriate pharmaceutically acceptable carrier
in a desired dosage, the pharmaceutical compositions of this
invention can be administered to humans and other animals orally,
rectally, parenterally, intracisternally, intravaginally,
intraperitoneally, subcutaneously, intradermally, intra-ocularly,
topically (as by powders, ointments, or drops), buccally, as an
oral or nasal spray, or the like, depending on the severity of the
disease or disorder being treated. In certain embodiments, the
compounds of the invention may be administered at dosage levels of
about 0.001 mg/kg to about 50 mg/kg, preferably from about 0.1
mg/kg to about 10 mg/kg for parenteral administration, or
preferably from about 1 mg/kg to about 50 mg/kg, more preferably
from about 10 mg/kg to about 50 mg/kg for oral administration, of
subject body weight per day, one or more times a day, to obtain the
desired therapeutic effect. It will also be appreciated that
dosages smaller than 0.001 mg/kg or greater than 50 mg/kg (for
example 50-100 mg/kg) can be administered to a subject. In certain
embodiments, compounds are administered orally or parenterally.
[0339] Moreover, pharmaceutical compositions comprising one or more
compounds of the invention may also contain other compounds or
agents for which co-administration with the compound(s) of the
invention is therapeutically advantageous. As many pharmaceutical
agents are used in the treatment of the diseases and disorders for
which the compounds of the invention are also beneficial, any may
be formulated together for administration. Synergistic formulations
are also embraced herein, where the combination of at least one
compound of the invention and at least one other compounds act more
beneficially than when each is given alone. Non-limiting examples
of pharmaceutical agents that may be combined therapeutically with
compounds of the invention include (non-limiting examples of
diseases or conditions treated with such combination are indicated
in parentheses): antivirals and antifibrotics, such as interferon
alpha (hepatitis B, and hepatitis C), combination of interferon
alpha and ribavirin (hepatitis C), Lamivudine (hepatitis B),
Adefovir dipivoxil (hepatitis B), interferon gamma (idiopathic
pulmonary fibrosis, liver fibrosis, and fibrosis in other organs);
anticoagulants, e.g., heparin and warfarin (ischemic stroke);
antiplatelets e.g., aspirin, ticlopidine and clopidogrel (ischemic
stroke); other growth factors involved in regeneration, e.g., VEGF
and FGF and mimetics of these growth factors; antiapoptotic agents;
and motility and morphogenic agents.
Treatment Kit
[0340] In other embodiments, the present invention relates to a kit
for conveniently and effectively carrying out the methods in
accordance with the present invention. In general, the
pharmaceutical pack or kit comprises one or more containers filled
with one or more of the ingredients of the pharmaceutical
compositions of the invention. Such kits are especially suited for
the delivery of solid oral forms such as tablets or capsules. Such
a kit preferably includes a number of unit dosages, and may also
include a card having the dosages oriented in the order of their
intended use. If desired, a memory aid can be provided, for example
in the form of numbers, letters, or other markings or with a
calendar insert, designating the days in the treatment schedule in
which the dosages can be administered. Alternatively, placebo
dosages, or calcium dietary supplements, either in a form similar
to or distinct from the dosages of the pharmaceutical compositions,
can be included to provide a kit in which a dosage is taken every
day. Optionally associated with such container(s) can be a notice
in the form prescribed by a governmental agency regulating the
manufacture, use or sale of pharmaceutical products, which notice
reflects approval by the agency of manufacture, use or sale for
human administration.
EQUIVALENTS
[0341] The representative examples that follow are intended to help
illustrate the invention, and are not intended to, nor should they
be construed to, limit the scope of the invention. Indeed, various
modifications of the invention and many further embodiments
thereof, in addition to those shown and described herein, will
become apparent to those skilled in the art from the full contents
of this document, including the examples which follow and the
references to the scientific and patent literature cited herein. It
should further be appreciated that the contents of those cited
references are incorporated herein by reference to help illustrate
the state of the art.
[0342] The following examples contain important additional
information, exemplification and guidance that can be adapted to
the practice of this invention in its various embodiments and the
equivalents thereof.
EXEMPLIFICATION
[0343] The compounds of this invention and their preparation can be
understood further by the examples that illustrate some of the
processes by which these compounds are prepared or used. It will be
appreciated, however, that these examples do not limit the
invention. Variations of the invention, now known or further
developed, are considered to fall within the scope of the present
invention as described herein and as hereinafter claimed.
[0344] 1) General Description of Synthetic Methods:
[0345] The practitioner has a well-established literature of small
molecule chemistry to draw upon, in combination with the
information contained herein, for guidance on synthetic strategies,
protecting groups, and other materials and methods useful for the
synthesis of the compounds of this invention.
[0346] The various references cited herein provide helpful
background information on preparing compounds similar to the
inventive compounds described herein or relevant intermediates, as
well as information on formulation, uses, and administration of
such compounds which may be of interest.
[0347] Moreover, the practitioner is directed to the specific
guidance and examples provided in this document relating to various
exemplary compounds and intermediates thereof.
[0348] The compounds of this invention and their preparation can be
understood further by the examples that illustrate some of the
processes by which these compounds are prepared or used. It will be
appreciated, however, that these examples do not limit the
invention. Variations of the invention, now known or further
developed, are considered to fall within the scope of the present
invention as described herein and as hereinafter claimed.
[0349] According to the present invention, any available techniques
can be used to make or prepare the inventive compounds or
compositions including them. For example, a variety of solution
phase synthetic methods such as those discussed in detail below may
be used. Alternatively or additionally, the inventive compounds may
be prepared using any of a variety combinatorial techniques,
parallel synthesis and/or solid phase synthetic methods known in
the art.
[0350] It will be appreciated as described below, that a variety of
inventive compounds can be synthesized according to the methods
described herein. The starting materials and reagents used in
preparing these compounds are either available from commercial
suppliers such as Aldrich Chemical Company (Milwaukee, Wis.),
Bachem (Torrance, Calif.), Sigma (St. Louis, Mo.), or are prepared
by methods well known to a person of ordinary skill in the art
following procedures described in such references as Fieser and
Fieser 1991, "Reagents for Organic Synthesis", vols 1-17, John
Wiley and Sons, New York, N.Y., 1991; Rodd 1989 "Chemistry of
Carbon Compounds", vols. 1-5 and supps, Elsevier Science
Publishers, 1989; "Organic Reactions", vols 1-40, John Wiley and
Sons, New York, N.Y., 1991; March 2001, "Advanced Organic
Chemistry", 5th ed. John Wiley and Sons, New York, N.Y.; and Larock
1990, "Comprehensive Organic Transformations: A Guide to Functional
Group Preparations", 2.sup.nd ed. VCH Publishers. These schemes are
merely illustrative of some methods by which the compounds of this
invention can be synthesized, and various modifications to these
schemes can be made and will be suggested to a person of ordinary
skill in the art having regard to this disclosure.
[0351] The starting materials, intermediates, and compounds of this
invention may be isolated and purified using conventional
techniques, including filtration, distillation, crystallization,
chromatography, and the like. They may be characterized using
conventional methods, including physical constants and spectral
data.
[0352] General Reaction Procedures:
[0353] Unless mentioned specifically, reaction mixtures were
stirred using a magnetically driven stirrer bar. An inert
atmosphere refers to either dry argon or dry nitrogen. Reactions
were monitored either by thin layer chromatography, by proton
nuclear magnetic resonance (NMR) or by high-pressure liquid
chromatography (HPLC), of a suitably worked up sample of the
reaction mixture.
[0354] General Work Up Procedures:
[0355] Unless mentioned specifically, reaction mixtures were cooled
to room temperature or below then quenched, when necessary, with
either water or a saturated aqueous solution of ammonium chloride.
Desired products were extracted by partitioning between water and a
suitable water-immiscible solvent (e.g. ethyl acetate,
dichloromethane, diethyl ether). The desired product containing
extracts were washed appropriately with water followed by a
saturated solution of brine. On occasions where the product
containing extract was deemed to contain residual oxidants, the
extract was washed with a 10% solution of sodium sulphite in
saturated aqueous sodium bicarbonate solution, prior to the
aforementioned washing procedure. On occasions where the product
containing extract was deemed to contain residual acids, the
extract was washed with saturated aqueous sodium bicarbonate
solution, prior to the aforementioned washing procedure (except in
those cases where the desired product itself had acidic character).
On occasions where the product containing extract was deemed to
contain residual bases, the extract was washed with 10% aqueous
citric acid solution, prior to the aforementioned washing procedure
(except in those cases where the desired product itself had basic
character). Post washing, the desired product containing extracts
were dried over anhydrous magnesium sulphate, and then filtered.
The crude products were then isolated by removal of solvent(s) by
rotary evaporation under reduced pressure, at an appropriate
temperature (generally less than 45.degree. C.).
[0356] General Purification Procedures:
[0357] Unless mentioned specifically, chromatographic purification
refers to flash column chromatography on silica, using a single
solvent or mixed solvent as eluent. Suitably purified desired
product containing elutes were combined and concentrated under
reduced pressure at an appropriate temperature (generally less than
45.degree. C.) to constant mass. Final compounds were dissolved in
50% aqueous acetonitrile, filtered and transferred to vials, then
freeze-dried under high vacuum before submission for biological
testing.
[0358] 1) Synthesis of Exemplary Compounds:
[0359] Compounds of Formula (I) and (II) of the invention can be
prepared following the reaction schemes 1-4 shown below.
Preparation of
4-[3-nitro-4-(1-homopiperidinyl)phenyl]-1-(2H)-phthalazinone
[0360] Step 1. Preparation of 2-(4'-chloro-3'-nitrobenzoyl)benzoic
acid. As shown in Scheme 1 below, 2-(4-chlorobenzoyl)benzoic acid
(80 g, 0.37 mol) was added to 98% sulphuric acid (210 ml) keeping
the temperature below 10.degree. C. A further volume of sulphuric
acid (60 ml) was then added. The mixture was stirred for 2 hrs
until complete solution. 90% Nitric acid (21 ml) was added to conc.
sulphuric acid (50 ml) with cooling and stirring. The nitrating
mixture was added to the benzoic acid solution dropwise keeping the
temperature below 15.degree. C. When the addition was complete the
mixture was stirred at 5-10.degree. C. for 1 hr, the mixture was
then poured onto crushed ice (2.51) and the ice allowed to melt.
The resulting white solid was filtered, washed with water (1.51)
and dried. Yield 92.7 g (98%). The combined products from 2 runs of
the above reaction were recrystallized by stirring in ethyl acetate
(1.51) at 70.degree. C. and adding IMS (.about.90 ml) to obtain a
clear solution. Pentane (1.51) was added to yield a white
crystalline solid (142.3 g)
##STR00041##
[0361] Step 2. Preparation of
2-(4-(homopiperidin-1-yl)-3-nitrobenzoyl)benzoic acid (Scheme 2).
To a solution of the 2-(4'-chloro-3'-nitrobenzoyl)benzoic acid
(prepared in Step 1; 142 g, 0.465 mol) in acetonitrile (1400 ml)
was added homopiperidine (138 g, 1.39 mol, 3 eq.). The mixture was
heated to 80.degree. C. for 4 hrs. The reaction mixture was then
concentrated to .about.500 ml and diluted with water to 2.5 l. The
aqueous was made acidic with conc. HCl solution and extracted with
a mixture of ether/ethyl acetate (1:1). This extract was separated
from water, dried (MgSO.sub.4), filtered and evaporated to dryness
at reduced pressure to yield a viscous orange oil. This was
triturated with a small volume of ether, then hexanes to yield a
yellow solid.
##STR00042##
[0362] Step 3. Preparation of
4-[3-nitro-4-(1-homopiperidinyl)phenyl]-1-(2H)-phthalazinone
(Scheme 3). To a suspension of
2-(4-(homopiperidin-1-yl)-3-nitrobenzoyl)benzoic acid (prepared in
Step 2; 80 g, 0217 mol) in ethanol (450 ml) was added hydrazine
hydrate (98%, 21.74 g, 0.435 mol, 2 eq.). After refluxing for 1.5
hrs, more hydrazine hydrate (8 ml) was added. It was refluxed for a
further 2 hrs. The reaction mixture was cooled to .about.15.degree.
C. and the orange crystals filtered, washed with IMS then ether
before drying. The yield was 66.88 g (83.9%) Melting point
187-189.degree. C.
##STR00043##
[0363] As an example, compounds of Formula (I) and (II) where m is
1 and R.sup.1 is nitro can be prepared in accordance with Scheme
4.
##STR00044##
[0364] These compounds can be further derivatized to yield the
corresponding amino derivatives (i.e., m=1 and
R.sup.1.dbd.NH.sub.2).
[0365] Compounds of Formula (I) and (II) of the invention where
R.sup.1 is hydrogen can be prepared following the reaction scheme
shown below.
Preparation of
4-[4-(1-homopiperidinyl)phenyl]-1-(2H)-phthalazinone
##STR00045##
[0367] Step 1. Preparation of
2-(4-(homopiperidin-1-yl)benzoyl)benzoic acid (Scheme 1). To a
solution of 2-(4'-chlorobenzoyl)benzoic acid in acetonitrile is
added homopiperidine. The mixture is heated to 80.degree. C. for 4
hrs. The reaction mixture is then concentrated and diluted with
water. The aqueous is made acidic with conc. HCl solution and is
extracted with a mixture of ether/ethyl acetate (1:1). This extract
is separated from water, dried (MgSO.sub.4), filtered and
evaporated to dryness at reduced pressure to yield the product.
##STR00046##
[0368] Step 2. Preparation of
4-[4-(1-homopiperidinyl)phenyl]-1-(2H)-phthalazinone (Scheme 3). To
a suspension of 2-(4-(homopiperidin-1-yl)benzoyl)benzoic acid in
ethanol is added hydrazine hydrate. After refluxing for 1.5 hrs,
more hydrazine hydrate is added. It is refluxed for a further 2
hrs. The reaction mixture is cooled to .about.15.degree. C. and the
product is filtered, washed with IMS then ether before drying.
[0369] It will be appreciated that the use of a substituted
homopiperidine in Scheme 5 would lead to the preparation of
corresponding substituted homopiperidinyl counterparts:
##STR00047##
[0370] In general, compounds of Formula (I) and (II) where m is 1
and R.sup.1 is hydrogen can be prepared in accordance with Scheme
7.
##STR00048##
[0371] Non-limiting examples of R.sup.2R.sup.3NH reagents shown in
Schemes 4 and 7 to produce compounds of the invention include:
dimethylamine, diethylamine, dipropylamine, di-tert-butylamine,
homopiperidine, 3-aminohomopiperidine, 2-methylhomopiperidine,
3-methylhomopiperidine, piperidine, 2-methylpiperidine,
3-methylpiperidine, diethylamine, 2,2,6,6,-tetramethylpiperidine,
4-benzylpiperidine, thiomorpholine, 4-methylpiperazine,
4-phenylpiperazine, 4-ethylsulfonylpiperazine, and pyrrolidone,
etc.
[0372] Other compounds of Formula (I) and (II) of the invention can
be prepared following the reaction scheme shown in Schemes 8 and 9
below.
##STR00049##
[0373] wherein r is 1-4 and R.sup.0 take the definition of
R.sup.1.
[0374] Reaction of appropriately substituted phthalic anhydrides
with aryl nucleophiles, such as Grignard reagents or aryl lithium
species, would provide the 2-acylbenzoic acid intermediates.
Treatment with hydrazine would afford ring closure to the desired
phthalazinone analogues. Synthetic methods for such transformations
are described in Yamaguchi, M. et al. J. Med. Chem. 1993, 36,
4052.
[0375] In cases where a nitro group is present, reduction of the
nitro group using standard methods would provide the corresponding
primary amine. Such amine could then be acylated using carboxylic
acids/acyl halides, isocyanates, or isothiocyanates, to form
amides, ureas, and thioureas, respectively.
##STR00050##
[0376] The foregoing schemes are merely exemplary of synthetic
routes to the compound of the invention. They may be readily
modified or varied to prepare the variety of compounds of the
invention.
[0377] The invention encompasses compounds described herein
substituted on the phthalazinone ring.
[0378] The foregoing compounds, compositions and methods of the
invention are illustrated by the following examples, which are
merely exemplary of aspects of the invention and are not
limiting.
[0379] 2) Biological Activity:
[0380] 1. Compounds of the Invention Induce Endothelial Cell
Proliferation and Migration:
[0381] a. Proliferation. The following assay was performed to
assess the activity of the compounds of the invention in cell
proliferation. Endothelial cells (HUVECs) were seeded in 96-well
plates at a density of 10,000 cells per well in the normal growth
medium (EGM-2-Clonetics) containing 2% fetal bovine serum, FGF,
VEGF, IGF, ascorbic acid, EGF, GA, heparin and hydrocortisone. The
cells were grown normally in the growth medium for 24 hr at
37.degree. C. and 5% CO.sub.2. The cells were then rinsed with
RPMI-1% BSA and starved for 1-2 hr. The stock solutions of the
compounds of the invention were made at a concentration of 10 mg/ml
in DMSO and diluted in RPMI-1% BSA at final concentrations of 0.001
uM to 50 uM. The cells were then washed and treated with the
compounds and incubated for another 20 hr at 37.degree. C. Then
.sup.3H thymidine (0.5 microgram/ml in RPMI-BSA) was added to the
cells and incubated at 37.degree. C. for 4 hr. The unincorporated
thymidine was removed by washing the cells four times with
1.times.PBS. Then the cells were lysed with 0.5M NaOH for 30 min
and the radioactivity counted in the beta counter. A similar
proliferation assay using monkey bronchial epithelial cells (4
MBR-5) and oligodendrocyte cells was also employed.
[0382] As shown in FIG. 1A, compound of the invention induced a
very large increase in HIVEC proliferation as measured by thymidine
incorporation. Typically, compounds of the invention show an 8 to
20 fold increase in HUVEC proliferation over control, a magnitude
not seen by HGF (shown in figure) or other molecules.
[0383] Dose response curves for three compounds of the invention
are shown in FIGS. 1B-D, and show stimulated HUVEC (endothelial
cell) proliferation at an ED.sub.50 (effective dose giving 50%
stimulation) of about 5-10 uM. Compounds also synergistically
stimulated HUVEC proliferation together with HGF, FGF, EGF, and
VEGF, which are angiogenic cytokines.
[0384] The following compounds induced proliferation of HUVEC and
oligodendrocytes with the ED.sub.50 shown in Tables 1 and 2,
respectively:
TABLE-US-00001 TABLE 1 HUVEC Compound ED.sub.50 ##STR00051## 2.5
.mu.M ##STR00052## 10 .mu.M ##STR00053## 5 .mu.M ##STR00054## 2
.mu.M ##STR00055## 3 .mu.M ##STR00056## 4 .mu.M ##STR00057## 3
.mu.M
TABLE-US-00002 TABLE 2 Oligodendrocyte Compound ED.sub.50
##STR00058## 0.2 .mu.M ##STR00059## 0.2 .mu.M ##STR00060## 0.2
.mu.M
[0385] 2. Compounds of the Invention Protect HUVEC Cells Against
Apoptosis. Inventive compounds also protect HUVEC against apoptosis
(programmed cell death) induced by serum starvation, hydrogen
peroxide or CRP. Endothelial cell (EC) apoptosis is an initiating
event in the pathogenesis of atherosclerosis. In fact, C-reactive
protein (CRP, which is elevated in atherosclerotic patients, is
known to induce endothelial apoptosis. It was determined whether
compounds of the invention protect ECs against apoptosis induced by
serum starvation, H.sub.2O.sub.2 (oxidative stress), or CRP. HUVECs
(Cambrex, Calif.) were grown to 80% confluence in 6-well plates in
complete serum medium. Cells were then washed with RPMI-1% BSA and
treated with vehicle, compound (10 uM) or SF/HGF (50 ng/ml) and
incubated for 24 hours. Apoptosis was induced by serum starvation
or addition of H.sub.2O.sub.2 (100 uM) or addition of CRP (10
ug/ml). Apoptotic cells were identified using the Vybrant Assay Kit
(Molecular Probes, Oreg.). As seen in FIGS. 2A, 2B and 2C,
inventive compound attenuated apoptosis in ECs Induced by serum
starvation, H.sub.2O.sub.2 and CRP, respectively.
[0386] 3. Compounds of the Invention Activate HGF/SF Signaling
Pathways.
[0387] One possible mechanism of the activities of the compounds of
the invention (which Applicants have no duty of disclosure thereof
and to which Applicants are not bound) is activation of the HGF
receptor, c-Met. Since the biological activity of HGF is mediated
through phosphorylation of its receptor, c-met, the ability of
compounds of the invention to phosphorylate c-met was tested.
[0388] a. Phosphorylation of c-met in Schwann cells. SF/HGF
bioactivity is mediated via phosphorylation and activation of its
receptor, c-Met. Schwann cells were purchased from ATCC, VA and
incubated with SF/HGF (50 ng/ml) or inventive compounds at 10 uM
concentration for one hour in serum-free medium. C-Met
phosphorylation was determined by performing SDS PAGE followed by
Western blotting using phosphor met antibody from Cell Signaling.
As seen in FIG. 3, compounds of the invention induced robust
phosphorylation of c-Met, indicating activation of the SF/HGF/c-Met
pathways.
[0389] b. Phosphorylation of c-met in HUVECs and MDCK cells. HUVECs
and MDCK cells were incubated with either HGF (80 ng) or instant
compounds (10 uM) for 1 hr, followed by Western blot analysis.
Compounds of the invention compounds phosphorylated c-Met in these
cells.
[0390] c. Intracellular Signaling Induced by Compounds and HGF. To
determine whether compound-mediated c-met phosphorylation induces
the same intracellular signaling cascades as HGF, endothelial cells
were stimulated with the instant compounds, and extracellular
receptor kinase (ERK) phosphorylation was then assayed by Western
blot analysis. Western blot analyses are then performed by probing
for total ERK using antibodies that do not distinguish between the
phosphorylated and non-phosphorylated forms; the membranes were
then stripped and re-probed with antibodies that recognize only
phosphorylated ERK. Unstimulated cells contain little
phosphorylated ERK. Under identical cell culture conditions,
however, instant compounds significantly increase the intracellular
levels of phosphorylated ERK, while total ERK remains unaffected.
These results are similar to phosphorylated ERK levels observed in
the presence of HGF.
[0391] 4. Schwann Cells, Neuronal Cells and Oligodendrocytes:
Proliferation, Migration and Myelin Production
[0392] a. Proliferation of Schwann Cells. Rat neuronal Schwann
cells (RSC96 from ATCC) were seeded in 96-well plate (10.sup.4
cell/well) in serum free medium for 16 hours. Cells were then
treated with compound of the invention or HGF (positive control) at
different concentrations for addition 16 hours. .sup.3H-thymidine
was added to the medium and incubation continued for another 4-5
hours. The cells were washed with PBS, harvested, and
.sup.3H-thymidine incorporation determined as a measure of
proliferation. Compound of the invention stimulates
[.sup.3H]-thymidine incorporation, indicating stimulation of
Schwann cell proliferation (FIG. 4A).
[0393] b. Proliferation of PC12 neuronal cells. In a similar assay
to that described above, it was shown that inventive compound
stimulated proliferation of PC12 cells, a neuronal cell type (FIG.
4B).
[0394] c. Migration of Schwann cells. In a cell migration assay
(Boyden Chamber, BD Bioscience), 50000 Schwann cells were seeded in
the inner chamber for 22 hours in the presence of 0.4% or 10% FBS,
in the presence of compounds of invention. The cell number was
quantified with 4.5 mg/ml Calcein following fluorescence reading.
Compounds of the invention stimulated Schwann cell migration in
this assay.
[0395] d. Myelin Production by Schwann Cells. Compounds of the
invention induce myelin production by Schwann cells in vitro.
Schwann cells at a density of 50,000 cells per well were seeded
into 3-well chamber slides in serum-free medium for 24 hours. Test
compounds (5 uM) or HGF/SF (50 ng/ml) were the added to the medium
and the incubation continued for an additional 4 hours. Cells were
washed and fluoromyelin (Molecular Probes) was added to each well.
Images were obtained using a confocal microscope. Inventive
compound induced over a three-fold increase in myelin production,
similar to the extent of induction by HGF/SF (FIG. 4C).
[0396] e. Stimulation of Oligodendrocyte Proliferation. Mouse
primary oligodendrocytes (Celprogen, Calif.) were seeded in 96-well
plates at 5000 cells/well in serum-free medium for 16 hours. Cells
were treated with test compound or HGF/SF (positive control) for
16-24 hours. WST1 cell proliferation reagent (Roche, N.J.) was
added to oligodendrocytes, and incubation was continued for another
4-5 hours. Cells with WST1 reagent were read using a plate reader
at an OD of 490 nm. Compounds and HGF/SF produced similar effects
on cell proliferation (FIG. 5).
[0397] 5. Axonal Growth.
[0398] Human HCN-2 neurons and Schwann cells were obtained from the
American Type Culture Collection (Manassas, Va.). Cells were
cultured in Dulbecco's modified Eagle's medium with 4 mM
L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5
g/L glucose supplemented with 0.2 mg/ml G418 and 0.001 mg/ml
puromycin, 90%, fetal bovine serum 10%. Cells are sub-cultured by
removing media, rinsing with 0.25% trypsin, 0.03% EDTA solution
followed by addition of fresh culture medium and plated onto
poly-1-lysine coated culture flasks.
[0399] Human neurons (HCN-2) were seeded in 6 well plates (1000
cell/well) and incubated in 1% FBS with HGF/SF (50 ng/ml) or
Compound of the invention (10 uM) over a 48 hour period. As shown
in FIG. 6A-D, compounds of the invention promote axonal growth. In
the presence of Schwann cells, the compounds stimulated axonal
growth to a greater extent.
[0400] 6. Peripheral Ischemia Model: Therapeutic Angiogenesis in
Mouse and Rat Hindlimb Ischemia (Peripheral Vascular Insufficiency)
Models.
[0401] a. Rats. To determine the time-dependent effects of
inventive compounds in augmenting distal flow, male Sprague-Dawley
rats (275-300 g) were subjected to left hindlimb ischemia and
treated with vehicle or compound (2 mg/kg, i.p.) daily until
sacrifice at day 14. Distal flow measurements using Laser Doppler
scanning (Moor Instruments, Inc.) were obtained and normalized to
pre-ischemic flow before and after the surgery. In the can, low
power laser light is directed across the tissue surface in a raster
pattern to construct a 2 dimensional image. Moving blood cells
shift the frequency of incident light according to the Doppler
principle. The back-scattered light at the detectors causes
constructive and destructive mixing of shifted light from moving
blood and non-shifted light from static tissue. Intensity
fluctuations are processed to give parameters of flux, which is
proportional to tissue blood flow. Flux values of the areas of
interest in the hindlimb are then compared between the left,
ischemic hindlimb and the right, non-ischemic hindlimb and
expressed as a fraction (ischemic/non-ischemic), with a value of 1
representing normal flow. As seen in FIG. 7, compared to the
vehicle-treated group, compound-treated animals exhibited an
enhanced recovery of blood flow.
[0402] 7. Stroke Model.
[0403] Ischemia was induced in rats by middle cerebral artery
occlusion (MCAO) for 24 hr. A compound of the invention, or placebo
(saline), was administered i.v. at 2 mg/kg at 1-2, and 20 hr or in
a delayed fashion at 4 hr and 20 hr post infarct induction. The
extent of cerebral infarction was determined by staining brain
sections with the mitochondrial activity indicator
2,3,5-triphenyltetrazolium chloride (TTC). As shown in FIG. 8, a
large portion of the untreated rat brain displayed cerebral
infarction, while inventive compound protects rats from such
injury. In addition, the compound also increased blood flow in the
injured area at day 7 and day 14 as detected by laser Doppler
imaging, indicating that it promotes neovascularization following
the injury.
[0404] 8. Cardiovascular Disease Model.
[0405] a. Atherosclerosis: Immediate Treatment. Male apolipoprotein
E (apoE)-deficient C57BL/6J mice were treated with a high fat diet
for 16 weeks. Vehicle (n=15) or compound of the invention (2 mg/kg)
(n=15) were administered daily, i.p. starting with the onset of
high-fat diet. Animals were sacrificed (week 16) and
atherosclerosis assays were performed on the aortic roots. Plaque
formation was assessed in transverse aortic sections using H&E
staining; Oil-Red-O staining was used to identify lipid deposits en
face. Blood samples were collected at the time of death and
analyzed for plasma cholesterol levels. ApoE deficient mice on
regular chow diet served as normal controls.
[0406] At the time of sacrifice, there was no difference in plasma
cholesterol levels (1200-1600 mg/dL) within the high-fat-treated
groups. Treatment with a compound of the invention significantly
reduced both plaque formation (p<0.05 vs. vehicle) and lipid
deposits (p<0.05 vs. vehicle) in these high-fat treated mice
(FIG. 9).
[0407] b. Delayed Treatment. In this series, mice were fed with
high fat diet for 10 weeks. Diet was then switched to regular chow
and animals were administered daily i.p injections of vehicle or
compound of the invention (2 mg/kg) for 8 weeks. Blood and aortic
sections were collected for analysis at the time of sacrifice (8
weeks into delayed treatment). There was no difference in the lipid
profile between the vehicle and the compound-treated groups.
Delayed treatment with inventive compound resulted in a significant
decrease in both plaque formation and lipid deposition.
[0408] c. Myocardial Ischemia/Reperfusion. Hearts from male
Sprague-Dawley rats were perfused in the Langendorrf Mode under
constant pressure. Normothermic, isovolumic contracting hearts were
subjected to 30 min global ischemia and 90 min reperfusion. Hearts
were treated 5 min prior to ischemia and 5 min into reperfusion
with vehicle or compound of the invention. As shown in FIG. 10, the
product of left ventricular diastolic pressure and heart rate
during reperfusion was normalized to pre-ischemic values.
[0409] 9. Fibrosis
[0410] a. Anti-fibrotic Effects In Vitro: Initiation of a fibrotic
cascade is crucial to atherosclerotic plaque formation.
Consequently, anti-fibrotic therapies may find use against
atherosclerotic plaque formation as well as in renal and pulmonary
fibrotic diseases. The effects of a compound of the invention on
TGFbeta1-induced alpha-smooth muscle actin (alpha SMA, fibrotic
marker) expression in rat kidney fibroblasts was evaluated. Rat
kidney fibroblasts cells (NRK-49F from ATCC) were activated by the
treatment with TGFbeta1 at 2 ng/ml for 2 days. Compound of the
invention was added and incubated for the same time period. Total
cellular RNA was isolated and alpha smooth muscle actin (alphaSMA)
mRNA was measured by reverse transcription-real time polymerase
chain reaction (RT-PCR). AlphaSMA is upregulated during fibroblast
activation and is a marker of fibrosis. The test compound inhibited
the alphaSMA level, indicating that it has an anti-fibrotic
potential (FIG. 11).
[0411] b. Anti-Fibrotic Effects In Vivo--Lung--Bleomycin Model:
Fibrogenesis in different tissues shares common features and
mechanisms including TGFbeta-stimulated pathological extracellular
matrix build up. The activity of inventive compounds in other
fibrosis models sheds light on an understanding in a fibrosis
condition in atherosclerotic plaque regression. The anti-fibrotic
activity of compounds of the invention was tested in a widely used
model of pulmonary fibrosis, viz. bleomycin-induced pulmonary
fibrosis. Briefly, C57BL/6 mice were anesthetized then administered
intra-tracheally 100 .mu.l of a solution containing bleomycin
hydrochloride (Sigma; 0.1 U/20 gm body weight). Mice were then
divided into vehicle or compound (2 mg/kg, i.p. daily)-treated
groups. Survival in these mice was recorded. After two weeks,
surviving mice were sacrificed and lungs harvested for
determination of hydroxyproline, a key index of fibrosis. Treatment
with inventive compound (1) to bleomycin-treated mice increased
survival rate (FIG. 12A), decreased lung collagen expression as
measured by RT-PCT (FIG. 12B), and decreased lung collagen content
as measured in the hydroxyproline assay (FIG. 12C). Quantitation of
histological analysis confirmed that treatment with inventive
compound effectively prevented the fibrotic injury seen in
bleomycin+vehicle-treated animals (FIG. 12D).
[0412] c. Anti-Fibrotic Effects In Vivo--Kidney--Doxorubicin Model.
Doxorubicin administration results in progressive renal dysfunction
and accumulation of interstitial collagen. This is a model for
chronic renal failure and fibrosis. Male Sprague-Dawley rats
(.about.300 g) were administered doxorubicin (10 mg/kg, iv).
Starting twenty-fours hours later, animals were treated daily with
vehicle or a compound of the invention (2 mg/kg, i.p., n=8/group).
Three weeks later animals were sacrificed for evaluation of renal
function and histopathology. As seen in FIG. 13, treatment compound
attenuates the renal dysfunction observed in the vehicle-treated
cohort as measured by BUN (FIG. 13A), serum creatinine (FIG. 13B)
and fibrotic score (FIG. 13C). Furthermore, interstitial collagen
accumulation (Masson's trichrome stain) was reduced.
[0413] d. Anti-fibrotic effects: liver. Since fibrogenesis in
different tissues shares common features and mechanisms including
TGFbeta1-stimulated pathological extracellular matrix build up,
compounds of the invention have been examined in the thioacetamide
(TAA) induced liver fibrosis model. Sprague Dawley rats were
treated with TAA at 200 mg/kg, i.p., three times a week for 8 weeks
to induce liver fibrosis. At the time of TAA treatment, rats were
also treated with compound of the invention via i.p route at 2
mg/kg body weight or vehicle (PEG300), daily, five times a week for
4 weeks, followed by sacrifice. In the co-treatment group, a panel
of fibrotic markers was measured including collagen content
(hydroproxyproline, FIG. 14A), and alpha SMA (FIG. 14B) and
collagen-1 gene expression (RT-PCR; FIG. 14C).
[0414] In a separate group of rats, compound (25 mg/kg) was
administered orally, with treatment onset delayed by 8 weeks
following TAA onset. Hydroxyproline, portal pressure, and fibrotic
score were measured to examine the oral efficacy of inventive
compound in reducing liver fibrosis by hydroxyproline content (FIG.
14D), portal pressure (FIG. 14E) and fibrotic score (FIG. 14F). A
fibrotic score of 4 indicates severe fibrosis while a score of 0
indicates no fibrosis.
[0415] 10. Renal Ischemia.
[0416] a. In a mouse model of transient unilateral renal artery
occlusion, male ICR mice are anesthetized and the left renal artery
occluded with a microvascular clamp. After 30 minutes, the clamp is
removed and the kidney allowed to reperfuse. Ten minutes into
reperfusion the nonischemic contralateral kidney is excised.
Animals are treated daily with vehicle or compound of the invention
(2 mg/kg, i.p.) until the day of sacrifice. Serum creatinine, BUN
and urine protein levels, measured at 1, 4 and 7 days post-ischemia
are used to determine the ability of compounds of the invention to
restore function to injured kidneys. In order to create a more
severe renal injury, animals are subjected to 45 minutes of
ischemia.
[0417] b. Protection against HgCl.sub.2-induced renal injury. In a
study mice are injected with a high dose of HgCl.sub.2 (7 mg/kg,
s.c.) and divided into treatment groups. Animals in the first group
receive vehicle or a compound of the invention (2 mg/kg, i.p.) on
the day of toxin injection and daily thereafter for 3 days, and are
euthanized on day 4. Blood samples collected prior to HgCl.sub.2
injection, on day 2 and on day 4 are analyzed for serum creatinine.
In the second group, treatment with vehicle or compound begins on
the day following toxin injection (i.e., 24 h delayed treatment)
and daily thereafter until day 6. Mice are euthanized on day 7.
Blood samples collected prior to HgCl.sub.2 injection, on day 4 and
day 7 are analyzed for serum creatinine and BUN. Serum creatinine,
BUN, and development of tubular necrosis are measured to indicate
positive clinical activity.
[0418] c. Protection against ureteral obstruction. The effects of
the compounds of invention on renal injury secondary to ureteral
obstruction are examined in a mouse model of transient unilateral
renal artery occlusion. Kidneys from mice subjected to unilateral
ureteral obstruction for 2 weeks are examined for histological
evidence of injury and protection by compound treatment.
Immunohistochemical staining is performed for fibronectin,
proliferating cell nuclear antigen, and TUNEL (for an assessment of
apoptosis). Trichrome staining is also performed to assess the
extent of collagen formation as an indication of interstitial
fibrosis.
[0419] 11. Diabetes.
[0420] Normal CD-1 mice were injected i.p. once with 100 mg/kg
streptozotocin (STZ) and then treated with a compound of the
invention at 2 mg/kg i.p. daily for seven days. At day 7, blood
samples were harvested and blood glucose determined. STZ treatment
resulted in hyperglycemia (high blood glucose). Treatment with
compound of the invention treatment ameliorated the hyperglycemia
in the diabetic mice (FIG. 15).
[0421] 12. Mouse model of multiple sclerosis. EAE was induced by
immunization of male C57 BL6 mice with 200 .mu.g MOG 35-55
emulsified in complete Freund's adjuvant (CFA) containing 5 mg/ml
M. tuberculosis (Difco, Mich.) on days 0 and 7 subcutaneously in
the hind flank as described by Ford, M. L. and B. D. Evavold,
Specificity, magnitude, and kinetics of MOG-specific CD8+ T cell
responses during experimental autoimmune encephalomyelitis. Eur. J.
Immunol. 35, 76-85, 2005. Compound of the invention (2 mg/kg) or
vehicle was administered i.p., daily for 3 weeks. Compound and
vehicle administration started on day 7, immediately after the
second injection of MOG 35-55. At the end of the 3-week treatment
period, disease severity with respect to locomotor deficit was
monitored in the blind scoring by two scientists according to the
following scale: 0, no disease; 1, flaccid tail; 2, hind limb
weakness; 3, hind limb paralysis; 4, forelimb weakness; 5,
moribund. As shown in FIG. 16, the test compound reduced locomotor
deficit.
[0422] 13. Inventive Compounds Decrease Monocytes Migration.
[0423] a. Compound treatment decreases monocyte migration and
binding to ECs. Elevated CRP has been shown to exert
pro-atherogenic effects on vascular cells exemplified by increasing
the secretion of monocyte chemoattractant protein (MCP-1) and
reducing nitric oxide bioactivity, and induce adhesion molecules,
such as vascular cell adhesion molecule-1 (VCAM-1), and increase
monocyte binding/migration to endothelial cells. HUVECs were
incubated with CRP (25 ug/ml), CRP together with test compound (10
uM) or HGF (50 ng/ml). The cellular extracts with lysis buffer and
analyzed for VCAM-1 and MCP-1 expression via Western blot analysis.
For Monocyte binding and migration experiments, monocytes were
labeled with a fluorescent dye Vybrant Did (Molecular Probes) and
treated to the HUVECs treated as above. Monocyte migration and
binding to ECs decreased with test compound treatment as shown in
FIG. 17. Test compound and HGF treatment also decreased VCAM-1
expression and MCP-1 expression and decreased monocyte migration
and binding to endothelial cells.
[0424] 14. Compound-mediated therapeutic angiogenesis. Compounds of
the invention induce angiogenesis in vivo, providing clear evidence
that compounds can at least mediate HGF-like biologic activity by
inducing c-met phosphorylation and activating specific
intracellular signaling cascades. To test whether this activity can
be used to therapeutic advantage, the ability of compounds to
induce blood vessel growth was tested in vivo. In this assay
compounds or vehicle (control, RPMI media+1% BSA) was mixed with
Matrigel, a matrix of reconstituted basement membrane. Samples were
injected subcutaneously into mice. After 10 days, mice were
sacrificed for histologic and morphometric analysis of Matrigel
plugs. Plugs containing compound show a greater density of cells.
These results are similar to above studies that demonstrated that
HGF dose-dependently increases the vessel area in this in vivo
assay. Such findings are also applicable to other cytokines and the
observations are generally applicable to other cytokines such as
but not limited to those described above.
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