U.S. patent application number 14/011341 was filed with the patent office on 2014-03-06 for rel inhibitors and methods of use thereof.
This patent application is currently assigned to The Trustees of the University of Pennsylvania. The applicant listed for this patent is The Trustees of the University of Pennsylvania. Invention is credited to Youhai H. CHEN, Ramachandran MURALI, Jing SUN.
Application Number | 20140066486 14/011341 |
Document ID | / |
Family ID | 40853748 |
Filed Date | 2014-03-06 |
United States Patent
Application |
20140066486 |
Kind Code |
A1 |
CHEN; Youhai H. ; et
al. |
March 6, 2014 |
REL INHIBITORS AND METHODS OF USE THEREOF
Abstract
This invention provides REL inhibitors which interfere with the
DNA binding capacity of a REL protein. Additionally this invention
provides methods of treating, abrogating, or preventing diseases
which respond with a positive clinical score to a REL inhibitor.
Methods of identifying REL inhibitor based on a REL protein three
dimensional model are described.
Inventors: |
CHEN; Youhai H.; (Newtown
Square, PA) ; MURALI; Ramachandran; (Swarthmore,
PA) ; SUN; Jing; (Upper Darby, PA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
The Trustees of the University of Pennsylvania |
Philadelphia |
PA |
US |
|
|
Assignee: |
The Trustees of the University of
Pennsylvania
Philadelphia
PA
|
Family ID: |
40853748 |
Appl. No.: |
14/011341 |
Filed: |
August 27, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12812224 |
Nov 8, 2010 |
8609730 |
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PCT/US09/30325 |
Jan 7, 2009 |
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14011341 |
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61006353 |
Jan 8, 2008 |
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61071374 |
Apr 24, 2008 |
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Current U.S.
Class: |
514/400 ;
514/508; 548/338.1; 558/4 |
Current CPC
Class: |
A61K 31/38 20130101;
A61K 31/4174 20130101; A61P 29/00 20180101; A61K 31/235 20130101;
A61P 19/02 20180101; A61K 31/426 20130101; A61P 25/00 20180101;
A61K 31/155 20130101; A61K 31/275 20130101; A61K 31/325 20130101;
A61P 37/06 20180101; C07C 335/32 20130101; A61P 3/10 20180101; C07D
233/64 20130101; A61K 31/519 20130101 |
Class at
Publication: |
514/400 ;
548/338.1; 558/4; 514/508 |
International
Class: |
C07D 233/64 20060101
C07D233/64; C07C 335/32 20060101 C07C335/32 |
Claims
1. A composition comprising a selective c-Rel: DNA binding
inhibitor represented by: the structure of formula (I) ##STR00167##
wherein Q.sub.1, Q.sub.2, Q.sub.3 are independently H, halogen,
CF.sub.3, OCH.sub.2Ph, O-alkyl, OCF.sub.3, alkyl, or Q.sub.1 and
Q.sub.2 form a saturated or unsaturated, substituted or
unsubstituted, carbocyclic or heterocyclic ring with the aniline
ring; X and Y are independently H, alkyl, or form a saturated or
unsaturated, substituted or unsubstituted, carbocyclic or
heterocyclic ring with N(R.sub.1)(R.sub.2). X.sub.1 and Y.sub.1 are
independently H, alkyl, or X and Y form together a double bond, or
form saturated or unsaturated, substituted or unsubstituted,
carbocyclic or heterocyclic ring with N(R.sub.1)(R.sub.2). R.sub.1
and R.sub.2 are independently H, NH.sub.2, --N=alkyl, -alkyl,
--CH(Ph).sub.2, substituted or unsubstituted aryl, carbocyclic or
heterocyclic aryl, substituted or unsubstituted phenyl, C(O)-alkyl,
or R.sub.1 and R.sub.2 form a saturated or unsaturated, substituted
or unsubstituted, carbocyclic or heterocyclic ring with the
nitrogen atom; or by the structure of formula (II): ##STR00168##
Wherein L.sub.1, L.sub.2, L.sub.3 and L.sub.4 are independently H,
halogen, alkyl, --NH.sub.2, --COOAlkyl, --NO.sub.2, pyrrolidine,
--O-alkyl, or L.sub.1 and L.sub.2 form a saturated or unsaturated,
substituted or unsubstituted, carbocyclic or heterocyclic fused
ring with the benzene ring; or L.sub.4 together with R.sub.2 forms
a 6 membered fused ring with the imidazole and benzene rings; R and
R.sub.1 are independently H, NHCO-alkyl, or form together a double
bond (.dbd.), or CO group (.dbd.O); R.sub.2 is H, SH, OH, alkyl,
-Ph-CF.sub.3, --CH.dbd.C(Ph)-OC(O)-Ph, CH.sub.2--S-Ph,
CH.sub.2--S-heterocyclic ring, CH.sub.2OC(O)NH-Ph,
--NHCH.sub.2CH.sub.2OH, -alkylene-OH, O-aryl, --O-alkyl,
O--CH.sub.2-Ph, O-phenyl, O-phenyl-alkyl, O-Ph-O-alkylene-Ph,
--OCH.sub.2Ph, --OCH.sub.2CH.dbd.CH-Ph, --S-Phenyl, NH-alkyl,
NH-phenyl, NH-aryl, --N(Me)-alkylene-phenyl, --NH-alkylene-phenyl,
--NH-alkylene-OMe, --NH--N.dbd.CH-Ph, --NH--N--C(O)-alkyl,
--NH-heterocyclic ring, NH-carbocyclic ring, --C(O)Ph, substituted
or unsubstituted, saturated or unsaturated hetrocyclic ring,
substituted or unsubstituted, saturated or unsaturated carbocyclic
ring, or R.sub.2 together with L.sub.4 forms a 6 membered fused
ring with the imidazole and benzene rings; R.sub.3 is H, COO-alkyl,
COOH, NO.sub.2, substituted or unsubstituted Ph,
C(O)--N.dbd.NC(O)Ph or C(O)NH.sub.2; and; R4 is H, Ph, alkyl,
NH.sub.2, OH, Ph-OH or CH.sub.2--OH; or by the structure of formula
(CXIX): ##STR00169## Wherein R.sub.1 is substituted phenyl or
unsubstituted phenyl; R.sub.2 is ##STR00170## wherein X.sub.1,
X.sub.2, or X.sub.3 are independently H, halogen, alkyl, CN, COOH,
or NH.sub.2; or X.sub.3 forms with the .dbd.N.sup.+H.sub.2 a five
membered fused ring; or R.sub.2 forms with .dbd.N.sup.+H.sub.2 a
five or six substituted or unsubstituted membered ring, or their
pharmaceutical salt.
2. The composition of claim 1, wherein the inhibitor interact with
L1 cavity on the surface of the c-Rel.
3. The composition of claim 1, wherein said L1 cavity comprises
amino acids Arg 21, Cys 26, Glu 27, Lys 110, and Lys 111.
4. The composition of claim 1, comprising the compound represented
by the structure of formula (III), or its pharmaceutical salt:
##STR00171##
5. The composition of claim 1, comprising the compound represented
by the structure of formula (IV), (V) or their pharmaceutical
salt.
6. The composition of claim 1, comprising the compound represented
by the structure of formula (CXX), or its pharmaceutical salt:
##STR00172##
7. Use of the composition of claim 1 for preventing, inhibiting,
suppressing or ameliorating symptoms associated with inflammatory
conditions that are multiple sclerosis, arthritis, diabetes,
colitis, lupus, autoimmunity, graft rejection, or a combination
thereof.
8. A method of inhibiting or suppressing the interaction between
c-Rel and a DNA, comprising the step of contacting the c-Rel with a
compound capable of masking the L1 cavity of the c-Rel, thereby
inhibiting or suppressing the interaction between c-Rel and a DNA
and inflammatory immune response.
9. The method of claim 8, whereby the compound capable of masking
the L1 cavity of the c-Rel is represented by: the compound set
forth by the structure of formula (I) ##STR00173## wherein Q.sub.1,
Q.sub.2, Q.sub.3 are independently H, halogen, CF.sub.3,
OCH.sub.2Ph, O-alkyl, OCF.sub.3, alkyl, or Q.sub.1 and Q.sub.2 form
a saturated or unsaturated, substituted or unsubstituted,
carbocyclic or heterocyclic ring with the aniline ring; X and Y are
independently H, alkyl, or form a saturated or unsaturated,
substituted or unsubstituted, carbocyclic or heterocyclic ring with
N(R.sub.1)(R.sub.2). X.sub.1 and Y.sub.1 are independently H,
alkyl, or X and Y form together a double bond, or form saturated or
unsaturated, substituted or unsubstituted, carbocyclic or
heterocyclic ring with N(R.sub.1)(R.sub.2). R.sub.1 and R.sub.2 are
independently H, NH.sub.2, --N=alkyl, -alkyl, --CH(Ph).sub.2,
substituted or unsubstituted aryl, carbocyclic or heterocyclic
aryl, substituted or unsubstituted phenyl, C(O)-alkyl, or R.sub.1
and R.sub.2 form a saturated or unsaturated, substituted or
unsubstituted, carbocyclic or heterocyclic ring with the nitrogen
atom; or by the compound set forth by the structure of formula
(II): ##STR00174## Wherein L.sub.1, L.sub.2, L.sub.3 and L.sub.4
are independently H, halogen, alkyl, --NH.sub.2, --COOAlkyl,
--NO.sub.2, pyrrolidine, --O-alkyl, or L.sub.1 and L.sub.2 form a
saturated or unsaturated, substituted or unsubstituted, carbocyclic
or heterocyclic fused ring with the benzene ring; or L.sub.4
together with R.sub.2 forms a 6 membered fused ring with the
imidazole and benzene rings; R and R.sub.1 are independently H,
NHCO-alkyl, or form together a double bond (.dbd.), or CO group
(.dbd.O); R.sub.2 is H, SH, OH, alkyl, -Ph-CF.sub.3,
--CH.dbd.C(Ph)-OC(O)-Ph, CH.sub.2--S-Ph, CH.sub.2--S-heterocyclic
ring, CH.sub.2OC(O)NH-Ph, --NHCH.sub.2CH.sub.2OH, -alkylene-OH,
O-aryl, --O-alkyl, O--CH.sub.2-Ph, O-phenyl, O-phenyl-alkyl,
O-Ph-O-alkylene-Ph, --OCH.sub.2Ph, --OCH.sub.2CH.dbd.CH-Ph,
--S-Phenyl, NH-alkyl, NH-phenyl, NH-aryl, --N(Me)-alkylene-phenyl,
--NH-alkylene-phenyl, --NH-alkylene-OMe, --NH--N.dbd.CH-Ph,
--NH--N--C(O)-alkyl, --NH-heterocyclic ring, NH-carbocyclic ring,
--C(O)Ph, substituted or unsubstituted, saturated or unsaturated
hetrocyclic ring, substituted or unsubstituted, saturated or
unsaturated carbocyclic ring, or R.sub.2 together with L.sub.4
forms a 6 membered fused ring with the imidazole and benzene rings;
R.sub.3 is H, COO-alkyl, COOH, NO.sub.2, substituted or
unsubstituted Ph, C(O)--N.dbd.NC(O)Ph or C(O)NH.sub.2; and; R4 is
H, Ph, alkyl, NH.sub.2, OH, Ph-OH or CH.sub.2--OH; or by the
compound set forth by the structure of formula (CXIX): ##STR00175##
Wherein R.sub.1 is substituted phenyl or unsubstituted phenyl;
R.sub.2 is ##STR00176## wherein X.sub.1, X.sub.2, or X.sub.3 are
independently H, halogen, alkyl, CN, COOH, or NH.sub.2; or X.sub.3
forms with the .dbd.N.sup.+H.sub.2 a five membered fused ring; or
R.sub.2 forms with .dbd.N.sup.+H.sub.2 a five or six substituted or
unsubstituted membered ring, or their pharmaceutical salt
10. The method of claim 9, wherein said inhibitor further inhibits
the production of interleukin-2, interferon-gamma, or the
combination thereof.
11. The method of claim 9, wherein said L1 cavity comprises the
amino acids Arg 21, Cys 26, Glu 27, Lys 110, and Lys 111.
12. The method of claim 9, whereby the compound capable of masking
the L1 cavity of the c-Rel is represented by the structure of
formula (III), or its pharmaceutical salt: ##STR00177##
13. The method of claim 9, whereby the compound capable of masking
the L1 cavity of the c-Rel is represented by the structure of
formula (IV), (V) or their pharmaceutical salt.
14. The method of claim 9, whereby the compound capable of masking
the L1 cavity of the c-Rel is represented by the structure of
formula (CXX), or its pharmaceutical salt: ##STR00178##
15. A method of treating, inhibiting or suppressing, or meliorating
symptoms associated with multiple sclerosis, arthritis, diabetes,
graft rejection, or a combination thereof in a subject, comprising
the step of contacting the subject with a composition comprising a
selective c-Rel:DNA binding inhibitor wherein said c-Rel DNA
binding inhibitor masks the L1 cavity of the c-Rel protein, thereby
treating, inhibiting or suppressing, or meliorating symptoms
associated with inflammatory conditions that are multiple
sclerosis, arthritis, diabetes, colitis, lupus, autoimmunity, graft
rejection, or a combination thereof in the subject.
16. The method of claim 15, whereby the inhibitor further inhibits
the production of interleukin-2, interferon-gamma, or both.
17. The method of claim 15, whereby said L1 cavity comprises amino
acids; Arg 21, Cys 26, Glu 27, Lys 110, and Lys 111.
18. The method of claim 15, whereby said composition comprises a
compound set forth by: the structure represented by formula (I):
##STR00179## wherein Q.sub.1, Q.sub.2, Q.sub.3 are independently H,
halogen, CF.sub.3, OCH.sub.2Ph, O-alkyl, OCF.sub.3, alkyl, or
Q.sub.1 and Q.sub.2 form a saturated or unsaturated, substituted or
unsubstituted, carbocyclic or heterocyclic ring with the aniline
ring; X and Y are independently H, alkyl, or form a saturated or
unsaturated, substituted or unsubstituted, carbocyclic or
heterocyclic ring with N(R.sub.1)(R.sub.2). X.sub.1 and Y.sub.1 are
independently H, alkyl, or X and Y form together a double bond, or
form saturated or unsaturated, substituted or unsubstituted,
carbocyclic or heterocyclic ring with N(R.sub.1)(R.sub.2). R.sub.1
and R.sub.2 are independently H, NH.sub.2, --N=alkyl, -alkyl,
--CH(Ph).sub.2, substituted or unsubstituted aryl, carbocyclic or
heterocyclic aryl, substituted or unsubstituted phenyl, C(O)-alkyl,
or R.sub.1 and R.sub.2 form a saturated or unsaturated, substituted
or unsubstituted, carbocyclic or heterocyclic ring with the
nitrogen atom; or by the structure of formula (II): ##STR00180##
wherein L.sub.1, L.sub.2, L.sub.3 and L.sub.4 are independently H,
halogen, alkyl, --NH.sub.2, --COOAlkyl, --NO.sub.2, pyrrolidine,
--O-alkyl, or L.sub.1 and L.sub.2 form a saturated or unsaturated,
substituted or unsubstituted, carbocyclic or heterocyclic fused
ring with the benzene ring; or L.sub.4 together with R.sub.2 forms
a 6 membered fused ring with the imidazole and benzene rings; R and
R.sub.1 are independently H, NHCO-alkyl, or form together a double
bond (.dbd.), or CO group (.dbd.O); R.sub.2 is H, SH, OH, alkyl,
-Ph-CF.sub.3, --CH.dbd.C(Ph)-OC(O)-Ph, CH.sub.2--S-Ph,
CH.sub.2--S-heterocyclic ring, CH.sub.2OC(O)NH-Ph,
--NHCH.sub.2CH.sub.2OH, -alkylene-OH, O-aryl, --O-alkyl,
O--CH.sub.2-Ph, O-phenyl, O-phenyl-alkyl, O-Ph-O-alkylene-Ph,
--OCH.sub.2Ph, --OCH.sub.2CH.dbd.CH-Ph, --S-Phenyl, NH-alkyl,
NH-phenyl, NH-aryl, --N(Me)-alkylene-phenyl, --NH-alkylene-phenyl,
--NH-alkylene-OMe, --NH--N.dbd.CH-Ph, --NH--N--C(O)-alkyl,
--NH-heterocyclic ring, NH-carbocyclic ring, --C(O)Ph, substituted
or unsubstituted, saturated or unsaturated hetrocyclic ring,
substituted or unsubstituted, saturated or unsaturated carbocyclic
ring, or R.sub.2 together with L.sub.4 forms a 6 membered fused
ring with the imidazole and benzene rings; R.sub.3 is H, COO-alkyl,
COOH, NO.sub.2, substituted or unsubstituted Ph,
C(O)--N.dbd.NC(O)Ph or C(O)NH.sub.2; and; R4 is H, Ph, alkyl,
NH.sub.2, OH, Ph-OH or CH.sub.2--OH; or by the structure of formula
(CXIX): ##STR00181## Wherein R.sub.1 is substituted phenyl or
unsubstituted phenyl; R.sub.2 is ##STR00182## wherein X.sub.1,
X.sub.2, or X.sub.3 are independently H, halogen, alkyl, CN, COOH,
or NH.sub.2; or X.sub.3 forms with the .dbd.N.sup.+H.sub.2 a five
membered fused ring; or R.sub.2 forms with .dbd.N.sup.+H.sub.2 a
five or six substituted or unsubstituted membered ring, or their
combination.
19. The method of claim 15, whereby said composition comprises a
compound set forth by formula (III): ##STR00183##
20. The method of claim 15, whereby said composition comprises a
compound set forth by formula (CXX): ##STR00184##
Description
FIELD OF INVENTION
[0001] This invention provides REL inhibitors and methods of using
and identifying the same. Specifically, the invention provides
c-Rel inhibitors and their use in the treatment of Multiple
Sclerosis.
BACKGROUND OF THE INVENTION
[0002] Experimental autoimmune encephalomyelitis (EAE) is an animal
model for multiple sclerosis (MS). While the genetic and
environmental factors that trigger MS vary, the common pathological
outcome of the disease is the destruction of myelin-producing
oligodendrocytes and their associated neuronal axons through a
process called encephalomyelitis. Development of encephalomyelitis
requires coordinated expression of a large number of genes that
mediate the activation, migration and effector function of
inflammatory cells (activated lymphoid and myeloid cells). These
include genes that encode inflammatory cytokines, chemokines, and
cytotoxic enzymes. Expression of these inflammatory genes is
tightly regulated at the transcriptional level by specific
transcription factors.
[0003] Recent studies indicate that, c-Rel, the lymphoid member of
the Rel/nuclear factor-.kappa.B (Rel/NF-.kappa.B) family, is a long
sought-after transcriptional regulator of EAE. The mammalian
Rel/NF-.kappa.B family consists of five members, each of which is
endowed with a distinct set of function not shared by other
members, although each member may also perform additional functions
common to the family.
[0004] Thus, unlike other members that are ubiquitously expressed,
c-Rel is preferentially expressed by inflammatory cells, and is
involved in regulating a special subset of immune responses.
c-Rel-deficient mice do not suffer from developmental problems or
infectious diseases, but are resistant to inflammatory diseases
(despite the normal expression of other Rel/NF-.kappa.B proteins);
c-Rel-deficient T cells are competent in survival and Th2 type
responses but are severely compromised in their Th1 and Th17 type
responses.
[0005] The Rel/NF-.kappa.B family of transcription factors
represents one of the most attractive targets for anti-inflammatory
therapy. Because Rel/NF-.kappa.B directly controls the expression
of multiple inflammatory genes, its blockade is more effective for
controlling inflammation than blocking one or a few downstream
inflammatory genes or proteins. The first generation
Rel/NF-.kappa.B drugs that block the entire Rel/NF-.kappa.B family
have already been tested in both humans and animals. These include
proteasome inhibitors (e.g., the FDA-approved PS-341),
Rel/NF-.kappa.B decoy oligodeoxynucleotides and the NBD
(neuro-binding domain) peptides, which are highly effective in
preventing and treating models of autoimmune diseases including
EAE. Additionally, glucocorticoids, which are currently used to
control acute inflammation in MS patients, mediate their
immunosuppressive effects, at least in part, through inhibiting
Rel/NF-.kappa.B (glucocorticoids upregulate I.kappa.B expression
and bind directly to Rel/NF-.kappa.B). However, because most
Rel/NF-.kappa.B proteins are ubiquitously expressed and are
involved in a variety of biological processes not related to
autoimmunity, these drugs have significant side effects. Therefore,
they can only be used for a short period of time to control acute
inflammation.
SUMMARY OF THE INVENTION
[0006] This invention provides, a selective c-Rel inhibitor having
the structure of formula (I):
##STR00001##
wherein Q.sub.1, Q.sub.2, Q.sub.3 are independently H, halogen,
CF.sub.3, OCH.sub.2Ph, O-alkyl, OCF.sub.3, alkyl, or Q.sub.1 and
Q.sub.2 form a saturated or unsaturated, substituted or
unsubstituted, carbocyclic or heterocyclic ring with the aniline
ring; X and Y are independently H, alkyl, or form a saturated or
unsaturated, substituted or unsubstituted, carbocyclic or
heterocyclic ring with N(R.sub.1)(R.sub.2). X.sub.1 and Y.sub.1 are
independently H, alkyl, or X and Y form together a double bond, or
form saturated or unsaturated, substituted or un-substituted,
carbocyclic or heterocyclic ring with N(R.sub.1) (R.sub.2). R.sub.1
and R.sub.2 are independently H, NH.sub.2, --N=alkyl, -alkyl,
--CH(Ph).sub.2, substituted or un-substituted aryl, carbocyclic or
heterocyclic aryl, substituted or un-substituted phenyl,
C(O)-alkyl, or R.sub.1 and R.sub.2 form a saturated or unsaturated,
substituted or un-substituted, carbocyclic or heterocyclic ring
with the nitrogen atom; or a selective c-Rel inhibitor having the
structure of formula (II):
##STR00002##
Wherein L.sub.1, L.sub.2, L.sub.3 and L.sub.4 are independently H,
halogen, alkyl, --NH.sub.2, --COO Alkyl, --NO.sub.2, pyrrolidine,
--O-alkyl, or L.sub.1 and L.sub.2 form a saturated or unsaturated,
substituted or un-substituted, carbocyclic or heterocyclic fused
ring with the benzene ring; or L.sub.4 together with R.sub.2 forms
a 6 membered fused ring with the imidazole and benzene rings; R and
R.sub.1 are independently H, NHCO-alkyl, or form together a double
bond (.dbd.), or CO group (.dbd.O); R.sub.2 is H, SH, OH, alkyl,
-Ph-CF.sub.3, --CH.dbd.C(Ph)-OC(O)-Ph, CH.sub.2--S-Ph,
CH.sub.2--S-heterocyclic ring, CH.sub.2OC(O)NH-Ph,
--NHCH.sub.2CH.sub.2OH, -alkylene-OH, O-aryl, --O-alkyl,
O--CH.sub.2-Ph, O-phenyl, O-phenyl-alkyl, O-Ph-O-alkylene-Ph,
--OCH.sub.2Ph, --OCH.sub.2CH.dbd.CH-Ph, --S-Phenyl, NH-alkyl,
NH-phenyl, NH-aryl, --N(Me)-alkylene-phenyl, --NH-alkylene-phenyl,
--NH-alkylene-OMe, --NH--N.dbd.CH-Ph, --NH--N--C(O)-alkyl,
--NH-heterocyclic ring, NH-carbocyclic ring, --C(O)Ph, substituted
or un-substituted, saturated or unsaturated hetrocyclic ring,
substituted or un-substituted, saturated or unsaturated carbocyclic
ring, or R.sub.2 together with L.sub.4 forms a 6 membered fused
ring with the imidazole and benzene rings; R.sub.3 is H, COO-alkyl,
COOH, NO.sub.2, substituted or un-substituted Ph,
C(O)--N.dbd.NC(O)Ph or C(O)NH.sub.2; and R.sub.4 is H, Ph, alkyl,
NH.sub.2, OH, Ph-OH or CH.sub.2--OH; or a selective c-Rel inhibitor
having the structure of formula (CXIX):
##STR00003##
Wherein R.sub.1 is substituted phenyl or unsubstituted phenyl,
whereby a substituted phenyl ring comprises CN, halogen, or alkyl
substituent;
R.sub.2 is
##STR00004##
[0007] wherein X.sub.1, X.sub.2, or X.sub.3 are independently H,
halogen, alkyl, CN, COOH, or NH.sub.2; or X.sub.3 forms with the
.dbd.N.sup.+H.sub.2 a five membered fused ring; or R.sub.2 forms
with .dbd.N.sup.+H.sub.2 a five or six substituted or unsubstituted
membered ring, whereby a five or six substituted membered ring
comprises CH.sub.2-Ph, aryl, or alkyl substituent.
[0008] In another embodiment, the present invention provides a
composition comprising a selective c-Rel inhibitor having the
structure of formula (I):
##STR00005##
wherein Q.sub.1, Q.sub.2, Q.sub.3 are independently H, halogen,
CF.sub.3, OCH.sub.2Ph, O-alkyl, OCF.sub.3, alkyl, or Q.sub.1 and
Q.sub.2 form a saturated or unsaturated, substituted or
un-substituted, carbocyclic or heterocyclic ring with the aniline
ring; X and Y are independently H, alkyl, or form a saturated or
unsaturated, substituted or un-substituted, carbocyclic or
heterocyclic ring with N(R.sub.1) (R.sub.2). X.sub.1 and Y.sub.1
are independently H, alkyl, or X and Y form together a double bond,
or form saturated or unsaturated, substituted or un-substituted,
carbocyclic or heterocyclic ring with N(R.sub.1) (R.sub.2). R.sub.1
and R.sub.2 are independently H, NH.sub.2, --N=alkyl, -alkyl,
--CH(Ph).sub.2, substituted or un-substituted aryl, carbocyclic or
heterocyclic aryl, substituted or un-substituted phenyl,
C(O)-alkyl, or R.sub.1 and R.sub.2 form a saturated or unsaturated,
substituted or un-substituted, carbocyclic or heterocyclic ring
with the nitrogen atom; or a selective c-Rel inhibitor having the
structure of formula (II):
##STR00006##
wherein L.sub.1, L.sub.2, L.sub.3 and L.sub.4 are independently H,
halogen, alkyl, --NH.sub.2, --COO Alkyl, --NO.sub.2, pyrrolidine,
--O-alkyl, or L.sub.1 and L.sub.2 form a saturated or unsaturated,
substituted or un-substituted, carbocyclic or heterocyclic fused
ring with the benzene ring; or L.sub.4 together with R.sub.2 forms
a 6 membered fused ring with the imidazole and benzene rings; R and
R.sub.1 are independently H, NHCO-alkyl, or form together a double
bond (.dbd.), or CO group (.dbd.O); R.sub.2 is H, SH, OH, alkyl,
-Ph-CF.sub.3, --CH.dbd.C(Ph)-OC(O)-Ph, CH.sub.2--S-Ph,
CH.sub.2--S-heterocyclic ring, CH.sub.2OC(O)NH-Ph,
--NHCH.sub.2CH.sub.2OH, -alkylene-OH, O-aryl, --O-alkyl,
O--CH.sub.2-Ph, O-phenyl, O-phenyl-alkyl, O-Ph-O-alkylene-Ph,
--OCH.sub.2Ph, --OCH.sub.2CH.dbd.CH-Ph, --S-Phenyl, NH-alkyl,
NH-phenyl, NH-aryl, --N(Me)-alkylene-phenyl, --NH-alkylene-phenyl,
--NH-alkylene-OMe, --NH--N.dbd.CH-Ph, --NH--N--C(O)-alkyl,
--NH-heterocyclic ring, NH-carbocyclic ring, --C(O)Ph, substituted
or un-substituted, saturated or unsaturated hetrocyclic ring,
substituted or un-substituted, saturated or unsaturated carbocyclic
ring, or R.sub.2 together with L.sub.4 forms a 6 membered fused
ring with the imidazole and benzene rings; R.sub.3 is H, COO-alkyl,
COOH, NO.sub.2, substituted or un-substituted Ph,
C(O)--N.dbd.NC(O)Ph or C(O)NH.sub.2; and; R4 is H, Ph, alkyl,
NH.sub.2, OH, Ph-OH or CH.sub.2--OH; or a selective c-Rel inhibitor
having the structure of formula (CXIX):
##STR00007##
Wherein R.sub.1 is substituted phenyl or unsubstituted phenyl,
whereby a substituted phenyl ring comprises CN, halogen, or alkyl
substituent;
R.sub.2 is
##STR00008##
[0009] wherein X.sub.1, X.sub.2, or X.sub.3 are independently H,
halogen, alkyl, CN, COOH, or NH.sub.2; or X.sub.3 forms with the
.dbd.N.sup.+H.sub.2 a five membered fused ring; or R.sub.2 forms
with .dbd.N.sup.+H.sub.2 a five or six substituted or unsubstituted
membered ring, whereby a five or six substituted membered ring
comprises CH.sub.2-Ph, aryl, or alkyl substituent.
[0010] In another embodiment, the present invention provides a
method of treating multiple sclerosis, arthritis, diabetes, graft
rejection, or a combination thereof in a subject comprising the
step of contacting an inflammatory cell with a selective c-Rel
inhibitor wherein the c-Rel inhibitor masks the L1 cavity of a
c-Rel protein, thereby treating or preventing multiple sclerosis,
arthritis, diabetes, graft rejection, or a combination thereof in a
subject.
[0011] In one embodiment, the invention provides a method of
inhibiting or suppressing the interaction between c-Rel and a DNA,
comprising the step of contacting the c-Rel with a compound capable
of masking the L1 cavity of the c-Rel, thereby inhibiting or
suppressing the interaction between c-Rel and a DNA and
inflammatory immune response.
[0012] In another embodiment, the present invention provides a
method of preventing multiple sclerosis, arthritis, diabetes, graft
rejection, or a combination thereof in a subject comprising the
step of contacting an inflammatory cell with a selective c-Rel
inhibitor wherein the c-Rel inhibitor masks the L1 cavity of a
c-Rel protein, thereby treating or preventing multiple sclerosis,
arthritis, diabetes, graft rejection, or a combination thereof in a
subject.
[0013] In another embodiment, the present invention provides a
method of inhibiting or suppressing multiple sclerosis, arthritis,
diabetes, graft rejection, or a combination thereof in a subject
comprising the step of contacting an inflammatory cell with a
selective c-Rel inhibitor wherein the c-Rel inhibitor masks the L1
cavity of a c-Rel protein, thereby treating or preventing multiple
sclerosis, arthritis, diabetes, graft rejection, or a combination
thereof in a subject.
[0014] In another embodiment, the present invention provides a
method of reducing the symptoms associated with multiple sclerosis,
arthritis, diabetes, graft rejection, or a combination thereof in a
subject comprising the step of contacting an inflammatory cell with
a selective c-Rel inhibitor wherein the c-Rel inhibitor masks the
L1 cavity of a c-Rel protein, thereby treating or preventing
multiple sclerosis, arthritis, diabetes, graft rejection, or a
combination thereof in a subject.
[0015] In another embodiment, the present invention provides a
method of identifying a selective c-Rel inhibitor comprising the
steps of: (a) constructing a c-Rel protein 3-D model; and (b)
minimizing the model to said identify said selective c-Rel
inhibitor, wherein said inhibitor interferes with L1 cavity,
thereby identifying a selective c-Rel DNA binding inhibitor.
BRIEF DESCRIPTION OF THE FIGURES
[0016] FIG. 1. FIG. 1A depicts a graph showing the IL-2 inhibitory
effect of R13D (compound III), R13096 (compound CXX), and R13015
(compound VI). FIG. 1A depicts a graph showing the IFN-gamma
inhibitory effect of R13D (compound III), R13096 (compound CXX),
and R13015 (compound VI).
[0017] FIG. 2. FIG. 2A depicts a surface model of c-Rel is shown in
pale green color. The DNA binding region is shown in brown shadow.
The L1 cavity, located between L1 and L2, is filled with an
inhibitor shown in multiple colors. FIG. 2B depicts the inhibitor
binding site together with the DNA (shown as a helix) binding site,
as seen in the surface model of c-Rel, the inhibitor occupies the
site where DNA binds. FIG. 2C is the chemical structure of the
active inhibitor, R13D (compound III).
[0018] FIG. 3. Depicts a graph showing Splenocytes that were
cultured under various conditions as indicated on the X-axis.
Twenty-four hours later, cytokines in the supernatant were
determined by ELISA (R13D is compound III, R31C is compound IV,
R32D is compound VII).
[0019] FIG. 4 depicts graphs of the results from C57BL/6 mice that
were immunized to induce EAE with 1) is a subcutaneous injection on
flanks of 300 .mu.g MOG38-50 peptide in 0.1 ml PBS emulsified in an
equal volume of complete Freund's adjuvant containing 500 .mu.g
mycobacterium tuberculosis H37RA, and 2) is an intravenous
injection of 100 ng pertussis toxin in 0.1 ml PBS. A second
injection of pertussis toxin (200 ng per mouse) was given 48 h
later. For FIG. 4A, mice were injected with 100 ug R13D (compound
III) daily from day 1 to day 13. For FIG. 4B, mice were injected
with 100 ug R13D (compound III) daily from day 25 (after onset) to
day 32. Mice were examined daily for signs of EAE and scored as
follows (6). 0--no disease; 1--tail paralysis; 2--hind limb
weakness; 3--hind limb paralysis; 4--hind limb plus forelimb
paralysis; 5--moribund or dead. The differences for all the
parameters between c-Rel.sup.+/+ and c-Rel.sup.-/- mice are
statistically significant (p<0.001). The experiment was repeated
twice with similar results.
[0020] FIG. 4 shows a cell-based assay for testing c-Rel blockers.
Splenocytes of C57BL/6 mice (n=4) were cultured with or without
plate-bound anti-CD3 (1 ug/ml) and soluble anti-CD28 (2 ug/ml), in
the presence of various concentrations of R96 as indicated. R96 was
dissolved in PBS, and the amount of the PBS added to each culture
was identical (5 ml per 200 ml culture). Twenty-four hours later,
cytokines in the supernatant were determined by ELISA (A-C). The
percentages of dead cells in the culture were determined by flow
cytometry following staining cells with annexin-V and propidium
iodide (PI) (D). No significant differences in the percentage of
annexin-V+ and PI+ cells (dead cells) were observed among different
cultures.
[0021] FIG. 5 shows prevention and treatment of EAE using c-Rel
blockers. C57BL/6 mice, 6-8 per group, were immunized to induce EAE
with MOG38-50 peptide. A) Mice were injected i.p. with vehicle or
100 ug/mouse/injection of R13 daily from day 1 to day 13. B) Mice
were injected i.p. with vehicle or 100 ug/mouse/injection of R13
daily from day 2 (after onset) to day 10. C) Mice were injected
i.p. with vehicle or 100 ug/mouse/injection of R96 daily from day 5
(after onset) to day 24. The differences between the two groups are
statistically significant as determined by Mann-Whitney U test
(p<0.001) for a after day 12, for B after day 9 and for C after
day 8. Results are representative of 3 experiments.
DETAILED DESCRIPTION OF THE INVENTION
[0022] In one embodiment, provided herein are REL inhibitors and
methods of using and identifying the same. In other embodiments,
provided herein are c-Rel inhibitors and their use in the treatment
of Multiple Sclerosis.
[0023] This invention provides a selective c-Rel inhibitor having
the structure of
##STR00009##
wherein Q.sub.1, Q.sub.2, Q.sub.3 are independently H, halogen,
CF.sub.3, OCH.sub.2Ph, O-alkyl, OCF.sub.3, alkyl, or Q.sub.1 and
Q.sub.2 form a saturated or unsaturated, substituted or
un-substituted, carbocyclic or heterocyclic ring with the aniline
ring; X and Y are independently H, alkyl, or form a saturated or
unsaturated, substituted or un-substituted, carbocyclic or
heterocyclic ring with N(R.sub.1)(R.sub.2). X.sub.1 and Y.sub.1 are
independently H, alkyl, or X and Y form together a double bond, or
form saturated or unsaturated, substituted or un-substituted,
carbocyclic or heterocyclic ring with N(R.sub.1) (R.sub.2). R.sub.1
and R.sub.2 are independently H, NH.sub.2, --N=alkyl, -alkyl,
--CH(Ph).sub.2, substituted or un-substituted aryl, carbocyclic or
heterocyclic aryl, substituted or un-substituted phenyl,
C(O)-alkyl, or R.sub.1 and R.sub.2 form a saturated or unsaturated,
substituted or un-substituted, carbocyclic or heterocyclic ring
with the nitrogen atom; or a selective c-Rel DNA binding inhibitor
having the structure of formula (II):
##STR00010##
Wherein L.sub.1, L.sub.2, L.sub.3 and L.sub.4 are independently H,
halogen, alkyl, --NH.sub.2, --COO Alkyl, --NO.sub.2, pyrrolidine,
--O-alkyl, or L.sub.1 and L.sub.2 form a saturated or unsaturated,
substituted or un-substituted, carbocyclic or heterocyclic fused
ring with the benzene ring; or L.sub.4 together with R.sub.2 forms
a 6 membered fused ring with the imidazole and benzene rings; R and
R.sub.1 are independently H, NHCO-alkyl, or form together a double
bond (.dbd.), or CO group (.dbd.O); R.sub.2 is H, SH, OH, alkyl,
-Ph-CF.sub.3, --CH.dbd.C(Ph)-OC(O)-Ph, CH.sub.2--S-Ph,
CH.sub.2--S-heterocyclic ring, CH.sub.2OC(O)NH-Ph,
--NHCH.sub.2CH.sub.2OH, -alkylene-OH, O-aryl, --O-alkyl,
O--CH.sub.2-Ph, O-phenyl, O-phenyl-alkyl, O-Ph-O-alkylene-Ph,
--OCH.sub.2Ph, --OCH.sub.2CH.dbd.CH-Ph, --S-Phenyl, NH-alkyl,
NH-phenyl, NH-aryl, --N(Me)-alkylene-phenyl, --NH-alkylene-phenyl,
--NH-alkylene-OMe, --NH--N.dbd.CH-Ph, --NH--N--C(O)-alkyl,
--NH-heterocyclic ring, NH-carbocyclic ring, --C(O)Ph, substituted
or un-substituted, saturated or unsaturated hetrocyclic ring,
substituted or un-substituted, saturated or unsaturated carbocyclic
ring, or R.sub.2 together with L.sub.4 forms a 6 membered fused
ring with the imidazole and benzene rings; R.sub.3 is H, COO-alkyl,
COOH, NO.sub.2, substituted or un-substituted Ph,
C(O)--N.dbd.NC(O)Ph or C(O)NH.sub.2; and; R4 is H, Ph, alkyl,
NH.sub.2, OH, Ph-OH or CH.sub.2--OH; or a selective c-Rel DNA
binding inhibitor having the structure of formula (CXIX):
##STR00011##
Wherein R.sub.1 is substituted phenyl or unsubstituted phenyl,
whereby a substituted phenyl ring comprises CN, halogen, or alkyl
substituent;
R.sub.2 is
##STR00012##
[0024] wherein X.sub.1, X.sub.2, or X.sub.3 are independently H,
halogen, alkyl, CN, COOH, or NH.sub.2; or X.sub.3 forms with the
.dbd.N.sup.+H.sub.2 a five membered fused ring; or R.sub.2 forms
with .dbd.N.sup.+H.sub.2 a five or six substituted or unsubstituted
membered ring, whereby a five or six substituted membered ring
comprises CH.sub.2-Ph, aryl, or alkyl substituent.
[0025] In another embodiment, the selective c-Rel inhibitor of the
present invention blocks a druggable cavity. In another embodiment,
the selective c-Rel DNA binding inhibitor of the present invention
blocks a small druggable cavity. In another embodiment, the
selective c-Rel DNA binding inhibitor of the present invention
blocks the biological activity of c-Rel. In another embodiment, the
selective c-Rel DNA binding inhibitor of the present invention
interferes with the residues in loops L1 and L2 of the c-Rel
protein (FIG. 2). In another embodiment, the selective c-Rel DNA
binding inhibitor of the present invention comprises a c-Rel
binding site. In another embodiment, upon binding of the selective
c-Rel DNA binding inhibitor to c-Rel, the L1 cavity of the c-Rel
protein is masked. In another embodiment, c-Rel:DNA contacts are
mediated by four loops, L1 and L2 loops (FIG. 2A) from the
N-terminal domain, and L3 and L4 loops from the C-terminal domain.
In another embodiment, a DNA fragment is sandwiched between these
loops.
[0026] In one embodiment, provided herein is a method of inhibiting
or suppressing the interaction between c-Rel and a DNA, comprising
the step of contacting the c-Rel with a compound capable of masking
the L1 cavity of the c-Rel, thereby inhibiting or suppressing the
interaction between c-Rel and a DNA and inflammatory immune
response. In another embodiment, the compounds described herein are
used in the methods of inhibiting or suppressing the interaction
between c-Rel and DNA. In one embodiment, the compound capable of
inhibiting the interaction between c-Rel and a DNA is the compound
represented by the structure of formula III, also referred to in
certain embodiments as R13:
##STR00013##
or a pharmaceutical salt thereof; or in another embodiment, by the
compound represented by the structure of formula CXX, also referred
to interchangeably is compound 096 hereinbelow:
##STR00014##
or a pharmaceutically acceptable salt thereof.
[0027] In another embodiment, the selective c-Rel DNA binding
inhibitor of the present invention binds the L1 cavity formed by
loops L1, L2 and a helix. In another embodiment, the L1 cavity
comprises the AA Arg 21, Cys 26, Glu 27, Lys 110 and Lys 111. In
another embodiment, the L1 cavity comprises the AA Arg 21 and Cys
26. In another embodiment, the L1 cavity comprises the AA Arg 21,
Cys 26 and Glu 27. In another embodiment, the L1 cavity comprises
the AA Arg 21, Cys 26, Glu 27 and Lys 110. In another embodiment,
the L1 cavity comprises the AA Arg 21, Cys 26, Glu 27, Lys 110 and
Lys 111.
[0028] In another embodiment, the AA Arg 21, Cys 26, Glu 27, Lys
110 and Lys 111 form the mouth of the L1 cavity (FIG. 2). In
another embodiment, the selective c-Rel DNA binding inhibitor of
the present invention is screened for blocking the DNA binding to
c-Rel. In another embodiment, the selective c-Rel DNA binding
inhibitor of the present invention is screened for blocking the DNA
binding to c-Rel by perturbing key contact residues in L1 loop.
[0029] In another embodiment, the inhibitor provided herein
comprises the structure represented by formula III:
##STR00015##
or its analog, metabolite or prodrug and their combination
thereof.
[0030] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula IV:
##STR00016##
or its analog, metabolite or prodrug and their combination
thereof.
[0031] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula V:
##STR00017##
or its analog, metabolite or prodrug and their combination
thereof.
[0032] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula VII:
##STR00018##
or its analog, metabolite or prodrug and their combination
thereof.
[0033] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula VIII:
##STR00019##
or its analog, metabolite or prodrug and their combination
thereof.
[0034] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula IX:
##STR00020##
or its analog, metabolite or prodrug and their combination
thereof.
[0035] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula X:
##STR00021##
or its analog, metabolite or prodrug and their combination
thereof.
[0036] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XI:
##STR00022##
or its analog, metabolite or prodrug and their combination
thereof.
[0037] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XII:
##STR00023##
or its analog, metabolite or prodrug and their combination
thereof.
[0038] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XIII:
##STR00024##
or its analog, metabolite or prodrug and their combination
thereof.
[0039] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XIV:
##STR00025##
or its analog, metabolite or prodrug and their combination
thereof.
[0040] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XV:
##STR00026##
or its analog, metabolite or prodrug and their combination
thereof.
[0041] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XVI:
##STR00027##
or its analog, metabolite or prodrug and their combination
thereof.
[0042] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XVII:
##STR00028##
or its analog, metabolite or prodrug and their combination
thereof.
[0043] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XVIII:
##STR00029##
or its analog, metabolite or prodrug and their combination
thereof.
[0044] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XIX:
##STR00030##
or its analog, metabolite or prodrug and their combination
thereof.
[0045] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XX:
##STR00031##
or its analog, metabolite or prodrug and their combination
thereof.
[0046] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXI:
##STR00032##
or its analog, metabolite or prodrug and their combination
thereof.
[0047] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXII:
##STR00033##
or its analog, metabolite or prodrug and their combination
thereof.
[0048] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXIII:
##STR00034##
or its analog, metabolite or prodrug and their combination
thereof.
[0049] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXIV:
##STR00035##
or its analog, metabolite or prodrug and their combination
thereof.
[0050] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXV:
##STR00036##
or its analog, metabolite or prodrug and their combination
thereof.
[0051] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXVI:
##STR00037##
or its analog, metabolite or prodrug and their combination
thereof.
[0052] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXVII:
##STR00038##
or its analog, metabolite or prodrug and their combination
thereof.
[0053] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXVIII:
##STR00039##
or its analog, metabolite or prodrug and their combination
thereof.
[0054] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXIX:
##STR00040##
or its analog, metabolite or prodrug and their combination
thereof.
[0055] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXX:
##STR00041##
or its analog, metabolite or prodrug and their combination
thereof.
[0056] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXXI:
##STR00042##
or its analog, metabolite or prodrug and their combination
thereof.
[0057] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXXII:
##STR00043##
or its analog, metabolite or prodrug and their combination
thereof.
[0058] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXXIII:
##STR00044##
or its analog, metabolite or prodrug and their combination
thereof.
[0059] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXXIV:
##STR00045##
or its analog, metabolite or prodrug and their combination
thereof.
[0060] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXXV:
##STR00046##
or its analog, metabolite or prodrug and their combination
thereof.
[0061] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXXVI:
##STR00047##
or its analog, metabolite or prodrug and their combination
thereof.
[0062] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXXVII:
##STR00048##
or its analog, metabolite or prodrug and their combination
thereof.
[0063] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXXVIII:
##STR00049##
or its analog, metabolite or prodrug and their combination
thereof.
[0064] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XXXIX:
##STR00050##
or its analog, metabolite or prodrug and their combination
thereof.
[0065] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XL:
##STR00051##
or its analog, metabolite or prodrug and their combination
thereof.
[0066] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XLI:
##STR00052##
or its analog, metabolite or prodrug and their combination
thereof.
[0067] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XLII:
##STR00053##
or its analog, metabolite or prodrug and their combination
thereof.
[0068] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XLIII:
##STR00054##
or its analog, metabolite or prodrug and their combination
thereof.
[0069] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XLIV:
##STR00055##
or its analog, metabolite or prodrug and their combination
thereof.
[0070] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XLV:
##STR00056##
or its analog, metabolite or prodrug and their combination
thereof.
[0071] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XLVI:
##STR00057##
or its analog, metabolite or prodrug and their combination
thereof.
[0072] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XLVII:
##STR00058##
or its analog, metabolite or prodrug and their combination
thereof.
[0073] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XLVIII:
##STR00059##
or its analog, metabolite or prodrug and their combination
thereof.
[0074] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XLIX:
##STR00060##
or its analog, metabolite or prodrug and their combination
thereof.
[0075] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula L:
##STR00061##
or its analog, metabolite or prodrug and their combination
thereof.
[0076] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LI:
##STR00062##
or its analog, metabolite or prodrug and their combination
thereof.
[0077] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LII:
##STR00063##
or its analog, metabolite or prodrug and their combination
thereof.
[0078] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LIII:
##STR00064##
or its analog, metabolite or prodrug and their combination
thereof.
[0079] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LIV:
##STR00065##
or its analog, metabolite or prodrug and their combination
thereof.
[0080] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LV:
##STR00066##
or its analog, metabolite or prodrug and their combination
thereof.
[0081] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LVI:
##STR00067##
or its analog, metabolite or prodrug and their combination
thereof.
[0082] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LVII:
##STR00068##
or its analog, metabolite or prodrug and their combination
thereof.
[0083] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LVIII:
##STR00069##
or its analog, metabolite or prodrug and their combination
thereof.
[0084] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LIX:
##STR00070##
or its analog, metabolite or prodrug and their combination
thereof.
[0085] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LX:
##STR00071##
or its analog, metabolite or prodrug and their combination
thereof.
[0086] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXI:
##STR00072##
or its analog, metabolite or prodrug and their combination
thereof.
[0087] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXII:
##STR00073##
or its analog, metabolite or prodrug and their combination
thereof.
[0088] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXIII:
##STR00074##
or its analog, metabolite or prodrug and their combination
thereof.
[0089] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXIV:
##STR00075##
or its analog, metabolite or prodrug and their combination
thereof.
[0090] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXV:
##STR00076##
or its analog, metabolite or prodrug and their combination
thereof.
[0091] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXVI:
##STR00077##
or its analog, metabolite or prodrug and their combination
thereof.
[0092] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXVII:
##STR00078##
or its analog, metabolite or prodrug and their combination
thereof.
[0093] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXVIII:
##STR00079##
or its analog, metabolite or prodrug and their combination
thereof.
[0094] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXIX:
##STR00080##
or its analog, metabolite or prodrug and their combination
thereof.
[0095] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXX:
##STR00081##
or its analog, metabolite or prodrug and their combination
thereof.
[0096] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXI:
##STR00082##
or its analog, metabolite or prodrug and their combination
thereof.
[0097] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXII:
##STR00083##
or its analog, metabolite or prodrug and their combination
thereof.
[0098] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXIII:
##STR00084##
or its analog, metabolite or prodrug and their combination
thereof.
[0099] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXIV:
##STR00085##
or its analog, metabolite or prodrug and their combination
thereof.
[0100] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXV:
##STR00086##
or its analog, metabolite or prodrug and their combination
thereof.
[0101] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXVI:
##STR00087##
or its analog, metabolite or prodrug and their combination
thereof.
[0102] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXVII:
##STR00088##
or its analog, metabolite or prodrug and their combination
thereof.
[0103] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXVIII:
##STR00089##
or its analog, metabolite or prodrug and their combination
thereof.
[0104] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXIX:
##STR00090##
or its analog, metabolite or prodrug and their combination
thereof.
[0105] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXX:
##STR00091##
or its analog, metabolite or prodrug and their combination
thereof.
[0106] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXXI:
##STR00092##
or its analog, metabolite or prodrug and their combination
thereof.
[0107] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXXII:
##STR00093##
or its analog, metabolite or prodrug and their combination
thereof.
[0108] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXXIII:
##STR00094##
or its analog, metabolite or prodrug and their combination
thereof.
[0109] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXXIV:
##STR00095##
or its analog, metabolite or prodrug and their combination
thereof.
[0110] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXV:
##STR00096##
or its analog, metabolite or prodrug and their combination
thereof.
[0111] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXXVI:
##STR00097##
or its analog, metabolite or prodrug and their combination
thereof.
[0112] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXXVII:
##STR00098##
or its analog, metabolite or prodrug and their combination
thereof.
[0113] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXVIII:
##STR00099##
or its analog, metabolite or prodrug and their combination
thereof.
[0114] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula LXXXIX:
##STR00100##
or its analog, metabolite or prodrug and their combination
thereof.
[0115] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XC:
##STR00101##
or its analog, metabolite or prodrug and their combination
thereof.
[0116] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XCI:
##STR00102##
or its analog, metabolite or prodrug and their combination
thereof.
[0117] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XCII:
##STR00103##
or its analog, metabolite or prodrug and their combination
thereof.
[0118] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula
##STR00104##
or its analog, metabolite or prodrug and their combination
thereof.
[0119] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XCIV:
##STR00105##
or its analog, metabolite or prodrug and their combination
thereof.
[0120] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XCV:
##STR00106##
or its analog, metabolite or prodrug and their combination
thereof.
[0121] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XCVI:
##STR00107##
or its analog, metabolite or prodrug and their combination
thereof.
[0122] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XCVII:
##STR00108##
or its analog, metabolite or prodrug and their combination
thereof.
[0123] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XCVIII:
##STR00109##
or its analog, metabolite or prodrug and their combination
thereof.
[0124] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula XCIX:
##STR00110##
or its analog, metabolite or prodrug and their combination
thereof.
[0125] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula C:
##STR00111##
or its analog, metabolite or prodrug and their combination
thereof.
[0126] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CI:
##STR00112##
or its analog, metabolite or prodrug and their combination
thereof.
[0127] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CII:
##STR00113##
or its analog, metabolite or prodrug and their combination
thereof.
[0128] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CIII:
##STR00114##
or its analog, metabolite or prodrug and their combination
thereof.
[0129] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CIV:
##STR00115##
or its analog, metabolite or prodrug and their combination
thereof.
[0130] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CV:
##STR00116##
or its analog, metabolite or prodrug and their combination
thereof.
[0131] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CVI:
##STR00117##
or its analog, metabolite or prodrug and their combination
thereof.
[0132] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CVII:
##STR00118##
or its analog, metabolite or prodrug and their combination
thereof.
[0133] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CVIII:
##STR00119##
or its analog, metabolite or prodrug and their combination
thereof.
[0134] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CIX:
##STR00120##
or its analog, metabolite or prodrug and their combination
thereof.
[0135] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CX:
##STR00121##
or its analog, metabolite or prodrug and their combination
thereof.
[0136] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXI:
##STR00122##
or its analog, metabolite or prodrug and their combination
thereof.
[0137] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXII:
##STR00123##
or its analog, metabolite or prodrug and their combination
thereof.
[0138] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula
##STR00124##
or its analog, metabolite or prodrug and their combination
thereof.
[0139] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXIV:
##STR00125##
or its analog, metabolite or prodrug and their combination
thereof.
[0140] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXV:
##STR00126##
or its analog, metabolite or prodrug and their combination
thereof.
[0141] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXVI:
##STR00127##
or its analog, metabolite or prodrug and their combination
thereof.
[0142] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXVII:
##STR00128##
or its analog, metabolite or prodrug and their combination
thereof.
[0143] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXVIII:
##STR00129##
or its analog, metabolite or prodrug and their combination
thereof.
[0144] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXIX
##STR00130##
Wherein R.sub.1 is substituted phenyl or unsubstituted phenyl,
whereby a substituted phenyl ring comprises CN, halogen, or alkyl
substituent;
R.sub.2 is
##STR00131##
[0145] wherein X.sub.1, X.sub.2, or X.sub.3 are independently H,
halogen, alkyl, CN, COOH, or NH.sub.2; or X.sub.3 forms with the
.dbd.N.sup.+H.sub.2 a five membered fused ring R.sub.2 forms with
.dbd.N.sup.+H.sub.2 a five or six substituted or unsubstituted
membered ring, whereby a five or six substituted membered ring
comprises CH.sub.2-Ph, aryl, or alkyl substituent.
[0146] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXX:
##STR00132##
or its analog, metabolite or prodrug and their combination
thereof.
[0147] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXI:
##STR00133##
or its analog, metabolite or prodrug and their combination
thereof.
[0148] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXII:
##STR00134##
or its analog, metabolite or prodrug and their combination
thereof.
[0149] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXIII:
##STR00135##
or its analog, metabolite or prodrug and their combination
thereof.
[0150] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXIV:
##STR00136##
or its analog, metabolite or prodrug and their combination
thereof.
[0151] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXV:
##STR00137##
or its analog, metabolite or prodrug and their combination
thereof.
[0152] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXVI:
##STR00138##
or its analog, metabolite or prodrug and their combination
thereof.
[0153] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXVII:
##STR00139##
or its analog, metabolite or prodrug and their combination
thereof.
[0154] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXVIII:
##STR00140##
or its analog, metabolite or prodrug and their combination
thereof.
[0155] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXIX:
##STR00141##
or its analog, metabolite or prodrug and their combination
thereof.
[0156] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXX:
##STR00142##
or its analog, metabolite or prodrug and their combination
thereof.
[0157] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXXI:
##STR00143##
or its analog, metabolite or prodrug and their combination
thereof.
[0158] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXXII:
##STR00144##
or its analog, metabolite or prodrug and their combination
thereof.
[0159] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXXIII:
##STR00145##
or its analog, metabolite or prodrug and their combination
thereof.
[0160] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXXIV:
##STR00146##
or its analog, metabolite or prodrug and their combination
thereof.
[0161] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXXV:
##STR00147##
or its analog, metabolite or prodrug and their combination
thereof.
[0162] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXXVI:
##STR00148##
or its analog, metabolite or prodrug and their combination
thereof.
[0163] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXXVII:
##STR00149##
or its analog, metabolite or prodrug and their combination
thereof.
[0164] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXXVIII:
##STR00150##
or its analog, metabolite or prodrug and their combination
thereof.
[0165] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXXXIX:
##STR00151##
or its analog, metabolite or prodrug and their combination
thereof.
[0166] In another embodiment, the inhibitor used in the
compositions and methods provided, comprises the structure
represented by formula CXL:
##STR00152##
or its analog, metabolite or prodrug and their combination
thereof.
[0167] In another embodiment, provided herein is that an inhibitor
provided herein inhibits the production of a pro-inflammatory
cytokine. In another embodiment, provided herein is that an
inhibitor provided herein inhibits the production of interleukin-2,
interferon-gamma, or the combination thereof.
[0168] In another embodiment, provided herein is a composition
comprising a selective c-Rel inhibitor that is any one of the
structures represented by formulas I-CXL. In another embodiment,
provided herein is a composition comprising a selective c-Rel DNA
binding inhibitor having the structure of formula (I):
##STR00153##
wherein Q.sub.1, Q.sub.2, Q.sub.3 are independently H, halogen,
CF.sub.3, OCH.sub.2Ph, O-alkyl, OCF.sub.3, alkyl, or Q.sub.1 and
Q.sub.2 form a saturated or unsaturated, substituted or
un-substituted, carbocyclic or heterocyclic ring with the aniline
ring; X and Y are independently H, alkyl, or form a saturated or
unsaturated, substituted or un-substituted, carbocyclic or
heterocyclic ring with N(R.sub.1)(R.sub.2). X.sub.1 and Y.sub.1 are
independently H, alkyl, or X and Y form together a double bond, or
form saturated or unsaturated, substituted or un-substituted,
carbocyclic or heterocyclic ring with N(R.sub.1) (R.sub.2). R.sub.1
and R.sub.2 are independently H, NH.sub.2, --N=alkyl, -alkyl,
--CH(Ph).sub.2, substituted or un-substituted aryl, carbocyclic or
heterocyclic aryl, substituted or un-substituted phenyl,
C(O)-alkyl, or R.sub.1 and R.sub.2 form a saturated or unsaturated,
substituted or un-substituted, carbocyclic or heterocyclic ring
with the nitrogen atom; or a selective c-Rel inhibitor having the
structure of formula (II):
##STR00154##
wherein L.sub.1, L.sub.2, L.sub.3 and L.sub.4 are independently H,
halogen, alkyl, --NH.sub.2, --COO Alkyl, --NO.sub.2, pyrrolidine,
--O-alkyl, or L.sub.1 and L.sub.2 form a saturated or unsaturated,
substituted or un-substituted, carbocyclic or heterocyclic fused
ring with the benzene ring; or L.sub.4 together with R.sub.2 forms
a 6 membered fused ring with the imidazole and benzene rings; R and
R.sub.1 are independently H, NHCO-alkyl, or form together a double
bond (.dbd.), or CO group (.dbd.O); R.sub.2 is H, SH, OH, alkyl,
-Ph-CF.sub.3, --CH.dbd.C(Ph)--OC(O)-Ph, CH.sub.2--S-Ph,
CH.sub.2--S-heterocyclic ring, CH.sub.2OC(O)NH-Ph,
--NHCH.sub.2CH.sub.2OH, -alkylene-OH, O-aryl, --O-alkyl,
O--CH.sub.2-Ph, O-phenyl, O-phenyl-alkyl, O-Ph-O-alkylene-Ph,
--OCH.sub.2Ph, --OCH.sub.2CH.dbd.CH-Ph, --S-Phenyl, NH-alkyl,
NH-phenyl, NH-aryl, --N(Me)-alkylene-phenyl, --NH-alkylene-phenyl,
--NH-alkylene-OMe, --NH--N.dbd.CH-Ph, --NH--N--C(O)-alkyl,
--NH-heterocyclic ring, NH-carbocyclic ring, --C(O)Ph, substituted
or un-substituted, saturated or unsaturated hetrocyclic ring,
substituted or un-substituted, saturated or unsaturated carbocyclic
ring, or R.sub.2 together with L.sub.4 forms a 6 membered fused
ring with the imidazole and benzene rings; R.sub.3 is H, COO-alkyl,
COOH, NO.sub.2, substituted or un-substituted Ph,
C(O)--N.dbd.NC(O)Ph or C(O)NH.sub.2; and; R.sub.4 is H, Ph, alkyl,
NH.sub.2, OH, Ph-OH or CH.sub.2--OH; or a selective c-Rel inhibitor
having the structure of formula (CXIX):
##STR00155##
Wherein R.sub.1 is a substituted phenyl or unsubstituted phenyl,
whereby a substituted phenyl ring comprises CN, halogen, or alkyl
substituent;
R.sub.2 is
##STR00156##
[0169] wherein X.sub.1, X.sub.2, or X.sub.3 are independently H,
halogen, alkyl, CN, COOH, or NH.sub.2; or X.sub.3 forms with the
.dbd.N.sup.+H.sub.2 a five membered fused ring; or R.sub.2 forms
with .dbd.N.sup.+H.sub.2 a five or six substituted or unsubstituted
membered ring, whereby a five or six substituted membered ring
comprises CH.sub.2-Ph, aryl, or alkyl substituent.
[0170] In another embodiment, the present invention provides a
composition comprising a selective c-Rel inhibitor having the
structure of formula (III):
##STR00157##
or its analog, metabolite or prodrug and their combination
thereof.
[0171] In another embodiment, provided herein is a composition
comprising a selective c-Rel inhibitor having the structure of
formula (IV):
##STR00158##
or its analog, metabolite or prodrug and their combination
thereof.
[0172] In another embodiment, provided herein is a composition
comprising a selective c-Rel inhibitor having the structure of
formula (V):
##STR00159##
or its analog, metabolite or prodrug and their combination
thereof.
[0173] In another embodiment, provided herein is a composition
comprising a selective c-Rel inhibitor having the structure of
formula (VII):
##STR00160##
[0174] In another embodiment, provided herein is a composition
comprising any inhibitor as described hereinabove or its analog,
metabolite or prodrug and their combination thereof. In another
embodiment, provided herein is a composition comprising
pharmaceutical excipients. In another embodiment, the compositions
provided herein provide a positive clinical score for preventing
multiple sclerosis. In another embodiment, the compositions
provided herein provide a positive clinical score for preventing
arthritis. In another embodiment, the compositions provided herein
provide a positive clinical score for preventing rheumatoid
arthritis. In another embodiment, the compositions provided herein
provide a positive clinical score for preventing diabetes. In
another embodiment, the compositions provided herein provide a
positive clinical score for preventing Diabetes Mellitus Type I. In
another embodiment, the compositions provided herein provide a
positive clinical score for preventing graft rejection. In another
embodiment, the compositions provided herein provide a positive
clinical score for preventing inflammation. In another embodiment,
the compositions provided herein provide a positive clinical score
for preventing neoplasma formation. In another embodiment, a
positive clinical score comprises slight but clearly defined
preventive effect for the above mentioned diseases.
[0175] In another embodiment, the compositions provided herein
provide a positive clinical score for treating multiple sclerosis.
In another embodiment, the compositions provided herein provide a
positive clinical score for treating arthritis. In another
embodiment, the compositions provided herein provide a positive
clinical score for treating rheumatoid arthritis. In another
embodiment, the compositions provided herein provide a positive
clinical score for treating diabetes. In another embodiment, the
compositions provided herein provide a positive clinical score for
treating type 1 diabetes. In another embodiment, the compositions
provided herein provide a positive clinical score for treating
graft rejection. In another embodiment, the compositions provided
herein provide a positive clinical score for treating inflammation.
In another embodiment, the compositions provided herein provide a
positive clinical score for treating neoplasma. In another
embodiment, a positive clinical score comprises slight but clearly
defined effect of treatment for the above mentioned diseases.
Pharmaceutical Compositions and Methods of Administration
[0176] The inhibitors used in the compositions and methods
provided, and pharmaceutical compositions comprising same can be,
in another embodiment, administered to a subject by any method
known to a person skilled in the art, such as parenterally,
paracancerally, transmucosally, transdermally, intramuscularly,
intravenously, intra-dermally, subcutaneously, intra-peritonealy,
intra-ventricularly, intra-cranially, intra-vaginally or
intra-tumorally.
[0177] In another embodiment of methods and compositions used in
the compositions and methods provided, the pharmaceutical
compositions are administered orally, and are thus formulated in a
form suitable for oral administration, i.e. as a solid or a liquid
preparation. Suitable solid oral formulations include tablets,
capsules, pills, granules, pellets and the like. Suitable liquid
oral formulations include solutions, suspensions, dispersions,
emulsions, oils and the like. In another embodiment used in the
compositions and methods provided, the active ingredient is
formulated in a capsule. In accordance with this embodiment, the
compositions used in the compositions and methods provided,
comprise, in addition to the active compound (e.g. the mimetic
compound, peptide or nucleotide molecule) and the inert carrier or
diluent, a hard gelating capsule.
[0178] In another embodiment, the pharmaceutical compositions are
administered by intravenous, intra-arterial, or intra-muscular
injection of a liquid preparation. Suitable liquid formulations
include solutions, suspensions, dispersions, emulsions, oils and
the like. In another embodiment, the pharmaceutical compositions
are administered intravenously and are thus formulated in a form
suitable for intravenous administration. In another embodiment, the
pharmaceutical compositions are administered intra-arterially and
are thus formulated in a form suitable for intra-arterial
administration. In another embodiment, the pharmaceutical
compositions are administered intra-muscularly and are thus
formulated in a form suitable for intra-muscular
administration.
[0179] In another embodiment, the pharmaceutical compositions are
administered topically to body surfaces and are thus formulated in
a form suitable for topical administration. Topical formulations
include, in another embodiment, gels, ointments, creams, lotions,
drops and the like.
[0180] In another embodiment, the pharmaceutical composition is
administered as a suppository, for example a rectal suppository or
a urethral suppository. In another embodiment, the pharmaceutical
composition is administered by subcutaneous implantation of a
pellet. In another embodiment, the pellet provides for controlled
release of active agent over a period of time.
[0181] In another embodiment, the active compound is delivered in a
vesicle, e.g. a liposome. In other embodiments, carriers or
diluents used in methods used in the compositions and methods
provided, include, but are not limited to, a gum, a starch (e.g.
corn starch, pregeletanized starch), a sugar (e.g., lactose,
mannitol, sucrose, dextrose), a cellulosic material (e.g.
microcrystalline cellulose), an acrylate (e.g. polymethylacrylate),
calcium carbonate, magnesium oxide, talc, or mixtures thereof.
[0182] In other embodiments, pharmaceutically acceptable carriers
for liquid formulations are aqueous or non-aqueous solutions,
suspensions, emulsions or oils. Examples of non-aqueous solvents
are propylene glycol, polyethylene glycol, and injectable organic
esters such as ethyl oleate. Aqueous carriers include water,
alcoholic/aqueous solutions, emulsions or suspensions, including
saline and buffered media. Examples of oils are those of animal,
vegetable, or synthetic origin, for example, peanut oil, soybean
oil, olive oil, sunflower oil, fish-liver oil, another marine oil,
or a lipid from milk or eggs.
[0183] In another embodiment, parenteral vehicles (for
subcutaneous, intravenous, intra-arterial, or intramuscular
injection) include sodium chloride solution, Ringer's dextrose,
dextrose and sodium chloride, lactated Ringer's and fixed oils.
Intravenous vehicles include fluid and nutrient replenishers,
electrolyte replenishers such as those based on Ringer's dextrose,
and the like. Examples are sterile liquids such as water and oils,
with or without the addition of a surfactant and other
pharmaceutically acceptable adjuvants. In general, water, saline,
aqueous dextrose and related sugar solutions, and glycols such as
propylene glycols or polyethylene glycol are preferred liquid
carriers, particularly for injectable solutions. Examples of oils
are those of animal, vegetable, or synthetic origin, for example,
peanut oil, soybean oil, olive oil, sunflower oil, fish-liver oil,
another marine oil, or a lipid from milk or eggs.
[0184] In other embodiments, the compositions further comprise
binders (e.g. acacia, cornstarch, gelatin, carbomer, ethyl
cellulose, guar gum, hydroxypropyl cellulose, hydroxypropyl methyl
cellulose, povidone), disintegrating agents (e.g. cornstarch,
potato starch, alginic acid, silicon dioxide, croscarmelose sodium,
crospovidone, guar gum, sodium starch glycolate), buffers (e.g.,
Tris-HCI., acetate, phosphate) of various pH and ionic strength,
additives such as albumin or gelatin to prevent absorption to
surfaces, detergents (e.g., Tween 20, Tween 80, Pluronic F68, bile
acid salts), protease inhibitors, surfactants (e.g. sodium lauryl
sulfate), permeation enhancers, solubilizing agents (e.g.,
glycerol, polyethylene glycerol), anti-oxidants (e.g., ascorbic
acid, sodium metabisulfite, butylated hydroxyanisole), stabilizers
(e.g. hydroxypropyl cellulose, hyroxypropylmethyl cellulose),
viscosity increasing agents (e.g. carbomer, colloidal silicon
dioxide, ethyl cellulose, guar gum), sweeteners (e.g. aspartame,
citric acid), preservatives (e.g., Thimerosal, benzyl alcohol,
parabens), lubricants (e.g. stearic acid, magnesium stearate,
polyethylene glycol, sodium lauryl sulfate), flow-aids (e.g.
colloidal silicon dioxide), plasticizers (e.g. diethyl phthalate,
triethyl citrate), emulsifiers (e.g. carbomer, hydroxypropyl
cellulose, sodium lauryl sulfate), polymer coatings (e.g.,
poloxamers or poloxamines), coating and film forming agents (e.g.
ethyl cellulose, acrylates, polymethacrylates) and/or adjuvants.
Each of the above excipients represents a separate embodiment used
in the compositions and methods provided.
[0185] In another embodiment, the pharmaceutical compositions
provided herein are controlled-release compositions, i.e.
compositions in which the active compound is released over a period
of time after administration. Controlled- or sustained-release
compositions include formulation in lipophilic depots (e.g. fatty
acids, waxes, oils). In another embodiment, the composition is an
immediate-release composition, i.e. a composition in which of the
active compound is released immediately after administration.
[0186] In another embodiment, the pharmaceutical composition is
delivered in a controlled release system. For example, the agent
may be administered using intravenous infusion, an implantable
osmotic pump, a transdermal patch, liposomes, or other modes of
administration. In one embodiment, a pump may be used. In another
embodiment, polymeric materials are used; e.g. in microspheres in
or an implant. In yet another embodiment, a controlled release
system is placed in proximity to the target cell, thus requiring
only a fraction of the systemic dose.
[0187] The compositions also include, in another embodiment,
incorporation of the active material into or onto particulate
preparations of polymeric compounds such as polylactic acid,
polglycolic acid, hydrogels, etc, or onto liposomes,
microemulsions, micelles, unilamellar or multilamellar vesicles,
erythrocyte ghosts, or spheroplasts.) Such compositions will
influence the physical state, solubility, stability, rate of in
vivo release, and rate of in vivo clearance.
[0188] Also included in the present invention are particulate
compositions coated with polymers (e.g. poloxamers or poloxamines)
and the compound coupled to antibodies directed against
tissue-specific receptors, ligands or antigens or coupled to
ligands of tissue-specific receptors.
[0189] Also comprehended by the invention are compounds modified by
the covalent attachment of water-soluble polymers such as
polyethylene glycol, copolymers of polyethylene glycol and
polypropylene glycol, carboxymethyl cellulose, dextran, polyvinyl
alcohol, polyvinylpyrrolidone or polyproline. The modified
compounds are known to exhibit substantially longer half-lives in
blood following intravenous injection than do the corresponding
unmodified compounds. Such modifications may also increase the
compounds solubility in aqueous solution, eliminate aggregation,
enhance the physical and chemical stability of the compound, and
greatly reduce the immunogenicity and reactivity of the compound.
As a result, the desired in vivo biological activity may be
achieved by the administration of such polymer-compound abducts
less frequently or in lower doses than with the unmodified
compound.
[0190] Each of the above additives, excipients, formulations and
methods of administration represents a separate embodiment used in
the compositions and methods provided.
[0191] In one embodiment, the methods used in the compositions and
methods provided, comprise administering an active compound as the
sole active ingredient. However, also encompassed within the scope
used in the compositions and methods provided, are methods for
treating diseases and disorders that comprise administering the
active compound in combination with one or more therapeutic agents.
These agents include, but are not limited to, insulin agents,
immunosuppressive agents, or drugs treating MS. In another
embodiment, these agents are appropriate for the disease or
disorder that is being treated, as is well known in the art.
[0192] In one embodiment, the methods used in the compositions and
methods provided, comprise administering an active compound as the
sole active ingredient. However, also encompassed within the scope
used in the compositions and methods provided, are methods for
treating diseases and disorders that comprise administering the
active compound in combination with one or more therapeutic agents.
These agents include, but are not limited to, insulin agents,
immunosuppressive agents, or drugs treating MS. In another
embodiment, these agents are appropriate for the disease or
disorder that is being treated, as is well known in the art.
[0193] In one embodiment, the invention further provides a method
of treating multiple sclerosis rejection in a subject comprising
the step of contacting an inflammatory cell with a selective c-Rel
DNA binding inhibitor. In one embodiment, the invention further
provides a method of treating arthritis in a subject comprising the
step of contacting an inflammatory cell with a selective c-Rel DNA
binding inhibitor. In one embodiment, the invention further
provides a method of treating diabetes rejection in a subject
comprising the step of contacting an inflammatory cell with a
selective c-Rel DNA binding inhibitor. In one embodiment, the
invention further provides a method of treating graft rejection in
a subject comprising the step of contacting an inflammatory cell
with a selective c-Rel DNA binding inhibitor. In one embodiment,
the invention provides that c-Rel DNA binding inhibitor masks the
L1 cavity of a c-Rel protein, thereby treating or preventing
multiple sclerosis, arthritis, diabetes, graft rejection, or a
combination thereof in a subject.
[0194] In another embodiment, methods of treating diseases of the
invention comprise contacting an inflammatory cell with an
inhibitor of the invention. In another embodiment, methods of
contacting an inflammatory cell with an inhibitor of the invention
are known to one of skill in the art. In another embodiment,
methods of contacting an inflammatory cell with an inhibitor of the
invention comprise administering a composition comprising an
inhibitor of the invention. In another embodiment, methods of
contacting an inflammatory cell with an inhibitor of the invention
comprise administering a composition which is targeted to a
particular inflammatory cell.
[0195] In one embodiment, the invention further provides a method
of preventing multiple sclerosis rejection in a subject comprising
the step of contacting an inflammatory cell with a selective c-Rel
DNA binding inhibitor. In one embodiment, the invention further
provides a method of preventing arthritis in a subject comprising
the step of contacting an inflammatory cell with a selective c-Rel
DNA binding inhibitor. In one embodiment, the invention further
provides a method of preventing diabetes rejection in a subject
comprising the step of contacting an inflammatory cell with a
selective c-Rel DNA binding inhibitor. In one embodiment, the
invention further provides a method of preventing graft rejection
in a subject comprising the step of contacting an inflammatory cell
with a selective c-Rel DNA binding inhibitor. In one embodiment,
the invention provides that c-Rel DNA binding inhibitor masks the
L1 cavity of a c-Rel protein, thereby preventing multiple
sclerosis, arthritis, diabetes, graft rejection, or a combination
thereof in a subject.
[0196] In another embodiment, methods of preventing diseases of the
invention comprise contacting an inflammatory cell with an
inhibitor of the invention.
[0197] In one embodiment, the invention further provides a method
of suppressing multiple sclerosis rejection in a subject comprising
the step of contacting an inflammatory cell with a selective c-Rel
DNA binding inhibitor. In one embodiment, the invention further
provides a method of suppressing arthritis in a subject comprising
the step of contacting an inflammatory cell with a selective c-Rel
DNA binding inhibitor. In one embodiment, the invention further
provides a method of suppressing diabetes rejection in a subject
comprising the step of contacting an inflammatory cell with a
selective c-Rel DNA binding inhibitor. In one embodiment, the
invention further provides a method of suppressing graft rejection
in a subject comprising the step of contacting an inflammatory cell
with a selective c-Rel DNA binding inhibitor. In one embodiment,
the invention provides that c-Rel DNA binding inhibitor masks the
L1 cavity of a c-Rel protein, thereby suppressing multiple
sclerosis, arthritis, diabetes, graft rejection, or a combination
thereof in a subject.
[0198] In another embodiment, methods of suppressing diseases of
the invention comprise contacting an inflammatory cell with an
inhibitor of the invention.
[0199] In one embodiment, the invention further provides a method
of reducing the symptoms associated with multiple sclerosis
rejection in a subject comprising the step of contacting an
inflammatory cell with a selective c-Rel DNA binding inhibitor. In
one embodiment, the invention further provides a method of reducing
the symptoms associated with arthritis in a subject comprising the
step of contacting an inflammatory cell with a selective c-Rel DNA
binding inhibitor. In one embodiment, the invention further
provides a method of reducing the symptoms associated with diabetes
rejection in a subject comprising the step of contacting an
inflammatory cell with a selective c-Rel DNA binding inhibitor. In
one embodiment, the invention further provides a method of reducing
the symptoms associated with graft rejection in a subject
comprising the step of contacting an inflammatory cell with a
selective c-Rel DNA binding inhibitor. In one embodiment, the
invention provides that c-Rel DNA binding inhibitor masks the L1
cavity of a c-Rel protein, thereby of reducing the symptoms
associated with multiple sclerosis, arthritis, diabetes, graft
rejection, or a combination thereof in a subject.
[0200] In another embodiment, methods of reducing the symptoms
associated with diseases of the invention comprise contacting an
inflammatory cell with an inhibitor of the invention.
[0201] In another embodiment, the invention further provides a
method of identifying a selective c-Rel DNA binding inhibitor
comprising the steps of: (a) constructing a c-Rel protein 3-D
model; and (b) minimizing the model to identify a selective c-Rel
DNA binding inhibitor, wherein the inhibitor interferes with L1
cavity thereby identifying a selective c-Rel DNA binding
inhibitor.
[0202] In another embodiment, the invention provides that an
inhibitor prevents Experimental Autoimmune Encephalomyelitis (EAE).
In another embodiment, the invention provides that an inhibitor
treats EAE. In another embodiment, the invention provides that an
inhibitor as described herein prevents and treats EAE.
[0203] In another embodiment, provided herein is a kit comprising a
reagent utilized in performing a method used in the compositions
and methods provided. In another embodiment, provided herein is a
kit comprising a composition, tool, or instrument used in the
compositions and methods provided.
EXPERIMENTAL DETAILS SECTION
Materials and Experimental Methods
Example 1
Treatment of Human Inflammatory Diseases Using C-Rel Inhibitors
[0204] Experimental autoimmune encephalomyelitis (EAE) is an animal
model for multiple sclerosis (MS). While the genetic and
environmental factors that trigger MS vary, the common pathological
outcome of the disease is the destruction of myelin-producing
oligodendrocytes and their associated neuronal axons through a
process called encephalomyelitis. Development of encephalomyelitis
requires coordinated expression of a large number of genes that
mediate the activation, migration and effector function of
inflammatory cells (activated lymphoid and myeloid cells). These
include genes that encode inflammatory cytokines, chemokines, and
cytotoxic enzymes. Expression of these inflammatory genes is
tightly regulated at the transcriptional level by specific
transcription factors. Recent studies indicate that, c-Rel, the
lymphoid member of the Rel/nuclear factor-.kappa.B
(Rel/NF-.kappa.B) family, is a long sought-after transcriptional
regulator of EAE. The mammalian Rel/NF-.kappa.B family consists of
five members, each of which is endowed with a distinct set of
function not shared by other members, although each member may also
perform additional functions common to the family. Thus, unlike
other members that are ubiquitously expressed, c-Rel is
preferentially expressed by inflammatory cells, and is involved in
regulating a special subset of immune responses. c-Rel-deficient
mice do not suffer from developmental problems or infectious
diseases, but are resistant to inflammatory diseases (despite the
normal expression of other Rel/NF-.kappa.B proteins);
c-Rel-deficient T cells are competent in survival and Th2 type
responses but are severely compromised in their Th1 and Th17 type
responses.
[0205] The Rel/NF-kB family of transcription factors represents one
of the most attractive targets for anti-inflammatory therapy.
Because Rel/NF-kB directly controls the expression of multiple
inflammatory genes, its blockade is more effective for controlling
inflammation than blocking one or a few downstream inflammatory
genes or proteins. The first generation Rel/NF-kB drugs that block
the entire Rel/NF-kB family have already been tested in both humans
and animals. These include proteasome inhibitors (e.g., the
FDA-approved PS-341), Rel/NF-kB decoy oligodeoxynucleotides and the
NBD (neuro-binding domain) peptides, which are highly effective in
preventing and treating models of autoimmune diseases including
EAE. Additionally, glucocorticoids, which are currently used to
control acute inflammation in MS patients, mediate their
immunosuppressive effects, at least in part, through inhibiting
Rel/NF-kB (glucocorticoids upregulate IkB expression and bind
directly to Rel/NF-kB). However, because most Rel/NF-kB proteins
are ubiquitously expressed and are involved in a variety of
biological processes not related to autoimmunity, these drugs have
significant side effects. Therefore, they can only be used for a
short period of time to control acute inflammation.
[0206] This experimental section describes a new generation of
drugs that are specific for c-Rel. Because c-Rel is expressed
preferentially in inflammatory cells (activated lymphoid and
myeloid cells) and is required for selected but not all immune
responses, drugs targeting it should have significantly less side
effects than drugs targeting the entire Rel/NF-.kappa.B family. As
detailed below, new classes of c-Rel inhibitors have now been
discovered that are effective in preventing and treating a
inflammatory diseases.
The L1 Cavity of c-Rel and Novel Inhibitors that Target it.
[0207] c-Rel:DNA contacts are mediated by four loops, L1 and L2
loops from the N-terminal domain, and L3 and L4 loops from the
C-terminal domain. DNA fragment is sandwiched between these loops.
The contact residues are highly conserved across species, including
human and mouse.
[0208] To design drugs that could disable human c-Rel binding to
DNA (therefore blocking the biological activity of c-Rel), the
chicken c-Rel 3-D model was humanized by replacing chicken residues
with their human counterparts. The model was then minimized using
the Insight II software (Accelrys, Inc). The minimized model was
used to discover small molecule inhibitors that can interfere with
the residues in loops L1 and L2. A small druggable cavity was
identified using programs available in Insight II. This structure
was designated as the L1 cavity. The L1 cavity is formed by loops
L1, L2 and a helix. Residues Arg 21, Cys 26, Glu 27, Lys 110 and
Lys 111 form the mouth of the L1 cavity (FIG. 2). A virtual
screening approach was used to identify small molecule inhibitors.
The inhibitors block the DNA binding to c-Rel by perturbing the key
contact residues mentioned above.
R13 Blocks Cytokine Production by Inflammatory Cells
[0209] To determine whether these inhibitors are effective in
suppressing cytokine production by inflammatory cells, spleen cells
of B6 mice were treated with anti-CD3 antibody (to activate the
inflammatory cells) with or without different concentrations of the
compounds. As shown in FIG. 3, R13 significantly inhibited both
Interleukin (IL)-2 and interferon-gamma production while the other
two compounds had an effect only on interferon-gamma The
proliferation was not significantly affected by these compounds
suggesting that they are not toxic to cells. As further shown in
FIG. 1 IL-2 and IFN-gamma were inhibited by compound III, compound
V, and compound VI at concentrations ranging from 5-20 .mu.M.
R13 is Effective in Both Preventing and Treating EAE
[0210] To determine the effect of R13 on EAE, the drug (inhibitor)
was injected either before or after the onset of the disease. As
shown in FIG. 5, R13D (compound III) is effective in both
preventing and treating EAE.
Example 2
Identification of c-Rel Blockers by Structure-Based Drug Design
(SBDD)
[0211] The three-dimensional structure of chicken c-Rel bound to
DNA has been determined (See e.g. Example 1). The structural
studies show that c-Rel:DNA contacts are mediated by four loops, L1
and L2 loops from the N-terminal domain, and L3 and L4 loops from
the C-terminal domain. DNA fragment is sandwiched between these
loops. The contact residues are highly conserved across species,
including chicken, human and mouse.
[0212] To design drugs that could disable human c-Rel binding to
DNA (therefore blocking the biological activity of c-Rel), chicken
c-Rel 3-D model was first humanized by replacing chicken residues
with their human counterparts. The model was then minimized using
the Insight II software (Accelrys, Inc). The minimized model was
used to discover small molecule inhibitors that can interfere with
the residues in loops L1 and L2. A small druggable cavity was
identified using programs available in Insight II. This structure
was designated as the L1 cavity. The L1 cavity is formed by loops
L1, L2 and a helix. Residues Arg 21, Cys 26, Glu 27, Lys 110 and
Lys 111 form the mouth of the L1 cavity (FIG. 2). A virtual
screening approach was used to identify small molecule inhibitors
that block DNA binding to the L1 cavity by perturbing these key
contact residues as we previously described. From .about.50,000
drug-like molecules of available chemical libraries, about 100 hits
were identified. Secondary screening using an IL-2 release assay
(described hereinbelow) led to the identification of 10 compounds
that significantly inhibited IL-2 production by T cells. These
compounds fall into two structural classes, the phenylcarbamates
(FIG. 2C) and sulfanyl-methaniminum-based compounds, represented by
R13:
##STR00161##
and R96 (the most potent compounds),
##STR00162##
respectively. The SBDD studies were conducted at the University of
Pennsylvania.
Example 3
c-Rel Blockers Selectively Regulate Cytokine Production but not
Survival of T Cells
[0213] To determine whether the c-Rel blockers identified above are
effective in regulating inflammatory cell function, splenocytes or
purified CD4+ splenic T cells of C57BL/6 mice were treated with
anti-CD3 and anti-CD28 in the presence of increasing concentrations
of the compounds. Cytokine expression was determined by ELISA, flow
cytometry or real-time PCR, whereas CD4 expression and cell
survival were determined by flow cytometry alone. Of 15 compounds
tested to date, including 5 from FP-HTS and 10 from SBDD, at least
10 significantly inhibited IL-2, IL-6, IL-17A, IL-21 and/or IFN-g
expression but not that of IL-4 and CD4 (which are not c-Rel
targets). Eight of these compounds showed no detectable
cytotoxicity to either resting or activated splenocytes in the
culture. A representative of these compounds, i.e., R96 identified
from SBDD, is shown in FIG. 4. The IC.sub.50 of R96 for IL-2
inhibition is .about.5 mM. Even at the highest concentration tested
(20 mM), R96 does not significantly affect the survival of CD4+ T
cells or total splenocytes. Because Th1 cytokine IFN-g inhibits the
activity of Th2 cells, the increased IL-4 in cultures containing 20
mM R96 could be due to the reduced IFN-g in the culture. Of note is
that although an increase in IL-4 was also observed in
c-Rel-deficient T cells in the culture, c-Rel-deficient mice do not
develop heightened humoral immunity or hypersensitivity because
c-Rel-deficiency also hinders B cell activation and antibody
production. In fact, c-Rel-deficient mice have reduced but not
increased IgG1, IgG3 and IgE production.
Example 4
c-Rel Blockers are Effective in Both Preventing and Treating
EAE
[0214] To determine the effect of c-Rel blockers on EAE, two lead
compounds were tested, R13 and R96, in both prophylaxis and
treatment regimens. For prophylaxis (which may be used to prevent
MS relapses during remission), the compounds were injected
intraperitoneally (i.p.) once a day for a total of 13 days starting
from the day of immunization. The amount of the compound used (100
mg/mouse/injection) was selected based on the IC.sub.50 of the
compound and the total volume of the mice (.about.20 ml), so that
the theoretical maximal mean concentration of the compound in the
whole animal is more than IC.sub.50 of the compound. The vehicle
used to dissolve the compound was PBS. Remarkably, injection of the
c-Rel blockers completely prevented the development of EAE in all
mice (FIG. 5A). To evaluate the disease pathology and anti-myelin
autoimmunity, mice were sacrificed 20 days after immunization and
tested as was previously described. As expected, anti-MOG Th1 (IL-2
and IFN-g) and Th17 (IL-17A) responses were significantly reduced
in mice treated with the c-Rel blocker. IL-4 was not detected in
splenocyte cultures of either group. Encephalomyelitis,
characterized by leukocyte infiltration of the white matter of
brain and spinal cord was evident in control but not c-Rel
blocker-treated mice. These results are similar to those shown
previously for c-Rel-deficient mice, indicating that blocking c-Rel
alone is sufficient to prevent EAE.
[0215] However, prophylaxis alone is not sufficient for controlling
MS, because most patients seek clinical care after the onset of the
disease. To determine the effectiveness of c-Rel blockers in
treating ongoing EAE, the c-Rel blockers were injected into mice
after the onset of the disease. Both R13 and R96 were found to
significantly ameliorated the symptoms of ongoing EAE (FIG. 5B,
5C). For example, the mean disease scores of mice before the
treatment were .about.1.0 in both groups (FIG. 5C). Seven days into
the treatment, the disease scores increased to 1.83.+-.0.16 in the
control group but decreased to 0.75.+-.0.43 in the R96-treated
group (p<0.001). Histological examination performed at the end
the experiment supported the clinical findings, with the c-Rel
blocker-treated group showed significantly reduced degree of
encephalomyelitis. Thus, c-Rel blockers are effective in
suppressing EAE after its onset.
Example 5
Synthesis of 096
[0216] The retrosynthetic analysis for the preparation of C-rel
inhibitor 096 is outlined in Scheme 1:
##STR00163##
[0217] Reaction of N-phenylthiourea 2 and benzyl chloride 3.sup.1
leads to the formation of 1. As shown in Scheme 2:
##STR00164##
[0218] The preparation of 3 is achieved by reaction of 2 with 4,
which leads to eight (8) novel analogs based on the nature of the
ester R group as indicated in Scheme 2 (eight new compounds,
including methyl ester, benzyl ester, and 3-substituted benzyl
esters, substituted with carboxylate and sulfonate, respectively,
are prepared).
[0219] Also examined is the use of amido functionalities, starting
from 6 as shown in 5 (Scheme 3), in lieu of the carboxyl group in
3. Using the same R groups indicated in Scheme 2, it should be
possible to prepare eight (8) additional amide-containing
analogs.
##STR00165##
[0220] The aromatic ring 3 are expanded to occupy a larger
conformational space in the binding pocket shown in FIG. 2.
Substitution of the A ring of 1 (Scheme 1) with .alpha.- and
.beta.-naphthylmethyl, 7 and 9, respectively, is shown in Scheme 4
and require the substitution of the commercially available .alpha.-
and .beta.-naphthylmethylchlorides 8 and 10 for 3. In each case,
the substitution of the naphthyl rings of 8 and 10 with negatively
charged groups, such as carboxylate, sulfonate, and also a
tetrazole, is carried out so that eight (8) additional analogs are
prepared in the expanded aryl series.
##STR00166##
[0221] Having described preferred embodiments of the invention with
reference to the accompanying drawings, it is to be understood that
the invention is not limited to the precise embodiments, and that
various changes and modifications may be effected therein by those
skilled in the art without departing from the scope or spirit of
the invention as defined in the appended claims.
* * * * *