U.S. patent application number 14/209449 was filed with the patent office on 2014-09-18 for pyrazolo compounds and uses thereof.
This patent application is currently assigned to Constellation Pharmaceuticals, Inc.. The applicant listed for this patent is Constellation Pharmaceuticals, Inc.. Invention is credited to Brian K. Albrecht, Steven F. Bellon, Victor S. Gehling, Jean-Christophe Harmange.
Application Number | 20140275092 14/209449 |
Document ID | / |
Family ID | 51529954 |
Filed Date | 2014-09-18 |
United States Patent
Application |
20140275092 |
Kind Code |
A1 |
Albrecht; Brian K. ; et
al. |
September 18, 2014 |
PYRAZOLO COMPOUNDS AND USES THEREOF
Abstract
The present invention relates to compounds useful as inhibitors
of one or more histone demethylses. The invention also provides
pharmaceutically acceptable compositions comprising compounds of
the present invention and methods of using said compositions in the
treatment of various disorders.
Inventors: |
Albrecht; Brian K.;
(Cambridge, MA) ; Bellon; Steven F.; (Wellesley,
MA) ; Gehling; Victor S.; (Somerville, MA) ;
Harmange; Jean-Christophe; (Andover, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Constellation Pharmaceuticals, Inc. |
Cambridge |
MA |
US |
|
|
Assignee: |
Constellation Pharmaceuticals,
Inc.
Cambridge
MA
|
Family ID: |
51529954 |
Appl. No.: |
14/209449 |
Filed: |
March 13, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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61778759 |
Mar 13, 2013 |
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Current U.S.
Class: |
514/246 ;
514/259.3; 514/267; 544/220; 544/250; 544/281; 544/91 |
Current CPC
Class: |
C07D 498/04 20130101;
A61K 31/519 20130101; C07D 487/04 20130101; C07D 487/14 20130101;
C12N 9/99 20130101; A61K 31/337 20130101; A61P 35/04 20180101; A61P
43/00 20180101; A61K 31/517 20130101; A61K 45/06 20130101; A61K
31/5377 20130101; A61P 35/00 20180101; A61K 31/437 20130101 |
Class at
Publication: |
514/246 ;
514/259.3; 514/267; 544/91; 544/220; 544/250; 544/281 |
International
Class: |
C07D 487/04 20060101
C07D487/04; C07D 498/04 20060101 C07D498/04 |
Claims
1. A compound of formula I: ##STR00186## or a pharmaceutically
acceptable salt thereof, wherein: R.sup.1 is --R, halogen, --OR,
--SR, --N(R').sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; each R
is independently hydrogen or an optionally substituted group
selected from C.sub.1-6 aliphatic, phenyl, a 3-7 membered saturated
or partially unsaturated carbocyclic ring, an 8-10 membered
bicyclic saturated, partially unsaturated or aryl ring, a 5-6
membered monocyclic heteroaryl ring having 1-3 heteroatoms
independently selected from nitrogen, oxygen, or sulfur, a 4-7
membered saturated or partially unsaturated heterocyclic ring
having 1-2 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, a 7-10 membered bicyclic saturated or partially
unsaturated heterocyclic ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur, or an 8-10 membered
bicyclic heteroaryl ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur; each R' is independently
--R, --C(O)R, --CO.sub.2R, or two R' on the same nitrogen are taken
together with their intervening atoms to form a 4-7 membered
heterocyclic ring having 1-2 heteroatoms independently selected
from nitrogen, oxygen, and sulfur; Ring A is ##STR00187## R.sup.2
and R.sup.3 are independently --R, halogen, --OR, --SR,
--N(R').sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; R.sup.2'
is --R, --OR, --SR, --N(R').sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.2' and R.sup.3 are taken together with their intervening
atoms to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 1-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; X is
--N(R.sup.4)--, --O--, or --S--; R.sup.4 is --R, --C(O)R,
--CO.sub.2R, or --S(O).sub.2R; or: R.sup.4 and R.sup.3 are taken
together with their intervening atoms to form an optionally
substituted 5-7 membered saturated, partially unsaturated, or
aromatic fused ring having 1-4 heteroatoms independently selected
from nitrogen, oxygen, and sulfur; R.sup.5 is R, --C(O)R,
--CO.sub.2R, --C(O)N(R').sub.2, --C(O)C(O)R, or
--C(O)CH.sub.2C(O)R; or: R.sup.5 and R.sup.2 are taken together
with their intervening atoms to form an optionally substituted 5-7
membered partially unsaturated or aromatic fused ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur; and R.sup.6 is --R, halogen, --OR, --SR, --N(R').sub.2,
--CN, --NO.sub.2, --C(O)R, --CO.sub.2R, --C(O)N(R').sub.2,
--C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R, --C(S)N(R').sub.2,
--C(S)OR, --S(O)R, --SO.sub.2R, --SO.sub.2N(R').sub.2,
--N(R')C(O)R, --N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.6 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; provided
that the compound is other than any one of the following:
##STR00188##
2. The compound according to claim 1, wherein said compound is of
formula II: ##STR00189## or a pharmaceutically acceptable salt
thereof.
3. The compound according to claim 2, wherein R.sup.4 is
hydrogen.
4. The compound according to claim 1, wherein R.sup.1 is
hydrogen.
5. The compound according to claim 1, wherein R.sup.2 is optionally
substituted C.sub.1-6 aliphatic.
6. The compound according to claim 5, wherein R.sup.2 is methyl,
ethyl, propyl, cyclopropyl, isopropyl, isobutyl, propargyl, or
allyl.
7. The compound according to claim 1, wherein R.sup.3 is optionally
substituted C.sub.1-6 aliphatic.
8. The compound according to claim 7, wherein the C.sub.1-6
aliphatic group is substituted with --OH or --OC.sub.1-6alkyl.
9. The compound according to claim 7, wherein the C.sub.1-6
aliphatic group is substituted with --NHC.sub.1-6alkyl or
--NH(C.sub.1-6alkyl).sub.2.
10. The compound according to claim 7, wherein R.sup.3 is
optionally substituted benzyl.
11. The compound according to claim 7, wherein R.sup.3 is one of
the following: ##STR00190##
12. The compound according to claim 7, wherein R.sup.3 is
--CF.sub.3.
13. The compound according to claim 1, wherein R.sup.3 is
--CO.sub.2R or --C(O)N(R').sub.2.
14. The compound according to claim 1, wherein R.sup.3 is a 5-6
membered monocyclic heteroaryl ring having 1-3 heteroatoms
independently selected from nitrogen, oxygen, or sulfur.
15. The compound according to claim 1, wherein neither of R.sup.2
and R.sup.3 is hydrogen.
16. The compound according to claim 1, wherein said compound is of
formula: ##STR00191## or a pharmaceutically acceptable salt
thereof.
17. The compound according to claim 16, wherein said compound is of
formula: ##STR00192##
18. A compound selected from the group consisting of: ##STR00193##
##STR00194## ##STR00195## ##STR00196## ##STR00197## ##STR00198##
##STR00199## ##STR00200## ##STR00201## or a pharmaceutically
acceptable salt thereof.
19. A composition comprising a compound of formula I: ##STR00202##
or a pharmaceutically acceptable salt thereof, wherein: R.sup.1 is
--R, halogen, --OR, --SR, --N(R').sub.2, --CN, --NO.sub.2, --C(O)R,
--CO.sub.2R, --C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R,
--C(O)CH.sub.2C(O)R, --C(S)N(R').sub.2, --C(S)OR, --S(O)R,
--SO.sub.2R, --SO.sub.2N(R').sub.2, --N(R')C(O)R,
--N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; each R
is independently hydrogen or an optionally substituted group
selected from C.sub.1 aliphatic, phenyl, a 3-7 membered saturated
or partially unsaturated carbocyclic ring, an 8-10 membered
bicyclic saturated, partially unsaturated or aryl ring, a 5-6
membered monocyclic heteroaryl ring having 1-3 heteroatoms
independently selected from nitrogen, oxygen, or sulfur, a 4-7
membered saturated or partially unsaturated heterocyclic ring
having 1-2 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, a 7-10 membered bicyclic saturated or partially
unsaturated heterocyclic ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur, or an 8-10 membered
bicyclic heteroaryl ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur; each R' is independently
--R, --C(O)R, --CO.sub.2R, or two R' on the same nitrogen are taken
together with the intervening nitrogen to form a 4-7 membered
heterocyclic ring having 1-2 heteroatoms independently selected
from nitrogen, oxygen, and sulfur; Ring A is ##STR00203## R.sup.2
and R.sup.3 are independently --R, halogen, --OR, --SR,
--N(R').sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; R.sup.2'
is --R, --OR, --SR, --N(R').sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.2' and R.sup.3 are taken together with their intervening
atoms to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 1-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; X is
--N(R.sup.4)--, --O--, or --S--; R.sup.4 is --R, --C(O)R,
--CO.sub.2R, or --S(O).sub.2R; or: R.sup.4 and R.sup.3 are taken
together with their intervening atoms to form an optionally
substituted 5-7 membered saturated, partially unsaturated, or
aromatic fused ring having 1-4 heteroatoms independently selected
from nitrogen, oxygen, and sulfur; R.sup.5 is R, --C(O)R,
--CO.sub.2R, --C(O)N(R').sub.2, --C(O)C(O)R, or
--C(O)CH.sub.2C(O)R; or: R.sup.5 and R.sup.2 are taken together
with their intervening atoms to form an optionally substituted 5-7
membered partially unsaturated or aromatic fused ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur; R.sup.6 is --R, halogen, --OR, --SR, --N(R').sub.2, --CN,
--NO.sub.2, --C(O)R, --CO.sub.2R, --C(O)N(R').sub.2, --C(O)SR,
--C(O)C(O)R, --C(O)CH.sub.2C(O)R, --C(S)N(R').sub.2, --C(S)OR,
--S(O)R, --SO.sub.2R, --SO.sub.2N(R').sub.2, --N(R')C(O)R,
--N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.6 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; and a
pharmaceutically acceptable adjuvant, carrier, or vehicle.
20. A method for inhibiting activity of a 2-oxoglutarate dependent
enzyme, or a mutant thereof, activity in a patient comprising the
step of administering to said patient a compound of formula I:
##STR00204## or a pharmaceutically acceptable salt thereof,
wherein: R.sup.1 is --R, halogen, --OR, --SR, --N(R').sub.2, --CN,
--NO.sub.2, --C(O)R, --CO.sub.2R, --C(O)N(R').sub.2, --C(O)SR,
--C(O)C(O)R, --C(O)CH.sub.2C(O)R, --C(S)N(R').sub.2, --C(S)OR,
--S(O)R, --SO.sub.2R, --SO.sub.2N(R').sub.2, --N(R')C(O)R,
--N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; each R
is independently hydrogen or an optionally substituted group
selected from C.sub.1-6 aliphatic, phenyl, a 3-7 membered saturated
or partially unsaturated carbocyclic ring, an 8-10 membered
bicyclic saturated, partially unsaturated or aryl ring, a 5-6
membered monocyclic heteroaryl ring having 1-3 heteroatoms
independently selected from nitrogen, oxygen, or sulfur, a 4-7
membered saturated or partially unsaturated heterocyclic ring
having 1-2 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, a 7-10 membered bicyclic saturated or partially
unsaturated heterocyclic ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur, or an 8-10 membered
bicyclic heteroaryl ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur; each R' is independently
--R, --C(O)R, --CO.sub.2R, or two R' on the same nitrogen are taken
together with the intervening nitrogen to form a 4-7 membered
heterocyclic ring having 1-2 heteroatoms independently selected
from nitrogen, oxygen, and sulfur; Ring A is ##STR00205## R.sup.2
and R.sup.3 are independently --R, halogen, --OR, --SR,
--N(R').sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; R.sup.2'
is --R, --OR, --SR, --N(R').sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.2' and R.sup.3 are taken together with their intervening
atoms to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 1-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; X is
--N(R.sup.4)--, --O--, or --S--; R.sup.4 is --R, --C(O)R,
--CO.sub.2R, or --S(O).sub.2R; or: R.sup.4 and R.sup.3 are taken
together with their intervening atoms to form an optionally
substituted 5-7 membered saturated, partially unsaturated, or
aromatic fused ring having 1-4 heteroatoms independently selected
from nitrogen, oxygen, and sulfur; R.sup.5 is R, --C(O)R,
--CO.sub.2R, --C(O)N(R').sub.2, --C(O)C(O)R, or
--C(O)CH.sub.2C(O)R; or: R.sup.5 and R.sup.2 are taken together
with their intervening atoms to form an optionally substituted 5-7
membered partially unsaturated or aromatic fused ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur; and R.sup.6 is --R, halogen, --OR, --SR, --N(R').sub.2,
--CN, --NO.sub.2, --C(O)R, --CO.sub.2R, --C(O)N(R').sub.2,
--C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R, --C(S)N(R').sub.2,
--C(S)OR, --S(O)R, --SO.sub.2R, --SO.sub.2N(R').sub.2,
--N(R')C(O)R, --N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.6 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; or a
composition according to claim 19.
21. A method for treating a GASC1-mediated disorder in a patient in
need thereof, comprising the step of administering to said patient
a compound of formula I: ##STR00206## or a pharmaceutically
acceptable salt thereof, wherein: R.sup.1 is --R, halogen, --OR,
--SR, --N(R').sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R') C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2,
--C.dbd.NOR, --C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or
--OC(O)N(R').sub.2; each R is independently hydrogen or an
optionally substituted group selected from C.sub.1-6 aliphatic,
phenyl, a 3-7 membered saturated or partially unsaturated
carbocyclic ring, an 8-10 membered bicyclic saturated, partially
unsaturated or aryl ring, a 5-6 membered monocyclic heteroaryl ring
having 1-3 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, a 4-7 membered saturated or partially
unsaturated heterocyclic ring having 1-2 heteroatoms independently
selected from nitrogen, oxygen, or sulfur, a 7-10 membered bicyclic
saturated or partially unsaturated heterocyclic ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, or
sulfur, or an 8-10 membered bicyclic heteroaryl ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, or
sulfur; each R' is independently --R, --C(O)R, --CO.sub.2R, or two
R' on the same nitrogen are taken together with the intervening
nitrogen to form a 4-7 membered heterocyclic ring having 1-2
heteroatoms independently selected from nitrogen, oxygen, and
sulfur; Ring A is ##STR00207## R.sup.2 and R.sup.3 are
independently --R, halogen, --OR, --SR, --N(R').sub.2, --CN,
--NO.sub.2, --C(O)R, --CO.sub.2R, --C(O)N(R').sub.2, --C(O)SR,
--C(O)C(O)R, --C(O)CH.sub.2C(O)R, --C(S)N(R').sub.2, --C(S)OR,
--S(O)R, --SO.sub.2R, --SO.sub.2N(R').sub.2, --N(R')C(O)R,
--N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; R.sup.2'
is --R, --OR, --SR, --N(R').sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R.sup.1).sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.2' and R.sup.3 are taken together with their intervening
atoms to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 1-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; X is
--N(R.sup.4)--, --O--, or --S--; R.sup.4 is --R, --C(O)R,
--CO.sub.2R, or --S(O).sub.2R; or: R.sup.4 and R.sup.3 are taken
together with their intervening atoms to form an optionally
substituted 5-7 membered saturated, partially unsaturated, or
aromatic fused ring having 1-4 heteroatoms independently selected
from nitrogen, oxygen, and sulfur; R.sup.5 is R, --C(O)R,
--CO.sub.2R, --C(O)N(R').sub.2, --C(O)C(O)R, or
--C(O)CH.sub.2C(O)R; or: R.sup.5 and R.sup.2 are taken together
with their intervening atoms to form an optionally substituted 5-7
membered partially unsaturated or aromatic fused ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur; and R.sup.6 is --R, halogen, --OR, --SR, --N(R').sub.2,
--CN, --NO.sub.2, --C(O)R, --CO.sub.2R, --C(O)N(R').sub.2,
--C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R, --C(S)N(R').sub.2,
--C(S)OR, --S(O)R, --SO.sub.2R, --SO.sub.2N(R').sub.2,
--N(R')C(O)R, --N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.6 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur. or a
composition according to claim 19.
22. The method of claim 20, wherein the 2-oxoglutarate dependent
enzyme is a JARID family enzyme.
23. A method for treating a JARID-mediated disorder in a patient in
need thereof, comprising the step of administering to said patient
a compound of formula I: ##STR00208## or a pharmaceutically
acceptable salt thereof, wherein: R.sup.1 is --R, halogen, --OR,
--SR, --N(R').sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; each R
is independently hydrogen or an optionally substituted group
selected from C.sub.1 aliphatic, phenyl, a 3-7 membered saturated
or partially unsaturated carbocyclic ring, an 8-10 membered
bicyclic saturated, partially unsaturated or aryl ring, a 5-6
membered monocyclic heteroaryl ring having 1-3 heteroatoms
independently selected from nitrogen, oxygen, or sulfur, a 4-7
membered saturated or partially unsaturated heterocyclic ring
having 1-2 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, a 7-10 membered bicyclic saturated or partially
unsaturated heterocyclic ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur, or an 8-10 membered
bicyclic heteroaryl ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur; each R' is independently
--R, --C(O)R, --CO.sub.2R, or two R' on the same nitrogen are taken
together with the intervening nitrogen to form a 4-7 membered
heterocyclic ring having 1-2 heteroatoms independently selected
from nitrogen, oxygen, and sulfur; Ring A is ##STR00209## R.sup.2
and R.sup.3 are independently --R, halogen, --OR, --SR,
--N(R').sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; R.sup.2'
is --R, --OR, --SR, --N(R').sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R.sup.1).sub.2;
or: R.sup.2' and R.sup.3 are taken together with their intervening
atoms to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 1-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; X is
--N(R.sup.4)--, --O--, or --S--; R.sup.4 is --R, --C(O)R,
--CO.sub.2R, or --S(O).sub.2R; or: R.sup.4 and R.sup.3 are taken
together with their intervening atoms to form an optionally
substituted 5-7 membered saturated, partially unsaturated, or
aromatic fused ring having 1-4 heteroatoms independently selected
from nitrogen, oxygen, and sulfur; R.sup.5 is R, --C(O)R,
--CO.sub.2R, --C(O)N(R').sub.2, --C(O)C(O)R, or
--C(O)CH.sub.2C(O)R; or: R.sup.5 and R.sup.2 are taken together
with their intervening atoms to form an optionally substituted 5-7
membered partially unsaturated or aromatic fused ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur; and R.sup.6 is --R, halogen, --OR, --SR, --N(R').sub.2,
--CN, --NO.sub.2, --C(O)R, --CO.sub.2R, --C(O)N(R').sub.2,
--C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R, --C(S)N(R').sub.2,
--C(S)OR, --S(O)R, --SO.sub.2R, --SO.sub.2N(R').sub.2,
--N(R')C(O)R, --N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
R.sup.6 and R.sup.3 are taken together with their intervening atoms
to form an optionally substituted 5-7 membered partially
unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur, or a
composition according to claim 19.
Description
PRIORITY OF INVENTION
[0001] This application claims priority from U.S. Provisional
Application No. 61/778,759, filed 13 Mar. 2013. The entire content
of this provisional application is hereby incorporated herein by
reference.
TECHNICAL FIELD OF THE INVENTION
[0002] The present invention relates to compounds useful as
inhibitors of histone demethylases.
BACKGROUND OF THE INVENTION
[0003] Packaging the 3 billion nucleotides of the human genome into
the nucleus of a cell requires tremendous compaction. To accomplish
this feat, DNA in our chromosomes is wrapped around spools of
proteins called histones to form dense repeating protein/DNA
polymers known as chromatin: the defining template for gene
regulation. Far from serving as mere packaging modules, chromatin
templates form the basis of a newly appreciated and fundamentally
important set of gene control mechanisms termed epigenetic
regulation. By conferring a wide range of specific chemical
modifications to histones and DNA, epigenetic regulators modulate
the structure, function, and accessibility of our genome, thereby
exerting a tremendous impact on gene expression. Hundreds of
epigenetic effectors have recently been identified, many of which
are chromatin-binding or chromatin-modifying enzymes.
Significantly, an increasing number of these enzymes have been
associated with a variety of disorders such as neurodegenerative
disorders, metabolic diseases, inflammation, and cancer. Thus,
therapeutic agents directed against this emerging class of gene
regulatory enzymes promise new approaches to the treatment of human
diseases.
SUMMARY OF THE INVENTION
[0004] It has now been found that compounds of this invention, and
pharmaceutically acceptable compositions thereof, are effective as
inhibitors of histone demethylases, including 2-oxoglutarate
dependent enzymes such as Jumonji domain containing proteins,
members of the H3K4 (histone 3 K4) demethylase family of proteins,
and/or members of the JARID subfamily of histone demethylases. Such
compounds are of formula I:
##STR00001##
or a pharmaceutically acceptable salt thereof, wherein R.sup.1 and
Ring A are as defined and described herein.
[0005] Provided compounds, and pharmaceutically acceptable
compositions thereof, are useful for treating a variety of
diseases, disorders or conditions associated with abnormal cellular
responses triggered by events mediated by histone demethylases such
as 2-oxoglutarate dependent enzymes, Jumonji domain containing
proteins, members of the H3K4 (histone 3K4) demethylase family of
proteins, and/or members of the JARID subfamily of enzymes. Such
diseases, disorders, or conditions include those described
herein.
[0006] Provided compounds are also useful for the study of histone
demethylases, such as 2-oxoglutarate dependent enzymes, Jumonji
domain containing proteins, members of the H3K4 (histone 3 K4)
demethylase family of proteins, and/or members of the JARID
subfamily of enzymes in biological and pathological phenomena, the
study of intracellular signal transduction pathways mediated by
such histone demethylases, and the comparative evaluation of new
inhibitors of these and other histone demethylases.
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS
1. General Description of Compounds of the Invention
[0007] In certain embodiments, the present invention provides a
compound of formula I:
##STR00002##
or a pharmaceutically acceptable salt thereof, wherein: [0008]
R.sup.1 is --R, halogen, --OR, --SR, --N(R').sub.2, --CN,
--NO.sub.2, --C(O)R, --CO.sub.2R, --C(O)N(R').sub.2, --C(O)SR,
--C(O)C(O)R, --C(O)CH.sub.2C(O)R, --C(S)N(R').sub.2, --C(S)OR,
--S(O)R, --SO.sub.2R, --SO.sub.2N(R').sub.2, --N(R')C(O)R,
--N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; [0009]
each R is independently hydrogen or an optionally substituted group
selected from C.sub.1-6 aliphatic, phenyl, a 3-7 membered saturated
or partially unsaturated carbocyclic ring, an 8-10 membered
bicyclic saturated, partially unsaturated or aryl ring, a 5-6
membered monocyclic heteroaryl ring having 1-3 heteroatoms
independently selected from nitrogen, oxygen, or sulfur, a 4-7
membered saturated or partially unsaturated heterocyclic ring
having 1-2 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, a 7-10 membered bicyclic saturated or partially
unsaturated heterocyclic ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur, or an 8-10 membered
bicyclic heteroaryl ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur; [0010] each R' is
independently --R, --C(O)R, --CO.sub.2R, or two R' on the same
nitrogen are taken together with their intervening atoms to form a
4-7 membered heterocyclic ring having 1-2 heteroatoms independently
selected from nitrogen, oxygen, and sulfur; [0011] Ring A is
[0011] ##STR00003## [0012] R.sup.2 and R.sup.3 are independently
--R, halogen, --OR, --SR, --N(R').sub.2, --CN, --NO.sub.2, --C(O)R,
--CO.sub.2R, --C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R,
--C(O)CH.sub.2C(O)R, --C(S)N(R').sub.2, --C(S)OR, --S(O)R,
--SO.sub.2R, --SO.sub.2N(R').sub.2, --N(R')C(O)R,
--N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
[0013] R.sup.2 and R.sup.3 are taken together with their
intervening atoms to form an optionally substituted 5-7 membered
partially unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; [0014]
R.sup.2' is --R, --OR, --SR, --N(R').sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
[0015] R.sup.2' and R.sup.3 are taken together with their
intervening atoms to form an optionally substituted 5-7 membered
partially unsaturated or aromatic fused ring having 1-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur; [0016] X
is --N(R.sup.4)--, --O--, or --S--; [0017] R.sup.4 is --R, --C(O)R,
--CO.sub.2R, or --S(O).sub.2R; or: [0018] R.sup.4 and R.sup.3 are
taken together with their intervening atoms to form an optionally
substituted 5-7 membered saturated, partially unsaturated, or
aromatic fused ring having 1-4 heteroatoms independently selected
from nitrogen, oxygen, and sulfur; [0019] R.sup.5 is R, --C(O)R,
--CO.sub.2R, --C(O)N(R').sub.2, --C(O)C(O)R, or
--C(O)CH.sub.2C(O)R; or: [0020] R.sup.5 and R.sup.2 are taken
together with their intervening atoms to form an optionally
substituted 5-7 membered partially unsaturated or aromatic fused
ring having 1-4 heteroatoms independently selected from nitrogen,
oxygen, and sulfur; and [0021] R.sup.6 is --R, halogen, --OR, --SR,
--N(R.sup.1).sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R.sup.1).sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2; or:
[0022] R.sup.6 and R.sup.3 are taken together with their
intervening atoms to form an optionally substituted 5-7 membered
partially unsaturated or aromatic fused ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur.
[0023] In some embodiments, the present invention provides a
compound of formula I other than any one of the following:
##STR00004## ##STR00005##
2. Compounds and Definitions
[0024] Definitions of specific functional groups and chemical terms
are described in more detail below. For purposes of this invention,
the chemical elements are identified in accordance with the
Periodic Table of the Elements, CAS version, Handbook of Chemistry
and Physics, 75.sup.th Ed., inside cover, and specific functional
groups are generally defined as described therein. Additionally,
general principles of organic chemistry, as well as specific
functional moieties and reactivity, are described in Organic
Chemistry, Thomas Sorrell, University Science Books, Sausalito,
1999; Smith and March March's Advanced Organic Chemistry, 5.sup.th
Edition, John Wiley & Sons, Inc., New York, 2001; Larock,
Comprehensive Organic Transformations, VCH Publishers, Inc., New
York, 1989; Carruthers, Some Modern Methods of Organic Synthesis,
3.sup.rd Edition, Cambridge University Press, Cambridge, 1987; the
entire contents of each of which are incorporated herein by
reference.
[0025] Unless otherwise stated, structures depicted herein are also
meant to include all isomeric (e.g., enantiomeric, diastereomeric,
and geometric (or conformational)) forms of the structure; for
example, the R and S configurations for each asymmetric center, Z
and E double bond isomers, and Z and E conformational isomers.
Therefore, single stereochemical isomers as well as enantiomeric,
diastereomeric, and geometric (or conformational) mixtures of the
present compounds are within the scope of the invention. Unless
otherwise stated, all tautomeric forms of the compounds of the
invention are within the scope of the invention. Additionally,
unless otherwise stated, structures depicted herein are also meant
to include compounds that differ only in the presence of one or
more isotopically enriched atoms. For example, compounds having the
present structures including the replacement of hydrogen by
deuterium or tritium, or the replacement of a carbon by a .sup.13C-
or .sup.14C-enriched carbon are within the scope of this invention.
Such compounds are useful, for example, as analytical tools, as
probes in biological assays, or as therapeutic agents in accordance
with the present invention.
[0026] Where a particular enantiomer is preferred, it may, in some
embodiments be provided substantially free of the corresponding
enantiomer, and may also be referred to as "optically enriched."
"Optically-enriched," as used herein, means that the compound is
made up of a significantly greater proportion of one enantiomer. In
certain embodiments the compound is made up of at least about 90%
by weight of a preferred enantiomer. In other embodiments the
compound is made up of at least about 95%, 98%, or 99% by weight of
a preferred enantiomer. Preferred enantiomers may be isolated from
racemic mixtures by any method known to those skilled in the art,
including chiral high pressure liquid chromatography (HPLC) and the
formation and crystallization of chiral salts or prepared by
asymmetric syntheses. See, for example, Jacques et al.,
Enantiomers, Racemates and Resolutions (Wiley Interscience, New
York, 1981); Wilen, et al., Tetrahedron 33:2725 (1977); Eliel, E.
L. Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962);
Wilen, S. H. Tables of Resolving Agents and Optical Resolutions p.
268 (E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind.
1972).
[0027] The term "heteroatom" means one or more of oxygen, sulfur,
nitrogen, phosphorus, or silicon (including, any oxidized form of
nitrogen, sulfur, phosphorus, or silicon; the quaternized form of
any basic nitrogen or; a substitutable nitrogen of a heterocyclic
ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in
pyrrolidinyl) or NR.sup.+ (as in N-substituted pyrrolidinyl)).
[0028] As used herein a "direct bond" or "covalent bond" refers to
a single, double or triple bond. In certain embodiments, a "direct
bond" or "covalent bond" refers to a single bond.
[0029] The terms "halo" and "halogen" as used herein refer to an
atom selected from fluorine (fluoro, --F), chlorine (chloro, --Cl),
bromine (bromo, --Br), and iodine (iodo, --I).
[0030] The term "aliphatic" or "aliphatic group", as used herein,
denotes a hydrocarbon moiety that may be straight-chain (i.e.,
unbranched), branched, or cyclic (including fused, bridging, and
spiro-fused polycyclic) and may be completely saturated or may
contain one or more units of unsaturation, but which is not
aromatic. Unless otherwise specified, aliphatic groups contain 1-6
carbon atoms. In some embodiments, aliphatic groups contain 1-4
carbon atoms, and in yet other embodiments aliphatic groups contain
1-3 carbon atoms. Suitable aliphatic groups include, but are not
limited to, linear or branched, alkyl, alkenyl, and alkynyl groups,
and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl
or (cycloalkyl)alkenyl.
[0031] The term "unsaturated", as used herein, means that a moiety
has one or more units of unsaturation.
[0032] The terms "cycloaliphatic", "carbocycle", "carbocyclyl",
"carbocyclo", or "carbocyclic", used alone or as part of a larger
moiety, refer to a saturated or partially unsaturated cyclic
aliphatic monocyclic or bicyclic ring systems, as described herein,
having from 3 to 10 members, wherein the aliphatic ring system is
optionally substituted as defined above and described herein.
Cycloaliphatic groups include, without limitation, cyclopropyl,
cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl,
cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, and
cyclooctadienyl. In some embodiments, the cycloalkyl has 3-6
carbons. The terms "cycloaliphatic", "carbocycle", "carbocyclyl",
"carbocyclo", or "carbocyclic" also include aliphatic rings that
are fused to one or more aromatic or nonaromatic rings, such as
decahydronaphthyl, tetrahydronaphthyl, decalin, or
bicyclo[2.2.2]octane, where the radical or point of attachment is
on an aliphatic ring.
[0033] As used herein, the term "cycloalkylene" refers to a
bivalent cycloalkyl group. In certain embodiments, a cycloalkylene
group is a 1,1-cycloalkylene group (i.e., a spiro-fused ring).
Exemplary 1,1-cycloalkylene groups include
##STR00006##
In other embodiments, a cycloalkylene group is a 1,2-cycloalkylene
group or a 1,3-cycloalkylene group. Exemplary 1,2-cycloalkylene
groups include
##STR00007##
[0034] The term "alkyl," as used herein, refers to a monovalent
saturated, straight- or branched-chain hydrocarbon radical derived
from an aliphatic moiety containing between one and six carbon
atoms by removal of a single hydrogen atom. In some embodiments,
alkyl contains 1-5 carbon atoms. In another embodiment, alkyl
contains 1-4 carbon atoms. In still other embodiments, alkyl
contains 1-3 carbon atoms. In yet another embodiment, alkyl
contains 1-2 carbons. Examples of alkyl radicals include, but are
not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl,
iso-butyl, sec-butyl, sec-pentyl, iso-pentyl, tert-butyl, n-pentyl,
neopentyl, n-hexyl, sec-hexyl, n-heptyl, n-octyl, n-decyl,
n-undecyl, dodecyl, and the like.
[0035] The term "alkenyl," as used herein, denotes a monovalent
group derived from a straight- or branched-chain aliphatic moiety
having at least one carbon-carbon double bond by the removal of a
single hydrogen atom. In certain embodiments, alkenyl contains 2-6
carbon atoms. In certain embodiments, alkenyl contains 2-5 carbon
atoms. In some embodiments, alkenyl contains 2-4 carbon atoms. In
another embodiment, alkenyl contains 2-3 carbon atoms. Alkenyl
groups include, for example, ethenyl ("vinyl"), propenyl ("allyl"),
butenyl, 1-methyl-2-buten-1-yl, and the like.
[0036] The term "alkynyl," as used herein, refers to a monovalent
group derived from a straight- or branched-chain aliphatic moiety
having at least one carbon-carbon triple bond by the removal of a
single hydrogen atom. In certain embodiments, alkynyl contains 2-6
carbon atoms. In certain embodiments, alkynyl contains 2-5 carbon
atoms. In some embodiments, alkynyl contains 2-4 carbon atoms. In
another embodiment, alkynyl contains 2-3 carbon atoms.
Representative alkynyl groups include, but are not limited to,
ethynyl, 2-propynyl ("propargyl"), 1-propynyl, and the like.
[0037] The term "aryl" used alone or as part of a larger moiety as
in "aralkyl", "aralkoxy", or "aryloxyalkyl", refers to monocyclic
and bicyclic ring systems having a total of five to 10 ring
members, wherein at least one ring in the system is aromatic and
wherein each ring in the system contains three to seven ring
members. The term "aryl" may be used interchangeably with the term
"aryl ring". In certain embodiments of the present invention,
"aryl" refers to an aromatic ring system which includes, but not
limited to, phenyl, biphenyl, naphthyl, anthracyl and the like,
which may bear one or more substituents. Also included within the
scope of the term "aryl", as it is used herein, is a group in which
an aromatic ring is fused to one or more non-aromatic rings, such
as indanyl, phthalimidyl, naphthimidyl, phenantriidinyl, or
tetrahydronaphthyl, and the like.
[0038] The terms "heteroaryl" and "heteroar-", used alone or as
part of a larger moiety, e.g., "heteroaralkyl", or
"heteroaralkoxy", refer to groups having 5 to 10 ring atoms,
preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 .pi.
electrons shared in a cyclic array; and having, in addition to
carbon atoms, from one to five heteroatoms. The term "heteroatom"
refers to nitrogen, oxygen, or sulfur, and includes any oxidized
form of nitrogen or sulfur, and any quaternized form of a basic
nitrogen. Heteroaryl groups include, without limitation, thienyl,
furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl,
oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl,
thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl,
indolizinyl, purinyl, naphthyridinyl, and pteridinyl. The terms
"heteroaryl" and "heteroar-", as used herein, also include groups
in which a heteroaromatic ring is fused to one or more aryl,
cycloaliphatic, or heterocyclyl rings, where the radical or point
of attachment is on the heteroaromatic ring. Nonlimiting examples
include indolyl, isoindolyl, benzothienyl, benzofuranyl,
dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl,
isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl,
4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl,
phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and
pyrido[2,3-b]-1,4-oxazin-3(4H)-one. A heteroaryl group may be mono-
or bicyclic. The term "heteroaryl" may be used interchangeably with
the terms "heteroaryl ring", "heteroaryl group", or
"heteroaromatic", any of which terms include rings that are
optionally substituted. The term "heteroaralkyl" refers to an alkyl
group substituted by a heteroaryl, wherein the alkyl and heteroaryl
portions independently are optionally substituted.
[0039] As used herein, the terms "heterocycle", "heterocyclyl",
"heterocyclic radical", and "heterocyclic ring" are used
interchangeably and refer to a stable 4- to 7-membered monocyclic
or 7-10-membered bicyclic heterocyclic moiety that is either
saturated or partially unsaturated, and having, in addition to
carbon atoms, one or more, preferably one to four, heteroatoms, as
defined above. When used in reference to a ring atom of a
heterocycle, the term "nitrogen" includes a substituted nitrogen.
As an example, in a saturated or partially unsaturated ring having
0-3 heteroatoms selected from oxygen, sulfur or nitrogen, the
nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in
pyrrolidinyl), or .sup.+NR (as in N-substituted pyrrolidinyl).
[0040] A heterocyclic ring can be attached to its pendant group at
any heteroatom or carbon atom that results in a stable structure
and any of the ring atoms can be optionally substituted. Examples
of such saturated or partially unsaturated heterocyclic radicals
include, without limitation, tetrahydrofuranyl, tetrahydrothienyl,
pyrrolidinyl, pyrrolidonyl, piperidinyl, pyrrolinyl,
tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl,
oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl,
oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms
"heterocycle", "heterocyclyl", "heterocyclyl ring", "heterocyclic
group", "heterocyclic moiety", and "heterocyclic radical", are used
interchangeably herein, and also include groups in which a
heterocyclyl ring is fused to one or more aryl, heteroaryl, or
cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl,
phenanthridinyl, 2-azabicyclo[2.2.1]heptanyl, octahydroindolyl, or
tetrahydroquinolinyl, where the radical or point of attachment is
on the heterocyclyl ring. A heterocyclyl group may be mono- or
bicyclic. The term "heterocyclylalkyl" refers to an alkyl group
substituted by a heterocyclyl, wherein the alkyl and heterocyclyl
portions independently are optionally substituted.
[0041] As used herein, the term "partially unsaturated" refers to a
ring moiety that includes at least one double or triple bond
between ring atoms but is not aromatic. The term "partially
unsaturated" is intended to encompass rings having multiple sites
of unsaturation, but is not intended to include aryl or heteroaryl
moieties, as herein defined.
[0042] The term "alkylene" refers to a bivalent alkyl group. An
"alkylene chain" is a polymethylene group, i.e.,
--(CH.sub.2).sub.n--, wherein n is a positive integer, preferably
from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3.
A substituted alkylene chain is a polymethylene group in which one
or more methylene hydrogen atoms are replaced with a substituent.
Suitable substituents include those described below for a
substituted aliphatic group.
[0043] As described herein, compounds of the invention may contain
"optionally substituted" moieties. In general, the term
"substituted", whether preceded by the term "optionally" or not,
means that one or more hydrogens of the designated moiety are
replaced with a suitable substituent. Unless otherwise indicated,
an "optionally substituted" group may have a suitable substituent
at each substitutable position of the group, and when more than one
position in any given structure may be substituted with more than
one substituent selected from a specified group, the substituent
may be either the same or different at each position. Combinations
of substituents envisioned under this invention are preferably
those that result in the formation of stable or chemically feasible
compounds. The term "stable", as used herein, refers to compounds
that are not substantially altered when subjected to conditions to
allow for their production, detection, and, in certain embodiments,
their recovery, purification, and use for one or more of the
purposes disclosed herein.
[0044] Suitable monovalent substituents on a substitutable carbon
atom of an "optionally substituted" group are independently
halogen; --(CH.sub.2).sub.0-4R.sup.o; --(CH.sub.2).sub.0-4OR.sup.o;
--O--(CH.sub.2).sub.0-4C(O)OR.sup.o;
--(CH.sub.2).sub.0-4CH(OR.sup.o).sub.2;
--(CH.sub.2).sub.0-4SR.sup.o; --(CH.sub.2).sub.0-4Ph, which may be
substituted with R.sup.o; --(CH.sub.2).sub.0-4O(CH.sub.2).sub.0-1Ph
which may be substituted with R.sup.o; --CH.dbd.CHPh, which may be
substituted with R.sup.o; --NO.sub.2; --CN; --N.sub.3;
--(CH.sub.2).sub.0-4N(R.sup.o).sub.2;
--(CH.sub.2).sub.0-4N(R.sup.oC(O)R.sup.o; --N(R.sup.o)C(S)R.sup.o;
--(CH.sub.2).sub.0-4N(R.sup.oC(O)NR.sup.o.sub.2;
--N(R.sup.oC(S)NR.sup.o.sub.2;
--(CH.sub.2).sub.0-4N(R.sup.o)C(O)OR.sup.o;
--N(R.sup.o)N(R.sup.oC(O)R.sup.o;
--N(R.sup.o)N(R.sup.oC(O)NR.sup.o.sub.2;
--N(R.sup.o)N(R.sup.oC(O)OR.sup.o; --(CH.sub.2).sub.0-4C(O)R.sup.o;
--C(S)R.sup.o; --(CH.sub.2).sub.0-4C(O)OR.sup.o;
--(CH.sub.2).sub.0-4C(O)SR.sup.o;
--(CH.sub.2).sub.0-4C(O)OSiR.sup.o.sub.3;
--(CH.sub.2).sub.0-4OC(O)R.sup.o; --OC(O)(CH.sub.2).sub.0-4SR--,
SC(S)SR.sup.o; --(CH.sub.2).sub.0-4 SC(O)R.sup.o;
--(CH.sub.2).sub.0-4C(O)NR.sup.o.sub.2; --C(S)NR.sup.o.sub.2;
--C(S)SR.sup.o; --SC(S)SR.sup.o,
--(CH.sub.2).sub.0-4OC(O)NR.sup.o.sub.2; --C(O)N(OR.sup.o)R.sup.o;
--C(O)C(O)R.sup.o; --C(O)CH.sub.2C(O)R.sup.o;
--C(NOR.sup.o)R.sup.o; --(CH.sub.2).sub.0-4SSR.sup.o;
--(CH.sub.2).sub.0-4S(O).sub.2R.sup.o;
--(CH.sub.2).sub.0-4S(O).sub.2OR.sup.o;
--(CH.sub.2).sub.0-4OS(O).sub.2R.sup.o; --S(O).sub.2NR.sup.o.sub.2;
--(CH.sub.2).sub.0-4S(O)R.sup.o;
--N(R.sup.o)S(O).sub.2NR.sup.o.sub.2; --N(R.sup.oS(O).sub.2R.sup.o;
--N(OR.sup.o)R.sup.o; --C(NH)NR.sup.o.sub.2; --P(O).sub.2R.sup.o;
--P(O)R.sup.o.sub.2; --OP(O)R.sup.o.sub.2; --OP(O)(OR.sup.o).sub.2;
--SiR.sup.o.sub.3; --(C.sub.1-4 straight or
branched)alkylene)O--N(R.sup.o).sub.2; or --(C.sub.1-4 straight or
branched)alkylene)C(O)O--N(R.sup.o).sub.2, wherein each R.sup.o may
be substituted as defined below and is independently hydrogen,
C.sub.1-6 aliphatic, --CH.sub.2Ph, --O(CH.sub.2).sub.0-1Ph, or a
5-6-membered saturated, partially unsaturated, or aryl ring having
0-4 heteroatoms independently selected from nitrogen, oxygen, or
sulfur, or, notwithstanding the definition above, two independent
occurrences of R.sup.o, taken together with their intervening
atom(s), form a 3-12-membered saturated, partially unsaturated, or
aryl mono- or bicyclic ring having 0-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur, which may be substituted
as defined below.
[0045] Suitable monovalent substituents on R.sup.o (or the ring
formed by taking two independent occurrences of R.sup.o together
with their intervening atoms), are independently halogen,
--(CH.sub.2).sub.0-2R.sup..cndot., -(haloR.sup..cndot.),
--(CH.sub.2).sub.0-2OH, --(CH.sub.2).sub.0-2OR.sup..cndot.,
--CH.sub.2).sub.0-2CH(OR.sup..cndot.).sub.2;
--O(haloR.sup..cndot.), --CN, --N.sub.3,
--(CH.sub.2).sub.0-2C(O)R.sup..cndot., --(CH.sub.2).sub.0-2C(O)OH,
--(CH.sub.2).sub.0-2C(O)OH, --(CH.sub.2).sub.0-2SR.sup..cndot.,
--(CH.sub.2).sub.0-2SH, --(CH.sub.2).sub.0-2NH.sub.2,
--(CH.sub.2).sub.0-2NHR.sup..cndot.,
--(CH.sub.2).sub.0-2NR.sup..cndot..sub.2, --NO.sub.2,
--SiR.sup..cndot..sub.3, --OSiR.sup..cndot..sub.3,
--C(O)SR.sup..cndot., --(C.sub.1-4 straight or branched
alkylene)C(O)OR.sup..cndot., or --SSR.sup..cndot. wherein each
R.sup..cndot. is unsubstituted or where preceded by "halo" is
substituted only with one or more halogens, and is independently
selected from C.sub.1-4 aliphatic, --CH.sub.2Ph,
--O(CH.sub.2).sub.0-1Ph, or a 5-6-membered saturated, partially
unsaturated, or aryl ring having 0-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur. Suitable divalent
substituents on a saturated carbon atom of R.sup.o include .dbd.O
and .dbd.S.
[0046] Suitable divalent substituents on a saturated carbon atom of
an "optionally substituted" group include the following: .dbd.O,
.dbd.S, .dbd.NNR*.sub.2, .dbd.NNHC(O)R*, .dbd.NNHC(O)OR*,
.dbd.NNHS(O).sub.2R*, .dbd.NR*, .dbd.NOR*,
--O(C(R*.sub.2)).sub.2-3O--, or --S(C(R*.sub.2)).sub.2-3S--,
wherein each independent occurrence of R* is selected from
hydrogen, C.sub.1-6 aliphatic which may be substituted as defined
below, or an unsubstituted 5-6-membered saturated, partially
unsaturated, or aryl ring having 0-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur. Suitable divalent
substituents that are bound to vicinal substitutable carbons of an
"optionally substituted" group include: --O(CR*.sub.2).sub.2-3O--,
wherein each independent occurrence of R* is selected from
hydrogen, C.sub.1-6 aliphatic which may be substituted as defined
below, or an unsubstituted 5-6-membered saturated, partially
unsaturated, or aryl ring having 0-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur.
[0047] Suitable substituents on the aliphatic group of R* include
halogen, --R.sup..cndot., -(haloR.sup..cndot.), --OH,
--OR.sup..cndot., --O(haloR.sup..cndot.), --CN, --C(O)OH,
--C(O)OR.sup..cndot., --NH.sub.2, --NHR.sup..cndot.,
--NR.sup..cndot..sub.2, or --NO.sub.2, wherein each R.sup..cndot.
is unsubstituted or where preceded by "halo" is substituted only
with one or more halogens, and is independently C.sub.1-4
aliphatic, --CH.sub.2Ph, --O(CH.sub.2).sub.0-1Ph, or a 5-6-membered
saturated, partially unsaturated, or aryl ring having 0-4
heteroatoms independently selected from nitrogen, oxygen, or
sulfur.
[0048] Suitable substituents on a substitutable nitrogen of an
"optionally substituted" group include --R.sup..dagger.,
--NR.sup..dagger..sub.2, --C(O)R.sup..dagger.,
--C(O)OR.sup..dagger., --C(O)C(O)R.sup..dagger.,
--C(O)CH.sub.2C(O)R.sup..dagger., --S(O).sub.2R.sup..dagger.,
--S(O).sub.2NR.sup..dagger..sub.2, --C(S)NR.sup..dagger..sub.2,
--C(NH)NR.sup..dagger..sub.2, or
--N(R.sup..dagger.S(O).sub.2R.sup..dagger.; wherein each
R.sup..dagger. is independently hydrogen, C.sub.1-6 aliphatic which
may be substituted as defined below, unsubstituted --OPh, or an
unsubstituted 5-6-membered saturated, partially unsaturated, or
aryl ring having 0-4 heteroatoms independently selected from
nitrogen, oxygen, or sulfur, or, notwithstanding the definition
above, two independent occurrences of R.sup..dagger., taken
together with their intervening atom(s) form an unsubstituted
3-12-membered saturated, partially unsaturated, or aryl mono- or
bicyclic ring having 0-4 heteroatoms independently selected from
nitrogen, oxygen, or sulfur.
[0049] Suitable substituents on the aliphatic group of
R.sup..dagger. are independently halogen, --R.sup..cndot.,
-(haloR.sup..cndot.), --OH, --OR.sup..cndot.,
--O(haloR.sup..cndot.), --CN, --C(O)OH, --C(O)OR.sup..cndot.,
--NH.sub.2, --NHR.sup..cndot., --NR.sup..cndot..sub.2, or
--NO.sub.2, wherein each R.sup..cndot. is unsubstituted or where
preceded by "halo" is substituted only with one or more halogens,
and is independently C.sub.1-4 aliphatic, --CH.sub.2Ph,
--O(CH.sub.2).sub.0-1Ph, or a 5-6-membered saturated, partially
unsaturated, or aryl ring having 0-4 heteroatoms independently
selected from nitrogen, oxygen, or sulfur.
[0050] As used herein, the term "inhibitor" is defined as a
compound that binds to and/or inhibits the target 2-oxoglutarated
dependent enzyme with measurable affinity. In certain embodiments,
an inhibitor has an IC.sub.50 and/or binding constant of less about
50 .mu.M, less than about 1 .mu.M less than about 500 nM, less than
about 100 nM, or less than about 10 nM.
[0051] The terms "measurable affinity" and "measurably inhibit," as
used herein, means a measurable change in activity of at least one
2-oxoglutarate dependent enzyme between a sample comprising a
provided compound, or composition thereof, and at least one
2-oxoglutarate. dependent enzyme, and an equivalent sample
comprising at least one 2-oxoglutarate dependent enzyme, in the
absence of said compound, or composition thereof.
3. Description of Exemplary Compounds
[0052] In certain embodiments, the present invention provides a
compound of formula I,
##STR00008##
or a pharmaceutically acceptable salt thereof, wherein R.sup.1 and
Ring A are as defined and described herein.
[0053] As defined generally above, R.sup.1 is --R, halogen, --OR,
--SR, --N(R').sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2,
wherein R and R' are as defined above and described herein. In some
embodiments, R.sup.1 is hydrogen. In some embodiments, R.sup.1 is
optionally substituted C.sub.1-6 aliphatic. In certain embodiments,
R.sup.1 is optionally substituted C.sub.1-6 alkyl, C.sub.1-6
alkenyl, or C.sub.1-6 alkynyl. In certain embodiments, R.sup.1 is
optionally substituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.1 is methyl. In certain other embodiments, R.sup.1 is ethyl
or tert-butyl. In some embodiments, R.sup.1 is --OR, --SR, or
--N(R').sub.2. In certain embodiments, R.sup.1 is --SR. In certain
embodiments, R.sup.1 is --NH.sub.2. In certain embodiments, R.sup.1
is --CN or --NO.sub.2. In some embodiments, R.sup.1 is halogen. In
certain embodiments, R.sup.1 is fluoro, chloro, bromo, or iodo. In
certain embodiments, R.sup.1 is fluoro. In some embodiments,
R.sup.1 is --C(O)R, --CO.sub.2R, --C(O)SR, --C(O)N(R').sub.2,
--C(O)C(O)R, or --C(O)CH.sub.2C(O)R. In certain embodiments,
R.sup.1 is --C(S)OR or --C(S)N(R').sub.2. In other embodiments,
R.sup.1 is --S(O)R, --SO.sub.2R, or --SO.sub.2N(R').sub.2. In some
embodiments, R.sup.1 is --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
or --N(R')C(.dbd.N(R'))N(R').sub.2. In certain embodiments, R.sup.1
is --N(R')N(R').sub.2. In some embodiments, R.sup.1 is
--C.dbd.NN(R').sub.2, --C.dbd.NOR, --C(.dbd.N(R'))N(R').sub.2,
--OC(O)R, or --OC(O)N(R').sub.2.
[0054] As defined generally above, Ring A is
##STR00009##
wherein X, R.sup.2, R.sup.2', R.sup.3, R.sup.5, and R.sup.6 are as
defined above and described herein. Thus, in certain embodiments, a
compound of the invention is of one of the following formulae:
##STR00010##
wherein R.sup.1, R.sup.2, R.sup.2', R.sup.3, R.sup.5, R.sup.6, and
X are as defined above and described herein.
[0055] As defined generally above, R.sup.2 is --R, halogen, --OR,
--SR, --N(R').sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2,
wherein R and R' are as defined above and described herein. In some
embodiments, R.sup.2 is hydrogen. In some embodiments, R.sup.2 is
optionally substituted C.sub.1-6 aliphatic. In certain embodiments,
R.sup.2 is optionally substituted C.sub.1-6 alkyl, C.sub.1-6
alkenyl, or C.sub.1-6 alkynyl. In certain embodiments, R.sup.2 is
optionally substituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.2 is ethyl. In certain other embodiments, R.sup.2 is methyl,
propyl, isopropyl, butyl, or isobutyl. In some embodiments, R.sup.2
is C.sub.1-6 alkyl substituted with an --OH or --OC.sub.1-6 alkyl
group. In certain embodiments, R.sup.2 is --CH.sub.2CH.sub.2OH or
--CH.sub.2CH.sub.2OCH.sub.3. In some embodiments, R.sup.2 is
cycloalkyl. In certain embodiments, R.sup.2 is cyclopropyl,
cyclobutyl, cyclopentyl, or cyclohexyl. In some embodiments,
R.sup.2 is optionally substituted C.sub.1-6 alkenyl. In certain
embodiments, R.sup.2 is allyl. In some embodiments, R.sup.2 is
optionally substituted C.sub.1-6 alkynyl. In certain embodiments,
R.sup.2 is 2-propynyl. In some embodiments, R.sup.2 is optionally
substituted benzyl. In certain embodiments, R.sup.2 is
unsubstituted benzyl. In certain other embodiments, R.sup.2 is
substituted benzyl. In some embodiments, R.sup.2 is C.sub.1-6 alkyl
substituted with an ester group. In certain embodiments, R.sup.2 is
--CH.sub.2CO.sub.2C.sub.1-6alkyl or --CH.sub.2CO.sub.2aryl. In
certain embodiments, R.sup.2 is --CH.sub.2CO.sub.2CH.sub.2CH.sub.3.
In some embodiments, R.sup.2 is --OR, --SR, or --N(R').sub.2. In
certain embodiments, R.sup.2 is --CN or --NO.sub.2. In some
embodiments, R.sup.2 is halogen. In certain embodiments, R.sup.2 is
fluoro, chloro, bromo, or iodo. In some embodiments, R.sup.2 is
--C(O)R, --CO.sub.2R, --C(O)SR, --C(O)N(R').sub.2, --C(O)C(O)R, or
--C(O)CH.sub.2C(O)R. In certain embodiments, R.sup.2 is --C(S)OR or
--C(S)N(R').sub.2. In other embodiments, R.sup.2 is --S(O)R,
--SO.sub.2R, or --SO.sub.2N(R').sub.2. In some embodiments, R.sup.2
is --N(R')C(O)R, --N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2, or
--N(R')C(.dbd.N(R'))N(R').sub.2. In some embodiments, R.sup.2 is
--C.dbd.NN(R').sub.2, --C.dbd.NOR, --C(.dbd.N(R'))N(R').sub.2,
--OC(O)R, or --OC(O)N(R').sub.2.
[0056] As defined generally above, R.sup.2' is --R, --OR, --SR,
--N(R').sub.2, --C(O)R, --CO.sub.2R, --C(O)N(R').sub.2, --C(O)SR,
--C(O)C(O)R, --C(O)CH.sub.2C(O)R, --C(S)N(R').sub.2, --C(S)OR,
--S(O)R, --SO.sub.2R, --SO.sub.2N(R').sub.2, --N(R')C(O)R,
--N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2,
wherein R and R' are as defined above and described herein. In some
embodiments, R.sup.2' is hydrogen. In some embodiments, R.sup.2' is
optionally substituted C.sub.1-6 aliphatic. In certain embodiments,
R.sup.2' is optionally substituted C.sub.1-6 alkyl, C.sub.1-6
alkenyl, or C.sub.1-6 alkynyl. In certain embodiments, R.sup.2' is
optionally substituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.2' is ethyl. In certain other embodiments, R.sup.2' is
methyl, propyl, isopropyl, butyl, or isobutyl. In some embodiments,
R.sup.2' is C.sub.1-6 alkyl substituted with an --OH or
--OC.sub.1-6alkyl group. In certain embodiments, R.sup.2' is
--CH.sub.2CH.sub.2OH or --CH.sub.2CH.sub.2OCH.sub.3. In some
embodiments, R.sup.2' is cycloalkyl. In certain embodiments,
R.sup.2' is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In
some embodiments, R.sup.2' is optionally substituted C.sub.1-6
alkenyl. In certain embodiments, R.sup.2' is allyl. In some
embodiments, R.sup.2' is optionally substituted C.sub.1-6 alkynyl.
In certain embodiments, R.sup.2' is 2-propynyl. In some
embodiments, R.sup.2' is optionally substituted benzyl. In certain
embodiments, R.sup.2' is unsubstituted benzyl. In certain other
embodiments, R.sup.2' is substituted benzyl. In some embodiments,
R.sup.2' is C.sub.1-6 alkyl substituted with an ester group. In
certain embodiments, R.sup.2' is --CH.sub.2CO.sub.2C.sub.1-6alkyl
or --CH.sub.2CO.sub.2aryl. In certain embodiments, R.sup.2' is
--CH.sub.2CO.sub.2CH.sub.2CH.sub.3. In some embodiments, R.sup.2'
is --OR, --SR, or --N(R').sub.2. In some embodiments, R.sup.2' is
--C(O)R, --CO.sub.2R, --C(O)SR, --C(O)N(R').sub.2, --C(O)C(O)R, or
--C(O)CH.sub.2C(O)R. In certain embodiments, R.sup.2' is --C(S)OR
or --C(S)N(R').sub.2. In other embodiments, R.sup.2' is --S(O)R,
--SO.sub.2R, or --SO.sub.2N(R').sub.2. In some embodiments,
R.sup.2' is --N(R')C(O)R, --N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2, or
--N(R')C(.dbd.N(R'))N(R').sub.2. In some embodiments, R.sup.2' is
--C.dbd.NN(R').sub.2, --C.dbd.NOR, --C(.dbd.N(R'))N(R').sub.2,
--OC(O)R, or --OC(O)N(R').sub.2.
[0057] As defined generally above, R.sup.3 is --R, halogen, --OR,
--SR, --N(R').sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)C(O)R, --C(O)CH.sub.2C(O)R, --S(O)R,
--SO.sub.2R, --SO.sub.2N(R').sub.2, --N(R')C(O)R,
--N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--C.dbd.NN(R').sub.2, --C.dbd.NOR, --OC(O)R, or --OC(O)N(R').sub.2,
wherein R and R' are as defined above and described herein. In some
embodiments, R.sup.3 is hydrogen. In some embodiments, R.sup.3 is
optionally substituted C.sub.1-6 aliphatic. In certain embodiments,
R.sup.3 is optionally substituted C.sub.1-6 alkyl, C.sub.1-6
alkenyl, or C.sub.1-6 alkynyl. In certain embodiments, R.sup.3 is
optionally substituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.3 is methyl. In certain other embodiments, R.sup.3 is ethyl,
propyl, isopropyl, butyl, or isobutyl. In certain embodiments,
R.sup.3 is --CF.sub.3. In some embodiments, R.sup.3 is C.sub.1-6
alkyl substituted with an --OH or --OC.sub.1-6alkyl group. In
certain embodiments, R.sup.3 is --CH.sub.2OH, --CH.sub.2CH.sub.2OH,
--CH.sub.2CH.sub.2CH.sub.2OH, --CH.sub.2OCH.sub.2CH.sub.3,
--CH.sub.2OCH.sub.3, --CH.sub.2CH.sub.2CH.sub.2OCH.sub.3,
--CH(OH)CH.sub.3, or --CH.sub.2CH.sub.2OCH.sub.3. In some
embodiments, R.sup.3 is C.sub.1-6 alkyl substituted with an
--NHC.sub.1-6alkyl or --N(C.sub.1-6alkyl).sub.2 group. In certain
embodiments, R.sup.3 is --CH.sub.2NHC.sub.1-6 alkyl. In certain
embodiments, R.sup.3 is --CH.sub.2NHCH.sub.3. In some embodiments,
R.sup.3 is C.sub.1-6 alkyl substituted with an aryl, heteroaryl,
carbocyclyl, or heterocyclyl ring. In some embodiments, R.sup.3 is
optionally substituted benzyl. In certain embodiments, R.sup.3 is
unsubstituted benzyl. In certain other embodiments, R.sup.3 is
substituted benzyl. In certain embodiments, R.sup.3 is
--C(R.sup.o.sub.2Ph. In certain embodiments, R.sup.3 is
--C(R.sup.o).sub.2Ph, wherein R.sup.o is hydrogen or methyl. In
certain embodiments, R.sup.3 is trifluoromethylbenzyl. In certain
embodiments, R.sup.3 is --C(R.sup.o).sub.2(heteroaryl). In certain
embodiments, R.sup.3 is --C(R.sup.o).sub.2(heteroaryl), wherein the
heteroaryl is pyridyl, pyrimidyl, pyrazinyl, pyridazinyl,
triazinyl, pyridinonyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl,
tetrazolyl, thienyl, furanyl, thiazolyl, isothiazolyl,
thiadiazolyl, oxazolyl, isoxazolyl, or oxadiazolyl. In certain
embodiments, R.sup.3 is --CH.sub.2(heteroaryl), wherein the
heteroaryl is pyridyl, pyrimidyl, pyrazinyl, pyridazinyl,
triazinyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl,
thienyl, furanyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl,
isoxazolyl, or oxadiazolyl. In certain embodiments, R.sup.3 is
--C(R.sup.o).sub.2(carbocyclyl). In certain embodiments, R.sup.3
is)-C(R.sup.o).sub.2(carbocyclyl), wherein the carbocyclyl is
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cycloheptyl.
In certain embodiments, R.sup.3 is --CH.sub.2(carbocyclyl), wherein
the carbocyclyl is cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, or cycloheptyl. In certain embodiments, R.sup.3
is)-C(R.sup.o).sub.2(heterocyclyl). In certain embodiments, R.sup.3
is --C(R.sup.o).sub.2(heterocyclyl), wherein the heterocyclyl is
tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl,
pyrrolidinyl, pyrrolidonyl, piperidinyl, pyrrolinyl,
tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl,
oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl,
oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. In certain
embodiments, R.sup.3 is --CH.sub.2(heterocyclyl), wherein the
heterocyclyl is tetrahydrofuranyl, tetrahydrothienyl,
tetrahydropyranyl, pyrrolidinyl, pyrrolidonyl, piperidinyl,
pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl,
decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl,
dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and
quinuclidinyl. In some embodiments, R.sup.3 is optionally
substituted C.sub.1-6 alkenyl. In certain embodiments, R.sup.3 is
allyl. In some embodiments, R.sup.3 is optionally substituted
C.sub.1-6 alkynyl. In certain embodiments, R.sup.3 is propargyl. In
some embodiments, R.sup.3 is an optionally substituted aryl or
heteroaryl group. In certain embodiments, R.sup.3 is phenyl. In
certain embodiments, R.sup.3 is substituted phenyl. In certain
embodiments, R.sup.3 is toluoyl. In certain other embodiments,
R.sup.3 is a 5-6 membered heteroaryl ring having 1-3 heteroatoms
selected from nitrogen, oxygen, and sulfur. In certain embodiments,
R.sup.3 is pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazinyl,
pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, thienyl,
furanyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl,
isoxazolyl, or oxadiazolyl. In some embodiments, R.sup.3 is --OR,
--SR, or --N(R').sub.2. In some embodiments, R.sup.3 is halogen. In
certain embodiments, R.sup.3 is fluoro, chloro, bromo, or iodo. In
some embodiments, R.sup.3 is --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, or --C(O)CH.sub.2C(O)R.
In certain embodiments, R.sup.3 is optionally substituted
--CO.sub.2C.sub.1-6alkyl. In certain embodiments, R.sup.3 is
--CO.sub.2Et or --CO.sub.2Bn. In certain embodiments, R.sup.3 is
--CONHC.sub.1-6alkyl. In certain embodiments, R.sup.3 is
--CONHCH.sub.3 or --CONHCH.sub.2CH.sub.3. In certain embodiments,
R.sup.3 is --C(S)OR or --C(S)N(R').sub.2. In other embodiments,
R.sup.3 is --S(O)R, --SO.sub.2R, or --SO.sub.2N(R').sub.2. In some
embodiments, R.sup.3 is --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
or --N(R')C(.dbd.N(R'))N(R').sub.2. In some embodiments, R.sup.3 is
--C.dbd.NN(R').sub.2, --C.dbd.NOR, --C(.dbd.N(R'))N(R').sub.2,
--OC(O)R, or --OC(O)N(R').sub.2.
[0058] In some embodiments, R.sup.2 and R.sup.3 are taken together
with their intervening atoms to form an optionally substituted 5-7
membered partially unsaturated or aromatic fused ring having 0-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur. In certain embodiments, R.sup.2 and R.sup.3 are taken
together with their intervening atoms to form a 5-membered fused
ring. In certain embodiments, R.sup.2 and R.sup.3 are taken
together with their intervening atoms to form a fused cyclopentene
ring. In certain embodiments, R.sup.2 and R.sup.3 are taken
together with their intervening atoms to form a 6-membered fused
ring. In certain embodiments, R.sup.2 and R.sup.3 are taken
together with their intervening atoms to form a fused cyclohexene
ring. In certain embodiments, R.sup.2 and R.sup.3 are taken
together with their intervening atoms to form a fused benzene ring.
In certain embodiments, R.sup.2 and R.sup.3 are taken together with
their intervening atoms to form a 5-7 membered partially
unsaturated fused ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, and sulfur. In certain embodiments,
R.sup.2 and R.sup.3 are taken together with their intervening atoms
to form a 5-7 membered aromatic fused ring having 1-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur.
[0059] In some embodiments, R.sup.2' and R.sup.3 are taken together
with their intervening atoms to form an optionally substituted 5-7
membered partially unsaturated or aromatic fused ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur. In certain embodiments, R.sup.2' and R.sup.3 are taken
together with their intervening atoms to form a 5-membered fused
ring. In certain embodiments, R.sup.2' and R.sup.3 are taken
together with their intervening atoms to form a 6-membered fused
ring. In certain embodiments, R.sup.2' and R.sup.3 are taken
together with their intervening atoms to form a fused pyridine
ring. In certain embodiments, R.sup.2' and R.sup.3 are taken
together with their intervening atoms to form a 5-7 membered
partially unsaturated fused ring having 1-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur. In
certain embodiments, R.sup.2' and R.sup.3 are taken together with
their intervening atoms to form a 5-7 membered aromatic fused ring
having 1-4 heteroatoms independently selected from nitrogen,
oxygen, and sulfur.
[0060] As defined generally above, X is --N(R.sup.4)--, --O--, or
--S--, wherein R.sup.4 is as defined above and described herein. In
certain embodiments, X is --O-- or --S--. In some embodiments, X is
--N(R.sup.4)--. In certain embodiments, X is --NH--. In certain
embodiments, X is --N(CH.sub.3)--.
[0061] As defined generally above, R.sup.4 is --R, --C(O)R,
--CO.sub.2R, or --S(O).sub.2R, or R.sup.4 and R.sup.3 are taken
together with their intervening atoms to form an optionally
substituted 5-7 membered saturated, partially unsaturated, or
aromatic fused ring having 1-4 heteroatoms independently selected
from nitrogen, oxygen, and sulfur. In certain embodiments, R.sup.4
is hydrogen. In some embodiments, R.sup.4 is optionally substituted
C.sub.1-6 alkyl. In certain embodiments, R.sup.4 is optionally
substituted C.sub.1-3 alkyl. In certain embodiments, R.sup.4 is
methyl. In certain embodiments, R.sup.4 is substituted C.sub.1-6
alkyl. In certain embodiments, R.sup.4 is benzyl. In certain
embodiments, R.sup.4 is --CH.sub.2CH.sub.2N(CH.sub.3).sub.2. In
some embodiments, R.sup.4 is aryl or heteroaryl. In certain
embodiments, R.sup.4 is phenyl. In some embodiments, R.sup.4 is
--C(O)R, --CO.sub.2R, or --S(O).sub.2R.
[0062] In some embodiments, R.sup.4 and R.sup.3 are taken together
with their intervening atoms to form an optionally substituted 5-7
membered saturated, partially unsaturated, or aromatic fused ring
having 1-4 heteroatoms independently selected from nitrogen,
oxygen, and sulfur. In certain embodiments, R.sup.4 and R.sup.3 are
taken together with their intervening atoms to form a 5-membered
fused ring. In certain embodiments, R.sup.4 and R.sup.3 are taken
together with their intervening atoms to form a fused pyrrolidine
ring. In certain embodiments, R.sup.4 and R.sup.3 are taken
together with their intervening atoms to form a 6-membered fused
ring. In certain embodiments, R.sup.4 and R.sup.3 are taken
together with their intervening atoms to form a fused piperidine
ring. In certain embodiments, R.sup.4 and R.sup.3 are taken
together with their intervening atoms to form a 5-7 membered
partially unsaturated fused ring having 1-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur. In
certain embodiments, R.sup.4 and R.sup.3 are taken together with
their intervening atoms to form a 5-7 membered aromatic fused ring
having 1-4 heteroatoms independently selected from nitrogen,
oxygen, and sulfur.
[0063] As defined generally above, R.sup.5 is R, --C(O)R,
--CO.sub.2R, --C(O)N(R').sub.2, --C(O)C(O)R, or
--C(O)CH.sub.2C(O)R, or R.sup.5 and R.sup.2 are taken together with
their intervening atoms to form an optionally substituted 5-7
membered partially unsaturated or aromatic fused ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur. In some embodiments, R.sup.5 is hydrogen. In some
embodiments, R.sup.5 is optionally substituted C.sub.1-6 alkyl. In
certain embodiments, R.sup.5 is methyl. In certain embodiments,
R.sup.5 is substituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.5 is C.sub.1-6 alkyl substituted with an --OH or
--OC.sub.1-6alkyl group. In certain embodiments, R.sup.5 is
--CH.sub.2CH.sub.2OCH.sub.3. In some embodiments, R.sup.4 is
--C(O)R, --CO.sub.2R, --C(O)N(R').sub.2, --C(O)C(O)R, or
--C(O)CH.sub.2C(O)R.
[0064] As defined generally above, R.sup.6 is --R, halogen, --OR,
--SR, --N(R').sub.2, --CN, --NO.sub.2, --C(O)R, --CO.sub.2R,
--C(O)N(R').sub.2, --C(O)SR, --C(O)C(O)R, --C(O)CH.sub.2C(O)R,
--C(S)N(R').sub.2, --C(S)OR, --S(O)R, --SO.sub.2R,
--SO.sub.2N(R').sub.2, --N(R')C(O)R, --N(R')C(O)N(R').sub.2,
--N(R')SO.sub.2R, --N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2,
--N(R')C(.dbd.N(R'))N(R').sub.2, --C.dbd.NN(R').sub.2, --C.dbd.NOR,
--C(.dbd.N(R'))N(R').sub.2, --OC(O)R, or --OC(O)N(R').sub.2,
wherein R and R' are as defined above and described herein. In some
embodiments, R.sup.6 is hydrogen. In some embodiments, R.sup.6 is
optionally substituted C.sub.1-6 aliphatic. In certain embodiments,
R.sup.6 is optionally substituted C.sub.1-6 alkyl, C.sub.1-6
alkenyl, or C.sub.1-6 alkynyl. In certain embodiments, R.sup.6 is
optionally substituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.6 is ethyl. In certain other embodiments, R.sup.6 is methyl,
propyl, isopropyl, butyl, or isobutyl. In some embodiments, R.sup.6
is C.sub.1-6 alkyl substituted with an --OH or --OC.sub.1-6alkyl
group. In certain embodiments, R.sup.6 is --CH.sub.2CH.sub.2OH or
--CH.sub.2CH.sub.2OCH.sub.3. In some embodiments, R.sup.6 is
cycloalkyl. In certain embodiments, R.sup.6 is cyclopropyl,
cyclobutyl, cyclopentyl, or cyclohexyl. In some embodiments,
R.sup.6 is optionally substituted C.sub.1-6 alkenyl. In certain
embodiments, R.sup.6 is allyl. In some embodiments, R.sup.6 is
optionally substituted C.sub.1-6 alkynyl. In certain embodiments,
R.sup.6 is 2-propynyl. In some embodiments, R.sup.6 is optionally
substituted benzyl. In certain embodiments, R.sup.6 is
unsubstituted benzyl. In certain other embodiments, R.sup.6 is
substituted benzyl. In some embodiments, R.sup.6 is C.sub.1-6 alkyl
substituted with an ester group. In certain embodiments, R.sup.6 is
--CH.sub.2CO.sub.2C.sub.1-6alkyl or --CH.sub.2CO.sub.2aryl. In
certain embodiments, R.sup.6 is --CH.sub.2CO.sub.2CH.sub.2CH.sub.3.
In some embodiments, R.sup.6 is --OR, --SR, or --N(R').sub.2. In
certain embodiments, R.sup.6 is --CN or --NO.sub.2. In some
embodiments, R.sup.6 is halogen. In certain embodiments, R.sup.6 is
fluoro, chloro, bromo, or iodo. In some embodiments, R.sup.6 is
--C(O)R, --CO.sub.2R, --C(O)SR, --C(O)N(R').sub.2, --C(O)C(O)R, or
--C(O)CH.sub.2C(O)R. In certain embodiments, R.sup.6 is --C(S)OR or
--C(S)N(R').sub.2. In other embodiments, R.sup.6 is --S(O)R,
--SO.sub.2R, or --SO.sub.2N(R').sub.2. In some embodiments, R.sup.6
is --N(R')C(O)R, --N(R')C(O)N(R').sub.2, --N(R')SO.sub.2R,
--N(R')SO.sub.2N(R').sub.2, --N(R')N(R').sub.2, or
--N(R')C(.dbd.N(R'))N(R').sub.2. In some embodiments, R.sup.6 is
--C.dbd.NN(R').sub.2, --C.dbd.NOR, --C(.dbd.N(R'))N(R').sub.2,
--OC(O)R, or --OC(O)N(R').sub.2.
[0065] In some embodiments, R.sup.6 and R.sup.3 are taken together
with their intervening atoms to form an optionally substituted 5-7
membered partially unsaturated or aromatic fused ring having 0-4
heteroatoms independently selected from nitrogen, oxygen, and
sulfur. In certain embodiments, R.sup.6 and R.sup.3 are taken
together with their intervening atoms to form a 5-membered fused
ring. In certain embodiments, R.sup.6 and R.sup.3 are taken
together with their intervening atoms to form a fused cyclopentene
ring. In certain embodiments, R.sup.6 and R.sup.3 are taken
together with their intervening atoms to form a 6-membered fused
ring. In certain embodiments, R.sup.6 and R.sup.3 are taken
together with their intervening atoms to form a fused cyclohexene
ring. In certain embodiments, R.sup.6 and R.sup.3 are taken
together with their intervening atoms to form a fused benzene ring.
In certain embodiments, R.sup.6 and R.sup.3 are taken together with
their intervening atoms to form a 5-7 membered partially
unsaturated fused ring having 1-4 heteroatoms independently
selected from nitrogen, oxygen, and sulfur. In certain embodiments,
R.sup.6 and R.sup.3 are taken together with their intervening atoms
to form a 5-7 membered aromatic fused ring having 1-4 heteroatoms
independently selected from nitrogen, oxygen, and sulfur.
[0066] As defined generally above, each R is independently hydrogen
or an optionally substituted group selected from C.sub.1-6
aliphatic, phenyl, a 3-7 membered saturated or partially
unsaturated carbocyclic ring, an 8-10 membered bicyclic saturated,
partially unsaturated or aryl ring, a 5-6 membered monocyclic
heteroaryl ring having 1-3 heteroatoms independently selected from
nitrogen, oxygen, or sulfur, a 4-7 membered saturated or partially
unsaturated heterocyclic ring having 1-2 heteroatoms independently
selected from nitrogen, oxygen, or sulfur, a 7-10 membered bicyclic
saturated or partially unsaturated heterocyclic ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, or
sulfur, or an 8-10 membered bicyclic heteroaryl ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, or
sulfur. In certain embodiments, R is hydrogen. In some embodiments,
R is optionally substituted C.sub.1-6 alkyl, alkenyl, or alkynyl.
In certain embodiments, R is optionally substituted C.sub.1-6
alkyl. In certain embodiments, R is unsubstituted C.sub.1-6 alkyl.
In certain embodiments, R is substituted C.sub.1-6 alkyl. In
certain embodiments, R is methyl, ethyl, propyl, butyl, isopropyl,
isobutyl, allyl, or benzyl.
[0067] In some embodiments, R is a 3-7 membered saturated or
partially unsaturated carbocyclic ring. In certain embodiments, R
is a 3-4 membered saturated carbocyclic ring. In other embodiments,
R is a 5-7 membered saturated or partially unsaturated carbocyclic
ring. In certain embodiments, R is cyclopropyl, cyclobutyl,
cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl,
or cycloheptenyl.
[0068] In some embodiments, R is a 4-7 membered saturated or
partially unsaturated heterocyclic ring having 1-2 heteroatoms
independently selected from nitrogen, oxygen, or sulfur. In certain
embodiments, R is a 4-7 membered saturated heterocyclic ring. In
other embodiments, R is a 5-7 membered partially unsaturated
heterocyclic ring. In certain embodiments, R is tetrahydrofuranyl,
tetrahydrothienyl, tetrahydropyranyl, pyrrolidinyl, pyrrolidonyl,
piperidinyl, pyrrolinyl, oxazolidinyl, piperazinyl, dioxanyl,
dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, or
morpholinyl.
[0069] In some embodiments, R is an 8-10 membered bicyclic
saturated or partially unsaturated carbocylic ring or a 7-10
membered bicyclic saturated or partially unsaturated heterocyclic
ring having 1-4 heteroatoms independently selected from nitrogen,
oxygen, or sulfur. In certain embodiments, R is decahydronaphthyl,
tetrahydronaphthyl, or decalin. In certain other embodiments, R is
tetrahydroquinolinyl, tetrahydroisoquinolinyl, or
decahydroquinolinyl. In some embodiments, R is a heterocyclyl ring
is fused to an aryl or heteroaryl ring. In certain embodiments, R
is indolinyl, 3H-indolyl, chromanyl, phenanthridinyl,
2-azabicyclo[2.2.1]heptanyl, octahydroindolyl, or
tetrahydroquinolinyl.
[0070] In some embodiments, R is phenyl or a 5-6 membered
heteroaryl ring having 1-3 heteroatoms independently selected from
nitrogen, oxygen, or sulfur. In certain embodiments, R is phenyl.
In certain other embodiments, R is a 5-membered heteroaryl ring
having 1-3 heteroatoms selected from nitrogen, oxygen, or sulfur.
In yet other embodiments, R is a 6-membered heteroaryl ring having
1-3 nitrogens. In certain embodiments, R is phenyl, pyridyl,
pyrimidyl, pyrazinyl, pyridazinyl, or triazinyl. In certain other
embodiments, R is pyrrolyl, pyrazolyl, imidazolyl, triazolyl,
tetrazolyl, thienyl, furanyl, thiazolyl, isothiazolyl,
thiadiazolyl, oxazolyl, isoxazolyl, or oxadiazolyl.
[0071] In some embodiments, R is bicyclic aromatic ring. In certain
embodiments, R is naphthyl. In other embodiments, R is an 8-10
membered bicyclic heteroaryl ring having 1-4 heteroatoms
independently selected from nitrogen, oxygen, or sulfur. In certain
embodiments, R is quinolinyl, quinoxalinyl, quinazolinyl,
pyridopyrazinyl, or pyridopyrimidyl. In certain other embodiments,
R is indolyl, benzimidazolyl, benzothiazolyl, benzofuranyl,
benzotriazolyl, benzoxazolyl, benzothiophenyl, indazolyl,
imidazopyridyl, imidazopyrimidyl, imidazopyrazinyl,
imidazopyridazinyl, pyrazolopyridyl, pyrazolopyrimidyl,
pyrazolopyrazinyl, pyrazolopyridazinyl, pyrrolothiazolyl,
imidazothiazolyl, thiazolopyridyl, thiazolopyrimidyl,
thiazolopypyrazinyl, thiazolopyridazinyl, oxazolopyridyl,
oxazolopyrimidyl, oxazolopyrazinyl, or oxazolopyridazinyl.
[0072] As defined generally above, each R' is independently --R,
--C(O)R, --CO.sub.2R, or two R' on the same nitrogen are taken
together with the intervening nitrogen to form a 4-7 membered
heterocyclic ring having 1-2 heteroatoms independently selected
from nitrogen, oxygen, and sulfur. In certain embodiments, R' is R
as defined and described above. In certain embodiments, R' is
--C(O)R or --CO.sub.2R. In some embodiments, two R' on the same
nitrogen are taken together with their intervening atoms to form a
4-7 membered heterocyclic ring having 1-2 heteroatoms independently
selected from nitrogen, oxygen, and sulfur. In certain embodiments,
two R' on the same nitrogen are taken together with their
intervening atoms to form an azetidine, pyrrolidine, piperidine,
morpholine, piperazine, homopiperidine, or homopiperazine ring.
[0073] According to one aspect, a provided compound is of formula
II:
##STR00011##
or a pharmaceutically acceptable salt thereof, wherein R.sup.1,
R.sup.2, R.sup.3, and R.sup.4 are as defined and described herein.
In certain embodiments, a compound of formula II has one of the
following formulae:
##STR00012##
[0074] According to another aspect, a provided compound is of
formula III:
##STR00013##
or a pharmaceutically acceptable salt thereof, wherein R.sup.1,
R.sup.2, R.sup.3, and R.sup.4 are as defined and described herein.
In certain embodiments, a compound of formula II has one of the
following formulae:
##STR00014##
[0075] According to another aspect, a provided compound is of
formula IV:
##STR00015##
or a pharmaceutically acceptable salt thereof, wherein R.sup.1,
R.sup.2, R.sup.3, and R.sup.5 are as defined and described herein.
In certain embodiments, R.sup.5 is optionally substituted C.sub.1-6
aliphatic. In certain embodiments, R.sup.5 is methyl. In some
embodiments, R.sup.5 is optionally substituted C.sub.1-6 alkyl. In
certain embodiments, R.sup.5 is substituted C.sub.1-6 alkyl. In
certain embodiments, R.sup.5 is C.sub.1-6alkyl substituted with
--OH or --OC.sub.1-6alkyl. In certain embodiments, R.sup.5 is
--CH.sub.2CH.sub.2OMe.
[0076] According to another aspect, a provided compound is of
formula V:
##STR00016##
or a pharmaceutically acceptable salt thereof, wherein R.sup.1,
R.sup.2', and R.sup.3 are as defined and described herein.
[0077] Exemplary compounds of formula I are set forth in Table 1
below.
TABLE-US-00001 TABLE 1 Exemplary Compounds of Formula I
##STR00017## I-1 ##STR00018## I-2 ##STR00019## I-3 ##STR00020## I-4
##STR00021## I-5 ##STR00022## I-6 ##STR00023## I-7 ##STR00024## I-8
##STR00025## I-9 ##STR00026## I-10 ##STR00027## I-11 ##STR00028##
I-12 ##STR00029## I-13 ##STR00030## I-14 ##STR00031## I-15
##STR00032## I-16 ##STR00033## I-17 ##STR00034## I-18 ##STR00035##
I-19 ##STR00036## I-20 ##STR00037## I-21 ##STR00038## I-22
##STR00039## I-23 ##STR00040## I-24 ##STR00041## I-25 ##STR00042##
I-26 ##STR00043## I-27 ##STR00044## I-28 ##STR00045## I-29
##STR00046## I-30 ##STR00047## I-31 ##STR00048## I-32 ##STR00049##
I-33 ##STR00050## I-34 ##STR00051## I-35 ##STR00052## I-36
##STR00053## I-37 ##STR00054## I-38 ##STR00055## I-39 ##STR00056##
I-40 ##STR00057## I-41 ##STR00058## I-42 ##STR00059## I-43
##STR00060## I-44 ##STR00061## I-45 ##STR00062## I-46 ##STR00063##
I-47 ##STR00064## I-48 ##STR00065## I-49 ##STR00066## I-50
##STR00067## I-51 ##STR00068## I-52 ##STR00069## I-53 ##STR00070##
I-54 ##STR00071## I-55 ##STR00072## I-56 ##STR00073## I-57
##STR00074## I-58 ##STR00075## I-59 ##STR00076## I-60 ##STR00077##
I-61 ##STR00078## I-62 ##STR00079## I-63 ##STR00080## I-64
##STR00081## I-65 ##STR00082## I-66 ##STR00083## I-67 ##STR00084##
I-68 ##STR00085## I-69 ##STR00086## I-70 ##STR00087## I-71
##STR00088## I-72 ##STR00089## I-73
[0078] In certain embodiments, the present invention provides any
compound depicted in Table 1, above, or a pharmaceutically
acceptable salt thereof.
[0079] In some embodiments, the present invention provides a
compound set forth in Table 1-a below.
TABLE-US-00002 TABLE 1-a Compounds of Formula I ##STR00090## I-4
##STR00091## I-21 ##STR00092## I-23 ##STR00093## I-25 ##STR00094##
I-29 ##STR00095## I-30 ##STR00096## I-49
[0080] In certain embodiments, the present invention provides any
compound depicted in Table 1-a, above, or a pharmaceutically
acceptable salt thereof.
4. Uses, Formulation and Administration
[0081] Pharmaceutically Acceptable Compositions
[0082] According to another embodiment, the invention provides a
composition comprising a provided compound or a pharmaceutically
acceptable derivative thereof and a pharmaceutically acceptable
carrier, adjuvant, or vehicle. The amount of compound in
compositions of this invention is such that is effective to
measurably inhibit a histone demethylase, or a mutant thereof, in a
biological sample or in a patient. In some embodiments, the histone
demethylase is a 2-oxoglutarate dependent enzyme. In some
embodiments, the histone demethylase is a Jumonji domain containing
protein. In some embodiments, the histone demethylase is a member
of the H3K4 (histone 3K4) demethylase family. In certain
embodiments, the histone demethylase is a JARID subfamily of
enzymes. In some embodiments, the histone demethylase is selected
from JARID1A, JARID1B, or a mutant thereof.
[0083] In certain embodiments, the amount of compound in
compositions of this invention is such that is effective to
measurably inhibit a 2-oxoglutarate dependent enzyme, or a mutant
thereof, in a biological sample or in a patient. In certain
embodiments, the 2-oxoglutarate dependent enzyme is a Jumonji
domain containing protein. In certain embodiments, the Jumonji
domain containing protein is a member of the JMJD2 subfamily. In
certain embodiments, the member of the JMJD2 subfamily is
GASC1.
[0084] In certain embodiments, the amount of compound in
compositions of this invention is such that is effective to
measurably inhibit a member of the H3K4 (histone 3K4) demethylase
family of proteins, or a mutant thereof, in a biological sample or
in a patient. In certain embodiments, the amount of compound in
compositions of this invention is such that is effective to
measurably inhibit a member of the JARID subfamily of proteins, or
a mutant thereof, in a biological sample or in a patient. In some
embodiments, the amount of compound in compositions of this
invention is such that is effective to measurably inhibit JARID1A,
JARID1B, or a mutant thereof, in a biological sample or in a
patient.
[0085] In certain embodiments, a composition of this invention is
formulated for administration to a patient in need of such
composition. In some embodiments, a composition of this invention
is formulated for oral administration to a patient.
[0086] The term "patient," as used herein, means an animal, such as
a mammal, such as a human.
[0087] The term "pharmaceutically acceptable carrier, adjuvant, or
vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that
does not destroy the pharmacological activity of the compound with
which it is formulated. Pharmaceutically acceptable carriers,
adjuvants or vehicles that may be used in the compositions of this
invention include, but are not limited to, ion exchangers, alumina,
aluminum stearate, lecithin, serum proteins, such as human serum
albumin, buffer substances such as phosphates, glycine, sorbic
acid, potassium sorbate, partial glyceride mixtures of saturated
vegetable fatty acids, water, salts or electrolytes, such as
protamine sulfate, disodium hydrogen phosphate, potassium hydrogen
phosphate, sodium chloride, zinc salts, colloidal silica, magnesium
trisilicate, polyvinyl pyrrolidone, cellulose-based substances,
polyethylene glycol, sodium carboxymethylcellulose, polyacrylates,
waxes, polyethylene-polyoxypropylene-block polymers, polyethylene
glycol and wool fat.
[0088] A "pharmaceutically acceptable derivative" means any
non-toxic salt, ester, salt of an ester or other derivative of a
compound of this invention that, upon administration to a
recipient, is capable of providing, either directly or indirectly,
a compound of this invention or an inhibitorily active metabolite
or residue thereof.
[0089] As used herein, the term "inhibitorily active metabolite or
residue thereof" means that a metabolite or residue thereof is also
an inhibitor of a histone demethylase enzyme, or a mutant
thereof.
[0090] Compositions of the present invention may be administered
orally, parenterally, by inhalation spray, topically, rectally,
nasally, buccally, vaginally or via an implanted reservoir. The
term "parenteral" as used herein includes subcutaneous,
intravenous, intramuscular, intra-articular, intra-synovial,
intrasternal, intrathecal, intrahepatic, intralesional and
intracranial injection or infusion techniques.
[0091] Liquid dosage forms for oral administration include, but are
not limited to, pharmaceutically acceptable emulsions,
microemulsions, solutions, suspensions, syrups and elixirs. In
addition to the active compounds, the liquid dosage forms may
contain inert diluents commonly used in the art such as, for
example, water or other solvents, solubilizing agents and
emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in
particular, cottonseed, groundnut, corn, germ, olive, castor, and
sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene
glycols and fatty acid esters of sorbitan, and mixtures thereof.
Besides inert diluents, the oral compositions can also include
adjuvants such as wetting agents, emulsifying and suspending
agents, sweetening, flavoring, and perfuming agents.
[0092] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution, suspension or emulsion in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, U.S.P.
and isotonic sodium chloride solution. In addition, sterile, fixed
oils are conventionally employed as a solvent or suspending medium.
For this purpose any bland fixed oil can be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid are used in the preparation of injectables.
[0093] Injectable formulations can be sterilized, for example, by
filtration through a bacterial-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use.
[0094] In order to prolong the effect of a compound of the present
invention, it is often desirable to slow the absorption of the
compound from subcutaneous or intramuscular injection. This may be
accomplished by the use of a liquid suspension of crystalline or
amorphous material with poor water solubility. The rate of
absorption of the compound then depends upon its rate of
dissolution that, in turn, may depend upon crystal size and
crystalline form. Alternatively, delayed absorption of a
parenterally administered compound form is accomplished by
dissolving or suspending the compound in an oil vehicle. Injectable
depot forms are made by forming microencapsule matrices of the
compound in biodegradable polymers such as
polylactide-polyglycolide. Depending upon the ratio of compound to
polymer and the nature of the particular polymer employed, the rate
of compound release can be controlled. Examples of other
biodegradable polymers include poly(orthoesters) and
poly(anhydrides). Depot injectable formulations are also prepared
by entrapping the compound in liposomes or microemulsions that are
compatible with body tissues.
[0095] Compositions for rectal or vaginal administration are
preferably suppositories which can be prepared by mixing the
compounds of this invention with suitable non-irritating excipients
or carriers such as cocoa butter, polyethylene glycol or a
suppository wax which are solid at ambient temperature but liquid
at body temperature and therefore melt in the rectum or vaginal
cavity and release the active compound.
[0096] Solid dosage forms for oral administration include capsules,
tablets, pills, powders, and granules. In such solid dosage forms,
the active compound is mixed with at least one inert,
pharmaceutically acceptable excipient or carrier such as sodium
citrate or dicalcium phosphate and/or a) fillers or extenders such
as starches, lactose, sucrose, glucose, mannitol, and silicic acid,
b) binders such as, for example, carboxymethylcellulose, alginates,
gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants
such as glycerol, d) disintegrating agents such as agar-agar,
calcium carbonate, potato or tapioca starch, alginic acid, certain
silicates, and sodium carbonate, e) solution retarding agents such
as paraffin, f) absorption accelerators such as quaternary ammonium
compounds, g) wetting agents such as, for example, cetyl alcohol
and glycerol monostearate, h) absorbents such as kaolin and
bentonite clay, and i) lubricants such as talc, calcium stearate,
magnesium stearate, solid polyethylene glycols, sodium lauryl
sulfate, and mixtures thereof. In the case of capsules, tablets and
pills, the dosage form may also comprise buffering agents.
[0097] Solid compositions of a similar type may also be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugar as well as high molecular
weight polyethylene glycols and the like. The solid dosage forms of
tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings and other
coatings well known in the pharmaceutical formulating art. They may
optionally contain opacifying agents and can also be of a
composition that they release the active ingredient(s) only, or
preferentially, in a certain part of the intestinal tract,
optionally, in a delayed manner. Examples of embedding compositions
that can be used include polymeric substances and waxes. Solid
compositions of a similar type may also be employed as fillers in
soft and hard-filled gelatin capsules using such excipients as
lactose or milk sugar as well as high molecular weight polethylene
glycols and the like.
[0098] The active compounds can also be in micro-encapsulated form
with one or more excipients as noted above. The solid dosage forms
of tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings, release
controlling coatings and other coatings well known in the
pharmaceutical formulating art. In such solid dosage forms the
active compound may be admixed with at least one inert diluent such
as sucrose, lactose or starch. Such dosage forms may also comprise,
as is normal practice, additional substances other than inert
diluents, e.g., tableting lubricants and other tableting aids such
a magnesium stearate and microcrystalline cellulose. In the case of
capsules, tablets and pills, the dosage forms may also comprise
buffering agents. They may optionally contain opacifying agents and
can also be of a composition that they release the active
ingredient(s) only, or preferentially, in a certain part of the
intestinal tract, optionally, in a delayed manner. Examples of
embedding compositions that can be used include polymeric
substances and waxes.
[0099] Dosage forms for topical or transdermal administration of a
compound of this invention include ointments, pastes, creams,
lotions, gels, powders, solutions, sprays, inhalants or patches.
The active component is admixed under sterile conditions with a
pharmaceutically acceptable carrier and any needed preservatives or
buffers as may be required. Ophthalmic formulation, ear drops, and
eye drops are also contemplated as being within the scope of this
invention. Additionally, the present invention contemplates the use
of transdermal patches, which have the added advantage of providing
controlled delivery of a compound to the body. Such dosage forms
can be made by dissolving or dispensing the compound in the proper
medium. Absorption enhancers can also be used to increase the flux
of the compound across the skin. The rate can be controlled by
either providing a rate controlling membrane or by dispersing the
compound in a polymer matrix or gel.
[0100] Pharmaceutically acceptable compositions provided by the
invention may also be administered by nasal aerosol or inhalation.
Such compositions are prepared according to techniques well-known
in the art of pharmaceutical formulation and may be prepared as
solutions in saline, employing benzyl alcohol or other suitable
preservatives, absorption promotors to enhance bioavailability,
fluorocarbons, and/or other conventional solubilizing or dispersing
agents.
[0101] Pharmaceutically acceptable compositions provided by the
invention may be formulated for oral administration. Such
formulations may be administered with or without food. In some
embodiments, pharmaceutically acceptable compositions of this
invention are administered without food. In other embodiments,
pharmaceutically acceptable compositions of this invention are
administered with food.
[0102] The amount of provided compounds that may be combined with
carrier materials to produce a composition in a single dosage form
will vary depending upon the patient to be treated and the
particular mode of administration. Provided compositions may be
formulate such that a dosage of between 0.01-100 mg/kg body
weight/day of the inhibitor can be administered to a patient
receiving these compositions.
[0103] It should also be understood that a specific dosage and
treatment regimen for any particular patient will depend upon a
variety of factors, including age, body weight, general health,
sex, diet, time of administration, rate of excretion, drug
combination, the judgment of the treating physician, and the
severity of the particular disease being treated. The amount of a
provided compound in the composition will also depend upon the
particular compound in the composition.
[0104] Uses of Compounds and Pharmaceutically Acceptable
Compositions
[0105] Compounds and compositions described herein are generally
useful for the inhibition of activity of one or more enzymes
involved in epigenetic regulation.
[0106] Epigenetics is the study of heritable changes in gene
expression caused by mechanisms other than changes in the
underlying DNA sequence. Molecular mechanisms that play a role in
epigenetic regulation include DNA methylation and chromatin/histone
modifications. Histone methylation, in particular, is critical in
many epigenetic phenomena.
[0107] Chromatin, the organized assemblage of nuclear DNA and
histone proteins, is the basis for a multitude of vital nuclear
processes including regulation of transcription, replication,
DNA-damage repair and progression through the cell cycle. A number
of factors, such as chromatin-modifying enzymes, have been
identified that play an important role in maintaining the dynamic
equilibrium of chromatin (Margueron, et al. (2005) Curr. Opin.
Genet. Dev. 15:163-176).
[0108] Histones are the chief protein components of chromatin. They
act as spools around which DNA winds, and they play a role in gene
regulation. There are a total of six classes of histones (H1, H2A,
H2B, H3, H4, and H5) organized into two super classes: core
histones (H2A, H2B, H3, and H4) and linker histones (H1 and H5).
The basic unit of chromatin is the nucleosome, which consists of
about 147 base pairs of DNA wrapped around the histone octamer,
consisting of two copies each of the core histones H2A, H2B, H3,
and H4 (Luger, et al. (1997) Nature 389:251-260).
[0109] Histones, particularly residues of the amino termini of
histones H3 and H4 and the amino and carboxyl termini of histones
H2A, H2B and H1, are susceptible to a variety of post-translational
modifications including acetylation, methylation, phosphorylation,
ribosylation sumoylation, ubiquitination, citrullination,
deimination, and biotinylation. The core of histones H2A and H3 can
also be modified. Histone modifications are integral to diverse
biological processes such as gene regulation, DNA repair, and
chromosome condensation.
[0110] One type of histone modification, demethylation, is
catalyzed by histone lysine demethylases (HKDM) or histone arginine
demethylases. The Jumonji domain containing family of
2-oxoglutarate dependent oxygenases represents a major class of
histone demethylases that are involved in epigenetic regulation.
Almost all Jumonji domain containing proteins described to date are
histone lysine demethylases, though JMJD6 has been found to be a
histone arginine demethylase. An important class of Jumonji domain
containing proteins is the JMJD2 (jumonji domain containing 2)
subfamily of JMJC-type lysine demethylases.
[0111] GASC1 (also known as JMJD2C) is a 2-oxoglutarate dependent
histone lysine demethylase in the JMJD2 subfamily. GASC1
demethylates trimethylated lysine 9 and lysine 36 on histone H3
(i.e., H3K9me3 and H3K36me3) (Whetstine, et al. (2006) Cell 125:
467-481). Trimethylation on lysine 9 of histone H3 is associated
with heterochromatin formation and transcriptional repression
(Cloos, et al. (2006) Nature 442: 307-311). GASC1 is also known to
bind to H3K4me3 and H4K20me3 (Huang, et al. (2006) Science 312:
748-751).
[0112] In some embodiments, enzymes that are inhibited by the
compounds and compositions described herein and against which the
methods described herein are useful include 2-oxoglutarate
dependent enzymes or an isoform or mutant thereof. In some
embodiments, the 2-oxoglutarate dependent enzyme is a Jumonji
domain containing protein. In certain embodiments, the Jumonji
domain containing protein is a member of the JMJD2 subfamily. In
certain embodiments, the member of the JMJD2 subfamily is
GASC1.
[0113] The activity of a provided compound as an inhibitor of a
2-oxoglutarate dependent enzyme (e.g. Jumonji domain containing
protein, e.g. JMJD2, e.g. GASC1) or an isoform or mutant thereof,
may be assayed in vitro, in vivo or in a cell line.
[0114] In vitro assays include assays that determine inhibition of
GASC1 or a mutant thereof. In some embodiments, inhibitor binding
may be determined by running a competition experiment where new
inhibitors are incubated with GASC1 bound to known radioligands.
Detailed conditions for assaying a provided compound as an
inhibitor of GASC1 or a mutant thereof are set forth in the
Examples below.
[0115] In some embodiments, detection of GASC1 activity is achieved
with in vitro histone lysine demethylase (HKDM) assays, which can
be either direct binding (non-catalytic) or enzymatic (catalytic)
assays. Types of substrates that are used in such assays may
include: short synthetic peptides corresponding to a number of
residues from the N-terminus of histone sequences comprising the
target lysine residue, single recombinant histone polypeptides,
histone octamers reconstituted with recombinant histone proteins,
and reconstituted nucleosomes (using reconstituted octamers and
specific recombinant DNA fragments). The reconstituted nucleosomes
may be mononucleosomes or oligonucleosomes. 2-Oxoglutarate, a
cofactor necessary for GASC1 function, can also be employed in a
competitive binding assay. Mass spectrometry and Western blot
analysis can also be used to detect GASC1 activity; see for example
Whetstine, et al. Cell 125: 467-481 (2006). For examples of HKDM
screening assays, see WO 2007/104314 and WO 2008/089883. It will be
understood that the assays described herein can be used for other
HKDM proteins in addition to GASC1. In certain embodiments, a
provided compound is competitive with 2-oxoglutarate.
[0116] GASC1 is implicated in proliferative diseases. The GASC1
gene was first identified in esophageal squamous cell carcinoma
cell lines, resulting in its designation as gene amplified in
squamous cell carcinoma 1 (GASC1) (Yang, et al. (2000) Cancer Res.
60: 4735-4739). Down regulation of GASC1 expression inhibits cell
proliferation, and histone methylation regulation is implicated in
tumorigenesis (Whetstine, et al. (2006) Cell 125: 467-481). GASC1
interacts with androgen receptor and another histone demethylase,
LSD1, in vitro and in vivo and increases androgen
receptor-dependent gene expression in prostate cells, implicating
GASC1 in prostate cancer (Wissmann, et al. (2007) Nat. Cell Biol.
9: 347-353). Furthermore, the GASC1 gene is amplified in basal like
breast tumors and in lung sarcomatoid carcinoma and is translocated
in MALT lymphomas (Han, et al. (2008) Genes Chromosomes Cancer 47:
490-499; Helias, et al. (2008) Cancer Genet. Cytogenet. 180: 51-55;
Italiano, et al. (2006) Cancer Genet. Cytogenet. 167: 122-130;
Vinatzer, et al. (2008) Clin Cancer Res 14: 6426-6431). GASC1 plays
an important role in cancer and other proliferative diseases.
[0117] As used herein, the terms "treatment," "treat," and
"treating" refer to reversing, alleviating, delaying the onset of,
or inhibiting the progress of a disease or disorder, or one or more
symptoms thereof, as described herein. In some embodiments,
treatment may be administered after one or more symptoms have
developed. In other embodiments, treatment may be administered in
the absence of symptoms. For example, treatment may be administered
to a susceptible individual prior to the onset of symptoms (e.g.,
in light of a history of symptoms and/or in light of genetic or
other susceptibility factors). Treatment may also be continued
after symptoms have resolved, for example to prevent or delay their
recurrence.
[0118] In certain embodiments, provided compounds inhibit one or
more 2-oxoglutarate dependent enzymes. In certain embodiments,
provided compounds inhibit one or more Jumonji domain containing
enzymes. In certain embodiments, provided compounds inhibit one or
more JMJD2 proteins. In certain embodiments, provided compounds
inhibit GASC1. Provided compounds are inhibitors of 2-oxoglutarate
dependent enzymes (e.g. GASC1) and are therefore useful for
treating one or more disorders associated with activity of a
2-oxoglutarate dependent enzyme (e.g. GASC1). In certain
embodiments, the present invention provides a method for treating a
GASC1-mediated disorder comprising the step of administering to a
patient in need thereof a provided compound, or a pharmaceutically
acceptable composition thereof.
[0119] As used herein, the term "GASC1-mediated" disorder or
condition means any disease or other deleterious condition in which
GASC1, or a mutant thereof, is known to play a role. Accordingly,
another embodiment of the present invention relates to treating or
lessening the severity of one or more diseases in which GASC1, or a
mutant thereof, is known to play a role.
[0120] Diseases and conditions treatable according to the methods
of this invention include, but are not limited to, cancer and other
proliferative disorders. In one embodiment, a human patient is
treated with a compound of the current invention and a
pharmaceutically acceptable carrier, adjuvant, or vehicle, wherein
said compound of is present in an amount to measurably inhibit
activity of a 2-oxoglutarate dependent enzyme (e.g. Jumonji domain
containing protein, e.g. JMJD2, e.g. GASC1).
[0121] Another important class of Jumonji domain containing
proteins is the H3K4 (histone 3K4) demethylases which are involved
in tissue development, cancer, and stem cell biology. (Roesch, et
al. (2010) Cell 141:283-594). Such H3K4 demethylases include the
JARID subfamily of histone demethylases (e.g., JARID1A and
JARID1B).
[0122] JARID1A (also known as KDM5A) is highly expressed in the
hematopoietic system. JARID1B (also known as KDM5B, PLU-1, and
RBP2-H1) is a member of the family of jumonji/ARID1 (JARID1)
histone 3K4 demethylases. In normal cells, JARID1B is marginally
expressed. However, JARID1B is highly expressed in regenerative
tissues such as testis and bone marrow. In cancer, JARID1B
functions as a transcriptional regulator of oncogenes, for example
BRCA1 in breast cancer (Yamane et al., (2007) Molecular Cell
25:801-812). Indeed, JARID is overexpressed in breast cancer. It
was also reported that JARID1B is highly expressed in slow-cycling
melanoma cells. Accordingly, inhibition of JARID1B is an important
target for eradicating all melanoma cells (rapidly proliferating
and slow-cycling) (Roesch, et al. (2010) Cell 141:283-594).
[0123] In some embodiments, enzymes that are inhibited by the
compounds and compositions described herein and against which the
methods described herein are useful include 2-oxoglutarate
dependent enzymes or an isoform or mutant thereof. In some
embodiments, the 2-oxoglutarate dependent enzyme is a Jumonji
domain containing protein. In certain embodiments, the Jumonji
domain containing protein is a member of the JMJD2 subfamily. In
certain embodiments, the member of the JMJD2 subfamily is GASC1. In
some embodiments, the enzyme is a member of the JARID subfamily. In
certain embodiments, the enzyme is JARID1A, PLU-1, or JMJD2B.
[0124] The activity of a provided compound as an inhibitor of a
histone demethylase enzyme (e.g. Jumonji domain containing protein,
e.g. JMJD2, JMJD2B, JARID1A, JARID1B, PLU-1, or GASC1) or an
isoform or mutant thereof, may be assayed in vitro, in vivo or in a
cell line.
[0125] In vitro assays include assays that determine inhibition of
an enzyme or a mutant thereof. In some embodiments, inhibitor
binding may be determined by running a competition experiment where
new inhibitors are incubated with the enzyme bound to known
radioligands. Detailed conditions for assaying a provided compound
as an inhibitor of enzyme or a mutant thereof are set forth in the
Examples below.
[0126] In some embodiments, detection of histone demethylase (e.g.
Jumonji domain containing protein, e.g. JMJD2, JMJD2B, JARID1A,
JARID1B, PLU-1, or GASC1) activity is achieved with in vitro
histone lysine demethylase (HKDM) assays, which can be either
direct binding (non-catalytic) or enzymatic (catalytic) assays.
Types of substrates that are used in such assays may include: short
synthetic peptides corresponding to a number of residues from the
N-terminus of histone sequences comprising the target lysine
residue, single recombinant histone polypeptides, histone octamers
reconstituted with recombinant histone proteins, and reconstituted
nucleosomes (using reconstituted octamers and specific recombinant
DNA fragments). The reconstituted nucleosomes may be
mononucleosomes or oligonucleosomes. 2-Oxoglutarate, a cofactor
necessary for GASC1 function, can also be employed in a competitive
binding assay. Mass spectrometry and Western blot analysis can also
be used to detect GASC1 activity; see for example Whetstine, et al.
Cell 125: 467-481 (2006). For examples of HKDM screening assays,
see WO 2007/104314 and WO 2008/089883. It will be understood that
the assays described herein can be used for other HKDM proteins in
addition to GASC1. In certain embodiments, a provided compound is
competitive with 2-oxoglutarate.
[0127] In certain embodiments, provided compounds inhibit one or
more 2-oxoglutarate dependent enzymes. In certain embodiments,
provided compounds inhibit one or more Jumonji domain containing
enzymes. In certain embodiments, provided compounds inhibit one or
more JMJD2 proteins. In certain embodiments, provided compounds
inhibit GASC1. In some embodiments, provided compounds inhibit one
or more of JARID1A, JARID1B, PLU-1, and/or JMJD2B. Provided
compounds are inhibitors of such histone demethylases and are
therefore useful for treating one or more disorders associated with
activity of one or more of JARID1A, JARID1B, PLU-1, and/or JMJD2B.
In certain embodiments, the present invention provides a method for
treating a JARID1A-, JARID1B-, PLU-1-, and/or JMJD2B-mediated
disorder comprising the step of administering to a patient in need
thereof a provided compound, or a pharmaceutically acceptable
composition thereof.
[0128] As used herein, the term "JARID1A-mediated" disorder or
condition means any disease or other deleterious condition in which
JARID1A, or a mutant thereof, is known to play a role. Accordingly,
another embodiment of the present invention relates to treating or
lessening the severity of one or more diseases in which JARID1A, or
a mutant thereof, is known to play a role.
[0129] As used herein, the term "JARID1B-mediated" disorder or
condition means any disease or other deleterious condition in which
JARID1B, or a mutant thereof, is known to play a role. Accordingly,
another embodiment of the present invention relates to treating or
lessening the severity of one or more diseases in which JARID1B, or
a mutant thereof, is known to play a role.
[0130] As used herein, the term "PLU-1-mediated" disorder or
condition means any disease or other deleterious condition in which
PLU-1, or a mutant thereof, is known to play a role. Accordingly,
another embodiment of the present invention relates to treating or
lessening the severity of one or more diseases in which PLU-1, or a
mutant thereof, is known to play a role.
[0131] As used herein, the term "JMJD2B-mediated" disorder or
condition means any disease or other deleterious condition in which
JMJD2B, or a mutant thereof, is known to play a role. Accordingly,
another embodiment of the present invention relates to treating or
lessening the severity of one or more diseases in which JMJD2B, or
a mutant thereof, is known to play a role.
[0132] The invention further relates to a method for treating,
ameliorating or preventing cancer or another proliferative disorder
by administration of an effective amount of a compound according to
this invention to a mammal, in particular a human in need of such
treatment. In some aspects of the invention, the disease to be
treated by the methods of the present invention may be cancer.
Examples of cancers that may be treated using the compounds and
methods described herein include, but are not limited to, adrenal
cancer, acinic cell carcinoma, acoustic neuroma, acral lentigious
melanoma, acrospiroma, acute eosinophilic leukemia, acute erythroid
leukemia, acute lymphoblastic leukemia, acute megakaryoblastic
leukemia, acute monocytic leukemia, acute promyelocytic leukemia,
adenocarcinoma, adenoid cystic carcinoma, adenoma, adenomatoid
odontogenic tumor, adenosquamous carcinoma, adipose tissue
neoplasm, adrenocortical carcinoma, adult T-cell leukemia/lymphoma,
aggressive NK-cell leukemia, AIDS-related lymphoma, alveolar
rhabdomyosarcoma, alveolar soft part sarcoma, ameloblastic fibroma,
anaplastic large cell lymphoma, anaplastic thyroid cancer, androgen
dependent cancer, angioimmunoblastic T-cell lymphoma,
angiomyolipoma, angiosarcoma, astrocytoma, atypical teratoid
rhabdoid tumor, B-cell chronic lymphocytic leukemia, B-cell
lymphoma, basal cell carcinoma, biliary tract cancer, bladder
cancer, blastoma, bone cancer, Brenner tumor, Brown tumor,
Burkitt's lymphoma, breast cancer, brain cancer, carcinoma,
carcinoma in situ, carcinosarcoma, cartilage tumor, cementoma,
myeloid sarcoma, chondroma, chordoma, choriocarcinoma, choroid
plexus papilloma, clear-cell sarcoma of the kidney,
craniopharyngioma, cutaneous T-cell lymphoma, cervical cancer,
colorectal cancer, Degos disease, desmoplastic small round cell
tumor, diffuse large B-cell lymphoma, dysembryoplastic
neuroepithelial tumor, dysgerminoma, embryonal carcinoma, endocrine
gland neoplasm, endodermal sinus tumor, enteropathy-associated
T-cell lymphoma, esophageal cancer, fetus in fetu, fibroma,
fibrosarcoma, follicular lymphoma, follicular thyroid cancer,
ganglioneuroma, gastrointestinal cancer, germ cell tumor,
gestational choriocarcinoma, giant cell fibroblastoma, giant cell
tumor of the bone, glial tumor, glioblastoma multiforme, glioma,
gliomatosis cerebri, glucagonoma, gonadoblastoma, granulosa cell
tumor, gynandroblastoma, gallbladder cancer, gastric cancer,
hemangioblastoma, head and neck cancer, hemangiopericytoma,
hematological malignancy, hepatoblastoma, hepatosplenic T-cell
lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, invasive
lobular carcinoma, intestinal cancer, kidney cancer, laryngeal
cancer, lentigo maligna, leukemia, leydig cell tumor, liposarcoma,
lung cancer, lymphangioma, lymphangiosarcoma, lymphoepithelioma,
lymphoma, acute lymphocytic leukemia, acute myelogeous leukemia,
chronic lymphocytic leukemia, liver cancer, small cell lung cancer,
non-small cell lung cancer, MALT lymphoma, malignant fibrous
histiocytoma, malignant peripheral nerve sheath tumor, malignant
triton tumor, mantle cell lymphoma, marginal zone B-cell lymphoma,
mast cell leukemia, mediastinal germ cell tumor, medullary
carcinoma of the breast, medullary thyroid cancer, medulloblastoma,
melanoma, meningioma, merkel cell cancer, mesothelioma, metastatic
urothelial carcinoma, mixed Mullerian tumor, mucinous tumor,
multiple myeloma, muscle tissue neoplasm, mycosis fungoides, myxoid
liposarcoma, myxoma, myxosarcoma, nasopharyngeal carcinoma,
neurinoma, neuroblastoma, neurofibroma, neuroma, nodular melanoma,
ocular cancer, oligoastrocytoma, oligodendroglioma, oncocytoma,
optic nerve sheath meningioma, optic nerve tumor, oral cancer,
osteosarcoma, ovarian cancer, Pancoast tumor, papillary thyroid
cancer, paraganglioma, pinealoblastoma, pineocytoma, pituicytoma,
pituitary adenoma, pituitary tumor, plasmacytoma, polyembryoma,
precursor T-lymphoblastic lymphoma, primary central nervous system
lymphoma, primary effusion lymphoma, preimary peritoneal cancer,
prostate cancer, pancreatic cancer, pharyngeal cancer, pseudomyxoma
periotonei, renal cell carcinoma, renal medullary carcinoma,
retinoblastoma, rhabdomyoma, rhabdomyosarcoma, Richter's
transformation, rectal cancer, sarcoma, Schwannomatosis, seminoma,
Sertoli cell tumor, sex cord-gonadal stromal tumor, signet ring
cell carcinoma, skin cancer, small blue round cell tumors, small
cell carcinoma, soft tissue sarcoma, somatostatinoma, soot wart,
spinal tumor, splenic marginal zone lymphoma, squamous cell
carcinoma, synovial sarcoma, Sezary's disease, small intestine
cancer, stomach cancer, T-cell lymphoma, testicular cancer,
thecoma, thyroid cancer, transitional cell carcinoma, throat
cancer, urachal cancer, urogenital cancer, urothelial carcinoma,
uveal melanoma, uterine cancer, verrucous carcinoma, visual pathway
glioma, vulvar cancer, vaginal cancer, Waldenstrom's
macroglobulinemia, Warthin's tumor, and Wilms' tumor.
[0133] In some embodiments, the present invention provides a method
for the treatment of benign proliferative disorder. Examples of
benign proliferative disorders treated with compounds according to
the present invention include, but are not limited to, benign soft
tissue tumors, bone tumors, brain and spinal tumors, eyelid and
orbital tumors, granuloma, lipoma, meningioma, multiple endocrine
neoplasia, nasal polyps, pituitary tumors, prolactinoma,
pseudotumor cerebri, seborrheic keratoses, stomach polyps, thyroid
nodules, cystic neoplasms of the pancreas, hemangiomas, vocal cord
nodules, polyps, and cysts, Castleman disease, chronic pilonidal
disease, dermatofibroma, pilar cyst, pyogenic granuloma, and
juvenile polyposis syndrome.
[0134] The invention further provides a method for the treatment a
subject, such as a human, suffering from one of the abovementioned
conditions, illnesses, disorders or diseases. The method comprises
administering a therapeutically effective amount of one or more of
the compounds according to this invention, which function by
inhibiting one or more 2-oxoglutarate dependent enzymes (e.g.
Jumonji domain containing protein, e.g. JMJD2, e.g. GASC1) and, in
general, by modulating protein methylation, to induce various
cellular effects, in particular induction or repression of gene
expression, arresting cell proliferation, inducing cell
differentiation and/or inducing apoptosis, to a subject in need of
such treatment.
[0135] The invention further provides a therapeutic method useful
for modulating protein methylation, gene expression, cell
proliferation, cell differentiation and/or apoptosis in vivo in
diseases mentioned above, in particular cancer, comprising
administering to a subject in need of such therapy a
pharmacologically active and therapeutically effective amount of
one or more of the compounds according to this invention.
[0136] The invention further provides a method for regulating
endogenous or heterologous promotor activity by contacting a cell
with a compound according to this invention.
[0137] The invention further relates to the use of provided
compounds for the production of pharmaceutical compositions which
are employed for the treatment and/or prophylaxis and/or
amelioration of the diseases, disorders, illnesses and/or
conditions as mentioned herein.
[0138] The invention further relates to the use of provided
compounds for the production of pharmaceutical compositions which
are employed for the treatment and/or prophylaxis of diseases
and/or disorders responsive or sensitive to the inhibition of
histone demethylases, particularly those diseases mentioned above,
such as e.g. cancer.
[0139] Provided compounds or compositions may be administered using
any amount and any route of administration effective for treating
or lessening the severity of cancer or other proliferative
disorder. The exact amount required will vary from subject to
subject, depending on the species, age, and general condition of
the subject, the severity of the infection, the particular agent,
its mode of administration, and the like. Compounds of the
invention are preferably formulated in unit dosage form for ease of
administration and uniformity of dosage. The expression "unit
dosage form" as used herein refers to a physically discrete unit of
agent appropriate for the patient to be treated. It will be
understood, however, that the total daily usage of the compounds
and compositions of the present invention will be decided by the
attending physician within the scope of sound medical judgment. The
specific effective dose level for any particular patient or
organism will depend upon a variety of factors including the
disorder being treated and the severity of the disorder; the
activity of the specific compound employed; the specific
composition employed; the age, body weight, general health, sex and
diet of the patient; the time of administration, route of
administration, and rate of excretion of the specific compound
employed; the duration of the treatment; drugs used in combination
or coincidental with the specific compound employed, and like
factors well known in the medical arts.
[0140] Pharmaceutically acceptable compositions of this invention
can be administered to humans and other animals orally, rectally,
parenterally, intracisternally, intravaginally, intraperitoneally,
topically (as by powders, ointments, or drops), bucally, as an oral
or nasal spray, or the like, depending on the severity of the
infection being treated. In certain embodiments, the compounds of
the invention may be administered orally or parenterally at dosage
levels of about 0.01 mg/kg to about 50 mg/kg and, in certain
embodiments, from about 1 mg/kg to about 25 mg/kg, of subject body
weight per day, one or more times a day, to obtain the desired
therapeutic effect.
[0141] According to one embodiment, the invention relates to a
method of inhibiting one or more histone demethylase (e.g. Jumonji
domain containing protein, e.g. JMJD2, JMJD2B, JARID1A, JARID1B,
PLU-1e.g. GASC1) activity in a biological sample comprising the
step of contacting said biological sample with a provided compound,
or a composition comprising said compound.
[0142] According to another embodiment, the invention relates to a
method of inhibiting a histone demethylase (e.g. Jumonji domain
containing protein, e.g. JMJD2, JMJD2B, JARID1A, JARID1B, PLU-1e.g.
GASC1), or a mutant thereof, activity in a biological sample
comprising the step of contacting said biological sample with a
provided compound, or a composition comprising said compound.
[0143] The term "biological sample", as used herein, includes,
without limitation, cell cultures or extracts thereof; biopsied
material obtained from a mammal or extracts thereof; and blood,
saliva, urine, feces, semen, tears, or other body fluids or
extracts thereof.
[0144] Inhibition of activity of a histone demethylase (e.g.
Jumonji domain containing protein, e.g. JMJD2, JMJD2B, JARID1A,
JARID1B, PLU-1e.g. GASC1) or a mutant thereof, in a biological
sample is useful for a variety of purposes that are known to one of
skill in the art. Examples of such purposes include, but are not
limited to, blood transfusion, organ-transplantation, biological
specimen storage, and biological assays.
[0145] According to another embodiment, the invention relates to a
method of inhibiting activity of a histone demethylase (e.g.
Jumonji domain containing protein, e.g. JMJD2, JMJD2B, JARID1A,
JARID1B, PLU-1e.g. GASC1), or a mutant thereof, in a patient
comprising the step of administering to said patient a provided
compound, or a composition comprising said compound. In certain
embodiments, the present invention provides a method for treating a
disorder mediated by a histone demethylase (e.g. Jumonji domain
containing protein, e.g. JMJD2, JMJD2B, JARID1A, JARID1B, PLU-1e.g.
GASC1), or a mutant thereof, in a patient in need thereof,
comprising the step of administering to said patient a compound
according to the present invention or pharmaceutically acceptable
composition thereof. Such disorders are described in detail
herein.
[0146] Depending upon the particular condition, or disease, to be
treated, additional therapeutic agents that are normally
administered to treat that condition may also be present in the
compositions of this invention or administered separately as a part
of a dosage regimen. As used herein, additional therapeutic agents
that are normally administered to treat a particular disease, or
condition, are known as "appropriate for the disease, or condition,
being treated."
[0147] In some embodiments, the additional therapeutic agent is an
epigenetic drug. As used herein, the term "epigenetic drug" refers
to a therapeutic agent that targets an epigenetic regulator.
Example of epigenetic regulators include the histone demethylase
(e.g. Jumonji domain containing protein, e.g. JMJD2, JMJD2B,
JARID1A, JARID1B, PLU-1e.g. GASC1) already described, as well as
other histone demethylases, histone lysine methyl transferases,
histone arginine methyl transferases, histone deacetylases, histone
acetylases, histone methylases, and DNA methyltransferases. Histone
deacetylase inhibitors include, but are not limited to,
vorinostat.
[0148] For example, compounds of the present invention, or a
pharmaceutically acceptable composition thereof, are administered
in combination with chemotherapeutic agents to treat proliferative
diseases and cancer. Examples of known chemotherapeutic agents
include, but are not limited to, doxorubicin, dexamethasone,
vincristine, cyclophosphamide, fluorouracil, topotecan,
interferons, platinum derivatives, taxanes (e.g., paclitaxel,
docetaxel), vinca alkaloids (e.g., vinblastine), anthracyclines
(e.g., doxorubicin), epipodophyllotoxins (e.g., etoposide),
cisplatin, an mTOR inhibitor (e.g., a rapamycin), methotrexate,
actinomycin D, dolastatin 10, colchicine, trimetrexate, metoprine,
cyclosporine, daunorubicin, teniposide, amphotericin, alkylating
agents (e.g., chlorambucil), 5-fluorouracil, camptothecin,
cisplatin, metronidazole, and imatinib mesylate, among others. In
other embodiments, a compound of the present invention is
administered in combination with a biologic agent, such as
bevacizumab or panitumumab.
[0149] In certain embodiments, compounds of the present invention,
or a pharmaceutically acceptable composition thereof, are
administered in combination with an antiproliferative or
chemotherapeutic agent selected from any one or more of abarelix,
aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine,
amifostine, anastrozole, arsenic trioxide, asparaginase,
azacitidine, BCG live, bevacuzimab, fluorouracil, bexarotene,
bleomycin, bortezomib, busulfan, calusterone, capecitabine,
camptothecin, carboplatin, carmustine, cetuximab, chlorambucil,
cladribine, clofarabine, cyclophosphamide, cytarabine,
dactinomycin, darbepoetin alfa, daunorubicin, denileukin,
dexrazoxane, docetaxel, doxorubicin (neutral), doxorubicin
hydrochloride, dromostanolone propionate, epirubicin, epoetin alfa,
elotinib, estramustine, etoposide phosphate, etoposide, exemestane,
filgrastim, floxuridine, fludarabine, fulvestrant, gefitinib,
gemcitabine, gemtuzumab, goserelin acetate, histrelin acetate,
hydroxyurea, ibritumomab, idarubicin, ifosfamide, imatinib
mesylate, interferon alfa-2a, interferon alfa-2b, irinotecan,
lenalidomide, letrozole, leucovorin, leuprolide acetate,
levamisole, lomustine, megestrol acetate, melphalan,
mercaptopurine, 6-MP, mesna, methotrexate, methoxsalen, mitomycin
C, mitotane, mitoxantrone, nandrolone, nelarabine, nofetumomab,
oprelvekin, oxaliplatin, paclitaxel, palifermin, pamidronate,
pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium,
pentostatin, pipobroman, plicamycin, porfimer sodium, procarbazine,
quinacrine, rasburicase, rituximab, sargramostim, sorafenib,
streptozocin, sunitinib maleate, talc, tamoxifen, temozolomide,
teniposide, VM-26, testolactone, thioguanine, 6-TG, thiotepa,
topotecan, toremifene, tositumomab, trastuzumab, tretinoin, ATRA,
uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine,
zoledronate, or zoledronic acid.
[0150] Other examples of agents the inhibitors of this invention
may also be combined with include, without limitation: treatments
for Alzheimer's Disease such as donepezil hydrochloride and
rivastigmine; treatments for Parkinson's Disease such as
L-DOPA/carbidopa, entacapone, ropinrole, pramipexole,
bromocriptine, pergolide, trihexephendyl, and amantadine; agents
for treating multiple sclerosis (MS) such as beta interferon (e.g.,
Avonex.RTM. and Rebie), glatiramer acetate, and mitoxantrone;
treatments for asthma such as albuterol and montelukast sodium;
agents for treating schizophrenia such as zyprexa, risperdal,
seroquel, and haloperidol; anti-inflammatory agents such as
corticosteroids, TNF blockers, IL-1 RA, azathioprine,
cyclophosphamide, and sulfasalazine; immunomodulatory and
immunosuppressive agents such as cyclosporin, tacrolimus,
rapamycin, mycophenolate mofetil, interferons, corticosteroids,
cyclophophamide, azathioprine, and sulfasalazine; neurotrophic
factors such as acetylcholinesterase inhibitors, MAO inhibitors,
interferons, anti-convulsants, ion channel blockers, riluzole, and
anti-Parkinsonian agents; agents for treating cardiovascular
disease such as beta-blockers, ACE inhibitors, diuretics, nitrates,
calcium channel blockers, and statins; agents for treating liver
disease such as corticosteroids, cholestyramine, interferons, and
anti-viral agents; agents for treating blood disorders such as
corticosteroids, anti-leukemic agents, and growth factors; and
agents for treating immunodeficiency disorders such as gamma
globulin.
[0151] In certain embodiments, compounds of the present invention,
or a pharmaceutically acceptable composition thereof, are
administered in combination with a monoclonal antibody or an siRNA
therapeutic.
[0152] Those additional agents may be administered separately from
an inventive compound-containing composition, as part of a multiple
dosage regimen. Alternatively, those agents may be part of a single
dosage form, mixed together with a compound of this invention in a
single composition. If administered as part of a multiple dosage
regime, the two active agents may be submitted simultaneously,
sequentially or within a period of time from one another normally
within five hours from one another.
[0153] As used herein, the term "combination," "combined," and
related terms refers to the simultaneous or sequential
administration of therapeutic agents in accordance with this
invention. For example, a compound of the present invention may be
administered with another therapeutic agent simultaneously or
sequentially in separate unit dosage forms or together in a single
unit dosage form. Accordingly, the present invention provides a
single unit dosage form comprising a compound of formula I, an
additional therapeutic agent, and a pharmaceutically acceptable
carrier, adjuvant, or vehicle.
[0154] The amount of both an inventive compound and additional
therapeutic agent (in those compositions which comprise an
additional therapeutic agent as described above) that may be
combined with the carrier materials to produce a single dosage form
will vary depending upon the host treated and the particular mode
of administration. In certain embodiments, compositions of this
invention are formulated such that a dosage of between 0.01-100
mg/kg body weight/day of an inventive can be administered.
[0155] In those compositions which comprise an additional
therapeutic agent, that additional therapeutic agent and the
compound of this invention may act synergistically. Therefore, the
amount of additional therapeutic agent in such compositions will be
less than that required in a monotherapy utilizing only that
therapeutic agent. In certain embodiments, in such compositions a
dosage of between 0.01-1,000 .mu.g/kg body weight/day of the
additional therapeutic agent can be administered.
[0156] The amount of additional therapeutic agent present in the
compositions of this invention will be no more than the amount that
would normally be administered in a composition comprising that
therapeutic agent as the only active agent. In certain embodiments,
the amount of additional therapeutic agent in the presently
disclosed compositions will range from about 50% to 100% of the
amount normally present in a composition comprising that agent as
the only therapeutically active agent.
[0157] The compounds of this invention, or pharmaceutical
compositions thereof, may also be incorporated into compositions
for coating an implantable medical device, such as prostheses,
artificial valves, vascular grafts, stents and catheters. Vascular
stents, for example, have been used to overcome restenosis
(re-narrowing of the vessel wall after injury). However, patients
using stents or other implantable devices risk clot formation or
platelet activation. These unwanted effects may be prevented or
mitigated by pre-coating the device with a pharmaceutically
acceptable composition comprising an inhibitor of a histone
demethylase (e.g. Jumonji domain containing protein, e.g. JMJD2,
JMJD2B, JARID1A, JARID1B, PLU-1e.g. GASC1). Implantable devices
coated with a compound of this invention are another embodiment of
the present invention.
EXEMPLIFICATION
[0158] As depicted in the Examples below, in certain exemplary
embodiments, compounds are prepared according to the following
general procedures. It will be appreciated that, although the
general methods depict the synthesis of certain compounds of the
present invention, the following general methods, and other methods
known to one of ordinary skill in the art, can be applied to all
compounds and subclasses and species of each of these compounds, as
described herein.
Example 1
Synthesis of ethyl 2-acetylpentanoate
##STR00097##
[0160] Ethyl 2-acetylpentanoate was synthesized according to Beddow
et al, Org. Biomol. Chem. 5: 2812-2825 (2007). To a solution of
t-BuOK (11.8 g, 0.11 mol) in THF (150 mL) was added ethyl
3-oxobutanoate (13 g, 0.1 mol) dropwise at 0.degree. C., after
stirred for 30 minutes, 1-bromopropane (12.3 g, 0.1 mmol) was added
dropwise and the mixture was refluxed for 16 h. The reaction
mixture was quenched by water, extracted with EtOAc (100
mL.times.2), combined organic layer was dried over anhydrous
Na.sub.2SO.sub.4, evaporated, purified by column chromatography to
give the expected compound ethyl 2-acetylpentaoate (8.5 g, 49%) as
colorless oil. m/z (ESI) 173 [M+H].sup.+.
Example 2
[0161] By a similar method to Example 1, using the appropriate
starting materials, the compounds in Table 2 were prepared.
TABLE-US-00003 TABLE 2 Compound name Structure Data ethyl
2-acetyl-4- methylpentanoate ##STR00098## m/z (ESI) 187 [M +
H].sup.+ ethyl 2- acetylpent-4- enoate ##STR00099## m/z (ESI) 171
[M + H].sup.+ ethyl 2-acetyl-4- methoxybutanoate ##STR00100## m/z
(ESI) 189 [M + H].sup.+ ethyl 2- acetylpent-4- ynoate ##STR00101##
m/z (ESI) 169 [M + H].sup.+ ethyl 2-ethyl- 4,4,4-trifluoro-3-
oxobutanoate ##STR00102## m/z (ESI) 213 [M + H].sup.+ ethyl
4-ethoxy-2- ethyl-3- oxobutanoate ##STR00103## m/z (ESI) 203 [M +
H].sup.+ ethyl 2-(2- methoxyacetyl) pent-4-enoate ##STR00104## m/z
(ESI) 201 [M + H].sup.+ ethyl 2- benzoylbutanoate ##STR00105## m/z
(ESI) 221 [M + H].sup.+ ethyl 2-(furan-2- carbonyl) butanoate
##STR00106## m/z (ESI) 211 [M + H].sup.+ 3-ethylpentane- 2,4-dione
##STR00107## m/z (ESI) 129 [M + H].sup.+
Example 3
Synthesis of ethyl 2-ethyl-4-methoxy-3-oxobutanoate
##STR00108##
[0163] Ethyl 2-ethyl-4-methoxy-3-oxobutanoate was prepared
according to WO 98/43968. Zinc (2 g, 30 mmol), methoxyacetonitrile
(1.42 g, 20 mmol) and a catalytic amount of mercuric chloride in
toluene (50 mL) were heated to reflux. Ethyl 2-bromobutanoate (5.85
g, 30 mmol) was added dropwise, then reflux continued for a hour,
and cooled to a room temperature. 10% Aqueous sulfuric solution
(16.5 mL) was added, and the organic layer was separated. The
aqueous layer was further extracted with ethyl ether and the
combined organic layers washed with water and saturated sodium
bicarbonate solution, then dried over anhydrous magnesium sulfate
and concentrated in vacuo. The residue was purified by column
chromatography to give the product as yellow oil (1.7 g, 45%).
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 4.14 (q, J=7.2 Hz, 2H),
4.07 (d, J=3.6 Hz, 2H), 3.45 (t, J=7.2 Hz, 1H), 3.37 (s, MI), 1.85
(m, 2H), 1.22 (t, J=7.2 Hz, 3H). 0.90 (t, J=7.5 Hz, 3H); m/z (ESI)
189 [M+H].sup.+.
Example 4
[0164] By a similar method to Example 3, using the appropriate
starting materials, the compounds in Table 3 were prepared and
isolated.
TABLE-US-00004 TABLE 3 Compound name Structure Data ethyl 2-(2-
methoxyacetyl) pentanoate ##STR00109## m/z (ESI) 225 [M + Na.sup.+]
ethyl 2-ethyl-3- oxo-4- phenylbutanoate ##STR00110## m/z (ESI) 257
[M + Na.sup.+]
Example 5
Synthesis of diethyl 2-ethyl-3-oxosuccinate
##STR00111##
[0166] Diethyl-2-ethyl-3-oxosuccinate was prepared according to
Soloway et al, J. Org. Chem. 69: 2677-2678 (1947). To a mixture of
NaH (60%, 12 g, 300 mmol) and diethyl oxalate (43.8 g, 300 mmol) in
ether (100 mL), ethyl butyrate (18 g, 150 mmol) was added. The
reaction mixture was refluxed over night. After cooling to room
temperature water was added, the mixture was extracted with EtOAc.
The organic layer was dried over with Na.sub.2SO.sub.4 and
evaporated in vacuo. The residue was purified by column
chromatography to give diethyl-2-ethyl-3-oxosuccinate (8 g, 24%) as
light oil. m/z (ESI) 217 [M+H].sup.+.
Example 6
Synthesis of ethyl 2-ethyl-6-methoxy-3-oxohexanoate
##STR00112##
[0168] Ethyl 2-ethyl-6-methoxy-3-oxohexanoate was prepared
according to WO2006124490.
Synthesis of ethyl 6-methoxy-3-oxohexanoate
[0169] To a solution of ethyl 3-oxobutanoate (1.3 g, 10 mol) in THF
(50 mL) was added NaH (60%, 480 mg, 12 mmol) at 0.degree. C. After
stirring under N.sub.2 at 0.degree. C. for 0.5 h, n-BuLi (4 mL, 10
mmol) was added at 0.degree. C. and then the solution of the
mixture was cooled to -25.degree. C. After 1-bromo-2-methoxyethane
(1.39 g, 10 mmol) was added, the solution of the mixture was
stirring for overnight at room temperature. The mixture was
evaporated in vacuo, purified by column chromatography to give
ethyl 6-methoxy-3-oxohexanoate (0.65 g, 34.5%). m/z (ESI) 211
[M+Na].sup.+.
Synthesis of ethyl 2-ethyl-6-methoxy-3-oxohexanoate
[0170] To a solution of ethyl 6-methoxy-3-oxohexanoate (650 mg,
3.45 mmol) in THF (50 mL), .sup.tBuOK (406 mg, 3.63 mmol) was added
at 0.degree. C. and then the solution of the mixture was stirring
for 30 min at 0.degree. C., followed by refluxing overnight. The
mixture was evaporated in vacuo, purified by column chromatography
to give ethyl 2-ethyl-6-methoxy-3-oxohexanoate (400 mg, 53.6%). m/z
(ESI) 217 [M+H].sup.+.
Example 7
Synthesis of
6-cyclopropyl-5-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-carbo-
nitrile
##STR00113##
[0171] Synthesis of 2-cyclopropyl-3-oxobutanenitrile
##STR00114##
[0173] To a solution of 2-cyclopropylacetonitrile (1.17 g, 14.4
mmol) in THF (10 mL) was added LDA (8.7 ml, 17.3 mmol) dropwise at
-78.degree. C. under N.sub.2. After stirred for 60 mins,
(CH.sub.3O).sub.2O (1.12 g, 14.4 mmol) was added dropwise at
-78.degree. C. and the mixture was stirred for 2 h at ambient
temperature. The reaction mixture was quenched by the aqueous of
HCl (2N), extracted with ethyl acetate (30 ml.times.3), combined
organic layer was dried over anhydrous Na.sub.2SO.sub.4 and
evaporated, purified by column chromatography to give
2-cyclopropyl-3-oxobutanenitrile as yellow oil. m/z (ESI) 124
[M+H].sup.+.
Synthesis of ethyl 2-cyclopropyl-3-oxobutanoate
##STR00115##
[0175] To a solution of 2-cyclopropyl-3-oxobutanenitrile (600 mg, 5
mmol) in EtOH (10 mL) was added acetyl chloride (3 mL) dropwise at
0.degree. C. After stirred for 16 h, EtOH was removed, the mixture
was added concentrated HCl (1 mL) and EtOH (10 mL), and stirred for
4 h at 40.degree. C. The mixture was quenched by water and
extracted with ethyl acetate (20 mL.times.3), combined organic
layer was dried over anhydrous Na.sub.2SO.sub.4 and evaporated,
purified by column chromatography to give ethyl
2-cyclopropyl-3-oxobutanoate (30 mg, 10%) as yellow oil. m/z (ESI)
171 [M+H].sup.+.
Example 8
Synthesis of 5-amino-3-ethyl-1H-pyrazole-4-carbonitrile
##STR00116##
[0177] 5-Amino-3-ethyl-1H-pyrazole-4-carbonitrile was prepared in a
manner substantially similar to that described in WO2005070916 and
US2006135526.
Synthesis of 2-(1-methoxypropylidene)malononitrile
##STR00117##
[0179] A mixture of malononitrile (180 g, 1.02 mol,) and triethyl
orthopropionate (66 g, 1 mol) was refluxed for 3 h. The reaction
mixture was distilled under vacuum to give the expected compound
12-1-a (60 g, 40%) as pale yellow oil. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 4.46 (q, J=6.9 Hz, 2H), 2.65 (q, J=7.5 Hz, 2H),
1.45 (t, J=6.9 Hz, 3H), 1.26 (t, J=7.5 Hz, 3H).
Synthesis of 5-amino-3-ethyl-1H-pyrazole-4-carbonitrile
##STR00118##
[0181] A solution of 2-(1-methoxypropylidene)malononitrile (10 g,
0.067 mol) in EtOH (50 mL) was added dropwise into the solution of
hydrazine monohydrate (6.8 ml, 0.134 mol) in EtOH (100 mL) at
0.degree. C. for 30 min After stirred for 3 h at 90.degree. C., the
mixture was concentrated and purified by column chromatography to
give the expected compound
5-amino-3-ethyl-1H-pyrazole-4-carbonitrile (5 g, 60% yield) as
yellow solid. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 4.40 (s,
2H), 2.69 (q, J=7.5 Hz, 2H), 1.29 (t, J=7.5 Hz, 3H).
Example 9
##STR00119##
[0182] General Procedure
[0183] A mixture of a cyanopyrazole (0.86 mmol), a beta-ketoester
(293 mg, 1.72 mmol) and acetic acid (3 mL) is stirred at 80.degree.
C. for 1.5 hours. The mixture is cooled to room temperature. The
solvent is removed in vacuo. The residue is purified by silica gel
column chromatography to yield the desired compound.
[0184] Using the general procedure above and the appropriate
starting materials, the compounds in Table 4 were prepared.
TABLE-US-00005 TABLE 4 Compound Name Structure Data 2-ethyl-9-oxo-
4,5,6,7,8,9- hexahydropyrazolo[5,1- b]quinazoline-3- carbonitrile
##STR00120## .sup.1H NMR (300 MHz, DMSO) .delta. 12.84 (brs, 1H),
2.74 (q, J = 6.9 Hz, 2H,), 2.62 (m, 2H), 2.40 (m, 2H) 1.71 (m, 4H),
1.26 (t, J = 6.9 Hz, 3H); m/z (ESI) 243 [M + H].sup.+.
2-methyl-9-oxo- 4,5,6,7,8,9- hexahydropyrazolo[5,1-
b]quinazoline-3- carbonitrile ##STR00121## .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 2.72 (m, 2H), 2.60 (m, 2H), 2.43 (s, 3H), 1.81
(m, 4H). m/z (ESI) 229 [M + H].sup.+. 9-oxo-4,5,6,7,8,9-
hexahydropyrazolo[5,1- b]quinazoline-3- carbonitrile ##STR00122##
.sup.1H NMR (300 MHz, DMSO-d6) .delta. 13.07 (s, 1H), 8.32 (s, 1H),
2.66 (m, 2H), 2.42 (m, 2H), 1.72 (m, 4H). 6-isopropyl-2,5-
dimethyl-7-oxo-4,7- dihydropyrazolo[1,5- a]pyrimidine-3-
carbonitrile ##STR00123## .sup.1H-NMR (300 MHz, CD.sub.3COCD.sub.3)
.delta. 3.11 (m, 1H), 2.49 (s, 3H), 2.36 ( s, 1H) 1.34 (d, J = 6.9
Hz, 6H); m/z (ESI) 231 [M + H].sup.+. 6-propyl-2,5-dimethyl-
7-oxo-4,7- dihydropyrazolo[1,5- a]pyrimidine-3- carbonitrile
##STR00124## .sup.1H NMR (300 MHz, CD.sub.3COCD.sub.3) .delta. 2.54
(t, J = 7.5 Hz, 2H), 2.49 (s, 3H), 2.36 (s, 3H), 2.06 (m, 2H), 1.55
(m, 2H), 0.96 (t, J = 7.5 Hz, 3H); m/z (ESI) 231 [M + H].sup.+.
6-isopropyl-5-methyl- 7-oxo-4,7- dihydropyrazolo[1,5-
a]pyrimidine-3- carbonitrile ##STR00125## .sup.1H NMR (300 MHz,
DMSO-d6) .delta. 7.93 (s, 1H), 3.02 (m, 1H), 2.26 (s, 3H), 1.27 (d,
J = 7.2 Hz, 6H); m/z (ESI) 217 [M + H].sup.+. 6-propyl-5-methyl-7-
oxo-4,7- dihydropyrazolo[1,5- a]pyrimidine-3- carbonitrile
##STR00126## .sup.1H NMR (300 MHz, DMSO-d6) .delta. 13.07 (s, 1H),
8.35 (s, 1H), 2.47 (m, 2H), 2.38 (s, 3H), 1.46 (q, J = 7.5 Hz, 2H),
0.92 (t, J = 7.5 Hz, 3H); m/z (ESI) 217 [M + H].sup.+. 6-ethyl-5-
(methoxymethyl)-2- methyl-7-oxo-4,7- dihydropyrazolo[1,5-
a]pyrimidine-3- carbonitrile ##STR00127## .sup.1H NMR (300 MHz,
DMSO) .delta. 4.26 (s, 2H), 3.27 (s, 3H), 2.48 (m, 2H), 2.29 (s,
3H), 1.01 (t, J = 7.2 Hz, 3H); m/z (ESI) 247 [M + H].sup.+.
6-ethyl-5- (methoxymethyl)-7- oxo-4,7- dihydropyrazolo[1,5-
a]pyrimidine-3- carbonitrile ##STR00128## .sup.1H NMR (300 MHz,
CD.sub.3COCD.sub.3) .delta. 7.93 (s, 1H), 4.40 (s, 2H), 3.34 (s,
3H), 2.65 (q, J = 7.5 Hz, 2H), 1.12 (t, J = 7.5 Hz, 3H); m/z (ESI)
233 [M + H].sup.+. ethyl 3-cyano-6-ethyl- 2-methyl-7-oxo-4,7-
dihydropyrazolo[1,5- a]pyrimidine-5- carboxylate ##STR00129##
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 4.52 (q, J = 7.2 Hz, 2H),
2.82 (q, J = 7.2 Hz, 2H), 2.50 (s, 3H), 1.47 (t, J = 7.2 Hz, 3H),
1.21 (t, J = 7.5 Hz, 3H); m/z (ESI) 275 [M + H].sup.+. ethyl
3-cyano-6-ethyl- 7-oxo-4,7- dihydropyrazolo[1,5- a]pyrimidine-5-
carboxylate ##STR00130## .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
8.26 (s, 1H), 4.44 (q, J = 7.2 Hz, 2H), 2.70 (q, J = 6.9 Hz, 2H),
1.42 (t, J = 7.2 Hz, 3H), 1.17 (t, J = 7.2 Hz, 3H); m/z (ESI) 262
[M + H].sup.+. 6-ethyl-5-methyl-7- oxo-4,7-dihydro-
[1,2,3]triazolo[1,5- a]pyrimidine-3- carbonitrile ##STR00131##
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 2.82 (s, 3H), 2.69 (q, J
= 7.5 Hz, 2H), 1.20 (t, J = 7.5 Hz, 3H); m/z (ESI) 204 [M +
H].sup.+. 5-methyl-7-oxo-6- propyl-4,7-dihydro-
[1,2,3]triazolo[1,5- a]pyrimidine-3- carbonitrile ##STR00132##
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 2.82 (s, 3H), 2.65 (t, J
= 7.5 Hz, 3H), 1.63 (m, 2H), 1.06 (t, J = 7.2 Hz, 3H); m/z (ESI)
218 [M + H].sup.+. 5-(methoxymethyl)-7- oxo-6-propyl-4,7-
dihydropyrazolo[1,5- a]pyrimidine-3- carbonitrile ##STR00133##
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.29 (s, 1H), 4.78 (s,
2H), 3.59 (s, 3H), 2.63 (m, 2H), 1.64 (m, 2I-I), 1.05 (t, J = 7.5
Hz, 3H); m/z (ESI) 247 [M + H].sup.+. 5-(ethoxymethyl)-6-
ethyl-7-oxo-4,7- dihydropyrazolo[1,5- a]pyrimidine-3- carbonitrile
##STR00134## .sup.1HNMR (300 MHz, CD.sub.3OD) .delta. 8.27 (s, 1H),
4.64 (s, 2H), 3.71 (q, J = 7.2 Hz, 2H), 2.69 (q, J = 7.5 Hz, 2H),
1.32 (t, J = 7.2 Hz, 3H), 1.20 (t, J = 7.5 Hz, 3H); m/z (ESI) 247
[M + H].sup.+. 6-(2-methoxyethyl)-5- methyl-7-oxo-4,7-
dihydropyrazolo[1,5- a]pyrimidine-3- carbonitrile ##STR00135##
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.26 (s, 1H), 3.62 (t, J
= 6.6 Hz, 2H), 3.39 (s, 3H), 2.93 (t, J = 6.6 Hz, 2H), 2.55 (s,
3H); m/z (ESI) 233 [M + H].sup.+. 6-ethyl-5-(3-
methoxypropyl)-7-oxo- 4,7- dihydropyrazolo[1,5- a]pyrimidine-3-
carbonitrile ##STR00136## .sup.1H NMR (300 MHz, CD.sub.3Cl) .delta.
11.87 (brs, 1H), 8.01 (s, 1H), 3.65 (m, 2H), 3.63 (s, 3H), 2.96 (m,
2H), 2.66 (q, J = 7.5 Hz, 2H), 2.08 (m, 2H), 1.13 (t, J = 7.5 Hz,
3H); m/z (ESI) 261 [M + H].sup.+. 5-methyl-7-oxo-6-
(prop-2-ynyl)-4,7- dihydropyrazolo[1,5- a]pyrimidine-3-
carbonitrile ##STR00137## .sup.1HNMR (300 MHz, CD.sub.3OD) .delta.
8.26 (s, 1H), 3.58 (d, J = 2.7 Hz, 2H), 2.59 (s, 3H), 2.37 (t, J =
2.7 Hz, 1H); m/z (ESI) 235 [M + Na].sup.+. 6-allyl-5-
(methoxymethyl)-7- oxo-4,7- dihydropyrazolo[1,5- a]pyrimidine-3-
carbonitrile ##STR00138## .sup.1HNMR (300 MHz, CD.sub.3Cl) .delta.
9.38 (brs, 1H), 8.07 (s, 1H), 5.83 (m, 1H), 5.08 (m, 2H), 4.55 (s,
2H), 3.63 (s, 3H), 3.29 (d, J = 6.3 Hz, 2H); m/z (ESI) 267 [M +
Na]+. 6-ethyl-7-oxo-5- phenyl-4,7- dihydropyrazolo[1,5-
a]pyrimidine-3- carbonitrile ##STR00139## .sup.1HNMR (300 MHz,
CD.sub.3OD) .delta. 8.30 (s, 1H), 7.64-7.57 (5H), 2.49 (q, J = 7.2
Hz, 2H), 1.14 (t, J = 7.2 Hz, 3H); m/z (ESI) 265 [M + H].sup.+.
5-benzyl-6-ethyl-7-oxo- 4,7- dihydropyrazolo[1,5- a]pyrimidine-3-
carbonitrile ##STR00140## .sup.1HNMR (300 MHz, CD.sub.3OD) .delta.
8.26 (s, 1H), 7.39-7.27 (5H), 4.20 (s, 2H), 2.65 (q, J = 7.5 Hz,
2H), 1.03 (t, J = 7.5 Hz, 3H); m/z (ESI) 279 [M + H].sup.+. benzyl
3-cyano-6-ethyl- 7-oxo-4,7- dihydropyrazolo[1,5- a]pyrimidine-5-
carboxylate ##STR00141## .sup.1HNMR (300 MHz, CD.sub.3OD) .delta.
8.22 (s, 1H), 7.56 ? 7.42 (5H), 5.51 (s, 2H), 2.79 (q, J = 7.2 Hz,
2H), 1.12 (t,J = 7.2 Hz, 3H); m/z (ESI) 245 [M + H].sup.+. ethyl
3-cyano-7-oxo-6- propyl-4,7- dihydropyrazolo[1,5- a]pyrimidine-5-
carboxylate ##STR00142## .sup.1HNMR (300 MI-lz, CD.sub.3OD) .delta.
8.17 (s, 1H), 4.43 (q, J = 7.2 Hz, 2H), 2.62 (t, J = 7.5 Hz, 2H),
1.62 (m, 2H), 1.44 (t, J = 7.2 Hz, 3H), 0.97 (t, J = 7.5 Hz, 3H);
m/z (ESI) 275 [M + H].sup.+. 6-ethyl-7-oxo-5-
(trifluoromethyl)-4,7- dihydropyrazolo[1,5- a]pyrimidine-3-
carbonitrile ##STR00143## .sup.1HNMR (300 MHz, CD.sub.3OD) .delta.
8.08 (s, 1H), 2.63 (q, J = 7.2 Hz, 2H), 1.06 (t, J = 7.2 Hz, 3H);
m/z (ESI) 257 [M + H].sup.+. 6-allyl-5-methyl-7-oxo- 4,7-
dihydropyrazolo[1,5- a]pyrimidine-3- carbonitrile ##STR00144##
.sup.1H NMR (300 MHz, DMSO) .delta. 13.18 1H), 8.37 (s, 1H), 5.83
(m, 1H), 5.02 (m, 2H), 3.29 (m, 2H), 2.35 (s, 3H); m/z (ESI) 215 [M
+ H].sup.+. 6-isobutyl-5-methyl-7- oxo-4,7- dihydropyrazolo[1,5-
a]pyrimidine-3- carbonitrile ##STR00145## .sup.1H NMR (300 MHz,
DMSO-d6) .delta. 13.16 (brs, 1H), 8.30 (s, 1H), 2.39-2.34 (5H),
1.86 (m, 1H), 0.85 (d, J = 6.6 Hz, 6H); m/z (ESI) 231 [M + H].
6-ethyl-5-(furan-3-yl)- 7-oxo-4,7- dihydropyrazolo[1,5-
a]pyrimidine-3- carbonitrile ##STR00146## .sup.1HNMR (300 MHz,
CD.sub.3OD) .delta. 8.28 (s, 1H), 8.05 (dd, J = 1.8 Hz, 0.9 Hz,
1H), 7.79 (m, 1H), 6.84 (dd, J = 1.8 Hz, 0.9 Hz, 1H), 2.66 (q, J =
7.5 Hz, 2H), 1.20 (t, J = 7.2 Hz, 3H); m/z (ESI) 277 [M +
Na].sup.+. 7-oxo-5- (trifluoromethyl)-4,7- dihydropyrazolo[1,5-
a]pyrimidine-3- carbonitrile ##STR00147## .sup.1HNMR (300 MHz,
CD.sub.3OD) .delta. 8.36 (s, 1H), 6.53 (s, 1H); m/z (ESI) 251 [M +
Na].sup.+. 6-cyclopropyl-5- methyl-7-oxo-4,7- dihydropyrazolo[1,5-
a]pyrimidine-3- carbonitrile ##STR00148## .sup.1HNMR (300 MHz,
CD.sub.3OD) .delta. 8.19 (s, 1H), 2.59 (s, 3H), 1.33 (m, 1H), 1.01
(m, 2H), 0.73 (m, 2); m/z (ESI) 237 [M + Na].sup.+.
Example 10
Synthesis of
4-methyl-9-oxo-4,5,6,7,8,9-hexahydropyrazolo[5,1-b]quinazoline-3-carbonit-
rile
##STR00149##
[0186]
9-oxo-4,5,6,7,8,9-hexahydropyrazolo[5,1-b]quinazoline-3-carbonitril-
e (50 mg, 0.23 mmol) was dissolved in DMF (1 mL), potassium
carbonate (63 mg, 0.46 mmol) was added followed by iodomethane (36
mg, 0.26 mmol). The mixture was stirred at room temperature
overnight, then diluted with water (10 mL) and extracted with EtOAc
(5 mL.times.3). The combined organic layer was dried and
concentrated to dryness. The residue was recrystallized from
methanol to afford 12-3 (25 mg, 47%). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.07 (s, 1H), 3.94 (s, 3H), 2.69 (m, 4H), 1.92
(m, 2H), 1.76 (m, 2H); m/z (ESI) 229 [M+H].sup.+.
[0187] By a similar method to Example 10, using the appropriate
starting material,
6-ethyl-4,5-dimethyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-
-3-carbonitrile was prepared and isolated.
TABLE-US-00006 Compound Name Structure Data 6-ethyl-4,5-
dimethyl-7-oxo-4,7- dihydropyrazolo[1,5- a]pyrimidine-3-
carbonitrile ##STR00150## .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
8.09 (s, 1H), 4.00 (s, 3H), 2.71 (q, J = 7.5 Hz, 2H), 2.48 (s, 3H),
1.14 (t, J = 7.5 Hz, 3H); m/z (ESI) 217 [M + H].sup.+.
Example 11
Synthesis of
4-(2-bromoethyl)-6-ethyl-5-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimid-
ine-3-carbonitrile
##STR00151##
[0189] To a solution of tetrabutylammoniunbromide (557 mg, 1.73
mmol) in water (20 mL), NaOH (76 mg, 1.9 mmol) was added at ambient
temperature, followed by
6-ethyl-5-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-carbonitril-
e (350 mg, 1.73 mmol) in CHCl.sub.3 (20 mL), the mixture continued
to stir for 10 min. The organic phase was separated, the aqueous
phase was extracted with CHCl.sub.3 (10 mL). The combined organic
phase was dried over anhydrous Na.sub.2SO.sub.4, evaporated to give
white solid. The solid was dissolved in CH.sub.3CN (10 mL),
followed by 1,2-dibromoethane (360 mg, 2 mmol). The mixture
continued to stir for 16 h at refluxing temperature, followed by
concentration and purification with column chromatography to give
4-(2-bromoethyl)-6-ethyl-5-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimid-
ine-3-carbonitrile (200 mg, 40%). m/z (ESI) 309 [M+H].sup.+.
Example 12
Synthesis of
4-(2-(dimethylamino)ethyl)-6-ethyl-5-methyl-7-oxo-4,7-dihydropyrazolo[1,5-
-a]pyrimidine-3-carbonitrile
##STR00152##
[0191] To a solution of
4-(2-bromoethyl)-6-ethyl-5-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimid-
ine-3-carbonitrile (200 mg, 0.65 mmol) in DMF (10 mL) were added
potassium carbonate (446 mg, 3.24 mmol) and dimethylamine
hydrochloride (155 mg, 1.94 mmol) consequently. The mixture was
stirred at room temperature overnight, then diluted with water (10
mL) and extracted with EtOAc (10 mL.times.3). The combined organic
layer was dried with Na.sub.2SO.sub.4, concentrated and purified by
column chromatography to afford
4-(2-(dimethylamino)ethyl)-6-ethyl-5-methyl-7-oxo-4,7-dihydropyrazolo[1,5-
-a]pyrimidine-3-carbonitrile (12 mg, 10%). .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.29 (s, 1H), 4.52 (t, J=7.2 Hz, 2H), 2.83 (t,
J=7.2 Hz, 2H), 2.72 (q, J=7.5 Hz, 2H), 2.61 (s, 3H), 2.39 (s, 6H)
1.15 (t, J=7.5 Hz, 3H); m/z (ESI) 274 [M+H].sup.+.
Example 13
Synthesis of
9-methoxy-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-3-carbonitrile
##STR00153##
[0192] Synthesis of
9-chloro-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-3-carbonitrile
##STR00154##
[0194] To a solution of
9-oxo-4,5,6,7,8,9-hexahydropyrazolo[5,1-b]quinazoline-3-carbonitrile
(1.0 g, 4.6 mmol) in dry POCl.sub.3 (20 mL) was added pyridine (0.2
mL) under a nitrogen atmosphere, the mixture was heated to
110.degree. C. overnight. After cooling down to room temperature,
the solvent was removed in vacuo, and the residue was purified by
silica gel column chromatography to yield the desired compound
9-chloro-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-3-carbonitrile
(0.6 g, yield 56%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.36
(s, 1H), 3.10 (m, 2H), 2.93 (m, 2H), 1.96 (m, 4H).
Synthesis of
9-methoxy-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-3-carbonitrile
##STR00155##
[0196] A mixture of
9-chloro-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-3-carbonitrile
(100 mg, 0.43 mmol) and MeONa (48 mg, 0.86 mmol) in MeOH (4 mL) was
stirred at room temperature for 1.5 hour, and the reaction was
quenched by saturated NH.sub.4Cl, and extracted with DCM (50
mL.times.3). The organic layer was washed by brine, dried over
Na.sub.2SO.sub.4, concentrated in vacuo, and crude product was
purified by preparation thick layer chromatography to yield the
desired compound
9-methoxy-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-3-carbonitrile
(20 mg, yield 21%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.25 (s, 1H), 4.44 (s, 3H), 3.02 (t, J=6.3 Hz,
2H), 2.80 (t, J=6.3 Hz, 2H), 1.89 (m, 4H); m/z (ESI) 229
[M+H].sup.+.
[0197] By a similar method to Example 14, using the appropriate
starting material,
6-ethyl-7-methoxy-5-methylpyrazolo[1,5-a]pyrimidine-3-carbonitr-
ile was prepared and isolated.
TABLE-US-00007 Compound Name Structure Data 6-ethyl-7-methoxy- 5-
methylpyrazolo[1,5- a]pyrimidine-3- carbonitrile ##STR00156##
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.28 (s, 1H), 4.43 (s,
3H), 2.77 (q, J = 7.8 Hz, 2H), 2.70 (s, 3H), 1.23 (t, J = 7.5 Hz,
3H); m/z (ESI) 217 [M].sup.+
Example 14
Synthesis of
9-(2-hydroxyethoxy)-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-3
carbonitrile
##STR00157##
[0199] A mixture of
9-chloro-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-3-carbonitrile
(100 mg, 0.43 mmol), Et.sub.3N (0.7 mL) in ethane-1,2-diol (2 mL)
was stirred at room temperature overnight. The reaction was diluted
with EtOAc (2.0 mL). The addition of hexane (2 mL) led to the
precipitation of the product. The product was filtered, washed with
EtOAc to yield the desired compound
9-(2-hydroxyethoxy)-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-
-3 carbonitrile (2.0 mg, yield 2%) as a white solid. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 8.69 (s, 1H), 4.94 (t, J=5.1 Hz, 1H),
4.79 (t, J=4.5 Hz, 2H), 3.73 (m, 2H), 2.94 (t, J=5.7 Hz, 2H), 2.82
(t, J=6.3 Hz, 2H), 1.83 (m, 4H); m/z (ESI) 259 [M+H].sup.+.
Example 15
Synthesis of
9-(2-methoxyethoxy)-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-3-carbon-
itrile
##STR00158##
[0201] A mixture of
9-chloro-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-3-carbonitrile
(100 mg, 0.43 mmol), Et.sub.3N (0.7 mL) in 2-methoxyethanol (2 mL)
was stirred at 70.degree. C. overnight. The solvent was removed in
vacuo, and the crude product was purified by preparation thin layer
chromatography to yield the desired compound
9-(2-methoxyethoxy)-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazoline-3-carbon-
itrile (10.0 mg, yield 9%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.24 (s, 1H), 4.92 (m, 2H), 3.75 (m, 2H), 3.37
(s, 3H), 3.03 (t, J=6.0 Hz, 2H), 2.84 (t, J=6.0 Hz, 2H), 1.91 (m,
41-1); m/z (ESI) 273 [M+H].sup.+.
Example 16
Synthesis of
3-cyano-6-ethyl-N-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-5-car-
boxamide
##STR00159##
[0202] Synthesis of
3-cyano-6-ethyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-5-carboxylic
acid
##STR00160##
[0204] To a solution of ethyl
3-cyano-6-ethyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-5-carboxylate
(500 mg, 1.92 mmol) in EtOH (50 mL) and H.sub.2O (5 mL) at
0.degree. C., LiOH (230 mg, 9.6 mmol) was added in portions. The
reaction mixture was stirred at room temperature for 2 h. After
removal of EtOH in vacuo, water (5 mL) was added, and the mixture
was filtered to produce
3-cyano-6-ethyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-5-carboxylic
acid (380 mg, 89%) as off-white solid. .sup.1H NMR (300 MHz,
DMSO-d6) .delta. 8.41 (s, 1H), 2.67 (q, J=6.9 Hz, 2H), 1.09 (t,
J=6.9 Hz, 3H).
Synthesis of
3-cyano-6-ethyl-N-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-5-car-
boxamide
##STR00161##
[0206] To a solution of
3-cyano-6-ethyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-5-carboxylic
acid (50 mg, 0.21 mmol), methanamine hydrochloride (15 mg, 0.21
mmol), PyBOP (105.20 mg, 0.21 mmol) and HOBT (41.85 mg, 0.31 mmol)
in DMF (2 mL) was added DIEA (129 mg, 1 mmol). The reaction mixture
was stirred at room temperature overnight. The mixture was diluted
with CH.sub.2Cl.sub.2 and purified by column chromatography to
produce
3-cyano-6-ethyl-N-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-5-car-
boxamide (16 mg, 31%) as off-white solid. .sup.1HNMR (300 MHz,
CD.sub.3OD) .delta. 8.16 (s, 1H), 2.95 (s, 3H), 2.79 (q, J=7.5 Hz,
2H), 1.20 (t, J=7.5 Hz, 3H); m/z (ESI) 246 [M+H].sup.+.
[0207] By a similar method as above, using the ethanamine
hydrochloride as reagent,
3-cyano-N,6-diethyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-5-
-carboxamide was prepared and isolated.
TABLE-US-00008 Compound Name Structure Data
3-cyano-N,6-diethyl-7-oxo- 4,7-dihydropyrazolo[1,5-
a]pyrimidine-5-carboxamide ##STR00162## .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.16 (s, 1H), 3.43 (q, J = 7.2 Hz, 2H), 2.77
(q, J = 7.5 Hz, 2H), 1.31 (t, J = 7.5 Hz, 3H), 1.21 (t, J = 7.2 Hz,
3H); m/z (ESI) 260 [M + H].sup.+.
Example 17
Synthesis of
4-benzyl-9-oxo-4,5,6,7,8,9-hexahydropyrazolo[5,1-b]quinazoline-3-carbonit-
rile
##STR00163##
[0209] To solution of
9-oxo-4,5,6,7,8,9-hexahydropyrazolo[5,1-b]quinazoline-3-carbonitrile
(50 mg, 0.23 mmol) in DMF (1 mL), potassium carbonate (63 mg, 0.46
mmol) and sodium iodide (5 mg, 3 mmol) were added followed by
benzyl bromide (43 mg, 0.25 mmol). The mixture was stirred at room
temperature overnight, then diluted with water (10 mL) and
extracted with EtOAc (5 mL.times.3). The combined organic layer was
dried and concentrated to dryness. The residue was purified by
thick layer chromatography on silica gel to afford
4-benzyl-9-oxo-4,5,6,7,8,9-hexahydropyrazolo[5,1-b]quinazoline-3-c-
arbonitrile (25 mg, 35%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
8.03 (s, 1H), 7.38 (m, 3H), 7.04 (d, J=5.6 Hz, 2H), 5.56 (s, 2H),
2.68 (m, 4H), 1.79 (m, 4H); m/z (ESI) 305 [M+H].sup.+.
Example 18
Synthesis of
9-oxo-4-phenyl-4,5,6,7,8,9-hexahydropyrazolo[5,1-b]quinazoline-3-carbonit-
rile
##STR00164##
[0211] To a solution of
9-oxo-4,5,6,7,8,9-hexahydropyrazolo[5,1-b]quinazoline-3-carbonitrile
(645 mg, 3 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added
Cu(OAc).sub.2, 4 A molecular sieve, Et.sub.3N (607 mg, 6 mmol) and
pyridine (474 mg, 6 mmol) consequently, the solution was stirred
for 2 days under O.sub.2. To the mixture was added NH.sub.3H.sub.2O
and adjusted to PH>8. After filtration, the solution was
concentrated and purified by column chromatography to give
9-oxo-4-phenyl-4,5,6,7,8,9-hexahydropyrazolo[5,1-b]quinazoline-3-carbonit-
rile (60 mg, 10%) as white solid. .sup.1H NMR (300 MHz, CD.sub.3Cl)
.delta. 7.99 (s, 1H), 7.52-7.36 (5H), 2.81 (m, 2H), 2.52 (m, 2H),
1.80 (m, 4H); m/z (ESI) 291 [M+H].sup.+.
Example 19
Synthesis of
6-ethyl-5-(hydroxymethyl)-2-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimi-
dine-3-carbonitrile
##STR00165##
[0213] To a solution of
6-ethyl-5-(methoxymethyl)-2-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimi-
dine-3-carbonitrile (50 mg, 0.2 mmol.) in CH.sub.2Cl.sub.2 (20 mL)
at -78.degree. C., BCl.sub.3 (1 mL, 1 mmol) was added dropwise, the
mixture continued to stir at -78.degree. C. for 2 h, then allowed
to room temperature for 16 h. The mixture was quenched by water,
and the aqueous phase was washed by CH.sub.2Cl.sub.2 (20
mL.times.3). After removal of water, the residue was washed with
CH.sub.2Cl.sub.2/MeOH (50 mL, V/V=20/1) and filtered. The solution
was concentrated and purified by column chromatography to give
6-ethyl-5-(hydroxymethyl)-2-methyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimi-
dine-3-carbonitrile (10 mg, 23% yield) as off-white solid. .sup.1H
NMR (300 MHz, CD.sub.3OD) .delta. 4.71 (s, 2H), 2.61 (m, 2H), 2.49
(s, 3H), 1.56 (t, J=7.2 Hz, 3H); m/z (ESI) 233 [M+H].sup.+.
[0214] By a similar method as above, using the appropriate starting
materials, the compounds in Table 5 were prepared and isolated.
TABLE-US-00009 TABLE 5 Compound Name Structure Data 6-ethyl-5-
(hydroxymethyl)-7- oxo-4,7- dihydropyrazolo[1,5- a]pyrimidine-3-
carbonitrile ##STR00166## .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
8.24 (s, 1H), 4.73 (s, 2H), 2.64 (m, 2H), 1.17 (t, J = 7.5 Hz, 3H);
m/z (ESI) 219 [M + H].sup.+ 5-(hydroxymethyl)- 7-oxo-6-propyl-4,7-
dihydropyrazolo[1,5- a]pyrimidine-3- carbonitrile ##STR00167##
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.30 (s, 1H), 4.78 (s,
2H), 2.63 (m, 2H), 1.64 (m, 2H), 1.06 (t, J = 7.5 Hz, 3H); m/z
(ESI) 233 [M + H].sup.+ 6-(2-hydroxyethyl)- 5-methyl-7-oxo-4,7-
dihydropyrazolo[1,5- a]pyrimidine-3- carbonitrile ##STR00168##
.sup.1H NMR (300 MHz, DMSO-d6) .delta. 13.08 (s, 1H), 8.35 (s, 1H),
4.61 (brs, 1H), 3.50 (m, 2H), 2.63 (t, J = 6.6 Hz, 2H), 2.39 (s,
3H); m/z (ESI) 219 [M + H].sup.+. 6-ethyl-5-(3- hydroxypropyl)-7-
oxo-4,7- dihydropyrazolo[1,5- a]pyrimidine-3- carbonitrile
##STR00169## .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.22 (s,
1H), 3.70 (t, J = 6.0 Hz, 2H), 2.86 (m, 2H), 2. 65 (q, J = 7.5 Hz,
2H), 1.93 (m, 2H), 1.19 (t, J = 7.5 Hz, 3H); m/z (ESI) 247 [M +
H].sup.+ 6-allyl-5- (hydroxymethyl)-7- oxo-4,7-
dihydropyrazolo[1,5- a]pyrimidine-3- carbonitrile ##STR00170##
.sup.1HNMR (300 MHz, CD.sub.3OD) .delta. 8.26 (s, 1H), 5.94 (m,
1H), 5.08 (m, 2H), 4.71 (s, 2H), 3.41 (d, J = 6.0 Hz, 2H); m/z
(ESI) 253 [M + Na].sup.+.
Example 20
Synthesis of
6-ethyl-5-oxo-5,7,8,9-tetrahydropyrazolo[1,5-a]pyrrolo[1,2-c]pyrimidine-1-
-carbonitrile
##STR00171##
[0216] To a solution of
6-ethyl-4,7-dihydro-5-(3-hydroxypropyl)-7-oxopyrazolo[1,5-a]pyrimidine-3--
carbonitrile (12 mg, 0.05 mmol) in THF (10 mL) was added DIAD (10
mg, 0.06 mmol) and PPh.sub.3 (16 mg, 0.06 mmol) at 0.degree. C. The
mixture was allowed to room temperature and stirred for overnight.
The mixture was evaporated in vacuo, purified by column
chromatography to give
6-ethyl-5-oxo-5,7,8,9-tetrahydropyrazolo[1,5-a]pyrrolo[1,2-c]pyrimidine-1-
-carbonitrile (5 mg, 43.8%). .sup.1H NMR (300 MHz, CD.sub.3OD)
.delta. 8.24 (s, 1H), 4.55 (t, J=7.5 Hz, 2H), 3.23 (t, J=7.8 Hz,
2H), 2.60 (q, J=7.5 Hz, 2H), 2.47 (m, 2H), 1.18 (t, J=7.5 Hz, 3H);
m/z (ESI) 229 [M+H].sup.+.
Example 21
Synthesis of
6-ethyl-5-formyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-carbonitril-
e
##STR00172##
[0218] To a solution of DMSO (780 mg, 5 mmol) in CH.sub.2Cl.sub.2
(20 mL) was added dropwise oxalyl dichloride (127 mg, 5 mmol) at
-78.degree. C., then continued to stir for 30 min.
6-ethyl-5-(hydroxymethyl)-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-ca-
rbonitrile (110 mg, 0.5 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added
dropwise. After stirring for 1 h, Et.sub.3N (1.5 g, 15 mmol) was
added dropwise and allowed to ambient temperature. The mixture was
quenched by water, extracted with CH.sub.2Cl.sub.2 (10 mL.times.3),
combined organic layer was dried over anhydrous Na.sub.2SO.sub.4
and evaporated. The crude was purified by column chromatography to
give
6-ethyl-5-formyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-carbonitril-
e (50 mg, 50%) as yellow solid. .sup.1H NMR (300 MHz, CD.sub.3Cl)
.delta. 10.12 (s, 1H), 8.32 (s, 1H), 3.04 (q, J=7.2 Hz, 2H), 1.23
(t, J=7.2 Hz, 3H); m/z (ESI) 217 [M+H].sup.+.
Example 22
Synthesis of
6-ethyl-5-(1-hydroxyethyl)-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-c-
arbonitrile
##STR00173##
[0220] To a solution of
6-ethyl-5-formyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-carbonitril-
e (50 mg, 0.23 mmol) in THF (10 mL) at -60.degree. C., CH.sub.3MgBr
(0.7 mL, 0.68 mmol) was added dropwise, and continued to stir for 1
h. The mixture was quenched by saturated NH.sub.4Cl, and extracted
by CH.sub.2Cl.sub.2 (20 mL.times.5). Combined organic layer was
dried over anhydrous Na.sub.2SO.sub.4 and evaporated. The residue
was purified by column chromatography to give
6-ethyl-5-(1-hydroxyethyl)-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-c-
arbonitrile (5 mg, 15%) as white solid. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.29 (s, 1H), 5.22 (m, 1H), 2.65 (m, 2H), 1.59
(d, J=6.6 Hz, 3H), 1.24 (t, J=7.5 Hz, 3H); m/z 233 [M+H].sup.+.
Example 23
Synthesis of
9-oxo-4,9-dihydropyrazolo[5,1-b]quinazoline-3-carbonitrile
##STR00174##
[0221] Synthesis of 2-acetamidobenzoic acid
##STR00175##
[0223] Anthranilic acid (100 g, 0.73 mol) in Ac.sub.2O (1000 mL)
was heated at 100.degree. C. for 2 h. The Ac.sub.2O was evaporated
in vacuo and the residue was washed with hexane to give the product
2-acetamidobenzoic acid (130 g, 100%) as off-white solid. .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 8.19 (m, 1H), 7.79 (m, 1H), 7.54
(m, 2H), 2.48 (s, 3H).
Synthesis of 3-amino-2-methylquinazolin-4(3H)-one
##STR00176##
[0225] A mixture of 2-acetamidobenzoic acid (6.0 g, 33.5 mmol) and
NH.sub.2NH.sub.2.H.sub.2O (5.9 g, 100 mmol) was stirred at
0.degree. C. for 10 min and heated to reflux for 30 min. After
removal of solvent, the residue was washed with ethanol to give
3-amino-2-methylquinazolin-4(3H)-one as off-white solid (1 g,
12.5%). .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.24 (m, 1H),
7.75 (m, 2H), 7.46 (m, 1H), 2.72 (s, 3H).
Synthesis of
9-oxo-4,9-dihydropyrazolo[5,1-b]quinazoline-3-carbaldehyde
##STR00177##
[0227] The Vilsmeier-Haack reaction was performed according to
Pandit, R. S.; Seshadri, S. Vilsmeier-Haack reaction. Indian. J.
Chem. 1973, 11(6), 532-537. To a solution of POCl.sub.3 (2.7 mL, 29
mmol) in DMF (5 mL), was added 3-amino-2-methylquinazolin-4(3H)-one
(1.0 g, 5.7 mmol) in DMF (10 mL) at 0.degree. C. Then the mixture
was heated at 70.degree. C. for 5 h and poured into crushed ice.
The resulting creamy solution was basified with NaHCO.sub.3 to pH=8
at 0.degree. C. when a bright yellow crystalline compound separated
out. It was filtered, washed with water. The solid was taken in
potassium carbonate solution (10%, 10 mL) and warmed at 60.degree.
C. for half an hour when a clear yellow solution was obtained. The
solution was neutralized to pH=5 with HCl (1N), and filtered to
give 9-oxo-4,9-dihydropyrazolo[5,1-b]quinazoline-3-carbaldehyde as
off-white solid (800 mg, 82.5%). .sup.1H NMR (300 MHz, DMSO-d6)
.delta. 9.90 (s, 1H), 8.44 (s, 1H), 8.24 (m, 1H), 7.86 (m, 2H),
7.42 (m, 1H).
Synthesis of
9-oxo-4,9-dihydropyrazolo[5,1-b]quinazoline-3-carbaldehyde
oxime
##STR00178##
[0229] 9-Oxo-4,9-dihydropyrazolo[5,1-b]quinazoline-3-carbaldehyde
(800 mg, 3.75 mmol) and hydroxylamine hydrochloride (250 mg, 3.6
mmol) were taken in EtOH (50 mL) and reflux for 3 h. The solvent
was removed in vacuo to give the crude product
9-oxo-4,9-dihydropyrazolo[5,1-b]quinazoline-3-carbaldehyde oxime
(700 mg, 82%) as off-white solid. .sup.1H NMR (300 MHz, DMSO-d6)
.delta. 12.48 (s, 1H), 11.35 (s, 1H), 8.57 (s, 1H), 8.21 (m, 1H),
7.82 (m, 1H), 7.52 (m, 1H), 7.34 (m, 1H).
Synthesis of
9-oxo-4,9-dihydropyrazolo[5,1-b]quinazoline-3-carbonitrile
##STR00179##
[0231] To a solution of
9-oxo-4,9-dihydropyrazolo[5,1-b]quinazoline-3-carbaldehyde oxime
(600 mg, 2.63 mmol) in dry CHCl.sub.3 (10 mL) was added phosphorus
oxychloride (0.5 mL, 5.5 mmol) and the mixture refluxed for 2 h.
After removal of CH.sub.3Cl, ice-cooled water was added followed by
sodium bicarbonate to adjust to pH around 8. Precipitate was
filtered, washed with water to give
9-oxo-4,9-dihydropyrazolo[5,1-b]quinazoline-3-carbonitrile (200 mg,
30%) as off-white solid. .sup.1H NMR (300 MHz, DMSO-d6) .delta.
13.33 (s, 1H), 8.43 (s, 1H), 8.23 (d, J=8.1 Hz, 1H), 7.86 (dd,
J=8.4 Hz, 6.9 Hz, 1H), 7.58 (d, J=8.1 Hz, 1H), 7.41 (dd, J=8.1 Hz,
7.2 Hz, 1H); m/z (ESI) 211 [M+H].sup.+.
Example 24
Synthesis of
3-ethyl-2-methyl-4-oxo-3,4-dihydropyrazolo[1,5-a][1,3,5]triazine-8-carbon-
itrile
##STR00180##
[0232] Synthesis of (E)-ethyl
N-4-cyano-1H-pyrazol-5-ylacetimidate
##STR00181##
[0234] Analogous to chemistry described in U.S. Pat. No. 4,892,576.
A solution of 5-amino-1H-pyrazole-4-carbonitrile (1.1 g, 10 mmol),
1,1,1-triethoxyethane (2 g, 12 mmol) and AcOH (3 drops) in MeCN (74
mL) was refluxed for 16 h. The resulting mixture was cooled down to
room temperature, evaporated under vacuo. The residue was purified
with column chromatography to yield (E)-ethyl
N-4-cyano-1H-pyrazol-5-ylacetimidate (400 mg, 22%). .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 7.80 (s, 1H), 4.33 (q, J=7.2 Hz, 2H),
2.10 (s, 3H), 1.38 (t, J=7.2 Hz, 3H).
Synthesis of
3-ethyl-2-methyl-4-oxo-3,4-dihydropyrazolo[1,5-a][1,3,5]triazine-8-carbon-
itrile
##STR00182##
[0236] To a solution of (E)-ethyl
N-4-cyano-1H-pyrazol-5-ylacetimidate (100 mg, 0.56 mmol) in
anhydrous THF (5 mL) was added TEA (57 mg, 0.56 mmol) and
isocyanatoethane (50 mg, 0.7 mmol) at 0.degree. C. The mixture was
warmed to ambient temperature and stirred for 18 h. The solvent was
removed at reduced pressure, and the residue was purified with
column chromatography to yield
3-ethyl-2-methyl-4-oxo-3,4-dihydropyrazolo[1,5-a][1,3,5]triazine-8-carbon-
itrile (10 mg, yield 8.8%). .sup.1H NMR (300 MHz, CD.sub.3OD)
.delta. 8.32 (s, 1H), 4.23 (q, J=7.2 Hz, 2H), 2.73 (s, 3H), 1.42
(t, J=7.2 Hz, 3H); m/z (ESI) 204 [M+H].sup.+.
Example 25
Synthesis of
6-ethyl-5-((methylamino)methyl)-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-
e-3-carbonitrile
##STR00183##
[0237] Synthesis of
5-(bromomethyl)-6-ethyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-carb-
onitrile
##STR00184##
[0239] To a solution of
6-ethyl-5-(hydroxymethyl)-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-ca-
rbonitrile (50 mg, 0.228 mmol) and PPh.sub.3 (120 mg, 0.456 mmol)
in CH.sub.2Cl.sub.2 (10 mL) was added CBr.sub.4 (152 mg, 0.456
mmol) at ambient temperature. The reaction mixture was stirred for
16 h. The residue was concentrated and purified by column
chromatography to give
5-(bromomethyl)-6-ethyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-carb-
onitrile (30 mg, 50%). m/z (ESI) 281/283 [M+H].sup.+.
Synthesis of
6-ethyl-5-((methylamino)methyl)-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-
e-3-carbonitrile
##STR00185##
[0240] To a solution of
5-(bromomethyl)-6-ethyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-carb-
onitrile (30 mg, 0.11 mmol) in 5 ml of DMF, Et.sub.3N (22 mg, 0.22
mmol) and methanamine hydrochloride (15 mg, 0.22 mmol) was added.
The mixture was stirred at ambient temperature for 16 h. The
residue was concentrated and purified by column chromatography to
afford
6-ethyl-5-((methylamino)methyl)-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-
e-3-carbonitrile (6 mg, 20%). .sup.1HNMR (300 MHz, CD.sub.3OD)
.delta. 8.18 (s, 1H), 4.34 (s, 2H), 2.89 (s, 3H), 2.66 (q, J=7.5
Hz, 2H), 1.20 (t, J=7.2 Hz, 3H); m/z (ESI) 232 [M+H].sup.+.
Example 26
Assessment of Inhibitory Effect of Test Compounds on GASC1
Demethylase Activity on Histone 3 Lysine 9 Trimethyl Peptide
(H3K9Me3)
GASC1 Demethylation Assay
[0241] 6.times.His tagged recombinant GASC1 (N 350aa) was purified
from E. Coli BL21(DE3) to near homogeneity. The demethylation
reaction buffer contained 50 mM TrisCl pH 7.5, 0.01% Triton X-100,
5% glycerol, 1 mM ascorbate (Cat# A4034, Sigma Aldrich), 5 .mu.M
.alpha.-ketoglutarate (# K2010, Sigma Aldrich) and 20 .mu.M
Fe.sub.2(NH.sub.4).sub.2(SO.sub.4).sub.2 (Cat# F1543, Sigma
Aldrich). In 25 .mu.L demethylation reaction system, 400 nM
recombinant GASC1 and 20 .mu.M H3K9me3 peptide (1-21 aa) were
incubated with compounds for 10 minutes, and then
.alpha.-ketoglutarate and Fe.sub.2(NH.sub.4).sub.2(SO.sub.4).sub.2
were added to initiate the reaction. All of the reactions were
incubated for 45 minutes at room temperature, and then 25 .mu.l of
1 N HCl was added to quench the reactions. After termination,
plates were sealed and frozen at -80.degree. C. and shipped on dry
ice to BioTrove Inc. (Woburn, Mass.) for anaylsis.
High Throughput Mass Spectrometry (HT-MS) Analysis
[0242] All the reactions were read by RapidFire.TM. HT-MS platform
developed in BioTrove Inc, and the method has been described in
detail previously (Assay and Drug Development Technologies, 2004;
2(4): 373-381). Briefly, at BioTrove, plates were thawed and
immediately analyzed using RapidFire.TM. system coupled to a Sciex
API4000 triple quadrapole mass spectrometer. The samples were
delivered directly from the plate to a clean-up cartridge (BioTrove
column A) to remove nonvolatile assay components with 0.1% formic
acid in a 3-sec wash cycle. The peptide substrate and demethylated
product were coeluted to the mass spectrometer with 80%
acetonitrile, 0.1% formic acid. Both the substrate and product
signals were read at their +5 charge species, and the conversion
from substrate to product is assessed by [H3K9me2 Read]/[H3K9me2
Read +H3K9me3 Read].
Example 27
[0243] Table 6 shows the activity of selected compounds of this
invention in the GASC1 inhibition assay. The compound numbers
correspond to the compound numbers in Table 1. Compounds having an
activity designated as "A" provided an IC.sub.50.ltoreq.1 .mu.M;
compounds having an activity designated as "B" provided an
IC.sub.50 1-10 .mu.M; compounds having an activity designated as
"C" provided an IC.sub.50 of 10-50 .mu.M; and compounds having an
activity designated as "D" provided an IC.sub.50.gtoreq.50
.mu.M.
TABLE-US-00010 TABLE 6 GASC1 Inhibition Data Compound # GASC1
Inhibition I-1 D I-2 D I-3 B I-4 A I-5 B I-6 D I-12 C I-13 D I-14 D
I-15 B I-16 B I-17 A I-18 D I-19 D I-20 B I-21 A I-22 B I-23 B I-24
B I-25 A I-26 B I-27 D I-28 D I-29 D I-30 B I-31 B I-32 B I-33 B
I-34 A I-35 B I-36 D I-37 D I-38 A I-39 B I-40 B I-41 A I-42 A I-43
B I-44 A I-45 A I-46 A I-47 A I-48 B I-49 B I-50 B I-51 A I-52 B
I-53 B I-54 B I-55 B
Example 28
Assessment of Inhibitory Effect of Test Compounds on JARID1A and
PLU-1 Demethylase Activity on Histone 3 Lysine 4 Trimethyl Peptide
(H3K4Me3)
JARID1A/PLU1 Demethylase Assays
[0244] FLAG tagged full length recombinant JARID1A and PLU1
proteins were purified from Sf9 insect cells to near homogeneity.
The demethylation reaction buffer contained 50 mM TrisCl pH 7.5,
0.01% Triton X-100, 0.005% BSA, 1 mM ascorbate (Cat# A4034, Sigma
Aldrich), 1.7 .mu.M .alpha.-ketoglutarate (# K2010, Sigma Aldrich)
and 20 .mu.M Fe.sub.2(NH.sub.4).sub.2(SO.sub.4).sub.2 (Cat# F1543,
Sigma Aldrich). In a 25 .mu.L demethylation reaction system, 20 nM
recombinant JARID1A or PLU1 proteins and 4 .mu.M H3K4me3 peptide
(1-21 aa), which can be biotinylated or unlabelled, were incubated
with compounds for 10 minutes, and then .alpha.-ketoglutarate and
Fe.sub.2(NH.sub.4).sub.2(SO.sub.4).sub.2 were added to initiate the
reaction. All of the reactions were incubated for 45 minutes at
room temperature, and then 25 .mu.l of 1 N HCl was added to quench
the reactions. After termination, plates were sealed and frozen at
-80.degree. C. for anaylsis.
High Throughput Mass Spectrometry (HT-MS) Analysis
[0245] All the reactions were read by RapidFire.TM. HT-MS platform
developed in BioCius Inc, and described in detail (Assay and Drug
Development Technologies, 2004; 2(4): 373-381). Briefly, plates
were thawed and immediately analyzed using RapidFire.TM. system
coupled to a Sciex API4000 triple quadrapole mass spectrometer. The
samples were delivered directly from the plate to a clean-up
cartridge (BioCius column A) to remove nonvolatile assay components
with 0.1% formic acid in a 3-sec wash cycle. The peptide substrate
and demethylated product were coeluted to the mass spectrometer
with 80% acetonitrile, 0.1% formic acid. Both the substrate and
product signals were read at their +5 charge species, and the
conversion from substrate to product assessed by [H3K4me2
Read]/[H3K4me2 Read +H3K4me3 Read].
Example 29
[0246] Table 7 shows the activity of selected compounds of this
invention in the JARID1A and PLU-1 inhibition assays. The compound
numbers correspond to the compound numbers in Table 1. Compounds
having an activity designated as "A" provided an IC.sub.50.ltoreq.1
.mu.M; compounds having an activity designated as "B" provided an
IC.sub.50.ltoreq.1-10 .mu.M; compounds having an activity
designated as "C" provided an IC.sub.50 of 10-50 .mu.M; and
compounds having an activity designated as "D" provided an
IC.sub.50.gtoreq.50 .mu.M.
TABLE-US-00011 TABLE 7 JARID1A and PLU-1 Inhibition Data Compound #
JARID1A PLU-1 I-4 B B I-21 A A I-23 A A I-25 A A I-29 B B I-30 A A
I-49 A B
[0247] While we have described a number of embodiments of this
invention, it is apparent that our basic examples may be altered to
provide other embodiments that utilize the compounds and methods of
this invention. Therefore, it will be appreciated that the scope of
this invention is to be defined by the appended claims rather than
by the specific embodiments that have been represented by way of
example.
* * * * *