U.S. patent application number 16/361069 was filed with the patent office on 2020-09-17 for inhibitors of bruton's tyrosine kinase for the treatment of solid tumors.
The applicant listed for this patent is Pharmacyclics LLC. Invention is credited to Joseph J. Buggy, Wei Chen, Lee Honigberg, David J. Loury, Erik J. Verner.
Application Number | 20200289515 16/361069 |
Document ID | / |
Family ID | 1000004866774 |
Filed Date | 2020-09-17 |
View All Diagrams
United States Patent
Application |
20200289515 |
Kind Code |
A1 |
Honigberg; Lee ; et
al. |
September 17, 2020 |
INHIBITORS OF BRUTON'S TYROSINE KINASE FOR THE TREATMENT OF SOLID
TUMORS
Abstract
Described herein are irreversible Btk inhibitor compounds, and
methods for using such irreversible inhibitors in the treatment of
diseases and disorders characterized by the presence or development
of solid tumors.
Inventors: |
Honigberg; Lee; (San
Francisco, CA) ; Verner; Erik J.; (Belmont, CA)
; Buggy; Joseph J.; (Mountain View, CA) ; Loury;
David J.; (Incline Village, NV) ; Chen; Wei;
(Saratoga, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Pharmacyclics LLC |
Sunnyvale |
CA |
US |
|
|
Family ID: |
1000004866774 |
Appl. No.: |
16/361069 |
Filed: |
March 21, 2019 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
15292614 |
Oct 13, 2016 |
|
|
|
16361069 |
|
|
|
|
13003811 |
May 25, 2011 |
|
|
|
PCT/US2009/050897 |
Jul 16, 2009 |
|
|
|
15292614 |
|
|
|
|
61081344 |
Jul 16, 2008 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 33/24 20130101;
A61K 31/704 20130101; A61K 31/675 20130101; A61K 45/06 20130101;
A61K 31/00 20130101; A61K 31/52 20130101; A61K 39/39558 20130101;
A61K 31/519 20130101; A61K 31/337 20130101; A61K 39/3955
20130101 |
International
Class: |
A61K 31/519 20060101
A61K031/519; A61K 31/00 20060101 A61K031/00; A61K 31/337 20060101
A61K031/337; A61K 31/675 20060101 A61K031/675; A61K 31/704 20060101
A61K031/704; A61K 33/24 20060101 A61K033/24; A61K 31/52 20060101
A61K031/52; A61K 39/395 20060101 A61K039/395; A61K 45/06 20060101
A61K045/06 |
Claims
1.-26. (canceled)
27. A method for treating a solid tumor in a subject comprising
administering to the subject a compound having the structure:
##STR00129## wherein the solid tumor is selected from colon cancer,
breast cancer, and non-small cell lung cancer.
28. The method of claim 27, wherein the solid tumor is breast
cancer and following administration of the compound, the subject
achieves a partial response or a complete response.
29. A method for treating colon cancer in a subject comprising
administering to the subject a compound having the structure:
##STR00130##
30. The method of claim 29, further comprising administering to the
subject an additional anti-cancer agent.
31. The method of claim 30, wherein the additional anti-cancer
agent is docetaxel.
32. The method of claim 29, wherein following administration of the
compound, the subject achieves a partial response or a complete
response.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 15/292,614, filed Oct. 13, 2016, which is a continuation of
U.S. patent application Ser. No. 13/003,811, filed May 25, 2011,
which is a United States National Phase Application of
International Application No. PCT/US2009/050897, entitled
"Inhibitors of Bruton's Tyrosine Kinase for the Treatment of Solid
Tumors" filed Jul. 16, 2009, which claims the benefit of U.S.
provisional patent application No. 61/081,344 entitled "Inhibitors
of Bruton's Tyrosine Kinase for the Treatment of Solid Tumors"
filed on Jul. 16, 2008, the contents of each of which are
incorporated herein by reference in their entirety.
SEQUENCE LISTING
[0002] The instant application contains a Sequence Listing which
has been submitted in ASCII format via EFS-Web and is hereby
incorporated by reference in its entirety. Said ASCII copy, created
on Mar. 21, 2017, is named PIR-78209_SL.txt and is 2804 bytes in
size.
BACKGROUND OF THE INVENTION
[0003] A kinase, alternatively known as a phosphotransferase, is a
type of enzyme that transfers phosphate groups from high-energy
donor molecules, such as ATP, to specific target molecules; the
process is termed phosphorylation. Protein kinases, which act on
and modify the activity of specific proteins, are used to transmit
signals and control complex processes in cells. Up to 518 different
kinases have been identified in humans. Their enormous diversity
and role in signaling makes them attractive targets for drug
design.
SUMMARY OF THE INVENTION
[0004] Described herein are inhibitors of Bruton's tyrosine kinase
(Btk). Also described herein are irreversible inhibitors of Btk.
Further described are irreversible inhibitors of Btk that form a
covalent bond with a cysteine residue on Btk.
[0005] Further described herein are inhibitors of other tyrosine
kinases, wherein the other tyrosine kinases share homology with Btk
by having a cysteine residue (including a Cys 481 residue) that
forms a covalent bond with the irreversible inhibitor (such
tyrosine kinases, are referred herein as "Btk tyrosine kinase
cysteine homologs"). Also described herein are irreversible
inhibitors of Btk tyrosine kinase cysteine homologs. In some
embodiments, the Btk tyrosine kinase cysteine homolog is HER4.
[0006] Additionally described herein are inhibitors of tyrosine
kinases that have an accessible cysteine residue near an active
site of the tyrosine kinase (referred herein as "Accessible
Cysteine Kinases" or ACKs). Also described herein are irreversible
inhibitors of ACKs. In some embodiments, the ACK is HER4.
[0007] Described herein are inhibitors of HER4. Also described
herein are irreversible inhibitors of HER4.
[0008] Also described herein are irreversible inhibitors of any of
the aforementioned tyrosine kinases, in which the irreversible
inhibitor includes a Michael acceptor moiety. Further described are
such irreversible inhibitors in which the Michael acceptor moiety
preferentially forms a covalent bond with the appropriate cysteine
residue on the desired tyrosine kinase relative to forming a
covalent bond with other biological molecules that contain an
accessible SH moiety.
[0009] Also described herein are methods for synthesizing such
irreversible inhibitors, methods for using such irreversible
inhibitors in the treatment of diseases (including diseases wherein
irreversible inhibition of Btk provides therapeutic benefit to an
individual having the disease).
[0010] Further described are pharmaceutical formulations that
include an irreversible inhibitor of Btk, an irreversible inhibitor
of an ACK, an irreversible inhibitor of HER4, an irreversible
inhibitor of a Btk tyrosine kinase cysteine homolog, or
combinations thereof.
[0011] Described herein, in certain embodiments, are methods for
treating a disorder characterized by the presence or development of
one or more solid tumors comprising administering to an individual
in need a pharmaceutical formulation comprising a compound of
Formula (I) having the structure:
##STR00001##
wherein: [0012] L.sub.a is CH.sub.2, O, NH or S; [0013] Ar is a
substituted or unsubstituted aryl, or a substituted or
unsubstituted heteroaryl; and either [0014] (i) Y is an optionally
substituted group selected from among alkylene, heteroalkylene,
arylene, heteroarylene, alkylenearylene, alkyleneheteroarylene,
alkylenecycloalkylene and alkyleneheterocycloalkylene; [0015] Z is
C(.dbd.O), NHC(.dbd.O), NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x,
where x is 1 or 2, and R.sup.a is H, substituted or unsubstituted
alkyl, substituted or unsubstituted cycloalkyl; and either [0016]
(a) R.sub.7 and R.sub.8 are H; R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0017] (b)
R.sub.6 and R.sub.8 are H; R.sub.7 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0018]
(c) R.sub.7 and R.sub.8 taken together form a bond; R.sub.6 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0019]
(ii) Y is an optionally substituted group selected from
cycloalkylene or heterocycloalkylene; [0020] Z is C(.dbd.O),
NHC(.dbd.O), NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x, where x is
1 or 2, and R.sup.a is H, substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl; and either [0021] (a)
R.sub.7 and R.sub.8 are H; R.sub.6 is substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0022] (b)
R.sub.6 and R.sub.8 are H; R.sub.7 is substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0023]
(c) R.sub.7 and R.sub.8 taken together form a bond; R.sub.6 is
substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and
pharmaceutically active metabolites, or pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof. In some embodiments, L.sub.a is O. In
some embodiments, Ar is phenyl. In some embodiments, Z is
C(.dbd.O), NHC(.dbd.O), or NCH.sub.3C(.dbd.O). In some embodiments,
Y is an optionally substituted group selected from among alkylene,
heteroalkylene, arylene, heteroarylene, alkylenearylene,
alkyleneheteroarylene, alkylenecycloalkylene and
alkyleneheterocycloalkylene; Z is C(.dbd.O), NHC(.dbd.O),
NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x, where x is 1 or 2, and
R.sup.a is H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl; and either (a) R.sub.7 and R.sub.8 are H;
R.sub.6 is H, substituted or unsubstituted C.sub.1-C.sub.4alkyl,
substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); (b) R.sub.6
and R.sub.8 are H; R.sub.7 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or (c)
R.sub.7 and R.sub.8 taken together form a bond; R.sub.6 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, R.sub.7 and R.sub.8 are H; and R.sub.6 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, Y is alkyleneheterocycloalkylene. In some embodiments,
R.sub.6 is H, substituted or unsubstituted C.sub.1-C.sub.4alkyl,
C.sub.1-C.sub.8alkylaminoalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, R.sub.6 and R.sub.8 are H; and R.sub.7 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, Y is alkyleneheterocycloalkylene. In some embodiments,
R.sub.7 is H, substituted or unsubstituted C.sub.1-C.sub.4alkyl,
C.sub.1-C.sub.8alkylaminoalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, R.sub.7 and R.sub.8 taken together form a bond; and
R.sub.6 is H, substituted or unsubstituted C.sub.1-C.sub.4alkyl,
substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, Y is alkyleneheterocycloalkylene. In some embodiments,
R.sub.6 is H, substituted or unsubstituted C.sub.1-C.sub.4alkyl,
C.sub.1-C.sub.8alkylaminoalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments Y is an optionally substituted group selected from
cycloalkylene or heterocycloalkylene; Z is C(.dbd.O), NHC(.dbd.O),
NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x, where x is 1 or 2, and
R.sup.a is H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl; and either (a) R.sub.7 and R.sub.8 are H;
R.sub.6 is substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); (b) R.sub.6
and R.sub.8 are H; R.sub.7 is substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or (c)
R.sub.7 and R.sub.8 taken together form a bond; R.sub.6 is
substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, R.sub.7 and R.sub.8 are H; and R.sub.6 is substituted
or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, R.sub.6 is substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, R.sub.6 and R.sub.8 are H; and R.sub.7 is substituted
or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, R.sub.7 is substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, R.sub.7 and R
.sub.8 taken together form a bond; and R.sub.6 is substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, R.sub.6 is substituted or unsubstituted
C.sub.1-C.sub.4alkyl, C.sub.1-C.sub.8alkylaminoalkyl, substituted
or unsubstituted C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, the disorder characterized by the presence or
development of one or more solid tumors is a sarcoma, lymphoma,
and/or carcinoma. In some embodiments, the disease is mammary
ductal carcinoma, lobular carcinoma, an adenocarcinoma (e.g.
pancreatic cancer and colon cancer), small cell lung carcinoma,
non-small cell lung carcinoma, and melanomas. In some embodiments,
the disease is breast cancer. In some embodiments, the disease is
mammary ductal carcinoma, lobular carcinoma, or a combination
thereof. In some embodiments, the breast cancer is ER positive. In
some embodiments, the breast cancer is ER negative. In some
embodiments, the breast cancer is progesterone receptor
(PgR)-positive. In some embodiments, the breast cancer is
PgR-negative. In some embodiments, the disease is pancreatic
cancer.
[0024] Described herein, in certain embodiments, are methods for
treating a disorder characterized by the presence or development of
one or more solid tumors comprising administering to an individual
in need a pharmaceutical formulation comprising a compound selected
from among:
(E)-4-(N-(2-hydroxyethyl)-N-methylamino)-1-(3-(4-phenoxyphenyl)-1H-pyrazo-
lo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)but-2-en-1-one (Compound 3);
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3-
-(1H-imidazol-4-yl)prop-2-en-1-one (Compound 4);
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-morpholinobut-2-en-1-one (Compound 5);
(E)-1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound 7);
(E)-N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-
-1-yl)cyclohexyl)-4-(dimethylamino)but-2-enamide (Compound 8);
N-((1r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)acrylamide (Compound 10);
(E)-1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)methyl)pyrolidin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound
11);
(E)-1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)methyl)pyrolidin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound
12);
1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 13);
1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 14);
1((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)me-
thyl)pyrrolidin-1-yl)but-2-yn-1-one (Compound 15);
1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)but-2-yn-1-one (Compound 16);
1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)but-2-yn-1-one (Compound 17);
(E)-N-((1,r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidi-
n-1-yl)cyclohexyl-4-(dimethylamino)but-2-enamide (Compound 18);
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
-N-methylacrylamide (Compound 19);
(E)-1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-4-
-morpholinobut-2-en-1-one (Compound 20);
(E)-1-((S-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)methyl)pyrrolidin-1-yl)-4-morpholinobut-2-en-1-one (Compound 21);
N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)but-2-ynamide (Compound 22);
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
acrylamide (Compound 23);
(E)-1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)piperidin-1-yl)-4-morpholinobut-2-en-1-one (Compound 24);
(E)-N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-
-1-yl)cyclohexyl)-4-morpholinobut-2-enamide (Compound 25). In some
embodiments, the disorder characterized by the presence or
development of one or more solid tumors is a sarcoma, lymphoma,
and/or carcinoma. In some embodiments, the disease is mammary
ductal carcinoma, lobular carcinoma, an adenocarcinoma (e.g.
pancreatic cancer and colon cancer), small cell lung carcinoma,
non-small cell lung carcinoma, and melanomas. In some embodiments,
the disease is breast cancer. In some embodiments, the disease is
mammary ductal carcinoma, lobular carcinoma, or a combination
thereof. In some embodiments, the breast cancer is ER positive. In
some embodiments, the breast cancer is ER negative. In some
embodiments, the breast cancer is progesterone receptor
(PgR)-positive. In some embodiments, the breast cancer is
PgR-negative. In some embodiments, the disease is pancreatic
cancer.
[0025] Described herein, in certain embodiments, are methods for
treating a disorder characterized by the presence or development of
one or more solid tumors comprising administering to an individual
in need thereof a composition containing a therapeutically
effective amount of a compound that forms a covalent bond with a
cysteine sidechain of a Bruton's tyrosine kinase, a Bruton's
tyrosine kinase homolog, an ACK, or a combination thereof. In some
embodiments, the disorder characterized by the presence or
development of one or more solid tumors is a sarcoma, lymphoma,
and/or carcinoma. In some embodiments, the disease is mammary
ductal carcinoma, lobular carcinoma, an adenocarcinoma (e.g.
pancreatic cancer and colon cancer), small cell lung carcinoma,
non-small cell lung carcinoma, and melanomas. In some embodiments,
the disease is breast cancer. In some embodiments, the disease is
mammary ductal carcinoma, lobular carcinoma, or a combination
thereof. In some embodiments, the breast cancer is ER positive. In
some embodiments, the breast cancer is ER negative. In some
embodiments, the breast cancer is progesterone receptor
(PgR)-positive. In some embodiments, the breast cancer is
PgR-negative. In some embodiments, the disease is pancreatic
cancer.
[0026] Described herein, in certain embodiments, are methods method
for treating a disorder characterized by the presence or
development of one or more solid tumors comprising administering to
an individual in need thereof a kinase inhibitor that selectively
and irreversibly binds to a protein tyrosine kinase selected from
Btk, a Btk homolog, a Btk kinase cysteine homolog, an ACK, and
HER4, in which the kinase inhibitor reversibly and non-selectively
binds to a multiplicity of protein tyrosine kinases, and further in
which the plasma half life of the kinase inhibitor is less than
about 4 hours. In some embodiments, the kinase inhibitor
selectively and irreversibly binds to at least one of Btk, Jak3,
Blk, Bmx, Tec, HER4, and Itk. In some embodiments, the kinase
inhibitor selectively and irreversibly binds to Btk. In some
embodiments, the kinase inhibitor selectively and irreversibly
binds to Btk and Tec. In some embodiments, the plasma half life of
the kinase inhibitor is less than about 3 hours. In some
embodiments, the kinase inhibitor has the structure of Formula
(VII):
##STR00002##
wherein:
##STR00003##
is a moiety that binds to the active site of a kinase, including a
tyrosine kinase, further including a Btk kinase cysteine homolog;
[0027] Y is an optionally substituted group selected from among
alkylene, heteroalkylene, arylene, heteroarylene,
heterocycloalkylene, cycloalkylene, alkylenearylene,
alkyleneheteroarylene, alkylenecycloalkylene, and
alkyleneheterocycloalkylene; [0028] Z is C(.dbd.O), OC(.dbd.O),
NHC(.dbd.O), NCH.sub.3C(.dbd.O), C(.dbd.S), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, NHS(.dbd.O).sub.x, where x is 1 or 2; [0029]
R.sub.7 and R.sub.8 are independently selected from among H,
unsubstituted C.sub.1-C.sub.4 alkyl, substituted
C.sub.1-C.sub.4alkyl, unsubstituted C.sub.1-C.sub.4heteroalkyl,
substituted C.sub.1-C.sub.4heteroalkyl, unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted C.sub.3-C.sub.6cycloalkyl,
unsubstituted C.sub.2-C.sub.6heterocycloalkyl, and substituted
C.sub.2-C.sub.6heterocycloalkyl; or [0030] R.sub.7 and R.sub.8
taken together form a bond; [0031] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted C.sub.2-C.sub.8heterocycloalkyl,
substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and
pharmaceutically active metabolites, or pharmaceutically acceptable
solvates, [0032] pharmaceutically acceptable salts, or
pharmaceutically acceptable prodrugs thereof.
[0033] In some embodiments,
##STR00004##
is a substituted fused biaryl moiety selected from
##STR00005##
In some embodiments, Z is C(.dbd.O), NHC(.dbd.O),
NCH.sub.3C(.dbd.O), or S(.dbd.O).sub.2. In some embodiments, each
of R.sub.7 and R.sub.8 is H; or R.sub.7 and R.sub.8 taken together
form a bond. In some embodiments, R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
embodiments, Y is a 4-, 5-, 6-, or 7-membered cycloalkylene ring;
or Y is a 4-, 5-, 6-, or 7-membered heterocycloalkylene ring; or Y
is a C.sub.1-C.sub.4 alkylene, or 4-, 5-, 6-, or 7-membered
heterocycloalkylene ring. In some embodiments, the disorder
characterized by the presence or development of one or more solid
tumors is a sarcoma, lymphoma, and/or carcinoma. In some
embodiments, the disease is mammary ductal carcinoma, lobular
carcinoma, an adenocarcinoma (e.g. pancreatic cancer and colon
cancer), small cell lung carcinoma, non-small cell lung carcinoma,
and melanomas. In some embodiments, the disease is breast cancer.
In some embodiments, the disease is mammary ductal carcinoma,
lobular carcinoma, or a combination thereof. In some embodiments,
the breast cancer is ER positive. In some embodiments, the breast
cancer is ER negative. In some embodiments, the breast cancer is
progesterone receptor (PgR)-positive. In some embodiments, the
breast cancer is PgR-negative. In some embodiments, the breast
cancer is ER negative. In some embodiments, the breast cancer is
progesterone receptor (PgR)-positive. In some embodiments, the
breast cancer is PgR-negative. In some embodiments, the disease is
pancreatic cancer.
[0034] Described herein, in certain embodiments, are methods for
treating a disorder characterized by the presence or development of
one or more solid tumors comprising administering to an individual
in need a composition containing a therapeutically effective amount
of a compound that forms a covalent bond with a cysteine sidechain
of a Blk or a Blk homolog. In some embodiments, the disorder
characterized by the presence or development of one or more solid
tumors is a sarcoma, lymphoma, and/or carcinoma. In some
embodiments, the disease is mammary ductal carcinoma, lobular
carcinoma, an adenocarcinoma (e.g. pancreatic cancer and colon
cancer), small cell lung carcinoma, non-small cell lung carcinoma,
and melanomas. In some embodiments, the disease is breast cancer.
In some embodiments, the disease is mammary ductal carcinoma,
lobular carcinoma, or a combination thereof. In some embodiments,
the breast cancer is ER positive. In some embodiments, the breast
cancer is ER negative. In some embodiments, the breast cancer is
progesterone receptor (PgR)-positive. In some embodiments, the
breast cancer is PgR-negative. In some embodiments, the disease is
pancreatic cancer.
[0035] Compounds described herein include those that have a
structure of any of Formula (A1-A6), Formula (B1-B6), Formula
(C1-C6), Formula (D1-D6), Formula (I), or Formula (VII), and
pharmaceutically acceptable salts, solvates, esters, acids and
prodrugs thereof. In certain embodiments, isomers and chemically
protected forms of compounds having a structure represented by any
of Formula (A1-A6), Formula (B1-B6), Formula (C1-C6), Formula
(D1-D6), Formula (I), or Formula (VII), are also provided.
[0036] In one aspect, provided herein are compounds of Formula (I).
Formula (I) is as follows:
##STR00006##
wherein [0037] L.sub.a is CH.sub.2, O, NH or S; [0038] Ar is a
substituted or unsubstituted aryl, or a substituted or
unsubstituted heteroaryl; and either [0039] (a) Y is an optionally
substituted group selected from among alkylene, heteroalkylene,
arylene, heteroarylene, alkylenearylene, alkyleneheteroarylene,
alkylenecycloalkylene and alkyleneheterocycloalkylene; [0040] Z is
C(.dbd.O), NHC(.dbd.O), NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x,
where x is 1 or 2, and R.sup.a is H, substituted or unsubstituted
alkyl, substituted or unsubstituted cycloalkyl; and either [0041]
(i) R.sub.7 and R.sub.8 are H; [0042] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8 hydroxyalkylaminoalkyl, C.sub.1-C.sub.8
alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0043] (ii)
R.sub.6 and R.sub.8 are H; [0044] R.sub.7 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8 hydroxyalkylaminoalkyl, C.sub.1-C.sub.8
alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0045]
(iii) R.sub.7 and R.sub.8 taken together form a bond; [0046]
R.sub.6 is H, substituted or unsubstituted C.sub.1-C.sub.4alkyl,
substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl, C.sub.1-C.sub.8
hydroxyalkylaminoalkyl, C.sub.1-C.sub.8 alkoxyalkylaminoalkyl,
substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl, substituted
or unsubstituted C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl,
substituted or unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0047]
(b) Y is an optionally substituted group selected from
cycloalkylene or heterocycloalkylene; [0048] Z is C(.dbd.O),
NHC(.dbd.O), NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x, where x is
1 or 2, and R.sup.a is H, substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl; and either [0049] (i)
R.sub.7 and R.sub.8 are H; [0050] R.sub.6 is substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8
hydroxyalkylaminoalkyl, C.sub.1-C.sub.8 alkoxyalkylaminoalkyl,
substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl, substituted
or unsubstituted C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl,
substituted or unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0051] (ii)
R.sub.6 and R.sub.8 are H; [0052] R.sub.7 is substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8
hydroxyalkylaminoalkyl, C.sub.1-C.sub.8 alkoxyalkylaminoalkyl,
substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl, substituted
or unsubstituted C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl,
substituted or unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0053]
(iii) R.sub.7 and R.sub.8 taken together form a bond; [0054]
R.sub.6 is substituted or unsubstituted C.sub.1-C.sub.4alkyl,
substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and
pharmaceutically active metabolites, or pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof.
[0055] In another embodiment are provided pharmaceutically
acceptable salts of compounds of Formula (I). By way of example
only, are salts of an amino group formed with inorganic acids such
as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid and perchloric acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid
or malonic acid. Further salts include those in which the
counterion is an anion, such as adipate, alginate, ascorbate,
aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, and valerate. Further salts
include those in which the counterion is an cation, such as sodium,
lithium, potassium, calcium, magnesium, ammonium, and quaternary
ammonium (substituted with at least one organic moiety)
cations.
[0056] In another embodiment are pharmaceutically acceptable esters
of compounds of Formula (I), including those in which the ester
group is selected from a formate, acetate, propionate, butyrate,
acrylate and ethylsuccinate.
[0057] In another embodiment are pharmaceutically acceptable
carbamates of compounds of Formula (I). In another embodiment are
pharmaceutically acceptable N-acyl derivatives of compounds of
Formula (I). Examples of N-acyl groups include N-acetyl and
N-ethoxycarbonyl groups.
[0058] For any and all of the embodiments, substituents are
optionally selected from among from a subset of the listed
alternatives. For example, in some embodiments, L.sub.a is
CH.sub.2, O, or NH. In other embodiments, L.sub.a is O or NH. In
yet other embodiments, L.sub.a is O.
[0059] In some embodiments, Ar is a substituted or unsubstituted
aryl. In yet other embodiments, Ar is a 6-membered aryl. In some
other embodiments, Ar is phenyl.
[0060] In some embodiments, x is 2. In yet other embodiments, Z is
C(.dbd.O), OC(.dbd.O), NHC(.dbd.O), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, or NHS(.dbd.O).sub.x. In some other embodiments,
Z is C(.dbd.O), NHC(.dbd.O), or NCH.sub.3C(.dbd.O).
[0061] In some embodiments Y is an optionally substituted group
selected from among alkylene, heteroalkylene, arylene,
heteroarylene, alkylenearylene, alkyleneheteroarylene, and
alkyleneheterocycloalkylene.
[0062] In some embodiments, Z is C(.dbd.O), NHC(.dbd.O),
NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x, where x is 1 or 2, and
R.sup.a is H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl.
[0063] In some embodiments, R.sub.7 and R.sub.8 are H; and R.sub.6
is H, substituted or unsubstituted C.sub.1-C.sub.4alkyl,
substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In other
embodiments, R.sub.6 and R.sub.8 are H; and R.sub.7 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In yet
further embodiments, R.sub.7 and R.sub.8 taken together form a
bond; and R.sub.6 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl).
[0064] In some embodiments, Y is an optionally substituted group
selected from cycloalkylene or heterocycloalkylene.
[0065] In some embodiments, Z is C(.dbd.O), NHC(.dbd.O),
NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x, where x is 1 or 2, and
R.sup.a is H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl.
[0066] In some embodiments, R.sub.7 and R.sub.8 are H; and R.sub.6
is substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In other
embodiments, R.sub.6 and R.sub.8 are H; and R.sub.7 is substituted
or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In further
embodiments, R.sub.7 and R.sub.8 taken together form a bond; and
R.sub.6 is substituted or unsubstituted C.sub.1-C.sub.4alkyl,
substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl).
[0067] Any combination of the groups described above for the
various variables is contemplated herein.
[0068] In one aspect, provided herein is a compound selected from
among:
(E)-4-(N-(2-hydroxyethyl)-N-methylamino)-1-(3-(4-phenoxyphenyl)-1H-pyrazo-
lo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)but-2-en-1-one (Compound 3);
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3-
-(1H-imidazol-4-yl)prop-2-en-1-one (Compound 4);
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-morpholinobut-2-en-1-one (Compound 5);
(E)-1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound 7);
(E)-N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-
-1-yl)cyclohexyl)-4-(dimethylamino)but-2-enamide (Compound 8);
N-((1r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)acrylamide (Compound 10);
(E)-1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)methyl)pyrolidin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound
11);
(E)-1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)methyl)pyrolidin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound
12);
1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 13);
1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 14);
1((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)me-
thyl)pyrrolidin-1-yl)but-2-yn-1-one (Compound 15);
1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)but-2-yn-1-one (Compound 16);
1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)but-2-yn-1-one (Compound 17);
(E)-N-((1,r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidi-
n-1-yl)cyclohexyl-4-(dimethylamino)but-2-enamide (Compound 18);
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
-N-methylacrylamide (Compound 19);
(E)-1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-4-
-morpholinobut-2-en-1-one (Compound 20);
(E)-1-((S-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)methyl)pyrrolidin-1-yl)-4-morpholinobut-2-en-1-one (Compound 21);
N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)but-2-ynamide (Compound 22);
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
acrylamide (Compound 23);
(E)-1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)piperidin-1-yl)-4-morpholinobut-2-en-1-one (Compound 24);
(E)-N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-
-1-yl)cyclohexyl)-4-morpholinobut-2-enamide (Compound 25).
[0069] In a further aspect are provided pharmaceutical
compositions, which include a therapeutically effective amount of
at least one of any of the compounds herein, or a pharmaceutically
acceptable salt, pharmaceutically active metabolite,
pharmaceutically acceptable prodrug, or pharmaceutically acceptable
solvate. In certain embodiments, compositions provided herein
further include a pharmaceutically acceptable diluent, excipient
and/or binder.
[0070] Pharmaceutical compositions formulated for administration by
an appropriate route and means containing effective concentrations
of one or more of the compounds provided herein, or
pharmaceutically effective derivatives thereof, that deliver
amounts effective for the treatment, prevention, or amelioration of
one or more symptoms of diseases, diseases or disorders that are
modulated or otherwise affected by tyrosine kinase activity, or in
which tyrosine kinase activity is implicated, are provided. The
effective amounts and concentrations are effective for ameliorating
any of the symptoms of any of the diseases, diseases or disorders
disclosed herein.
[0071] In certain embodiments, provided herein is a pharmaceutical
composition containing: i) a physiologically acceptable carrier,
diluent, and/or excipient; and ii) one or more compounds provided
herein.
[0072] In one aspect, provided herein are methods for treating an
individual with a disease treatable by a compound disclosed herein,
the method comprising administering a compound provided herein. In
some embodiments, provided herein is a method of inhibiting the
activity of tyrosine kinase(s) (e.g., Btk, HER4, an ACK, or a Btk
tyrosine kinase cysteine homolog), or of treating a disorder, which
benefits from inhibition of tyrosine kinase(s) (e.g., Btk, HER4, an
ACK, or a Btk tyrosine kinase cysteine homolog), in an individual,
which includes administering to the patient a therapeutically
effective amount of at least one of any of the compounds herein, or
pharmaceutically acceptable salt, pharmaceutically active
metabolite, pharmaceutically acceptable prodrug, or
pharmaceutically acceptable solvate. In some embodiments, the
disease is a sarcoma, lymphoma, and/or carcinoma. In some
embodiments, the disease is mammary ductal carcinoma, lobular
carcinoma, an adenocarcinoma (e.g. pancreatic cancer and colon
cancer), small cell lung carcinoma, non-small cell lung carcinoma,
and melanomas. In some embodiments, the disease is breast cancer.
In some embodiments, the disease is mammary ductal carcinoma,
lobular carcinoma, or a combination thereof. In some embodiments,
the breast cancer is ER positive. In some embodiments, the breast
cancer is ER negative. In some embodiments, the breast cancer is
progesterone receptor (PgR)-positive. In some embodiments, the
breast cancer is PgR-negative. In some embodiments, the disease is
pancreatic cancer.
[0073] In another aspect, provided herein is the use of a compound
disclosed herein for inhibiting the activity of a Bruton's tyrosine
kinase (Btk), the activity of an ACK, the activity of HER4, or the
activity of a Btk tyrosine kinase cysteine homolog, or for the
treatment of a disorder, which benefits from inhibiting the
activity of a Bruton's tyrosine kinase (Btk), the activity of an
ACK, the activity of HER4, or the activity of a Btk tyrosine kinase
cysteine homolog. In some embodiments, the disease is a sarcoma,
lymphoma, and/or carcinoma. In some embodiments, the disease is
mammary ductal carcinoma, lobular carcinoma, an adenocarcinoma
(e.g. pancreatic cancer and colon cancer), small cell lung
carcinoma, non-small cell lung carcinoma, and melanomas. In some
embodiments, the disease is breast cancer. In some embodiments, the
disease is mammary ductal carcinoma, lobular carcinoma, or a
combination thereof. In some embodiments, the breast cancer is ER
positive. In some embodiments, the breast cancer is ER negative. In
some embodiments, the breast cancer is progesterone receptor
(PgR)-positive. In some embodiments, the breast cancer is
PgR-negative. In some embodiments, the disease is pancreatic
cancer.
[0074] In some embodiments, compounds provided herein are
administered to a mammal. In some embodiments, the mammal is a
human. In some embodiments, the mammal is a non-human. In some
embodiments, compounds provided herein are orally administered. In
other embodiments, the pharmaceutical formulation that is
formulated for a route of administration is selected from oral
administration, parenteral administration, buccal administration,
nasal administration, topical administration, or rectal
administration.
[0075] In other embodiments, compounds provided herein are used for
the formulation of a medicament for the inhibition of tyrosine
kinase activity. In some other embodiments, compounds provided
herein are used for the formulation of a medicament for the
inhibition of Bruton's tyrosine kinase (Btk) activity. In some
other embodiments, compounds provided herein are used for the
formulation of a medicament for the inhibition of the activity of
an ACK. In some other embodiments, compounds provided herein are
used for the formulation of a medicament for the inhibition of the
activity of HER4. In some other embodiments, compounds provided
herein are used for the formulation of a medicament for the
inhibition of the activity of a Btk tyrosine kinase cysteine
homolog.
[0076] Articles of manufacture including packaging material, a
compound or composition or pharmaceutically acceptable derivative
thereof provided herein, which is effective for inhibiting the
activity of tyrosine kinase(s), such as Btk, within the packaging
material, and a label that indicates that the compound or
composition, or pharmaceutically acceptable salt, pharmaceutically
active metabolite, pharmaceutically acceptable prodrug, or
pharmaceutically acceptable solvate thereof, is used for inhibiting
the activity of tyrosine kinase(s) (e.g., Btk, HER4, an ACK, or a
Btk tyrosine kinase cysteine homolog) are provided.
[0077] In another aspect are inhibited tyrosine kinases comprising
a Bruton's tyrosine kinase, a Bruton's tyrosine kinase homolog, a
Btk tyrosine kinase cysteine homolog thereof, an ACK covalently
bound to an inhibitor, or HER4 covalently bound to an inhibitor
having the structures:
##STR00007## ##STR00008##
wherein indicates the point of attachment between the inhibitor and
the tyrosine kinase. In a further embodiment, the inhibitor is
covalently bound to a cysteine residue on the tyrosine kinase.
[0078] In a further aspect, provided herein is a method for
treating mammary ductal carcinoma, lobular carcinoma, pancreatic
cancer, diffuse large B cell lymphoma or follicular lymphoma by
administering to an individual in need thereof a composition
containing a therapeutically effective amount of a compound that
forms a covalent bond with a cysteine sidechain of a Bruton's
tyrosine kinase, a Bruton's tyrosine homolog, an ACK, HER4, or a
Btk tyrosine kinase cysteine homolog. In one embodiment, the
compound forms a covalent bound with the activated form of a
Bruton's tyrosine kinase, a Bruton's tyrosine homolog, an ACK,
HER4, or a Btk tyrosine kinase cysteine homolog. In further or
alternative embodiments, the compound irreversibly inhibits the
Bruton's tyrosine kinase, the Bruton's tyrosine homolog, the ACK,
the HER4, or the Btk tyrosine kinase cysteine homolog to which it
is covalently bound. In a further or alternative embodiment, the
compound forms a covalent bond with a cysteine residue on a
Bruton's tyrosine kinase, a Bruton's tyrosine homolog, an ACK,
HER4, or Btk tyrosine kinase cysteine homolog.
[0079] Further described herein are methods, assays and systems for
identifying an irreversible inhibitor of a kinase, including a
protein kinase, further including a tyrosine kinase. Further
described are methods, assays and systems for determining an
appropriate irreversible inhibitor of a kinase, including a
tyrosine kinase, in which the inhibitor forms a covalent bond with
a cysteine residue on the kinase, further wherein the cysteine
residue is near an active site of the kinase. In further
embodiments, the inhibitor also has a moiety that binds an active
site of the kinase. In some embodiments, the kinases share homology
with Btk by having a cysteine residue (including a Cys 481 residue)
that forms a covalent bond with the irreversible inhibitor (such
tyrosine kinases, are referred herein as "Btk kinase cysteine
homologs"). In some embodiments the Btk kinase cysteine homolog(s)
are selected from the Tec family of kinases, the EGFR family of
kinases, the Jak3 family of kinases and/or the Btk-Src family of
kinases.
[0080] In some embodiments, the irreversible inhibitor is a
selective irreversible inhibitor, including selectivity for a
particular Btk kinase cysteine homolog over other Btk kinase
cysteine homologs. In some embodiments the selective and
irreversible inhibitor is an effective inhibitor for a kinase
selected from Btk, a Btk homolog, and ACK, HER4, or a Btk kinase
cysteine homolog, but is not an effective inhibitor for at least
one other different kinase selected from kinase selected from Btk,
a Btk homolog, and ACK, HER4, or a Btk kinase cysteine homolog.
[0081] Also described herein are kinase inhibitors that selectively
and irreversibly bind to a protein tyrosine kinase selected from
Btk, a Btk homolog, an ACK, HER4, and a Btk kinase cysteine
homolog, in which the kinase inhibitor reversibly and
non-selectively binds to a multiplicity of protein tyrosine
kinases. In one embodiment the plasma half life of the kinase
inhibitor is less than about 4 hours. In another embodiment the
plasma half life of the kinase inhibitor is less than about 3
hours.
[0082] In a further embodiment are kinase inhibitors that
selectively and irreversibly bind to at least one of Btk, Jak3,
Blk, Bmx, Tec, and Itk. In another embodiment are kinase inhibitors
that selectively and irreversibly bind to Btk. In another
embodiment are kinase inhibitors that selectively and irreversibly
bind to Jak3. In another embodiment are kinase inhibitors that
selectively and irreversibly bind to Tec. In another embodiment are
kinase inhibitors that selectively and irreversibly bind to Itk. In
another embodiment are kinase inhibitors that selectively and
irreversibly bind to Btk and Tec. In another embodiment are kinase
inhibitors that selectively and irreversibly bind to Blk. In yet a
further embodiment are kinase inhibitors that reversibly and
non-selectively bind to a multiplicity of src-family protein kinase
inhibitors.
[0083] Also described herein are irreversible inhibitors that are
identified using such methods, assays and systems. Such
irreversible inhibitor comprise an active site binding moiety that
binds to an active site of a kinase, including a tyrosine kinase,
further including a Btk kinase cysteine homolog, further including
an ACK, further including HER4; a Michael acceptor moiety; and a
moiety that links the active site binding moiety to the Michael
acceptor moiety. In some embodiments, the Michael acceptor moiety
comprises and alkene and/or an alkyne moiety. In some embodiments,
the irreversible inhibitor is a selective irreversible inhibitor,
including selectivity for a particular Btk kinase cysteine homolog
over other Btk kinase cysteine homologs.
[0084] In any of the aforementioned embodiments, the irreversible
inhibitors have the structure of Formula (VII):
##STR00009##
wherein: [0085] wherein
##STR00010##
[0085] is a moiety that binds to the active site of a kinase,
including a tyrosine kinase, further including a Btk kinase
cysteine homolog; [0086] Y is an optionally substituted group
selected from among alkylene, heteroalkylene, arylene,
heteroarylene, heterocycloalkylene, cycloalkylene, alkylenearylene,
alkyleneheteroarylene, alkylenecycloalkylene, and
alkyleneheterocycloalkylene; [0087] Z is C(.dbd.O), OC(.dbd.O),
NHC(.dbd.O), NCH.sub.3C(.dbd.O), C(.dbd.S), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, NHS(.dbd.O).sub.x, where x is 1 or 2; [0088]
R.sub.7 and R.sub.8 are independently selected from among H,
unsubstituted C.sub.1-C.sub.4 alkyl, substituted
C.sub.1-C.sub.4alkyl, unsubstituted C.sub.1-C.sub.4heteroalkyl,
substituted C.sub.1-C.sub.4heteroalkyl, unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted C.sub.3-C.sub.6cycloalkyl,
unsubstituted C.sub.2-C.sub.6heterocycloalkyl, and substituted
C.sub.2-C.sub.6heterocycloalkyl; or [0089] R.sub.7 and R.sub.8
taken together form a bond; [0090] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted C.sub.2-C.sub.8heterocycloalkyl,
substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and [0091]
pharmaceutically active metabolites, or pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof.
[0092] In another embodiment are provided pharmaceutically
acceptable salts of compounds of Formula (VII). By way of example
only, are salts of an amino group formed with inorganic acids such
as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid and perchloric acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid
or malonic acid. Further salts include those in which the
counterion is an anion, such as adipate, alginate, ascorbate,
aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, and valerate. Further salts
include those in which the counterion is an cation, such as sodium,
lithium, potassium, calcium, magnesium, ammonium, and quaternary
ammonium (substituted with at least one organic moiety)
cations.
[0093] In another embodiment are pharmaceutically acceptable esters
of compounds of Formula (VII), including those in which the ester
group is selected from a formate, acetate, propionate, butyrate,
acrylate and ethylsuccinate.
[0094] In another embodiment are pharmaceutically acceptable
carbamates of compounds of Formula (VII). In another embodiment are
pharmaceutically acceptable N-acyl derivatives of compounds of
Formula (VII). Examples of N-acyl groups include N-acetyl and
N-ethoxycarbonyl groups.
[0095] In some embodiments,
##STR00011##
is a substituted fused biaryl moiety selected from
##STR00012##
[0096] In some embodiments Z is C(.dbd.O), NHC(.dbd.O),
NCH.sub.3C(.dbd.O), or S(.dbd.O).sub.2. In other embodiments, x is
2. In yet other embodiments, Z is C(.dbd.O), OC(.dbd.O),
NHC(.dbd.O), S(.dbd.O).sub.x, OS(.dbd.O).sub.x, or
NHS(.dbd.O).sub.x. In some other embodiments, Z is C(.dbd.O),
NHC(.dbd.O), or S(.dbd.O).sub.2.
[0097] In some embodiments, R.sub.7 and R.sub.8 are independently
selected from among H, unsubstituted C.sub.1-C.sub.4 alkyl,
substituted C.sub.1-C.sub.4alkyl, unsubstituted
C.sub.1-C.sub.4heteroalkyl, and substituted
C.sub.1-C.sub.4heteroalkyl; or R.sub.7 and R.sub.8 taken together
form a bond. In yet other embodiments, each of R.sub.7 and R.sub.8
is H; or R.sub.7 and R.sub.8 taken together form a bond.
[0098] In some embodiments, R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
aryl, substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
other embodiments, R.sub.6 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.2alkyl-N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In yet other
embodiments, R.sub.6 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, --CH.sub.2--O--(C.sub.1-C.sub.3alkyl),
--CH.sub.2--N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(phenyl), or C.sub.1-C.sub.4alkyl(5- or
6-membered heteroaryl). In yet other embodiments, R.sub.6 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl,
--CH.sub.2--O--(C.sub.1-C.sub.3alkyl),
--CH.sub.2--(C.sub.1-C.sub.6alkylamino),
C.sub.1-C.sub.4alkyl(phenyl), or C.sub.1-C.sub.4alkyl(5- or
6-membered heteroaryl). In some embodiments, R.sub.6 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl,
--CH.sub.2--O--(C.sub.1-C.sub.3alkyl),
--CH.sub.2--N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(phenyl), or C.sub.1-C.sub.4alkyl(5- or
6-membered heteroaryl containing 1 or 2 N atoms), or
C.sub.1-C.sub.4alkyl(5- or 6-membered heterocycloalkyl containing 1
or 2 N atoms).
[0099] In some embodiments, Y is an optionally substituted group
selected from among alkylene, heteroalkylene, cycloalkylene, and
heterocycloalkylene. In other embodiments, Y is an optionally
substituted group selected from among C.sub.1-C.sub.6alkylene,
C.sub.1-C.sub.6heteroalkylene, 4-, 5-, 6-, or 7-membered
cycloalkylene, and 4-, 5-, 6-, or 7-membered heterocycloalkylene.
In yet other embodiments, Y is an optionally substituted group
selected from among C.sub.1-C.sub.6alkylene,
C.sub.1-C.sub.6heteroalkylene, 5- or 6-membered cycloalkylene, and
5- or 6-membered heterocycloalkylene containing 1 or 2 N atoms. In
some other embodiments, Y is a 5- or 6-membered cycloalkylene, or a
5- or 6-membered heterocycloalkylene containing 1 or 2 N atoms. In
some embodiments, Y is a 4-, 5-, 6-, or 7-membered cycloalkylene
ring; or Y is a 4-, 5-, 6-, or 7-membered heterocycloalkylene
ring.
[0100] Any combination of the groups described above for the
various variables is contemplated herein.
[0101] In any of the aforementioned methods, assays and systems:
such methods, assays and systems comprise a multiplicity of test
irreversible inhibitors, in which the test irreversible inhibitors
each have the same
##STR00013##
moiety, but differ in at least one of Y, Z, R.sub.6, R.sub.7, or
R.sub.8. In further embodiments, the multiplicity of test
irreversible inhibitors is a panel of test irreversible inhibitors.
In further embodiments, the binding of the panel of test
irreversible inhibitors to at least one kinase is determined
(including a panel of kinases, further including a panel of kinases
selected from Btk, Btk homologs, and Btk kinase cysteine homologs).
In further embodiments, the determined binding data is used to
select and/or further design a selective irreversible
inhibitor.
[0102] Irreversible inhibitors described herein include those that
have a structure of any of Formula (A1-A6), Formula (B1-B6),
Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula (VII),
and pharmaceutically acceptable salts, solvates, esters, acids and
prodrugs thereof. In certain embodiments, isomers and chemically
protected forms of compounds having a structure represented by any
of Formula (A1-A6), Formula (B1-B6), Formula (C1-C6), Formula
(D1-D6), Formula (I), or Formula (VII), are also provided.
[0103] In one aspect, provided herein is an irreversible inhibitor
compound selected from among:
1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperi-
din-1-yl)prop-2-en-1-one;
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)but-2-en-1-one;
1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperi-
din-1-yl)sulfonylethene;
1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperi-
din-1-yl)prop-2-yn-1-one;
1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperi-
din-1-yl)prop-2-en-1-one;
N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)acrylamide;
1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)py-
rrolidin-1-yl)prop-2-en-1-one;
1-((S)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)py-
rrolidin-1-yl)prop-2-en-1-one;
1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)prop-2-en-1-one;
1-((S)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)prop-2-en-1-one; and
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-(dimethylamino)but-2-en-1-one;
(E)-4-(N-(2-hydroxyethyl)-N-methylamino)-1-(3-(4-phenoxyphenyl)-1H-pyrazo-
lo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)but-2-en-i-one (Compound 3);
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3-
-(1H-imidazol-4-yl)prop-2-en-1-one (Compound 4);
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-morpholinobut-2-en-1-one (Compound 5);
(E)-1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound 7);
(E)-N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-
-1-yl)cyclohexyl)-4-(dimethylamino)but-2-enamide (Compound 8);
N-((1r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)acrylamide (Compound 10);
(E)-1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)methyl)pyrolidin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound
11);
(E)-1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)methyl)pyrolidin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound
12);
1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 13);
1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 14);
1((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)me-
thyl)pyrrolidin-1-yl)but-2-yn-1-one (Compound 15);
1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)but-2-yn-1-one (Compound 16);
1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)but-2-yn-1-one (Compound 17);
(E)-N-((1,r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidi-
n-1-yl)cyclohexyl-4-(dimethylamino)but-2-enamide (Compound 18);
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
-N-methylacrylamide (Compound 19);
(E)-1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-4-
-morpholinobut-2-en-1-one (Compound 20);
(E)-1-((S-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)methyl)pyrrolidin-1-yl)-4-morpholinobut-2-en-1-one (Compound 21);
N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)but-2-ynamide (Compound 22);
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
acrylamide (Compound 23);
(E)-1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)piperidin-1-yl)-4-morpholinobut-2-en-1-one (Compound 24);
(E)-N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-
-1-yl)cyclohexyl)-4-morpholinobut-2-enamide (Compound 25).
[0104] Further described herein are pharmaceutical formulations
comprising the kinase inhibitors of any kinase inhibitor compound
previously listed. In one embodiment the pharmaceutical formulation
includes a pharmaceutical acceptable excipient. In some
embodiments, pharmaceutical formulations provided herein are
administered to a human. In some embodiments, the irreversible
and/or selective kinase inhibitors provided herein are orally
administered. In other embodiments, the irreversible and/or
selective kinase inhibitors provided herein are used for the
formulation of a medicament for the inhibition of tyrosine kinase
activity. In some other embodiments, the irreversible and/or
selective kinase inhibitors provided herein are used for the
formulation of a medicament for the inhibition of a kinase
activity, including a tyrosine kinase activity, including a Btk
activity, including a Btk homolog activity, including a Btk kinase
cysteine homolog activity, including an ACK activity, including
HER4.
[0105] In any of the aforementioned aspects are further embodiments
in which administration is enteral, parenteral, or both, and
wherein (a) the effective amount of the compound is systemically
administered to the mammal; (b) the effective amount of the
compound is administered orally to the mammal; (c) the effective
amount of the compound is intravenously administered to the mammal;
(d) the effective amount of the compound administered by
inhalation; (e) the effective amount of the compound is
administered by nasal administration; or (f) the effective amount
of the compound is administered by injection to the mammal; (g) the
effective amount of the compound is administered topically (dermal)
to the mammal; (h) the effective amount of the compound is
administered by ophthalmic administration; or (i) the effective
amount of the compound is administered rectally to the mammal. In
further embodiments the pharmaceutical formulation is formulated
for a route of administration selected from oral administration,
parenteral administration, buccal administration, nasal
administration, topical administration, or rectal
administration.
[0106] In any of the aforementioned aspects are further embodiments
comprising single administrations of the effective amount of the
pharmaceutical formulation, including further embodiments in which
(i) the pharmaceutical formulations is administered once; (ii) the
pharmaceutical formulations is administered to the mammal once a
day; (iii) the pharmaceutical formulations is administered to the
mammal multiple times over the span of one day; (iv) continually;
or (v) continuously.
[0107] In any of the aforementioned aspects are further embodiments
comprising multiple administrations of the effective amount of the
pharmaceutical formulations, including further embodiments in which
(i) the pharmaceutical formulations is administered in a single
dose; (ii) the time between multiple administrations is every 6
hours; (iii) the pharmaceutical formulations is administered to the
mammal every 8 hours. In further or alternative embodiments, the
method comprises a drug holiday, wherein the administration of the
pharmaceutical formulations is temporarily suspended or the dose of
the pharmaceutical formulations being administered is temporarily
reduced; at the end of the drug holiday, dosing of the
pharmaceutical formulations is resumed. The length of the drug
holiday varies from 2 days to 1 year.
[0108] Further described herein is a method for increasing the
selectivity of a test protein kinase inhibitor that irreversibly
and selectively binds to at least one protein kinase inhibitor
selected from Btk, a Btk homolog, a Btk kinase cysteine homolog, an
ACK, or HER4an ACK, or HER4HER4. In one embodiment the test protein
tyrosine kinase inhibitor is chemically modified to decrease the
plasma half life to less than about 4 hours. In another embodiment
the test protein tyrosine kinase inhibitor is chemically modified
to decrease the plasma half life to about 3 hours. In yet another
embodiment the test protein tyrosine kinase inhibitor
non-selectively and reversibly binds to a multiplicity of
src-family protein tyrosine kinases.
[0109] In one embodiment the test protein kinase inhibitor has the
structure of Formula (VII):
##STR00014##
wherein: [0110] wherein
##STR00015##
[0110] is a moiety that binds to the active site of a kinase,
including a tyrosine kinase, further including a Btk kinase
cysteine homolog; [0111] Y is an optionally substituted group
selected from among alkylene, heteroalkylene, arylene,
heteroarylene, heterocycloalkylene, cycloalkylene, alkylenearylene,
alkyleneheteroarylene, alkylenecycloalkylene, and
alkyleneheterocycloalkylene; [0112] Z is C(.dbd.O), OC(.dbd.O),
NHC(.dbd.O), NCH.sub.3C(.dbd.O), C(.dbd.S), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, NHS(.dbd.O).sub.x, where x is 1 or 2; [0113]
R.sub.7 and R.sub.8 are independently selected from among H,
unsubstituted C.sub.1-C.sub.4 alkyl, substituted
C.sub.1-C.sub.4alkyl, unsubstituted C.sub.1-C.sub.4heteroalkyl,
substituted C.sub.1-C.sub.4heteroalkyl, unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted C.sub.3-C.sub.6cycloalkyl,
unsubstituted C.sub.2-C.sub.6heterocycloalkyl, and substituted
C.sub.2-C.sub.6heterocycloalkyl; or [0114] R.sub.7 and R.sub.8
taken together form a bond; [0115] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted C.sub.2-C.sub.8heterocycloalkyl,
substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and [0116]
pharmaceutically active metabolites, or pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof.
[0117] In a further aspect, provided herein is a method for
treating a solid tumor by administering to an individual in need
thereof a test protein kinase inhibitor composition containing a
therapeutically effective amount of a compound that forms a
covalent bond (including an irreversible and/or selective covalent
bond) with Btk, a Btk homolog, a Btk kinase cysteine homolog, an
ACK, or HER4an ACK, or HER4. In one embodiment, the compound forms
a covalent bound with the activated form of Btk, a Btk homolog, a
Btk kinase cysteine homolog, an ACK, or HER4an ACK, or HER4. In
further or alternative embodiments, the compound irreversibly
inhibits Btk, a Btk homolog, a Btk kinase cysteine homolog, an ACK,
or HER4an ACK, or HER4 to which it is covalently bound. In a
further or alternative embodiment, the compound forms a covalent
bond (including an irreversible and/or selective covalent bond)
with a cysteine residue on Btk, a Btk homolog, a Btk kinase
cysteine homolog, an ACK, or HER4, or HER4.
[0118] In any of the aforementioned aspects are further embodiments
comprising single administrations of the effective amount of the
pharmaceutical formulation, including further embodiments in which
(i) the pharmaceutical formulations is administered once; (ii) the
pharmaceutical formulations is administered to the mammal once a
day; (iii) the pharmaceutical formulations is administered to the
mammal multiple times over the span of one day; (iv) continually;
or (v) continuously.
[0119] Also described herein is a method of identifying an
irreversible inhibitor of a kinase selected from Btk, a Btk
homolog, a Btk kinase cysteine homolog, an ACK, or HER4, or HER4
comprising: [0120] (1) contacting a multiplicity of kinases
selected from Btk, a Btk homolog, a Btk kinase cysteine homolog, an
ACK, or HER4, or HER4 with a compound that comprises a Michael
acceptor moiety; [0121] (2) contacting at least one non-kinase
molecule having at least one accessible SH group with the compound
that comprises a Michael acceptor moiety; and [0122] (3)
determining the covalent binding of the compound that comprises a
Michael acceptor with the multiplicity of kinases and the at least
one non-kinase molecule; and repeating steps (1), (2), and (3) for
at least one other compound that comprises a Michael acceptor
moiety.
[0123] Further described herein is a method of identifying an
irreversible inhibitor of a kinase selected from Btk, a Btk
homolog, a Btk kinase cysteine homolog, an ACK, or HER4 comprising:
[0124] (1) contacting a multiplicity of kinases selected from Btk,
a Btk homolog, a Btk kinase cysteine homolog, an ACK, or HER4 with
a compound that comprises a Michael acceptor moiety; [0125] (2)
contacting at least one non-kinase molecule having at least one
accessible SH group with the compound that comprises a Michael
acceptor moiety; and [0126] (3) determining the covalent binding of
the compound that comprises a Michael acceptor with the
multiplicity of kinases and the at least one non-kinase molecule;
and repeating steps (1), (2), and (3) for at least one other
compound that comprises a Michael acceptor moiety; and [0127] (4)
comparing the covalent binding of the compound that comprises a
Michael acceptor with the multiplicity of kinases and the at least
one non-kinase molecule; and repeating steps (1), (2), (3) and (4)
for at least one other compound that comprises a Michael acceptor
moiety.
[0128] In one embodiment the at least one non-kinase molecule
having at least one accessible SH group includes glutathione and/or
hemoglobin. In another embodiment the desired irreversible
inhibitor is selective for a particular kinase relative to other
kinases, glutathione and hemoglobin.
[0129] In some embodiments, the methods, assays and systems for
identifying an irreversible inhibitor of a kinase comprise
contacting each kinase with an Activity Probe. In further
embodiments, the methods, assays and systems for identifying an
irreversible inhibitor of a kinase further comprise a panel of
kinases comprising at least two kinases selected from Btk, a Btk
homolog, an ACK, HER4, and a Btk kinase cysteine homolog. In
further embodiments, the panel of kinases comprises at least three
such kinases, at least four such kinases, at least five such
kinases, at least six such kinases, at least seven such kinases, at
least eight such kinases, at least nine such kinases, or at least
ten such kinases.
[0130] In one embodiment steps (1) and (2) of the method of
identifying an irreversible inhibitor of a kinase selected from
Btk, a Btk homolog, a Btk kinase cysteine homolog, an ACK, or HER4
is conducted in vivo. In another embodiment step (3) of the method
of identifying an irreversible inhibitor of a kinase selected from
Btk, a Btk homolog, a Btk kinase cysteine homolog, an ACK, or HER4
is conducted in part using an Activity Probe.
[0131] In one embodiment, contacting a multiplicity of kinases
selected from Btk, a Btk homolog, a Btk kinase cysteine homolog, an
ACK, or HER4 with a compound that comprises a Michael acceptor
moiety is conducted in vivo. In another embodiment contacting at
least one non-kinase molecule having at least one accessible SH
group with the compound that comprises a Michael acceptor moiety is
conducted in vivo. In a further embodiment determining the covalent
binding of the compound that comprises a Michael acceptor with the
multiplicity of kinases and the at least one non-kinase molecule is
conducted in part using an Activity Probe. In a further embodiment
the determining step uses mass spectrometry. In yet further
embodiments the determining step uses fluorescence.
[0132] In further embodiments of methods and assays for identifying
an irreversible inhibitor of a kinase, including a protein kinase,
including a tyrosine kinase, a panel of kinases is contacted with
at least one irreversible inhibitor. In a further embodiment, the
panel of kinases is also contacted with an Activity Probe. In a
further embodiment, the binding of an irreversible inhibitor to a
kinase is determined from the binding of the Activity Probe to the
kinase. In a further embodiment, the binding of the Activity Probe
to a kinase is determined using fluorescence technique. In further
or alternative methods and assays, the Activity Probe is compatible
with flow cytometry. In further embodiments, the binding of the
irreversible inhibitor to one kinase is compared to the binding of
the irreversible inhibitor to at least one other kinase. In any of
the aforementioned embodiments, the panel of kinases is selected
from Btk, Btk homologs, and Btk kinase cysteine homologs. In a
further or alternative embodiment, the binding of an irreversible
inhibitor to a kinase is determined by mass spectrometry.
[0133] Also described herein are activity probes of Bruton's
tyrosine kinase (Btk), Btk homologs, and Btk kinase cysteine
homologs (collectively "Activity Probes"). Further described are
Activity Probes that include an irreversible inhibitor of Btk, a
Btk homolog, a Btk kinase cysteine homolog, an ACK, or HER4; a
linker moiety; and a reporter moiety. Further described are
Activity Probes that include a Michael addition acceptor moiety in
the structure of the Activity Probe. Further described are Activity
Probes that form a covalent bond with a cysteine residue on Btk, a
Btk homolog and/or a Btk kinase cysteine homolog. Also described
herein are Activity Probes that form a non-covalent bond with a
cysteine residue on Btk, a Btk homolog and/or a Btk kinase cysteine
homolog. Also described herein are methods for synthesizing such
Activity Probes, methods for using such Activity Probes in the
study of the activity of Btk, a Btk homolog, a Btk kinase cysteine
homolog, an ACK, or HER4, methods for using such Activity Probes in
the study of inhibitors (including the development of new
inhibitors) of Btk, a Btk homolog, a Btk kinase cysteine homolog,
an ACK, or HER4, and methods for using such Activity Probes in the
study of the pharmacodynamics of inhibitors of Btk, a Btk homolog,
a Btk kinase cysteine homolog, an ACK, or HER4.
[0134] In one embodiment are Activity Probes wherein the linker
moiety is selected from a bond, an optionally substituted alkyl
moiety, an optionally substituted heterocycle moiety, an optionally
substituted amide moiety, a ketone moiety, an optionally
substituted carbamate moiety, an ester moiety, or a combination
thereof. In another embodiment are Activity Probes wherein the
linker moiety comprises an optionally substituted heterocycle
moiety. In a further embodiment are Activity Probes wherein the
optionally substituted heterocycle moiety comprises a
piperazinyl-based moiety.
[0135] Also described herein are Activity Probes wherein the
reporter moiety is selected from the group consisting of a label, a
dye, a photocrosslinker, a cytotoxic compound, a drug, an affinity
label, a photoaffinity label, a reactive compound, an antibody or
antibody fragment, a biomaterial, a nanoparticle, a spin label, a
fluorophore, a metal-containing moiety, a radioactive moiety, a
novel functional group, a group that covalently or noncovalently
interacts with other molecules, a photocaged moiety, an actinic
radiation excitable moiety, a ligand, a photoisomerizable moiety,
biotin, a biotin analogue, a moiety incorporating a heavy atom, a
chemically cleavable group, a photocleavable group, a redox-active
agent, an isotopically labeled moiety, a biophysical probe, a
phosphorescent group, a chemiluminescent group, an electron dense
group, a magnetic group, an intercalating group, a chromophore, an
energy transfer agent, a biologically active agent, a detectable
label, or a combination thereof. In another embodiment are Activity
Probes wherein the reporter moiety is a fluorophore. In yet another
embodiment are Activity Probes wherein the fluorophore is a Bodipy
fluorophore. In yet a further embodiment are Activity Probes
wherein the Bodipy fluorophore is a Bodipy FL fluorophore.
[0136] Presented herein are Activity Probes wherein the inhibitor
moiety is derived from an irreversible inhibitor of Btk, a Btk
homolog, a Btk kinase cysteine homolog, an ACK, or HER4. In one
embodiment, are Activity Probes wherein the irreversible inhibitor
is:
##STR00016##
[0137] In another embodiment are Activity Probes having the
structure:
##STR00017##
[0138] In a further embodiment are Activity Probes wherein the
probe selectively labels a phosphorylated conformation of Btk, a
Btk homolog, a Btk kinase cysteine homolog, an ACK, or HER4. In
another embodiment are Activity Probes wherein the phosphorylated
conformation of Btk, a Btk homolog, a Btk kinase cysteine homolog,
an ACK, or HER4 is either an active or inactive form of Btk, a Btk
homolog, a Btk kinase cysteine homolog, an ACK, or HER4. In a
further embodiment are Activity Probes wherein the phosphorylated
conformation of Btk, a Btk homolog, a Btk kinase cysteine homolog,
an ACK, or HER4 is an active form of Btk, a Btk homolog, a Btk
kinase cysteine homolog, an ACK, or HER4. In one embodiment are
Activity Probes of wherein the probe is cell permeable.
[0139] In one aspect is a method for assessing the efficacy of a
potential Btk, Btk homolog and/or Btk kinase cysteine homolog
inhibitor in a mammal, comprising administering a potential Btk,
Btk homolog and/or Btk kinase cysteine homolog inhibitor to a
mammal, administering the Activity Probe described herein to the
mammal or to cells isolated from the mammal; measuring the activity
of the reporter moiety of the Activity Probe, and comparing the
activity of the reporter moiety to a standard.
[0140] In another aspect is a method for assessing the
pharmacodynamics of BTK, Btk homolog and/or Btk kinase cysteine
homolog inhibitor in a mammal, comprising administering BTK, Btk
homolog and/or Btk kinase cysteine homolog inhibitor to the mammal,
administering the Activity Probe presented herein to the mammal or
to cells isolated from the mammal, and measuring the activity of
the reporter moiety of the Activity Probe at different time points
following the administration of the inhibitor.
[0141] In a further aspect is a method for in vitro labeling of
Btk, a Btk homolog, a Btk kinase cysteine homolog, an ACK, or HER4
comprising contacting an active Btk, Btk homolog and/or Btk kinase
cysteine homolog with the Activity Probe described herein. In one
embodiment is a method for in vitro labeling of Btk, a Btk homolog,
a Btk kinase cysteine homolog, an ACK, or HER4 wherein the
contacting step comprises incubating the active Btk, Btk homolog
and/or Btk kinase cysteine homolog with the Activity Probe
presented herein.
[0142] In another aspect is a method for in vitro labeling of Btk,
a Btk homolog, a Btk kinase cysteine homolog, an ACK, or HER4
comprising contacting cells or tissues expressing the Btk, Btk
homolog and/or Btk kinase cysteine homolog with an Activity Probe
described herein.
[0143] In one aspect is a method for detecting a labeled Btk, Btk
homolog and/or Btk kinase cysteine homolog comprising separating
proteins, the proteins comprising Btk, a Btk homolog and/or a Btk
kinase cysteine homolog labeled by an Activity Probe described
herein, by electrophoresis and detecting the Activity Probe by
fluorescence.
[0144] In further embodiments the irreversible inhibitor of a
kinase further comprises an active site binding moiety. In yet
further embodiments the irreversible inhibitor of a kinase further
comprises a linker moiety that links the Michael acceptor moiety to
the active binding moiety.
[0145] In one embodiment the irreversible inhibitor of a kinase has
the structure of Formula (VII):
##STR00018##
wherein: [0146] wherein
##STR00019##
[0146] is a moiety that binds to the active site of a kinase,
including a tyrosine kinase, further including a Btk kinase
cysteine homolog; [0147] Y is an optionally substituted group
selected from among alkylene, heteroalkylene, arylene,
heteroarylene, heterocycloalkylene, cycloalkylene, alkylenearylene,
alkyleneheteroarylene, alkylenecycloalkylene, and
alkyleneheterocycloalkylene; [0148] Z is C(.dbd.O), OC(.dbd.O),
NHC(.dbd.O), NCH.sub.3C(.dbd.O), C(.dbd.S), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, NHS(.dbd.O).sub.x, where x is 1 or 2; [0149]
R.sub.7 and R.sub.8 are independently selected from among H,
unsubstituted C.sub.1-C.sub.4 alkyl, substituted
C.sub.1-C.sub.4alkyl, unsubstituted C.sub.1-C.sub.4heteroalkyl,
substituted C.sub.1-C.sub.4heteroalkyl, unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted C.sub.3-C.sub.6cycloalkyl,
unsubstituted C.sub.2-C.sub.6heterocycloalkyl, and substituted
C.sub.2-C.sub.6heterocycloalkyl; or [0150] R.sub.7 and R.sub.8
taken together form a bond; [0151] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted C.sub.2-C.sub.8heterocycloalkyl,
substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.5cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and [0152]
pharmaceutically active metabolites, or pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof.
[0153] In one embodiment the method of identifying an irreversible
inhibitor of a kinase selected from Btk, a Btk homolog, a Btk
kinase cysteine homolog, an ACK, or HER4 comprising steps (1), (2),
(3), and (4) further comprises analyzing the structure-function
activity relationship between the structure of the linker moiety
and/or the Michael acceptor moiety of each compound, and the
binding and/or selectivity of each compound to at least one kinase.
In another embodiment the method of identifying an irreversible
inhibitor of a kinase selected from Btk, a Btk homolog, a Btk
kinase cysteine homolog, an ACK, or HER4 comprising steps (1), (2),
(3), and (4) further comprises analyzing the structure-function
activity relationship between the structure of Y--Z and/or
##STR00020##
of each compound, and the binding and/or selectivity of each
compound to at least one kinase.
[0154] In one embodiment the structure of the active site binding
moiety of each compound is not varied. In another embodiment the
structure of
##STR00021##
of each compound is not varied.
[0155] Also described herein is a method for improving the kinase
selectivity of an inhibitor comprising use of any method previously
listed.
[0156] One aspect described herein is an assay comprising any of
the methods previously listed. Another aspect described herein is
system comprising any of the methods previously listed. In a
further aspect described herein is an irreversible inhibitor of a
kinase selected from Btk, a Btk homolog, an ACK, HER4, and a Btk
kinase cysteine homolog, wherein the inhibitor is identified using
any methods described herein.
[0157] In some aspects described herein the irreversible inhibitor
is selective for one kinase selected from Btk, a Btk homolog, an
ACK, HER4, and a Btk kinase cysteine homolog over at least one
other kinase selected from Btk, a Btk homolog, an ACK, HER4, and a
Btk kinase cysteine homolog. In other aspects described herein the
irreversible inhibitor is selective for at least one kinase
selected from Btk, a Btk homolog, an ACK, HER4, and a Btk kinase
cysteine homolog over at least one other non-kinase molecule having
an accessible SH group.
[0158] In certain embodiments, provided herein is a pharmaceutical
composition containing: i) a physiologically acceptable carrier,
diluent, and/or excipient; and ii) one or more compounds provided
herein.
[0159] In a further aspect, provided herein is a method for
treating a solid tumor comprising administering to an individual in
need thereof a composition containing a therapeutically effective
amount of a compound that forms a covalent bond with a cysteine
sidechain of a Bruton's tyrosine kinase or Bruton's tyrosine
homolog. In some embodiments, the solid tumor is a sarcoma,
lymphoma, and/or carcinoma. In some embodiments, the solid tumor is
a mammary ductal carcinoma, a lobular carcinoma, an adenocarcinoma
(e.g. pancreatic cancer and colon cancer), a small cell lung
carcinoma, a non-small cell lung carcinoma, or a melanoma. In some
embodiments, the solid tumor is a mammary ductal carcinoma, a
lobular carcinoma, or a combination thereof. In some embodiments,
the solid tumor is pancreatic cancer.
[0160] Also described herein are methods to identify biomarkers for
patient selection or patient monitoring prior to or during
treatment with any kinase inhibitor compound described herein. In
one embodiment, an individual that has lymphoma is administered a
pharmaceutical composition of any kinase inhibitor compound
described herein which inhibits B cell receptor (BCR) signaling. In
another embodiment, the inhibition of the BCR signaling by any
kinase inhibitor compound described herein is correlated with the
induction of apoptosis. In another embodiment, an individual with
lymphoma is selected for treatment with a pharmaceutical
composition of any kinase inhibitor compound described herein based
on a biomarker that indicates that the lymphoma in that patient has
high levels of pErk or Erk transcriptional targets. In another
embodiment, the response to treatment with a pharmaceutical
composition of any kinase inhibitor compound described herein is
measured by a reduction in levels of pErk or Erk transcriptional
targets.
[0161] Other objects, features and advantages of the methods and
compositions described herein will become apparent from the
following detailed description. It should be understood, however,
that the detailed description and the specific examples, while
indicating specific embodiments, are given by way of illustration
only. The section headings used herein are for organizational
purposes only and are not to be construed as limiting the subject
matter described.
Certain Terminology
[0162] It is to be understood that the foregoing general
description and the following detailed description are exemplary
and explanatory only and are not restrictive of any subject matter
claimed. In this application, the use of the singular includes the
plural unless specifically stated otherwise. It must be noted that,
as used in the specification and the appended claims, the singular
forms "a," "an" and "the" include plural referents unless the
context clearly dictates otherwise. In this application, the use of
"or" means "and/or" unless stated otherwise. Furthermore, use of
the term "including" as well as other forms, such as "include",
"includes," and "included," is not limiting.
[0163] Definition of standard chemistry terms are found in
reference works, including Carey and Sundberg "ADVANCED ORGANIC
CHEMISTRY 4.sup.TH ED." Vols. A (2000) and B (2001), Plenum Press,
New York. Unless otherwise indicated, conventional methods of mass
spectroscopy, NMR, HPLC, protein chemistry, biochemistry,
recombinant DNA techniques and pharmacology, within the skill of
the art are employed. Unless specific definitions are provided, the
nomenclature employed in connection with, and the laboratory
procedures and techniques of, analytical chemistry, synthetic
organic chemistry, and medicinal and pharmaceutical chemistry
described herein are those known in the art. Standard techniques
are optionally used for chemical syntheses, chemical analyses,
pharmaceutical preparation, formulation, and delivery, and
treatment of patients. Standard techniques are optionally used for
recombinant DNA, oligonucleotide synthesis, and tissue culture and
transformation (e.g., electroporation, lipofection). Reactions and
purification techniques are performed using documented
methodologies or as described herein.
[0164] It is to be understood that the methods and compositions
described herein are not limited to the particular methodology,
protocols, cell lines, constructs, and reagents described herein
and as such optionally vary. It is also to be understood that the
terminology used herein is for the purpose of describing particular
embodiments only, and is not intended to limit the scope of the
methods and compositions described herein, which will be limited
only by the appended claims.
[0165] Unless stated otherwise, the terms used for complex moieties
(i.e., multiple chains of moieties) are to be read equivalently
either from left to right or right to left. For example, the group
alkylenecycloalkylene refers both to an alkylene group followed by
a cycloalkylene group or as a cycloalkylene group followed by an
alkylene group.
[0166] The suffix "ene" appended to a group indicates that such a
group is a diradical. By way of example only, a methylene is a
diradical of a methyl group, that is, it is a --CH.sub.2-- group;
and an ethylene is a diradical of an ethyl group, i.e.,
--CH.sub.2CH.sub.2--.
[0167] An "alkyl" group refers to an aliphatic hydrocarbon group.
The alkyl moiety includes a "saturated alkyl" group, which means
that it does not contain any alkene or alkyne moieties. The alkyl
moiety also includes an "unsaturated alkyl" moiety, which means
that it contains at least one alkene or alkyne moiety. An "alkene"
moiety refers to a group that has at least one carbon-carbon double
bond, and an "alkyne" moiety refers to a group that has at least
one carbon-carbon triple bond. The alkyl moiety, whether saturated
or unsaturated, includes branched, straight chain, or cyclic
moieties. Depending on the structure, an alkyl group includes a
monoradical or a diradical (i.e., an alkylene group), and if a
"lower alkyl" having 1 to 6 carbon atoms.
[0168] As used herein, C.sub.1-C.sub.x includes C.sub.1-C.sub.2,
C.sub.1-C.sub.3 . . . C.sub.1-C.sub.x.
[0169] The "alkyl" moiety optionally has 1 to 10 carbon atoms
(whenever it appears herein, a numerical range such as "1 to 10"
refers to each integer in the given range; e.g., "1 to 10 carbon
atoms" means that the alkyl group is selected from a moiety having
1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and
including 10 carbon atoms, although the present definition also
covers the occurrence of the term "alkyl" where no numerical range
is designated). The alkyl group of the compounds described herein
may be designated as "C.sub.1-C.sub.4 alkyl" or similar
designations. By way of example only, "C.sub.1-C.sub.4 alkyl"
indicates that there are one to four carbon atoms in the alkyl
chain, i.e., the alkyl chain is selected from among methyl, ethyl,
propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, and t-butyl.
Thus C.sub.1-C.sub.4 alkyl includes C.sub.1-C.sub.2 alkyl and
C.sub.1-C.sub.3 alkyl. Alkyl groups are optionally substituted or
unsubstituted. Typical alkyl groups include, but are in no way
limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl,
tertiary butyl, pentyl, hexyl, ethenyl, propenyl, butenyl,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
[0170] The term "alkenyl" refers to a type of alkyl group in which
the first two atoms of the alkyl group form a double bond that is
not part of an aromatic group. That is, an alkenyl group begins
with the atoms --C(R).dbd.C(R)--R, wherein R refers to the
remaining portions of the alkenyl group, which are either the same
or different. The alkenyl moiety is optionally branched, straight
chain, or cyclic (in which case, it is also known as a
"cycloalkenyl" group). Depending on the structure, an alkenyl group
includes a monoradical or a diradical (i.e., an alkenylene group).
Alkenyl groups are optionally substituted. Non-limiting examples of
an alkenyl group include --CH.dbd.CH.sub.2,
--C(CH.sub.3).dbd.CH.sub.2, --CH.dbd.CHCH.sub.3,
--C(CH.sub.3).dbd.CHCH.sub.3. Alkenylene groups include, but are
not limited to, --CH.dbd.CH--, --C(CH.sub.3).dbd.CH--,
--CH.dbd.CHCH.sub.2--, --CH.dbd.CHCH.sub.2CH.sub.2-- and
--C(CH.sub.3).dbd.CHCH.sub.2--. Alkenyl groups optionally have 2 to
10 carbons, and if a "lower alkenyl" having 2 to 6 carbon
atoms.
[0171] The term "alkynyl" refers to a type of alkyl group in which
the first two atoms of the alkyl group form a triple bond. That is,
an alkynyl group begins with the atoms --C.ident.C--R, wherein R
refers to the remaining portions of the alkynyl group, which is
either the same or different. The "R" portion of the alkynyl moiety
may be branched, straight chain, or cyclic. Depending on the
structure, an alkynyl group includes a monoradical or a diradical
(i.e., an alkynylene group). Alkynyl groups are optionally
substituted. Non-limiting examples of an alkynyl group include, but
are not limited to, --C.ident.CH, --C.ident.CCH.sub.3,
--C.ident.CCH.sub.2CH.sub.3, --C.ident.C--, and
--C.ident.CCH.sub.2--. Alkynyl groups optionally have 2 to 10
carbons, and if a "lower alkynyl" having 2 to 6 carbon atoms.
[0172] An "alkoxy" group refers to a (alkyl)O-- group, where alkyl
is as defined herein.
[0173] "Hydroxyalkyl" refers to an alkyl radical, as defined
herein, substituted with at least one hydroxy group. Non-limiting
examples of a hydroxyalkyl include, but are not limited to,
hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl,
1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl,
4-hydroxybutyl, 2,3-dihydroxypropyl,
1-(hydroxymethyl)-2-hydroxyethyl, 2,3-dihydroxybutyl,
3,4-dihydroxybutyl and 2-(hydroxymethyl)-3-hydroxypropyl.
[0174] "Alkoxyalkyl" refers to an alkyl radical, as defined herein,
substituted with an alkoxy group, as defined herein.
[0175] The term "alkylamine" refers to the --N(alkyl).sub.xH.sub.y
group, where x and y are selected from among x=1, y=1 and x=2, y=O.
When x=2, the alkyl groups, taken together with the N atom to which
they are attached, optionally form a cyclic ring system.
[0176] "Alkylaminoalkyl" refers to an alkyl radical, as defined
herein, substituted with an alkylamine, as defined herein.
[0177] "Hydroxyalkylaminoalkyl" refers to an alkyl radical, as
defined herein, substituted with an alkylamine, and alkylhydroxy,
as defined herein.
[0178] "Alkoxyalkylaminoalkyl" refers to an alkyl radical, as
defined herein, substituted with an alkylamine and substituted with
an alkylalkoxy, as defined herein.
[0179] An "amide" is a chemical moiety with the formula --C(O)NHR
or --NHC(O)R, where R is selected from among alkyl, cycloalkyl,
aryl, heteroaryl (bonded through a ring carbon) and heteroalicyclic
(bonded through a ring carbon). In some embodiments, an amide
moiety forms a linkage between an amino acid or a peptide molecule
and a compound described herein, thereby forming a prodrug. Any
amine, or carboxyl side chain on the compounds described herein can
be amidified. The procedures and specific groups to make such
amides are found in sources such as Greene and Wuts, Protective
Groups in Organic Synthesis, 3.sup.rd Ed., John Wiley & Sons,
New York, N.Y., 1999, which is incorporated herein by reference for
this disclosure.
[0180] The term "ester" refers to a chemical moiety with formula
--COOR, where R is selected from among alkyl, cycloalkyl, aryl,
heteroaryl (bonded through a ring carbon) and heteroalicyclic
(bonded through a ring carbon). Any hydroxy, or carboxyl side chain
on the compounds described herein can be esterified. The procedures
and specific groups to make such esters are found in sources such
as Greene and Wuts, Protective Groups in Organic Synthesis,
3.sup.rd Ed., John Wiley & Sons, New York, N.Y., 1999, which is
incorporated herein by reference for this disclosure.
[0181] As used herein, the term "ring" refers to any covalently
closed structure. Rings include, for example, carbocycles (e.g.,
aryls and cycloalkyls), heterocycles (e.g., heteroaryls and
non-aromatic heterocycles), aromatics (e.g. aryls and heteroaryls),
and non-aromatics (e.g., cycloalkyls and non-aromatic
heterocycles). Rings can be optionally substituted. Rings can be
monocyclic or polycyclic.
[0182] As used herein, the term "ring system" refers to one, or
more than one ring.
[0183] The term "membered ring" can embrace any cyclic structure.
The term "membered" is meant to denote the number of skeletal atoms
that constitute the ring. Thus, for example, cyclohexyl, pyridine,
pyran and thiopyran are 6-membered rings and cyclopentyl, pyrrole,
furan, and thiophene are 5-membered rings.
[0184] The term "fused" refers to structures in which two or more
rings share one or more bonds.
[0185] The term "carbocyclic" or "carbocycle" refers to a ring
wherein each of the atoms forming the ring is a carbon atom.
Carbocycle includes aryl and cycloalkyl. The term thus
distinguishes carbocycle from heterocycle ("heterocyclic") in which
the ring backbone contains at least one atom which is different
from carbon (i.e. a heteroatom). Heterocycle includes heteroaryl
and heterocycloalkyl. Carbocycles and heterocycles can be
optionally substituted.
[0186] The term "aromatic" refers to a planar ring having a
delocalized .pi.-electron system containing 4n+2 .pi. electrons,
where n is an integer. Aromatic rings can be formed from five, six,
seven, eight, nine, or more than nine atoms. Aromatics can be
optionally substituted. The term "aromatic" includes both
carbocyclic aryl (e.g., phenyl) and heterocyclic aryl (or
"heteroaryl" or "heteroaromatic") groups (e.g., pyridine). The term
includes monocyclic or fused-ring polycyclic (i.e., rings which
share adjacent pairs of carbon atoms) groups.
[0187] As used herein, the term "aryl" refers to an aromatic ring
wherein each of the atoms forming the ring is a carbon atom. Aryl
rings can be formed by five, six, seven, eight, nine, or more than
nine carbon atoms. Aryl groups can be optionally substituted.
Examples of aryl groups include, but are not limited to phenyl,
naphthalenyl, phenanthrenyl, anthracenyl, fluorenyl, and indenyl.
Depending on the structure, an aryl group can be a monoradical or a
diradical (i.e., an arylene group).
[0188] An "aryloxy" group refers to an (aryl)O-- group, where aryl
is as defined herein.
[0189] The term "carbonyl" as used herein refers to a group
containing a moiety selected from the group consisting of --C(O)--,
--S(O)--, --S(O)2-, and --C(S)--, including, but not limited to,
groups containing a least one ketone group, and/or at least one
aldehyde group, and/or at least one ester group, and/or at least
one carboxylic acid group, and/or at least one thioester group.
Such carbonyl groups include ketones, aldehydes, carboxylic acids,
esters, and thioesters. In some embodiments, such groups are a part
of linear, branched, or cyclic molecules.
[0190] The term "cycloalkyl" refers to a monocyclic or polycyclic
radical that contains only carbon and hydrogen, and is optionally
saturated, partially unsaturated, or fully unsaturated. Cycloalkyl
groups include groups having from 3 to 10 ring atoms. Illustrative
examples of cycloalkyl groups include the following moieties:
##STR00022##
and the like. Depending on the structure, a cycloalkyl group is
either a monoradical or a diradical (e.g., an cycloalkylene group),
and if a "lower cycloalkyl" having 3 to 8 carbon atoms.
[0191] "Cycloalkylalkyl" means an alkyl radical, as defined herein,
substituted with a cycloalkyl group. Non-limiting cycloalkylalkyl
groups include cyclopropylmethyl, cyclobutylmethyl,
cyclopentylmethyl, cyclohexylmethyl, and the like.
[0192] The term "heterocycle" refers to heteroaromatic and
heteroalicyclic groups containing one to four heteroatoms each
selected from O, S and N, wherein each heterocyclic group has from
4 to 10 atoms in its ring system, and with the proviso that the
ring of said group does not contain two adjacent O or S atoms.
Herein, whenever the number of carbon atoms in a heterocycle is
indicated (e.g., C.sub.1-C.sub.6 heterocycle), at least one other
atom (the heteroatom) must be present in the ring. Designations
such as "C.sub.1-C.sub.6 heterocycle" refer only to the number of
carbon atoms in the ring and do not refer to the total number of
atoms in the ring. It is understood that the heterocylic ring can
have additional heteroatoms in the ring. Designations such as "4-6
membered heterocycle" refer to the total number of atoms that are
contained in the ring (i.e., a four, five, or six membered ring, in
which at least one atom is a carbon atom, at least one atom is a
heteroatom and the remaining two to four atoms are either carbon
atoms or heteroatoms). In heterocycles that have two or more
heteroatoms, those two or more heteroatoms can be the same or
different from one another. Heterocycles can be optionally
substituted. Binding to a heterocycle can be at a heteroatom or via
a carbon atom. Non-aromatic heterocyclic groups include groups
having only 4 atoms in their ring system, but aromatic heterocyclic
groups must have at least 5 atoms in their ring system. The
heterocyclic groups include benzo-fused ring systems. An example of
a 4-membered heterocyclic group is azetidinyl (derived from
azetidine). An example of a 5-membered heterocyclic group is
thiazolyl. An example of a 6-membered heterocyclic group is
pyridyl, and an example of a 10-membered heterocyclic group is
quinolinyl. Examples of non-aromatic heterocyclic groups are
pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl,
tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl,
piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl,
azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl,
thiepanyl, oxazepinyl, diazepinyl, thiazepinyl,
1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl,
2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl,
dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl,
dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl,
3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, 3H-indolyl
and quinolizinyl. Examples of aromatic heterocyclic groups are
pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl,
pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl,
oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl,
indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl,
indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl,
pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl,
benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl,
quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. The
foregoing groups, as derived from the groups listed above, are
optionally C-attached or N-attached where such is possible. For
instance, a group derived from pyrrole includes pyrrol-1-yl
(N-attached) or pyrrol-3-yl (C-attached). Further, a group derived
from imidazole includes imidazol-1-yl or imidazol-3-yl (both
N-attached) or imidazol-2-yl, imidazol-4-yl or imidazol-5-yl (all
C-attached). The heterocyclic groups include benzo-fused ring
systems and ring systems substituted with one or two oxo (.dbd.O)
moieties such as pyrrolidin-2-one. Depending on the structure, a
heterocycle group can be a monoradical or a diradical (i.e., a
heterocyclene group).
[0193] The terms "heteroaryl" or, alternatively, "heteroaromatic"
refers to an aromatic group that includes one or more ring
heteroatoms selected from nitrogen, oxygen and sulfur. An
N-containing "heteroaromatic" or "heteroaryl" moiety refers to an
aromatic group in which at least one of the skeletal atoms of the
ring is a nitrogen atom. Illustrative examples of heteroaryl groups
include the following moieties:
##STR00023##
and the like. Depending on the structure, a heteroaryl group can be
a monoradical or a diradical (i.e., a heteroarylene group).
[0194] As used herein, the term "non-aromatic heterocycle",
"heterocycloalkyl" or "heteroalicyclic" refers to a non-aromatic
ring wherein one or more atoms forming the ring is a heteroatom. A
"non-aromatic heterocycle" or "heterocycloalkyl" group refers to a
cycloalkyl group that includes at least one heteroatom selected
from nitrogen, oxygen and sulfur. In some embodiments, the radicals
are fused with an aryl or heteroaryl. Heterocycloalkyl rings can be
formed by three, four, five, six, seven, eight, nine, or more than
nine atoms. Heterocycloalkyl rings can be optionally substituted.
In certain embodiments, non-aromatic heterocycles contain one or
more carbonyl or thiocarbonyl groups such as, for example, oxo- and
thio-containing groups. Examples of heterocycloalkyls include, but
are not limited to, lactams, lactones, cyclic imides, cyclic
thioimides, cyclic carbamates, tetrahydrothiopyran, 4H-pyran,
tetrahydropyran, piperidine, 1,3-dioxin, 1,3-dioxane, 1,4-dioxin,
1,4-dioxane, piperazine, 1,3-oxathiane, 1,4-oxathiin,
1,4-oxathiane, tetrahydro-1,4-thiazine, 2H-1,2-oxazine, maleimide,
succinimide, barbituric acid, thiobarbituric acid, dioxopiperazine,
hydantoin, dihydrouracil, morpholine, trioxane,
hexahydro-1,3,5-triazine, tetrahydrothiophene, tetrahydrofuran,
pyrroline, pyrrolidine, pyrrolidone, pyrrolidone, pyrazoline,
pyrazolidine, imidazoline, imidazolidine, 1,3-dioxole,
1,3-dioxolane, 1,3-dithiole, 1,3-dithiolane, isoxazoline,
isoxazolidine, oxazoline, oxazolidine, oxazolidinone, thiazoline,
thiazolidine, and 1,3-oxathiolane. Illustrative examples of
heterocycloalkyl groups, also referred to as non-aromatic
heterocycles, include:
##STR00024##
and the like. The term heteroalicyclic also includes all ring forms
of the carbohydrates, including but not limited to the
monosaccharides, the disaccharides and the oligosaccharides.
Depending on the structure, a heterocycloalkyl group can be a
monoradical or a diradical (i.e., a heterocycloalkylene group).
[0195] The term "halo" or, alternatively, "halogen" or "halide"
means fluoro, chloro, bromo and iodo.
[0196] The term "haloalkyl," refers to alkyl structures in which at
least one hydrogen is replaced with a halogen atom. In certain
embodiments in which two or more hydrogen atoms are replaced with
halogen atoms, the halogen atoms are all the same as one another.
In other embodiments in which two or more hydrogen atoms are
replaced with halogen atoms, the halogen atoms are not all the same
as one another.
[0197] The term "fluoroalkyl," as used herein, refers to alkyl
group in which at least one hydrogen is replaced with a fluorine
atom. Examples of fluoroalkyl groups include, but are not limited
to, --CF.sub.3, --CH.sub.2CF.sub.3, --CF.sub.2CF.sub.3,
--CH.sub.2CH.sub.2CF.sub.3 and the like.
[0198] As used herein, the term "heteroalkyl" refers to optionally
substituted alkyl radicals in which one or more skeletal chain
atoms is a heteroatom, e.g., oxygen, nitrogen, sulfur, silicon,
phosphorus or combinations thereof. The heteroatom(s) are placed at
any interior position of the heteroalkyl group or at the position
at which the heteroalkyl group is attached to the remainder of the
molecule. Examples include, but are not limited to,
--CH.sub.2--O--CH.sub.3, --CH.sub.2--CH.sub.2--O--CH.sub.3,
--CH.sub.2--NH--CH.sub.3, --CH.sub.2--CH.sub.2--NH--CH.sub.3,
--CH.sub.2--N(CH.sub.3)--CH.sub.3,
--CH.sub.2--CH.sub.2--NH--CH.sub.3,
--CH.sub.2--CH.sub.2--N(CH.sub.3)--CH.sub.3,
--CH.sub.2--S--CH.sub.2--CH.sub.3, --CH.sub.2--CH.sub.2,
--S(O)--CH.sub.3, --CH.sub.2--CH.sub.2--S(O).sub.2--CH.sub.3,
--CH.dbd.CH--O--CH.sub.3, --Si(CH.sub.3).sub.3,
--CH.sub.2--CH.dbd.N--OCH.sub.3, and
--CH.dbd.CH--N(CH.sub.3)--CH.sub.3. In addition, in some
embodiments, up to two heteroatoms are consecutive, such as, by way
of example, --CH.sub.2--NH--OCH.sub.3 and
--CH.sub.2--O--Si(CH.sub.3).sub.3.
[0199] The term "heteroatom" refers to an atom other than carbon or
hydrogen. Heteroatoms are typically independently selected from
among oxygen, sulfur, nitrogen, silicon and phosphorus, but are not
limited to these atoms. In embodiments in which two or more
heteroatoms are present, the two or more heteroatoms can all be the
same as one another, or some or all of the two or more heteroatoms
can each be different from the others.
[0200] The term "bond" or "single bond" refers to a chemical bond
between two atoms, or two moieties when the atoms joined by the
bond are considered to be part of larger sub structure.
[0201] The term "moiety" refers to a specific segment or functional
group of a molecule. Chemical moieties are often recognized
chemical entities embedded in or appended to a molecule.
[0202] A "thioalkoxy" or "alkylthio" group refers to a --S-alkyl
group.
[0203] A "SH" group is also referred to either as a thiol group or
a sulfhydryl group.
[0204] The term "optionally substituted" or "substituted" means
that the referenced group may be substituted with one or more
additional group(s) individually and independently selected from
alkyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy,
alkoxy, aryloxy, alkylthio, arylthio, alkylsulfoxide,
arylsulfoxide, alkylsulfone, arylsulfone, cyano, halo, acyl, nitro,
haloalkyl, fluoroalkyl, amino, including mono- and di-substituted
amino groups, and the protected derivatives thereof. By way of
example an optional substituents may be L.sub.sR.sub.s, wherein
each L.sub.s is independently selected from a bond, --O--,
--C(.dbd.O)--, --S--, --S(.dbd.O)--, --S(.dbd.O).sub.2--, --NH--,
--NHC(O)--, --C(O)NH--, S(.dbd.O).sub.2NH--, --NHS(.dbd.O).sub.2,
--OC(O)NH--, --NHC(O)O--, -(substituted or unsubstituted
C.sub.1-C.sub.6 alkyl), or -(substituted or unsubstituted
C.sub.2-C.sub.6 alkenyl); and each R.sub.s is independently
selected from H, (substituted or unsubstituted
C.sub.1-C.sub.4alkyl), (substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl), heteroaryl, or heteroalkyl. The
protecting groups that forms the protective derivatives of the
above substituents include those found in sources such as Greene
and Wuts, above.
[0205] The term "Michael acceptor moiety" refers to a functional
group that can participate in a Michael reaction, wherein a new
covalent bond is formed between a portion of the Michael acceptor
moiety and the donor moiety. The Michael acceptor moiety is an
electrophile and the "donor moiety" is a nucleophile. The "G"
groups presented in any of Formula (A1-A6), Formula (B1-B6),
Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula (VII) are
non-limiting examples of Michael acceptor moieties.
[0206] The term "nucleophile" or "nucleophilic" refers to an
electron rich compound, or moiety thereof. An example of a
nucleophile includes, but in no way is limited to, a cysteine
residue of a molecule, such as, for example Cys 481 of Btk.
[0207] The term "electrophile", or "electrophilic" refers to an
electron poor or electron deficient molecule, or moiety thereof.
Examples of electrophiles include, but in no way are limited to,
Michael acceptor moieties.
[0208] The term "acceptable" or "pharmaceutically acceptable", with
respect to a formulation, composition or ingredient, as used
herein, means having no persistent detrimental effect on the
general health of the subject being treated or does not abrogate
the biological activity or properties of the compound, and is
relatively nontoxic.
[0209] As used herein, the term "agonist" refers to a compound, the
presence of which results in a biological activity of a protein
that is the same as the biological activity resulting from the
presence of a naturally occurring ligand for the protein, such as,
for example, Btk.
[0210] As used herein, "ACK" and "Accessible Cysteine Kinase" are
synonyms. They mean a kinase with an accessible cysteine residue.
ACKS include, but are not limited to, BTK, ITK, Bmx/ETK, TEC, EFGR,
HER4, HER4, LCK, BLK, C-src, FGR, Fyn, HCK, Lyn, YES, ABL, Brk,
CSK, FER, JAK3, SYK. In some embodiments, the ACK is HER4.
[0211] As used herein, the term "partial agonist" refers to a
compound the presence of which results in a biological activity of
a protein that is of the same type as that resulting from the
presence of a naturally occurring ligand for the protein, but of a
lower magnitude.
[0212] As used herein, the term "antagonist" refers to a compound,
the presence of which results in a decrease in the magnitude of a
biological activity of a protein. In certain embodiments, the
presence of an antagonist results in complete inhibition of a
biological activity of a protein, such as, for example, Btk. In
certain embodiments, an antagonist is an inhibitor.
[0213] As used herein, "amelioration" of the symptoms of a
particular disorder by administration of a particular compound or
pharmaceutical composition refers to any lessening of severity,
delay in onset, slowing of progression, or shortening of duration,
whether permanent or temporary, lasting or transient that can be
attributed to or associated with administration of the compound or
composition.
[0214] "Bioavailability" refers to the percentage of the weight of
compounds disclosed herein, such as, compounds of any of Formula
(A1-A6), Formula (B1-B6), Formula (C1-C6), Formula (D1-D6), Formula
(I), or Formula (VII), dosed that is delivered into the general
circulation of the animal or human being studied. The total
exposure (AUC.sub.(0-.infin.)) of a drug when administered
intravenously is usually defined as 100% bioavailable (F %). "Oral
bioavailability" refers to the extent to which compounds disclosed
herein, such as, compounds of any of Formula (A1-A6), Formula
(B1-B6), Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula
(VII), are absorbed into the general circulation when the
pharmaceutical composition is taken orally as compared to
intravenous injection.
[0215] The term "biophysical probe," as used herein, refers to
probes which detect or monitor structural changes in molecules
(including biomolecules) in biological systems or in the presence
of other biomolecules (e.g., ex vivo, in vivo or in vitro). In some
embodiments, such molecules include, but are not limited to,
proteins and the "biophysical probe" is used to detect or monitor
interaction of proteins with other macromolecules. In other
embodiments, examples of biophysical probes include, but are not
limited to, spin-labels, fluorophores, and photoactivatable
groups.
[0216] "Blood plasma concentration" refers to the concentration of
compounds disclosed herein, such as, compounds of any of Formula
(A1-A6), Formula (B1-B6), Formula (C1-C6), Formula (D1-D6), Formula
(I), or Formula (VII), in the plasma component of blood of an
individual. It is understood that the plasma concentration of
compounds of any of Formula (A1-A6), Formula (B1-B6), Formula
(C1-C6), Formula (D1-D6), Formula (I), or Formula (VII), may vary
significantly between subjects, due to variability with respect to
metabolism and/or possible interactions with other therapeutic
agents. In accordance with one embodiment disclosed herein, the
blood plasma concentration of the compounds of any of Formula
(A1-A6), Formula (B1-B6), Formula (C1-C6), Formula (D1-D6), Formula
(I), or Formula (VII), does vary from subject to subject. Likewise,
values such as maximum plasma concentration (C.sub.max) or time to
reach maximum plasma concentration (T.sub.max), or total area under
the plasma concentration time curve (AUC.sub.(0-.infin.)) may vary
from subject to subject. Due to this variability, the amount
necessary to constitute "a therapeutically effective amount" of a
compound of any of Formula (A1-A6), Formula (B1-B6), Formula
(C1-C6), Formula (D1-D6), Formula (I), or Formula (VII), is
expected to vary from subject to subject.
[0217] The term "Bruton's tyrosine kinase," as used herein, refers
to Bruton's tyrosine kinase from Homo sapiens, as disclosed in,
e.g., U.S. Pat. No. 6,326,469 (GenBank Accession No.
NP_000052).
[0218] The term "Bruton's tyrosine kinase homolog," as used herein,
refers to orthologs of Bruton's tyrosine kinase, e.g., the
orthologs from mouse (GenBank Accession No. AAB47246), dog (GenBank
Accession No. XP_549139.), rat (GenBank Accession No.
NP_001007799), chicken (GenBank Accession No. NP_989564), or zebra
fish (GenBank Accession No. XP_698117), and fusion proteins of any
of the foregoing that exhibit kinase activity towards one or more
substrates of Bruton's tyrosine kinase (e.g. a peptide substrate
having the amino acid sequence "AVLESEEELYSSARQ" (SEQ ID NO:
1)).
[0219] The term "HER4", also known as ERBB4, also known as "V-erb-a
erythroblastic leukemia viral oncogene homolog 4" means either (a)
the nucleic acid sequence encoding a receptor tyrosine kinase that
is a member of the epidermal growth factor receptor subfamily, or
(b) the protein thereof. For the nucleic acid sequence that
comprises the human HER4 gene see GenBank Accession No.
NM_001042599. For the amino acid sequence that comprises the human
HER4 protein see GenBank Accession No. NP_001036064.
[0220] The term "chemiluminescent group," as used herein, refers to
a group which emits light as a result of a chemical reaction
without the addition of heat. By way of example only, luminol
(5-amino-2,3-dihydro-1,4-phthalazinedione) reacts with oxidants
like hydrogen peroxide (H.sub.2O.sub.2) in the presence of a base
and a metal catalyst to produce an excited state product
(3-aminophthalate, 3-APA).
[0221] The term "chromophore," as used herein, refers to a molecule
which absorbs light of visible wavelengths, UV wavelengths or IR
wavelengths.
[0222] The terms "co-administration" or the like, as used herein,
are meant to encompass administration of the selected therapeutic
agents to a single patient, and are intended to include treatment
regimens in which the agents are administered by the same or
different route of administration or at the same or different
time.
[0223] In other embodiments, the term "detectable label," as used
herein, refers to a label which is observable using analytical
techniques including, but not limited to, fluorescence,
chemiluminescence, electron-spin resonance, ultraviolet/visible
absorbance spectroscopy, mass spectrometry, nuclear magnetic
resonance, magnetic resonance, and electrochemical methods.
[0224] The term "dye," as used herein, refers to a soluble,
coloring substance which contains a chromophore.
[0225] The terms "effective amount" or "therapeutically effective
amount," as used herein, refer to a sufficient amount of an agent
or a compound being administered which will relieve to some extent
one or more of the symptoms of the disorder being treated. The
result can be reduction and/or alleviation of the signs, symptoms,
or causes of a disease, or any other desired alteration of a
biological system. For example, an "effective amount" for
therapeutic uses is the amount of the composition including a
compound as disclosed herein required to provide a clinically
significant decrease in disease symptoms without undue adverse side
effects. An appropriate "effective amount" in any individual case
is optionally determined using techniques, such as a dose
escalation study. The term "therapeutically effective amount"
includes, for example, a prophylactically effective amount. An
"effective amount" of a compound disclosed herein is an amount
effective to achieve a desired pharmacologic effect or therapeutic
improvement without undue adverse side effects. It is understood
that "an effect amount" or "a therapeutically effective amount" can
vary from subject to subject, due to variation in metabolism of the
compound of any of Formula (A1-A6), Formula (B1-B6), Formula
(C1-C6), Formula (D1-D6), Formula (I), or Formula (VII), age,
weight, general condition of the subject, the condition being
treated, the severity of the condition being treated, and the
judgment of the prescribing physician.
[0226] The term "electron dense group," as used herein, refers to a
group which scatters electrons when irradiated with an electron
beam. Such groups include, but are not limited to, ammonium
molybdate, bismuth subnitrate cadmium iodide, 99%, carbohydrazide,
ferric chloride hexahydrate, hexamethylene tetramine, 98.5%, indium
trichloride anhydrous, lanthanum nitrate, lead acetate trihydrate,
lead citrate trihydrate, lead nitrate, periodic acid,
phosphomolybdic acid, phosphotungstic acid, potassium ferricyanide,
potassium ferrocyanide, ruthenium red, silver nitrate, silver
proteinate (Ag Assay: 8.0-8.5%) "Strong", silver tetraphenylporphin
(S-TPPS), sodium chloroaurate, sodium tungstate, thallium nitrate,
thiosemicarbazide (TSC), uranyl acetate, uranyl nitrate, and
vanadyl sulfate.
[0227] In other embodiments, the term "energy transfer agent," as
used herein, refers to a molecule which either donates or accepts
energy from another molecule. By way of example only, fluorescence
resonance energy transfer (FRET) is a dipole-dipole coupling
process by which the excited-state energy of a fluorescence donor
molecule is non-radioactively transferred to an unexcited acceptor
molecule which then fluorescently emits the donated energy at a
longer wavelength.
[0228] The terms "enhance" or "enhancing" means to increase or
prolong either in potency or duration a desired effect. By way of
example, "enhancing" the effect of therapeutic agents refers to the
ability to increase or prolong, either in potency or duration, the
effect of therapeutic agents on during treatment of a disorder. An
"enhancing-effective amount," as used herein, refers to an amount
adequate to enhance the effect of a therapeutic agent in the
treatment of a disorder. When used in an individual, amounts
effective for this use will depend on the severity and course of
the disorder, previous therapy, the individual's health status and
response to the drugs, and the judgment of the treating
physician.
[0229] The term "fluorophore," as used herein, refers to a molecule
which upon excitation emits photons and is thereby fluorescent.
[0230] The term "homologous cysteine," as used herein refers to a
cysteine residue found with in a sequence position that is
homologous to that of cysteine 481 of Bruton's tyrosine kinase, as
defined herein. For example, cysteine 482 is the homologous
cysteine of the rat ortholog of Bruton's tyrosine kinase; cysteine
479 is the homologous cysteine of the chicken ortholog; and
cysteine 481 is the homologous cysteine in the zebra fish ortholog.
In another example, the homologous cysteine of TXK, a Tec kinase
family member related to Bruton's tyrosine, is Cys 350. Other
examples of kinases having homologous cysteines are shown in FIG.
7. See also the sequence alignments of tyrosine kinases (TK)
published on the world wide web at
kinase.com/human/kinome/phylogeny.html.
[0231] The term "identical," as used herein, refers to two or more
sequences or subsequences which are the same. In addition, the term
"substantially identical," as used herein, refers to two or more
sequences which have a percentage of sequential units which are the
same when compared and aligned for maximum correspondence over a
comparison window, or designated region as measured using
comparison algorithms or by manual alignment and visual inspection.
By way of example only, two or more sequences are "substantially
identical" if the sequential units are about 60% identical, about
65% identical, about 70% identical, about 75% identical, about 80%
identical, about 85% identical, about 90% identical, or about 95%
identical over a specified region. Such percentages to describe the
"percent identity" of two or more sequences. The identity of a
sequence can exist over a region that is at least about 75-100
sequential units in length, over a region that is about 50
sequential units in length, or, where not specified, across the
entire sequence. This definition also refers to the complement of a
test sequence. By way of example only, two or more polypeptide
sequences are identical when the amino acid residues are the same,
while two or more polypeptide sequences are "substantially
identical" if the amino acid residues are about 60% identical,
about 65% identical, about 70% identical, about 75% identical,
about 80% identical, about 85% identical, about 90% identical, or
about 95% identical over a specified region. The identity can exist
over a region that is at least about 75-100 amino acids in length,
over a region that is about 50 amino acids in length, or, where not
specified, across the entire sequence of a polypeptide sequence. In
addition, by way of example only, two or more polynucleotide
sequences are identical when the nucleic acid residues are the
same, while two or more polynucleotide sequences are "substantially
identical" if the nucleic acid residues are about 60% identical,
about 65% identical, about 70% identical, about 75% identical,
about 80% identical, about 85% identical, about 90% identical, or
about 95% identical over a specified region. The identity can exist
over a region that is at least about 75-100 nucleic acids in
length, over a region that is about 50 nucleic acids in length, or,
where not specified, across the entire sequence of a polynucleotide
sequence.
[0232] The terms "inhibits", "inhibiting", or "inhibitor" of a
kinase, as used herein, refer to inhibition of enzymatic
phosphotransferase activity.
[0233] The term "irreversible inhibitor," as used herein, refers to
a compound that, upon contact with a target protein (e.g., a
kinase) causes the formation of a new covalent bond with or within
the protein, whereby one or more of the target protein's biological
activities (e.g., phosphotransferase activity) is diminished or
abolished notwithstanding the subsequent presence or absence of the
irreversible inhibitor.
[0234] The term "irreversible Btk inhibitor," as used herein,
refers to an inhibitor of Btk that can form a covalent bond with an
amino acid residue of Btk. In one embodiment, the irreversible
inhibitor of Btk can form a covalent bond with a Cys residue of
Btk; in particular embodiments, the irreversible inhibitor can form
a covalent bond with a Cys 481 residue (or a homolog thereof) of
Btk or a cysteine residue in the homologous corresponding position
of another tyrosine kinase, as shown in FIG. 7.
[0235] The term "isolated," as used herein, refers to separating
and removing a component of interest from at least some portion of
components not of interest. Isolated substances can be in either a
dry or semi-dry state, or in solution, including but not limited to
an aqueous solution. The isolated component can be in a homogeneous
state or the isolated component can be a part of a pharmaceutical
composition that comprises additional pharmaceutically acceptable
carriers and/or excipients. By way of example only, nucleic acids
or proteins are "isolated" when such nucleic acids or proteins are
free of at least some of the cellular components with which it is
associated in the natural state, or that the nucleic acid or
protein has been concentrated to a level greater than the
concentration of its in vivo or in vitro production. Also, by way
of example, a gene is isolated when separated from open reading
frames which flank the gene and encode a protein other than the
gene of interest.
[0236] In some embodiments, the term "label," as used herein,
refers to a substance which is incorporated into a compound and is
readily detected, whereby its physical distribution is detected
and/or monitored.
[0237] The term "linkage," as used herein to refer to bonds or a
chemical moiety formed from a chemical reaction between the
functional group of a linker and another molecule. In some
embodiments, such bonds include, but are not limited to, covalent
linkages and non-covalent bonds, while such chemical moieties
include, but are not limited to, esters, carbonates, imines,
phosphate esters, hydrazones, acetals, orthoesters, peptide
linkages, and oligonucleotide linkages. Hydrolytically stable
linkages means that the linkages are substantially stable in water
and do not react with water at useful pH values, including but not
limited to, under physiological conditions for an extended period
of time, perhaps even indefinitely. Hydrolytically unstable or
degradable linkages means that the linkages are degradable in water
or in aqueous solutions, including for example, blood. In other
embodiments, enzymatically unstable or degradable linkages means
that the linkage is degraded by one or more enzymes. By way of
example only, PEG and related polymers include degradable linkages
in the polymer backbone or in the linker group between the polymer
backbone and one or more of the terminal functional groups of the
polymer molecule. Such degradable linkages include, but are not
limited to, ester linkages formed by the reaction of PEG carboxylic
acids or activated PEG carboxylic acids with alcohol groups on a
biologically active agent, wherein such ester groups generally
hydrolyze under physiological conditions to release the
biologically active agent. Other hydrolytically degradable linkages
include but are not limited to carbonate linkages; imine linkages
resulted from reaction of an amine and an aldehyde; phosphate ester
linkages formed by reacting an alcohol with a phosphate group;
hydrazone linkages which are reaction product of a hydrazide and an
aldehyde; acetal linkages that are the reaction product of an
aldehyde and an alcohol; orthoester linkages that are the reaction
product of a formate and an alcohol; peptide linkages formed by an
amine group, including but not limited to, at an end of a polymer
such as PEG, and a carboxyl group of a peptide; and oligonucleotide
linkages formed by a phosphoramidite group, including but not
limited to, at the end of a polymer, and a 5' hydroxyl group of an
oligonucleotide.
[0238] The phrase "measuring the activity of the reporter moiety"
(or a similarly worded phrase) refers to methods for quantifying
(in absolute, approximate or relative terms) the reporter moiety in
a system under study. In some embodiments, such methods include any
methods that quantify a reporter moiety that is a dye; a
photocrosslinker; a cytotoxic compound; a drug; an affinity label;
a photoaffinity label; a reactive compound; an antibody or antibody
fragment; a biomaterial; a nanoparticle; a spin label; a
fluorophore, a metal-containing moiety; a radioactive moiety; a
novel functional group; a group that covalently or noncovalently
interacts with other molecules; a photocaged moiety; an actinic
radiation excitable moiety; a ligand; a photoisomerizable moiety;
biotin; a biotin analogue; a moiety incorporating a heavy atom; a
chemically cleavable group; a photocleavable group; a redox-active
agent; an isotopically labeled moiety; a biophysical probe; a
phosphorescent group; a chemiluminescent group; an electron dense
group; a magnetic group; an intercalating group; a chromophore; an
energy transfer agent; a biologically active agent; a detectable
label; and any combination of the above.
[0239] A "metabolite" of a compound disclosed herein is a
derivative of that compound that is formed when the compound is
metabolized. The term "active metabolite" refers to a biologically
active derivative of a compound that is formed when the compound is
metabolized. The term "metabolized," as used herein, refers to the
sum of the processes (including, but not limited to, hydrolysis
reactions and reactions catalyzed by enzymes, such as, oxidation
reactions) by which a particular substance is changed by an
organism. Thus, enzymes produce specific structural alterations to
a compound. For example, cytochrome P450 catalyzes a variety of
oxidative and reductive reactions while uridine diphosphate
glucuronyl transferases catalyze the transfer of an activated
glucuronic-acid molecule to aromatic alcohols, aliphatic alcohols,
carboxylic acids, amines and free sulfhydryl groups. Further
information on metabolism is obtained from The Pharmacological
Basis of Therapeutics, 9th Edition, McGraw-Hill (1996). Metabolites
of the compounds disclosed herein are optionally identified either
by administration of compounds to a host and analysis of tissue
samples from the host, or by incubation of compounds with hepatic
cells in vitro and analysis of the resulting compounds. In some
embodiments, metabolites of a compound are formed by oxidative
processes and correspond to the corresponding hydroxy-containing
compound. In some embodiments, a compound is metabolized to
pharmacologically active metabolites.
[0240] The term "modulate," as used herein, means to interact with
a target either directly or indirectly so as to alter the activity
of the target, including, by way of example only, to enhance the
activity of the target, to inhibit the activity of the target, to
limit the activity of the target, or to extend the activity of the
target.
[0241] As used herein, the term "modulator" refers to a compound
that alters an activity of a molecule. For example, a modulator can
cause an increase or decrease in the magnitude of a certain
activity of a molecule compared to the magnitude of the activity in
the absence of the modulator. In certain embodiments, a modulator
is an inhibitor, which decreases the magnitude of one or more
activities of a molecule. In certain embodiments, an inhibitor
completely prevents one or more activities of a molecule. In
certain embodiments, a modulator is an activator, which increases
the magnitude of at least one activity of a molecule. In certain
embodiments the presence of a modulator results in an activity that
does not occur in the absence of the modulator.
[0242] The term "moiety incorporating a heavy atom," as used
herein, refers to a group which incorporates an ion of atom which
is usually heavier than carbon. In some embodiments, such ions or
atoms include, but are not limited to, silicon, tungsten, gold,
lead, and uranium.
[0243] The term "nanoparticle," as used herein, refers to a
particle which has a particle size between about 500 nm to about 1
nm.
[0244] As used herein, the term "pERK" refers to phosphorylated
ERK1 and ERK2 at Thr202/Tyr 204 as detected by commercially
available phospho-specific antibodies (e.g. Cell Signaling
Technologies #4377).
[0245] The term "photoaffinity label," as used herein, refers to a
label with a group, which, upon exposure to light, forms a linkage
with a molecule for which the label has an affinity. By way of
example only, in some embodiments, such a linkage is covalent or
non-covalent.
[0246] The term "photocaged moiety," as used herein, refers to a
group which, upon illumination at certain wavelengths, covalently
or non-covalently binds other ions or molecules.
[0247] The term "photoisomerizable moiety," as used herein, refers
to a group wherein upon illumination with light changes from one
isomeric form to another.
[0248] The term "plasma half life," as used herein refers to
half-life in rat, dog or human as determined by measure drug
concentration over time in plasma following a single dose and
fitting data to standard pharmacokinetic models using software such
as WinNonLin to determine the time at which drug has been 50%
eliminated from plasma.
[0249] The term "prophylactically effective amount," as used
herein, refers that amount of a composition applied to an
individual which will relieve to some extent one or more of the
symptoms of a disease, disorder being treated. In such prophylactic
applications, such amounts may depend on the patient's state of
health, weight, and the like.
[0250] The term "radioactive moiety," as used herein, refers to a
group whose nuclei spontaneously give off nuclear radiation, such
as alpha, beta, or gamma particles; wherein, alpha particles are
helium nuclei, beta particles are electrons, and gamma particles
are high energy photons.
[0251] As used herein, the term "selective binding compound" refers
to a compound that selectively binds to any portion of one or more
target proteins.
[0252] As used herein, the term "selectively binds" refers to the
ability of a selective binding compound to bind to a target
protein, such as, for example, Btk, with greater affinity than it
binds to a non-target protein. In certain embodiments, specific
binding refers to binding to a target with an affinity that is at
least 10, 50, 100, 250, 500, 1000 or more times greater than the
affinity for a non-target.
[0253] As used herein, the term "selective modulator" refers to a
compound that selectively modulates a target activity relative to a
non-target activity. In certain embodiments, specific modulator
refers to modulating a target activity at least 10, 50, 100, 250,
500, 1000 times more than a non-target activity.
[0254] The term "spin label," as used herein, refers to molecules
which contain an atom or a group of atoms exhibiting an unpaired
electron spin (i.e. a stable paramagnetic group) that in some
embodiments are detected by electron spin resonance spectroscopy
and in other embodiments are attached to another molecule. Such
spin-label molecules include, but are not limited to, nitryl
radicals and nitroxides, and in some embodiments are single
spin-labels or double spin-labels.
[0255] The term "substantially purified," as used herein, refers to
a component of interest that may be substantially or essentially
free of other components which normally accompany or interact with
the component of interest prior to purification. By way of example
only, a component of interest may be "substantially purified" when
the preparation of the component of interest contains less than
about 30%, less than about 25%, less than about 20%, less than
about 15%, less than about 10%, less than about 5%, less than about
4%, less than about 3%, less than about 2%, or less than about 1%
(by dry weight) of contaminating components. Thus, a "substantially
purified" component of interest may have a purity level of about
70%, about 75%, about 80%, about 85%, about 90%, about 95%, about
96%, about 97%, about 98%, about 99% or greater.
[0256] The term "individual" as used herein, refers to a mammal
which is the object of treatment, observation or experiment. The
term is not to be construed as requiring the supervision of a
medical practitioner (e.g., a physician, physician's assistant,
nurse, orderly, hospice care worker).
[0257] As used herein, the term "target activity" refers to a
biological activity capable of being modulated by a selective
modulator. Certain exemplary target activities include, but are not
limited to, binding affinity, signal transduction, enzymatic
activity, tumor growth, inflammation or inflammation-related
processes, and amelioration of one or more symptoms associated with
a disorder.
[0258] As used herein, the term "target protein" refers to a
molecule or a portion of a protein capable of being bound by a
selective binding compound. In certain embodiments, a target
protein is Btk.
[0259] The terms "treat," "treating" or "treatment", as used
herein, include alleviating, abating or ameliorating a symptom of a
disorder, preventing additional symptoms, ameliorating or
preventing the underlying metabolic causes of symptoms, inhibiting
the disorder, e.g., arresting the development of the disorder,
relieving the disorder, causing regression of the disorder,
relieving a condition caused by the disorder, or stopping the
symptoms of the disorder. The terms "treat," "treating" or
"treatment", include, but are not limited to, prophylactic and/or
therapeutic treatments.
[0260] As used herein, the IC.sub.50 refers to an amount,
concentration or dosage of a particular test compound that achieves
a 50% inhibition of a maximal response, such as inhibition of Btk,
in an assay that measures such response.
[0261] As used herein, EC.sub.50 refers to a dosage, concentration
or amount of a particular test compound that elicits a
dose-dependent response at 50% of maximal expression of a
particular response that is induced, provoked or potentiated by the
particular test compound.
BRIEF DESCRIPTION OF THE FIGURES
[0262] FIG. 1A presents an illustrative table of GI.sub.50
concentrations of Compound 1 that results in 50% decrease in cell
proliferation. A variety of lymphoma cell lines incubated with a
range of concentrations of Compound 1.
[0263] FIG. 1B presents an illustrative line graph showing
inhibition of tumor growth in DLCL2 xenograft models.
[0264] FIG. 1C presents an illustrative line graph showing
inhibition of tumor growth in DOHH2 xenograft models. For in vivo
lymphoma xenograft studies, 5E6 DOHH2 or DLCL2 cells in 50%
matrigel were implanted subcutaneously in SCID mice and dosed
orally with Compound 1 beginning when tumor size reached 100
mm.sup.2.
[0265] FIG. 2 presents an illustrative line graph showing
inhibition of collagen-induced arthritis in male DBA/1OlaHsd mice.
Compound 1 or vehicle was dosed orally once per day starting at day
1. Dexamethasone was included as a positive control. Paw
inflammation was scored from 0-5 and averaged across all paws from
all animals for each group in the study. Compound 1 at 12.5 mg/kg
and 50 mg/kg regressed inflammation through the end of the study
(day 11) while 3.125 mg/kg significantly reduced the increase in
paw inflammation.
[0266] FIG. 3 presents an illustrative line graph showing
inhibition of disease progression in a mouse MRL/lpr model of
lupus. MRL/lpr mice (Jax strain 000485) were dosed orally once per
day from 8 weeks of age until 20 weeks of age and urine protein
levels were measured weekly. Compound 1 at 3.125 mg/kg, 12.5 mg/kg,
and 50 mg/kg significantly reduced proteinuria, indicating
amelioration of the progressive autoimmune renal failure seen in
this mouse strain.
[0267] FIG. 4 presents an illustrative bar graph showing inhibition
of mast cell degranulation in a mouse passive cutaneous anaphylaxis
model. 23 hours after mice were sensitized with an intradermal
injection of monoclonal anti-DNP-IgE in the back, they received a
single oral dose of Compound 1 or vehicle. After one hour, animals
were challenged with an intravenous injection of DNP-BSA and Evans
Blue dye and the area of extravasation was measured. Increasing
doses of Compound 1 significantly decreased the amount of Evans
Blue release, indicating decreased mast cell activation and
vascular permeabilization.
[0268] FIG. 5 presents an illustrative line graph showing in vivo
plasma concentrations post-dosing of male jugular vein cannulated
rats with Compounds 1, 7, 8, and 12. Blood samples were collected
at 0.0833 (5 minutes), 0.333 (20 minutes), 1, 3, 6, 9, and 24 hours
post-dosing from orally dosed rats. Compound 1 and Compound 12 have
a short half-life in vivo. In contrast, Compound 7 and Compound 8
have a significantly longer in vivo half-life. Compounds like 1 and
12 are predicted to have enhanced kinase selectivity in vivo
because inhibition will be sustained only for those kinases that
are irreversibly inhibited.
[0269] FIG. 6 presents an illustrative bar graph showing brief
exposure to Compound 1 in vitro is sufficient to inhibit B cell
activation in normal human B cells. B cells were purified from
blood from healthy donors by negative selecting using the
RosetteSep Human B cell enrichment cocktail. Cells were plated in
growth media and indicated concentrations of Compound 1 were added.
After incubation for 1 hour at 37.degree. C., cells were washed
three times using an 8-fold dilution in growth media for each wash.
Cells were then stimulated with IgM F(ab')2 for 18 hours at
37.degree. C., stained with anti-CD69-PE antibody and analyzed by
flow cytometry. This protocol mimics the predicted exposure of
cells to Compound 1 in vivo and demonstrates that inhibition of B
cells is sustained despite washing out of Compound 1.
[0270] FIG. 7 presents illustrative ACKs, including Btk and Btk
cysteine homologs. FIG. 7 discloses SEQ ID NOS: 2-14, respectively,
in order of appearance.
[0271] FIG. 8 shows efficacy of HER4 inhibitor Compound 1 in
MDA-MB-453 grown as a xenograft in nude mice.
DETAILED DESCRIPTION OF THE INVENTION
Solid Tumors
[0272] In some embodiments, the compounds and formulations
described herein are utilized to treat one or more disorders
characterized by the presence or development of a solid tumor. As
used herein, "solid tumors" are neoplasms characterized by an
absence of liquid areas. In some embodiments, the solid tumor is
benign. In some embodiments, the solid tumor is malignant. In some
embodiments, the cancer is characterized by the presence of one or
more solid tumor is a sarcoma, carcinoma, and/or lymphoma.
[0273] In some embodiments, the disorder characterized by the
presence or development of one or more solid tumors is a sarcoma.
Sarcomas are cancers of the bone, cartilage, fat, muscle, blood
vessels, or other connective or supportive tissue. Sarcomas
include, but are not limited to, chondrosarcoma, Ewing's sarcoma,
malignant hemangioendothelioma, malignant schwannoma, osteosarcoma,
soft tissue sarcomas (e.g. alveolar soft part sarcoma,
angiosarcoma, cystosarcoma phylloides, dermatofibrosarcoma, desmoid
tumor, epithelioid sarcoma, extraskeletal osteosarcoma,
fibrosarcoma, hemangiopericytoma, hemangiosarcoma, Kaposi's
sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma,
lymphosarcoma, malignant fibrous histiocytoma, neurofibrosarcoma,
rhabdomyosarcoma, and synovial sarcoma).
[0274] In some embodiments, the disorder characterized by the
presence or development of one or more solid tumors is a lymphoma.
Lymphomas are solid neoplasms that originate in lymphocytes.
Hodgkin lymphoma is marked by the presence of the Reed-Sternberg
cell. Non-Hodgkin lymphomas are all lymphomas which are not
Hodgkin's lymphoma. Non-Hodgkin lymphomas are further divided into
indolent lymphomas and aggressive lymphomas. Non-Hodgkin's
lymphomas include, but are not limited to, diffuse large B cell
lymphoma; follicular lymphoma, Mucosa-Associated Lymphatic Tissue
lymphoma (MALT), small cell lymphocytic lymphoma, mantle cell
lymphoma, Burkitt's lymphoma, mediastinal large B cell lymphoma,
Waldenstrom macroglobulinemia, nodal marginal zone B cell lymphoma
(NMZL), splenic marginal zone lymphoma (SMZL), extranodal marginal
zone B cell lymphoma, intravascular large B cell lymphoma, primary
effusion lymphoma, and Lymphomatoid granulomatosis.
[0275] In some embodiments, the disorder characterized by the
presence or development of one or more solid tumors is a carcinoma.
Carcinomas are cancers that begin in the epithelial cells. By way
of non-limiting example, carcinomas include most breast cancers
(e.g. mammary ductal carcinoma and lobular carcinoma), most
pancreatic cancers, most lung cancers (e.g. small cell lung
carcinoma, and non-small cell lung carcinoma), most colon cancers,
most kidney cancers, and melanomas. In some embodiments, the
disease is mammary ductal carcinoma, lobular carcinoma, an
adenocarcinoma (e.g. pancreatic cancer and colon cancer), small
cell lung carcinoma, non-small cell lung carcinoma, and melanomas.
In some embodiments, the disease is breast cancer. In some
embodiments, the disease is mammary ductal carcinoma, lobular
carcinoma, or a combination thereof. In some embodiments, the
breast cancer is ER positive. In some embodiments, the breast
cancer is ER negative. In some embodiments, the breast cancer is
progesterone receptor (PgR)-positive. In some embodiments, the
breast cancer is PgR-negative. In some embodiments, the disease is
pancreatic cancer.
[0276] Pancreatic cancer is defined as the presence of malignant
tumors of the pancreas. The prognosis for individuals with
pancreatic cancer is generally regarded as poor. In general only
about 10 to 15% of patients diagnosed with the disorder will
survive for 1 year or more; only about 3% live for 5 years or more;
and only about 2% live for 10 years or more. The majority of
pancreatic tumors are classified as adenocarcinomas.
[0277] Mammary ductal carcinoma is a type of breast cancer. It
comes in two forms. Infiltrating ductal carcinoma (IDC) is an
invasive, malignant and abnormal proliferation of neoplastic cells
in the breast tissue. Ductal carcinoma in situ (DCIS), is a
noninvasive, possibly malignant neoplasm that is still confined to
the lactiferous ducts, where breast cancer most often
originates.
[0278] Lobular carcinoma is a neoplasm primarily found in the
lobules of a gland. It comes in two forms. Lobular carcinoma in
situ (LCIS) is a condition caused by neoplastic (but not
necessarily cancerous) cells in the lobules of a breast. Invasive
lobular carcinoma (aka infiltrating lobular carcinoma) is a type of
breast cancer that begins in the lobules and then invades
surrounding tissues.
[0279] The growth and development (e.g. into malignant tumors) of a
solid tumor requires the growth of new blood vessels (i.e.
angiogenesis). The transcription factor MYC is often overexpressed
in cancerous cells. In certain instances, MYC facilitates
angiogenesis in tumors by recruiting mast cells to the tumor. In
certain instances, tumor cells will undergo hypoxia and cell death
if mast cell recruitment is inhibited. In some embodiments, mast
cell recruitment is inhibited by the use of a Btk inhibitor. In
some embodiments, mast cells are killed (e.g. by necrosis or
apoptosis) by the use of a Btk inhibitor.
Irreversible Inhibitor Compounds
[0280] In the following description of irreversible kinase
inhibitor compounds suitable for use in the methods described
herein, definitions of referred-to standard chemistry terms may be
found in reference works (if not otherwise defined herein),
including Carey and Sundberg "Advanced Organic Chemistry 4th Ed."
Vols. A (2000) and B (2001), Plenum Press, New York. In addition,
nucleic acid and amino acid sequences for Btk (e.g., human Btk) are
disclosed in, e.g., U.S. Pat. No. 6,326,469. Unless specific
definitions are provided, the nomenclature employed in connection
with, and the laboratory procedures and techniques of, analytical
chemistry, synthetic organic chemistry, and medicinal and
pharmaceutical chemistry described herein are those known in the
art. Standard techniques can be used for chemical syntheses,
chemical analyses, pharmaceutical preparation, formulation, and
delivery, and treatment of patients
[0281] The inhibitor compounds described herein are selective for
kinases having an accessible cysteine residue (such kinases are
also known as Accessible Cysteine Kinases, or ACKs) that is able to
form a covalent bond with a Michael acceptor moiety on the
inhibitor compound. In some embodiments, the cysteine residue is
accessible or becomes accessible when the binding site moiety of
the irreversible inhibitor binds to the kinase. That is, the
binding site moiety of the irreversible inhibitor binds to an
active site of the ACK and the Michael acceptor moiety of
irreversible inhibitor gains access (in one embodiment the step of
binding leads to a conformational change in the ACK, thus exposing
the cysteine) or is otherwise exposed to the cysteine residue of
the ACK; as a result a covalent bond is formed between the "S" of
the cysteine residue and the Michael acceptor of the irreversible
inhibitor. Consequently, the binding site moiety of the
irreversible inhibitor remains bound or otherwise blocks the active
site of the ACK.
[0282] In one embodiment, the ACK is Btk, a homolog of Btk or a
tyrosine kinase having a cysteine residue in an amino acid sequence
position that is homologous to the amino acid sequence position of
cysteine 481 in Btk. See, e.g., kinases in FIG. 7. In some
embodiments, the ACK is HER4. Inhibitor compounds described herein
include a Michael acceptor moiety, a binding site moiety and a
linker that links the binding site moiety and the Michael acceptor
moiety (and in some embodiments, the structure of the linker
provides a conformation, or otherwise directs the Michael acceptor
moiety, so as to improve the selectivity of the irreversible
inhibitor for a particular ACK).
[0283] Generally, an irreversible inhibitor compound used in the
methods described herein is identified or characterized in an in
vitro assay, e.g., a cellular biochemical assay or a cellular
functional assay. Such assays are useful to determine an in vitro
IC.sub.50 for an irreversible inhibitor compound.
[0284] For example, a cellular kinase assay is used to determine
kinase activity after incubation of the kinase in the absence or
presence of a range of concentrations of a candidate irreversible
inhibitor compound. If the candidate compound is in fact an
irreversible inhibitor, kinase activity will not be recovered by
repeat washing with inhibitor-free medium. See, e.g., J. B. Smaill,
et al. (1999), J. Med. Chem. 42(10):1803-1815. Further, covalent
complex formation between a Kinase and a candidate irreversible
inhibitor is a useful indicator of irreversible inhibition of the
Kinase that is readily determined by a number of methods (e.g.,
mass spectrometry). For example, some irreversible Kinase-inhibitor
compounds form a covalent bond with the aforenoted cysteine residue
(e.g., via a Michael reaction).
[0285] High throughput assays for many a cellular biochemical
assays (e.g., kinase assays) and cellular functional assays (e.g.,
calcium flux) are documented methodologies. In addition, high
throughput screening systems are commercially available (see, e.g.,
Zymark Corp., Hopkinton, Mass.; Air Technical Industries, Mentor,
OH; Beckman Instruments, Inc. Fullerton, Calif.; Precision Systems,
Inc., Natick, Mass., etc.). These systems typically automate entire
procedures including all sample and reagent pipetting, liquid
dispensing, timed incubations, and final readings of the microplate
in detector(s) appropriate for the assay. Automated systems thereby
allow the identification and characterization of a large number of
irreversible compounds.
[0286] In some embodiments, irreversible inhibitor compounds are
used for the manufacture of a medicament for treating any of the
foregoing conditions (e.g. lymphomas, carcinomas, and/or
sarcomas).
[0287] In some embodiments, the irreversible inhibitor compound
used for the methods described herein inhibits a Kinase activity
with an in vitro IC.sub.50 of less than 10 .mu.M. (e.g., less than
1 .mu.M, less than 0.5 .mu.M, less than 0.4 .mu.M, less than 0.3
.mu.M, less than 0.1, less than 0.08 .mu.M, less than 0.06 .mu.M,
less than 0.05 .mu.M, less than 0.04 .mu.M, less than 0.03 .mu.M,
less than less than 0.02 .mu.M, less than 0.01, less than 0.008
.mu.M, less than 0.006 .mu.M, less than 0.005 .mu.M, less than
0.004 .mu.M, less than 0.003 .mu.M, less than less than 0.002
.mu.M, less than 0.001, less than 0.00099 .mu.M, less than 0.00098
.mu.M, less than 0.00097 .mu.M, less than 0.00096 .mu.M, less than
0.00095 .mu.M, less than 0.00094 .mu.M, less than 0.00093 .mu.M,
less than 0.00092, or less than 0.00090 .mu.M).
[0288] In one embodiment, the irreversible inhibitor compound
selectively and irreversibly inhibits an activated form of its
target tyrosine kinase (e.g., a phosphorylated form of the tyrosine
kinase). For example, activated Btk is transphosphorylated at
tyrosine 551. Thus, in these embodiments the irreversible Btk
inhibitor inhibits the target kinase in cells only once the target
kinase is activated by the signaling events.
Particular Irreversible Inhibitor Compounds for ACKs
[0289] Described herein are compounds of any of Formula (A1-A6),
Formula (B1-B6), Formula (C1-C6), Formula (D1-D6), Formula (I), or
Formula (VII). Also described herein are pharmaceutically
acceptable salts, pharmaceutically acceptable solvates,
pharmaceutically active metabolites, and pharmaceutically
acceptable prodrugs of such compounds. Pharmaceutical compositions
that include at least one such compound or a pharmaceutically
acceptable salt, pharmaceutically acceptable solvate,
pharmaceutically active metabolite or pharmaceutically acceptable
prodrug of such compound, are provided. In some embodiments, when
compounds disclosed herein contain an oxidizable nitrogen atom, the
nitrogen atom is optionally converted to an N-oxide. In certain
embodiments, isomers and chemically protected forms of compounds
having a structure represented by any of Formula (A1-A6), Formula
(B1-B6), Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula
(VII), are also provided.
[0290] In one aspect are compounds (including irreversible
inhibitors of ACKs, including Btk and its cysteine homologs) having
the structure of Formula (I):
##STR00025##
wherein [0291] L.sub.a is CH.sub.2, O, NH or S; [0292] Ar is a
substituted or unsubstituted aryl, or a substituted or
unsubstituted heteroaryl; and either [0293] (a) Y is an optionally
substituted group selected from among alkylene, heteroalkylene,
arylene, heteroarylene, alkylenearylene, alkyleneheteroarylene,
alkylenecycloalkylene and alkyleneheterocycloalkylene; [0294] Z is
C(.dbd.O), NHC(.dbd.O), NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x,
where x is 1 or 2, and R.sup.a is H, substituted or unsubstituted
alkyl, substituted or unsubstituted cycloalkyl; and either [0295]
(i) R.sub.7 and R.sub.8 are H; [0296] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8 hydroxyalkylaminoalkyl, C.sub.1-C.sub.8
alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0297] (ii)
R.sub.6 and R.sub.8 are H; [0298] R.sub.7 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8 hydroxyalkylaminoalkyl, C.sub.1-C.sub.8
alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0299]
(iii) R.sub.7 and R.sub.8 taken together form a bond; [0300]
R.sub.6 is H, substituted or unsubstituted C.sub.1-C.sub.4alkyl,
substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl, C.sub.1-C.sub.8
hydroxyalkylaminoalkyl, C.sub.1-C.sub.8 alkoxyalkylaminoalkyl,
substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl, substituted
or unsubstituted C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl,
substituted or unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0301]
(b) Y is an optionally substituted group selected from
cycloalkylene or heterocycloalkylene; [0302] Z is C(.dbd.O),
NHC(.dbd.O), NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x, where x is
1 or 2, and R.sup.a is H, substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl; and either [0303] (i)
R.sub.7 and R.sub.8 are H; [0304] R.sub.6 is substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8
hydroxyalkylaminoalkyl, C.sub.1-C.sub.8 alkoxyalkylaminoalkyl,
substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl, substituted
or unsubstituted C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl,
substituted or unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0305] (ii)
R.sub.6 and R.sub.8 are H; [0306] R.sub.7 is substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8
hydroxyalkylaminoalkyl, C.sub.1-C.sub.8 alkoxyalkylaminoalkyl,
substituted or unsubstituted C.sub.3-C.sub.6cycloalkyl, substituted
or unsubstituted C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl,
substituted or unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0307]
(iii) R.sub.7 and R.sub.8 taken together form a bond; [0308]
R.sub.6 is substituted or unsubstituted C.sub.1-C.sub.4alkyl,
substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and
pharmaceutically active metabolites, or pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof.
[0309] In another embodiment are provided pharmaceutically
acceptable salts of compounds of Formula (I). By way of example
only, are salts of an amino group formed with inorganic acids such
as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid and perchloric acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid
or malonic acid. Further salts include those in which the
counterion is an anion, such as adipate, alginate, ascorbate,
aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, and valerate. Further salts
include those in which the counterion is an cation, such as sodium,
lithium, potassium, calcium, magnesium, ammonium, and quaternary
ammonium (substituted with at least one organic moiety)
cations.
[0310] In another embodiment are pharmaceutically acceptable esters
of compounds of Formula (I), including those in which the ester
group is selected from a formate, acetate, propionate, butyrate,
acrylate and ethylsuccinate.
[0311] In another embodiment are pharmaceutically acceptable
carbamates of compounds of Formula (I). In another embodiment are
pharmaceutically acceptable N-acyl derivatives of compounds of
Formula (I). Examples of N-acyl groups include N-acetyl and
N-ethoxycarbonyl groups.
[0312] For any and all of the embodiments, substituents can be
selected from among from a subset of the listed alternatives. For
example, in some embodiments, L.sub.a is CH.sub.2, O, or NH. In
other embodiments, L.sub.a is O or NH. In yet other embodiments,
L.sub.a is O.
[0313] In some embodiments, Ar is a substituted or unsubstituted
aryl. In yet other embodiments, Ar is a 6-membered aryl. In some
other embodiments, Ar is phenyl.
[0314] In some embodiments, x is 2. In yet other embodiments, Z is
C(.dbd.O), OC(.dbd.O), NHC(.dbd.O), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, or NHS(.dbd.O).sub.x. In some other embodiments,
Z is C(.dbd.O), NHC(.dbd.O), or NCH.sub.3C(.dbd.O).
[0315] In some embodiments Y is an optionally substituted group
selected from among alkylene, heteroalkylene, arylene,
heteroarylene, alkylenearylene, alkyleneheteroarylene, and
alkyleneheterocycloalkylene.
[0316] In some embodiments, Z is C(.dbd.O), NHC(.dbd.O),
NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x, where x is 1 or 2, and
R.sup.a is H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl.
[0317] In some embodiments, R.sub.7 and R.sub.8 are H; and R.sub.6
is H, substituted or unsubstituted C.sub.1-C.sub.4alkyl,
substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In other
embodiments, R.sub.6 and R.sub.8 are H; and R.sub.7 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In yet
further embodiments, R.sub.7 and R.sub.8 taken together form a
bond; and R.sub.6 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl).
[0318] In some embodiments, Y is an optionally substituted group
selected from cycloalkylene or heterocycloalkylene.
[0319] In some embodiments, Z is C(.dbd.O), NHC(.dbd.O),
NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x, where x is 1 or 2, and
R.sup.a is H, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl.
[0320] In some embodiments, R.sub.7 and R.sub.8 are H; and R.sub.6
is substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In other
embodiments, R.sub.6 and R.sub.8 are H; and R.sub.7 is substituted
or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In further
embodiments, R.sub.7 and R.sub.8 taken together form a bond; and
R.sub.6 is substituted or unsubstituted C.sub.1-C.sub.4alkyl,
substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl).
[0321] In one aspect are compounds (including irreversible
inhibitors of ACKs, including Btk and its cysteine homologs) having
the structure of Formula (VII):
##STR00026## [0322] wherein
##STR00027##
[0322] is a moiety that binds to the active site of a kinase,
including a tyrosine kinase, further including a Btk kinase
cysteine homolog; [0323] Y is an optionally substituted group
selected from among alkylene, heteroalkylene, arylene,
heteroarylene, heterocycloalkylene, cycloalkylene, alkylenearylene,
alkyleneheteroarylene, alkylenecycloalkylene, and
alkyleneheterocycloalkylene; [0324] Z is C(.dbd.O), OC(.dbd.O),
NHC(.dbd.O), NCH.sub.3C(.dbd.O), C(.dbd.S), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, NHS(.dbd.O).sub.x, where x is 1 or 2; [0325]
R.sub.7 and R.sub.8 are independently selected from among H,
unsubstituted C.sub.1-C.sub.4 alkyl, substituted
C.sub.1-C.sub.4alkyl, unsubstituted C.sub.1-C.sub.4heteroalkyl,
substituted C.sub.1-C.sub.4heteroalkyl, unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted C.sub.3-C.sub.6cycloalkyl,
unsubstituted C.sub.2-C.sub.6heterocycloalkyl, and substituted
C.sub.2-C.sub.6heterocycloalkyl; or [0326] R.sub.7 and R.sub.8
taken together form a bond; and [0327] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted C.sub.2-C.sub.8heterocycloalkyl,
substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and
pharmaceutically active metabolites, or pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof.
[0328] In another embodiment are provided pharmaceutically
acceptable salts of compounds of Formula (VII). By way of example
only, are salts of an amino group formed with inorganic acids such
as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid and perchloric acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid
or malonic acid. Further salts include those in which the
counterion is an anion, such as adipate, alginate, ascorbate,
aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, and valerate. Further salts
include those in which the counterion is an cation, such as sodium,
lithium, potassium, calcium, magnesium, ammonium, and quaternary
ammonium (substituted with at least one organic moiety)
cations.
[0329] In another embodiment are pharmaceutically acceptable esters
of compounds of Formula (VII), including those in which the ester
group is selected from a formate, acetate, propionate, butyrate,
acrylate and ethylsuccinate.
[0330] In another embodiment are pharmaceutically acceptable
carbamates of compounds of Formula (VII). In another embodiment are
pharmaceutically acceptable N-acyl derivatives of compounds of
Formula (VII). Examples of N-acyl groups include N-acetyl and
N-ethoxycarbonyl groups.
[0331] In some embodiments, x is 2. In yet other embodiments, Z is
C(.dbd.O), OC(.dbd.O), NHC(.dbd.O), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, or NHS(.dbd.O).sub.x. In some other embodiments,
Z is C(.dbd.O), NHC(.dbd.O), or S(.dbd.O).sub.2.
[0332] In some embodiments, R.sub.7 and R.sub.8 are independently
selected from among H, unsubstituted C.sub.1-C.sub.4 alkyl,
substituted C.sub.1-C.sub.4alkyl, unsubstituted
C.sub.1-C.sub.4heteroalkyl, and substituted
C.sub.1-C.sub.4heteroalkyl; or R.sub.7 and R.sub.8 taken together
form a bond. In yet other embodiments, each of R.sub.7 and R.sub.8
is H; or R.sub.7 and R.sub.8 taken together form a bond.
[0333] In some embodiments, R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
other embodiments, R.sub.6 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.2alkyl-N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In yet other
embodiments, R.sub.6 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, --CH.sub.2--O--(C.sub.1-C.sub.3alkyl),
--CH.sub.2--N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(phenyl), or C.sub.1-C.sub.4alkyl(5- or
6-membered heteroaryl). In yet other embodiments, R.sub.6 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl,
--CH.sub.2--O--(C.sub.1-C.sub.3alkyl),
--CH.sub.2--(C.sub.1-C.sub.6alkylamino),
C.sub.1-C.sub.4alkyl(phenyl), or C.sub.1-C.sub.4alkyl(5- or
6-membered heteroaryl). In some embodiments, R.sub.6 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl,
--CH.sub.2--O--(C.sub.1-C.sub.3alkyl),
--CH.sub.2--N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(phenyl), or C.sub.1-C.sub.4alkyl(5- or
6-membered heteroaryl containing 1 or 2 N atoms), or
C.sub.1-C.sub.4alkyl(5- or 6-membered heterocycloalkyl containing 1
or 2 N atoms).
[0334] In some embodiments, Y is an optionally substituted group
selected from among alkylene, heteroalkylene, arylene,
heteroarylene, heterocycloalkylene, cycloalkylene, alkylenearylene,
alkyleneheteroarylene, alkylenecycloalkylene, and
alkyleneheterocycloalkylene. In other embodiments, Y is an
optionally substituted group selected from among
C.sub.1-C.sub.6alkylene, C.sub.1-C.sub.6heteroalkylene, 4-, 5-, 6-,
or 7-membered cycloalkylene, and 4-, 5-, 6-, or 7-membered
heterocycloalkylene. In yet other embodiments, Y is an optionally
substituted group selected from among C.sub.1-C.sub.6alkylene,
C.sub.1-C.sub.6heteroalkylene, 5- or 6-membered cycloalkylene, and
5- or 6-membered heterocycloalkylene containing 1 or 2 N atoms. In
some other embodiments, Y is a 5- or 6-membered cycloalkylene, or a
5- or 6-membered heterocycloalkylene containing 1 or 2 N atoms. In
some embodiments, Y is a 4-, 5-, 6-, or 7-membered cycloalkylene
ring; or Y is a 4-, 5-, 6-, or 7-membered heterocycloalkylene
ring.
[0335] In one aspect are compounds (including irreversible
inhibitors of ACKs, including Btk and its cysteine homologs) having
the structure of Formula (A1):
##STR00028##
wherein [0336] A is independently selected from N or CR.sub.5;
[0337] R.sub.1 is H, L.sub.2-(substituted or unsubstituted alkyl),
L.sub.2-(substituted or unsubstituted cycloalkyl),
L.sub.2-(substituted or unsubstituted alkenyl),
L.sub.2-(substituted or unsubstituted cycloalkenyl),
L.sub.2-(substituted or unsubstituted heterocycle),
L.sub.2-(substituted or unsubstituted heteroaryl), or
L.sub.2-(substituted or unsubstituted aryl), where L.sub.2 is a
bond, O, S, --S(.dbd.O), --S(.dbd.O).sub.2, C(.dbd.O),
-(substituted or unsubstituted C.sub.1-C.sub.6 alkyl), or
-(substituted or unsubstituted C.sub.2-C.sub.6 alkenyl); [0338]
R.sub.2 and R.sub.3 are independently selected from H, lower alkyl
and substituted lower alkyl; [0339] R.sub.4 is L.sub.3-X-L.sub.4-G,
wherein, [0340] L.sub.3 is optional, and when present is a bond, or
an optionally substituted group selected from alkyl, heteroalkyl,
aryl, heteroaryl, alkylaryl, alkylheteroaryl, or
alkylheterocycloalkyl; [0341] X is optional, and when present is a
bond, O, --C(.dbd.O), S, --S(.dbd.O), --S(.dbd.O).sub.2, --NH,
--NR.sub.9, --NHC(O), --C(O)NH, --NR.sub.9C(O), --C(O)NR.sub.9,
--S(.dbd.O).sub.2NH, --NHS(.dbd.O).sub.2,
--S(.dbd.O).sub.2NR.sub.9--, --NR.sub.9S(.dbd.O).sub.2,
--OC(O)NH--, --NHC(O)O--, --OC(O)NR.sub.9--, --NR.sub.9C(O)O--,
--CH.dbd.NO--, --ON.dbd.CH--, --NR.sub.10C(O)NR.sub.10--,
heteroaryl, aryl, --NR.sub.10C(.dbd.NR.sub.11)NR.sub.10--,
--NR.sub.10C(.dbd.NR.sub.11)--, --C(.dbd.NR.sub.11)NR.sub.10--,
--OC(.dbd.NR.sub.11)--, or --C(.dbd.NR.sub.11)O--; [0342] L.sub.4
is optional, and when present is a bond, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted alkenyl, substituted or unsubstituted
alkynyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, substituted or unsubstituted heterocycle;
[0343] or L.sub.3, X and L.sub.4 taken together form a nitrogen
containing heterocyclic ring, or an optionally substituted group
selected from alkyl, heteroalkyl, aryl, heteroaryl, alkylaryl,
alkylheteroaryl, or alkylheterocycloalkyl; [0344] G is
[0344] ##STR00029## where R.sup.a is H, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl; and
either [0345] R.sup.7 and R.sub.8 are H; [0346] R.sub.6 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted or
unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0347]
R.sub.6 and R.sub.8 are H; [0348] R.sub.7 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0349]
R.sub.7 and R.sub.8 taken together form a bond; [0350] R.sub.6 is
H, substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted
or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0351]
R.sub.5 is H, halogen, -L.sub.6-(substituted or unsubstituted
C.sub.1-C.sub.3 alkyl), -L.sub.6-(substituted or unsubstituted
C.sub.2-C.sub.4 alkenyl), -L.sub.6-(substituted or unsubstituted
heteroaryl), or -L.sub.6-(substituted or unsubstituted aryl),
wherein L.sub.6 is a bond, O, S, --S(.dbd.O), S(.dbd.O).sub.2, NH,
C(O), --NHC(O)O, --OC(O)NH, --NHC(O), or --C(O)NH; [0352] each
R.sub.9 is independently selected from among H, substituted or
unsubstituted lower alkyl, and substituted or unsubstituted lower
cycloalkyl; [0353] each R.sub.10 is independently H, substituted or
unsubstituted lower alkyl, or substituted or unsubstituted lower
cycloalkyl; or [0354] two R.sub.10 groups can together form a 5-,
6-, 7-, or 8-membered heterocyclic ring; or [0355] R.sub.9 and
R.sub.10 can together form a 5-, 6-, 7-, or 8-membered heterocyclic
ring; or [0356] each R.sub.11 is independently selected from H,
--S(.dbd.O).sub.2R.sub.8, --S(.dbd.O).sub.2NH.sub.2, --C(O)R.sub.8,
--CN, --NO.sub.2, heteroaryl, or heteroalkyl; and pharmaceutically
active metabolites, pharmaceutically acceptable solvates,
pharmaceutically acceptable salts, or pharmaceutically acceptable
prodrugs thereof.
[0357] In another embodiment are provided pharmaceutically
acceptable salts of compounds of Formula (A1). By way of example
only, are salts of an amino group formed with inorganic acids such
as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid and perchloric acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid
or malonic acid. Further salts include those in which the
counterion is an anion, such as adipate, alginate, ascorbate,
aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, and valerate. Further salts
include those in which the counterion is an cation, such as sodium,
lithium, potassium, calcium, magnesium, ammonium, and quaternary
ammonium (substituted with at least one organic moiety)
cations.
[0358] In another embodiment are pharmaceutically acceptable esters
of compounds of Formula (A1), including those in which the ester
group is selected from a formate, acetate, propionate, butyrate,
acrylate and ethylsuccinate.
[0359] In another embodiment are pharmaceutically acceptable
carbamates of compounds of Formula (A1). In another embodiment are
pharmaceutically acceptable N-acyl derivatives of compounds of
Formula (A1). Examples of N-acyl groups include N-acetyl and
N-ethoxycarbonyl groups.
[0360] In a further or alternative embodiment, the compound of
Formula (A1) has the following structure of Formula (B1):
##STR00030##
wherein: [0361] Y is an optionally substituted group selected from
among alkylene, heteroalkylene, arylene, heteroarylene,
alkylenearylene, alkyleneheteroarylene, and
alkyleneheterocycloalkylene; [0362] each R.sub.a is independently
H, halogen, --CF.sub.3, --CN, --NO.sub.2, OH, NH.sub.2,
-L.sub.a-(substituted or unsubstituted alkyl),
-L.sub.a-(substituted or unsubstituted alkenyl),
-L.sub.a-(substituted or unsubstituted heteroaryl), or
-L.sub.a-(substituted or unsubstituted aryl), wherein L.sub.a is a
bond, O, S, --S(.dbd.O), --S(.dbd.O).sub.2, NH, C(O), CH.sub.2,
--NHC(O)O, --NHC(O), or --C(O)NH; [0363] G is
##STR00031##
[0363] where R.sup.a is H, substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl; and either [0364] R.sub.7
and R.sub.8 are H; [0365] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0366]
R.sub.6 and R.sub.8 are H; [0367] R.sub.7 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0368]
R.sub.7 and R.sub.8 taken together form a bond; [0369] R.sub.6 is
H, substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted
or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0370]
R.sub.12 is H or lower alkyl; or [0371] Y and R.sub.12 taken
together form a 4-, 5-, or 6-membered heterocyclic ring; and [0372]
pharmaceutically acceptable active metabolites, pharmaceutically
acceptable solvates, pharmaceutically acceptable salts, or
pharmaceutically acceptable prodrugs thereof.
[0373] In further or alternative embodiments, G is selected from
among
##STR00032##
where R is H, alkyl, alkylhydroxy, heterocycloalkyl, heteroaryl,
alkylalkoxy, alkylalkoxyalkyl.
[0374] In further or alternative embodiments,
##STR00033##
is selected from among
##STR00034##
[0375] In further or alternative embodiment, the compound of
Formula (B1) has the following structure of Formula (C1):
##STR00035## [0376] Y is an optionally substituted group selected
from among alkyl, heteroalkyl, aryl, heteroaryl, alkylaryl,
alkylheteroaryl, and alkylheterocycloalkyl; [0377] R.sub.12 is H or
lower alkyl; or [0378] Y and R.sub.12 taken together form a 4-, 5-,
or 6-membered heterocyclic ring; [0379] G is
##STR00036##
[0379] where R.sup.a is H, substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl; and either [0380] R.sub.7
and R.sub.8 are H; [0381] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0382]
R.sub.6 and R.sub.8 are H; [0383] R.sub.7 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0384]
R.sub.7 and R.sub.8 taken together form a bond; [0385] R.sub.6 is
H, substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted
or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and [0386]
pharmaceutically acceptable active metabolites, pharmaceutically
acceptable solvates, pharmaceutically acceptable salts, or
pharmaceutically acceptable prodrugs thereof.
[0387] In a further or alternative embodiment, the "G" group of any
of Formula (A1), Formula (B1), or Formula (C1) is any group that is
used to tailor the physical and biological properties of the
molecule. Such tailoring/modifications are achieved using groups
which modulate Michael acceptor chemical reactivity, acidity,
basicity, lipophilicity, solubility and other physical properties
of the molecule. The physical and biological properties modulated
by such modifications to G include, by way of example only,
enhancing chemical reactivity of Michael acceptor group,
solubility, in vivo absorption, and in vivo metabolism. In
addition, in vivo metabolism includes, by way of example only,
controlling in vivo PK properties, off-target activities, potential
toxicities associated with cypP450 interactions, drug-drug
interactions, and the like. Further, modifications to G allow for
the tailoring of the in vivo efficacy of the compound through the
modulation of, by way of example, specific and non-specific protein
binding to plasma proteins and lipids and tissue distribution in
vivo.
[0388] In one aspect are compounds (including irreversible
inhibitors of ACKs, including Btk and its cysteine homologs) having
the structure of Formula (D1):
##STR00037##
wherein [0389] L.sub.a is CH.sub.2, O, NH or S; [0390] Ar is an
optionally substituted aromatic carbocycle or an aromatic
heterocycle; [0391] Y is an optionally substituted group selected
from among alkylene, heteroalkylene, arylene, heteroarylene,
alkylenearylene, alkyleneheteroarylene, and
alkyleneheterocycloalkylene, or combination thereof; [0392] Z is
C(.dbd.O), NHC(.dbd.O), NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O).sub.x,
where x is 1 or 2, and R.sup.a is H, substituted or unsubstituted
alkyl, substituted or unsubstituted cycloalkyl; and either [0393]
R.sub.7 and R.sub.8 are H; [0394] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0395]
R.sub.6 and R.sub.8 are H; [0396] R.sub.7 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0397]
R.sub.7 and R.sub.8 taken together form a bond; [0398] R.sub.6 is
H, substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted
or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0399] or
combinations thereof; and pharmaceutically active metabolites, or
pharmaceutically acceptable solvates, pharmaceutically acceptable
salts, or pharmaceutically acceptable prodrugs thereof.
[0400] In another embodiment are provided pharmaceutically
acceptable salts of compounds of Formula (D1). By way of example
only, are salts of an amino group formed with inorganic acids such
as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid and perchloric acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid
or malonic acid. Further salts include those in which the
counterion is an anion, such as adipate, alginate, ascorbate,
aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, and valerate. Further salts
include those in which the counterion is an cation, such as sodium,
lithium, potassium, calcium, magnesium, ammonium, and quaternary
ammonium (substituted with at least one organic moiety)
cations.
[0401] In another embodiment are pharmaceutically acceptable esters
of compounds of Formula (D1), including those in which the ester
group is selected from a formate, acetate, propionate, butyrate,
acrylate and ethylsuccinate.
[0402] In another embodiment are pharmaceutically acceptable
carbamates of compounds of Formula (D1). In another embodiment are
pharmaceutically acceptable N-acyl derivatives of compounds of
Formula (D1). Examples of N-acyl groups include N-acetyl and
N-ethoxycarbonyl groups.
[0403] In a further or alternative embodiment, L.sub.a is O.
[0404] In a further or alternative embodiment, Ar is phenyl.
[0405] In a further or alternative embodiment, Z is C(.dbd.O),
NHC(.dbd.O), or NCH.sub.3C(.dbd.O).
[0406] In a further or alternative embodiment, each of R.sub.1,
R.sub.2, and R.sub.3 is H.
[0407] In one aspect are compounds (including irreversible
inhibitors of ACKs, including Btk and its cysteine homologs) having
the structure of Formula (D1):
##STR00038##
wherein: [0408] L.sub.a is CH.sub.2, O, NH or S; [0409] Ar is a
substituted or unsubstituted aryl, or a substituted or
unsubstituted heteroaryl; [0410] Y is an optionally substituted
group selected from among alkylene, heteroalkylene, arylene,
heteroarylene, alkylenearylene, alkylenehetroarylene,
alkylenecycloalkylene and alkyleneheterocycloalkylene; [0411] Z is
C(.dbd.O), NHC(.dbd.O), NR.sup.aC(.dbd.O), NR.sup.aS(.dbd.O), where
x is 1 or 2, and R.sup.a is substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl; and either [0412] R.sub.7
and R.sub.8 are H; [0413] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); [0414]
R.sub.6 and R.sub.8 are H; [0415] R.sub.7 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); or [0416]
R.sub.7 and R.sub.8 taken together form a bond; [0417] R.sub.6 is
H, substituted or unsubstituted C.sub.1-C.sub.4alkyl, substituted
or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted
C.sub.1-C.sub.8alkylC.sub.3-C.sub.6cycloalkyl, substituted or
unsubstituted aryl, substituted or unsubstituted
C.sub.2-C.sub.8heterocycloalkyl, substituted or unsubstituted
heteroaryl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl), C.sub.1-C.sub.8alkylethers,
C.sub.1-C.sub.8alkylamides, or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and
pharmaceutically active metabolites, or pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof.
[0418] In another embodiment are provided pharmaceutically
acceptable salts of compounds of Formula (D1). By way of example
only, are salts of an amino group formed with inorganic acids such
as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid and perchloric acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid
or malonic acid. Further salts include those in which the
counterion is an anion, such as adipate, alginate, ascorbate,
aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, and valerate. Further salts
include those in which the counterion is an cation, such as sodium,
lithium, potassium, calcium, magnesium, ammonium, and quaternary
ammonium (substituted with at least one organic moiety)
cations.
[0419] In another embodiment are pharmaceutically acceptable esters
of compounds of Formula (D1), including those in which the ester
group is selected from a formate, acetate, propionate, butyrate,
acrylate and ethylsuccinate.
[0420] In another embodiment are pharmaceutically acceptable
carbamates of compounds of Formula (D1). In another embodiment are
pharmaceutically acceptable N-acyl derivatives of compounds of
Formula (D1). Examples of N-acyl groups include N-acetyl and
N-ethoxycarbonyl groups.
[0421] For any and all of the embodiments, substituents can be
selected from among from a subset of the listed alternatives. For
example, in some embodiments, L.sub.a is CH.sub.2, O, or NH. In
other embodiments, L.sub.a is O or NH. In yet other embodiments,
L.sub.a is O.
[0422] In some embodiments, Ar is a substituted or unsubstituted
aryl. In yet other embodiments, Ar is a 6-membered aryl. In some
other embodiments, Ar is phenyl.
[0423] In some embodiments, x is 2. In yet other embodiments, Z is
C(.dbd.O), OC(.dbd.O), NHC(.dbd.O), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, or NHS(.dbd.O).sub.x. In some other embodiments,
Z is C(.dbd.O), NHC(.dbd.O), or S(.dbd.O).sub.2.
[0424] In some embodiments, R.sub.7 and R.sub.8 are independently
selected from among H, unsubstituted C.sub.1-C.sub.4 alkyl,
substituted C.sub.1-C.sub.4alkyl, unsubstituted
C.sub.1-C.sub.4heteroalkyl, and substituted
C.sub.1-C.sub.4heteroalkyl; or R.sub.7 and R.sub.8 taken together
form a bond. In yet other embodiments, each of R.sub.7 and R.sub.8
is H; or R.sub.7 and R.sub.8 taken together form a bond.
[0425] In some embodiments, R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.2alkyl-N(C.sub.1-C.sub.3alkyl).sub.2, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
other embodiments, R.sub.6 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.2alkyl-N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In yet other
embodiments, R.sub.6 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, --CH.sub.2--O--(C.sub.1-C.sub.3alkyl),
--CH.sub.2--N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(phenyl), or C.sub.1-C.sub.4alkyl(5- or
6-membered heteroaryl). In some embodiments, R.sub.6 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl,
--CH.sub.2--O--(C.sub.1-C.sub.3alkyl),
--CH.sub.2--N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(phenyl), or C.sub.1-C.sub.4alkyl(5- or
6-membered heteroaryl containing 1 or 2 N atoms), or
C.sub.1-C.sub.4alkyl(5- or 6-membered heterocycloalkyl containing 1
or 2 N atoms).
[0426] In some embodiments, Y is an optionally substituted group
selected from among alkylene, heteroalkylene, cycloalkylene, and
heterocycloalkylene. In other embodiments, Y is an optionally
substituted group selected from among C.sub.1-C.sub.6alkylene,
C.sub.1-C.sub.6heteroalkylene, 4-, 5-, 6- or 7-membered
cycloalkylene, and 4-, 5-, 6- or 7-membered heterocycloalkylene. In
yet other embodiments, Y is an optionally substituted group
selected from among C.sub.1-C.sub.6alkylene,
C.sub.1-C.sub.6heteroalkylene, 5-, or 6-membered cycloalkylene, and
5-, or 6-membered heterocycloalkylene containing 1 or 2 N atoms. In
some other embodiments, Y is a 5-, or 6-membered cycloalkylene, or
a 5-, or 6-membered heterocycloalkylene containing 1 or 2 N
atoms.
[0427] In one aspect are compounds (including irreversible
inhibitors of ACKs, including Btk and its cysteine homologs) having
the structure of Formula (A2-A6):
##STR00039##
wherein [0428] A is independently selected from N or CR.sub.5;
[0429] R.sub.1 is H, L.sub.2-(substituted or unsubstituted alkyl),
L.sub.2-(substituted or unsubstituted cycloalkyl),
L.sub.2-(substituted or unsubstituted alkenyl),
L.sub.2-(substituted or unsubstituted cycloalkenyl),
L.sub.2-(substituted or unsubstituted heterocycle),
L.sub.2-(substituted or unsubstituted heteroaryl), or
L.sub.2-(substituted or unsubstituted aryl), where L.sub.2 is a
bond, O, S, --S(.dbd.O), --S(.dbd.O).sub.2, C(.dbd.O),
-(substituted or unsubstituted C.sub.1-C.sub.6 alkyl), or
-(substituted or unsubstituted C.sub.2-C.sub.6 alkenyl); [0430]
R.sub.2 and R.sub.3 are independently selected from H, lower alkyl
and substituted lower alkyl; [0431] R.sub.4 is L.sub.3-X-L.sub.4-G,
wherein, [0432] L.sub.3 is optional, and when present is a bond,
optionally substituted or unsubstituted alkyl, optionally
substituted or unsubstituted cycloalkyl, optionally substituted or
unsubstituted alkenyl, optionally substituted or unsubstituted
alkynyl; [0433] X is optional, and when present is a bond, O,
--C(.dbd.O), S, --S(.dbd.O), --S(.dbd.O).sub.2, --NH, --NR.sub.9,
--NHC(O), --C(O)NH, --NR.sub.9C(O), --C(O)NR.sub.9,
--S(.dbd.O).sub.2NH, --NHS(.dbd.O).sub.2,
--S(.dbd.O).sub.2NR.sub.9--, --NR.sub.9S(.dbd.O).sub.2,
--OC(O)NH--, --NHC(O)O--, --OC(O)NR.sub.9--, --NR.sub.9C(O)O--,
--CH.dbd.NO--, --ON.dbd.CH--, --NR.sub.10C(O)NR.sub.10--,
heteroaryl, aryl, --NR.sub.10C(.dbd.NR.sub.11)NR.sub.10--,
--NR.sub.10C(.dbd.NR.sub.11)--, --C(.dbd.NR.sub.11)NR.sub.10--,
--OC(.dbd.NR.sub.11)--, or --C(.dbd.NR.sub.10)O--; [0434] L.sub.4
is optional, and when present is a bond, substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted alkenyl, substituted or unsubstituted
alkynyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, substituted or unsubstituted heterocycle;
[0435] or L.sub.3, X and L.sub.4 taken together form a nitrogen
containing heterocyclic ring; [0436] G is
[0436] ##STR00040## wherein, [0437] R.sub.6, R.sub.7 and R.sub.8
are independently selected from among H, lower alkyl or substituted
lower alkyl, lower heteroalkyl or substituted lower heteroalkyl,
substituted or unsubstituted lower cycloalkyl, and substituted or
unsubstituted lower heterocycloalkyl; [0438] R.sub.5 is H, halogen,
-L.sub.6-(substituted or unsubstituted C.sub.1-C.sub.3 alkyl),
-L.sub.6-(substituted or unsubstituted C.sub.2-C.sub.4 alkenyl),
-L.sub.6-(substituted or unsubstituted heteroaryl), or
-L.sub.6-(substituted or unsubstituted aryl), wherein L.sub.6 is a
bond, O, S, --S(.dbd.O), S(.dbd.O).sub.2, NH, C(O), --NHC(O)O,
--OC(O)NH, --NHC(O), or --C(O)NH; [0439] each R.sub.9 is
independently selected from among H, substituted or unsubstituted
lower alkyl, and substituted or unsubstituted lower cycloalkyl;
[0440] each R.sub.10 is independently H, substituted or
unsubstituted lower alkyl, or substituted or unsubstituted lower
cycloalkyl; or [0441] two R.sub.10 groups can together form a 5-,
6-, 7-, or 8-membered heterocyclic ring; or [0442] R.sub.9 and
R.sub.10 can together form a 5-, 6-, 7-, or 8-membered heterocyclic
ring; or [0443] each R.sub.11 is independently selected from H,
--S(.dbd.O).sub.2R.sub.8, --S(.dbd.O).sub.2NH.sub.2, --C(O)R.sub.8,
--CN, --NO.sub.2, heteroaryl, or heteroalkyl; and pharmaceutically
active metabolites, or pharmaceutically acceptable solvates,
pharmaceutically acceptable salts, or pharmaceutically acceptable
prodrugs thereof.
[0444] In another embodiment are provided pharmaceutically
acceptable salts of compounds of Formula (A2-A6). By way of example
only, are salts of an amino group formed with inorganic acids such
as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid and perchloric acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid
or malonic acid. Further salts include those in which the
counterion is an anion, such as adipate, alginate, ascorbate,
aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, and valerate. Further salts
include those in which the counterion is an cation, such as sodium,
lithium, potassium, calcium, magnesium, ammonium, and quaternary
ammonium (substituted with at least one organic moiety)
cations.
[0445] In another embodiment are pharmaceutically acceptable esters
of compounds of Formula (A2-A6), including those in which the ester
group is selected from a formate, acetate, propionate, butyrate,
acrylate and ethylsuccinate.
[0446] In another embodiment are pharmaceutically acceptable
carbamates of compounds of Formula (A2-A6). In another embodiment
are pharmaceutically acceptable N-acyl derivatives of compounds of
Formula (A2-A6). Examples of N-acyl groups include N-acetyl and
N-ethoxycarbonyl groups.
[0447] In a further or alternative embodiment, the compound of
Formula (A2-A6) has the following structure of Formula (B2-B6):
##STR00041## ##STR00042##
wherein: [0448] Y is alkylene or substituted alkylene, or a 4-, 5-,
or 6-membered cycloalkylene ring; [0449] each R.sub.a is
independently H, halogen, --CF.sub.3, --CN, --NO.sub.2, OH,
NH.sub.2, -L.sub.a-(substituted or unsubstituted alkyl),
-L.sub.a-(substituted or unsubstituted alkenyl),
-L.sub.a-(substituted or unsubstituted heteroaryl), or
-L.sub.a-(substituted or unsubstituted aryl), wherein L.sub.a is a
bond, O, S, --S(.dbd.O), --S(.dbd.O).sub.2, NH, C(O), CH.sub.2,
--NHC(O)O, --NHC(O), or --C(O)NH; [0450] G is
##STR00043##
[0450] wherein, [0451] R.sub.6, R.sub.7 and R.sub.8 are
independently selected from among H, lower alkyl or substituted
lower alkyl, lower heteroalkyl or substituted lower heteroalkyl,
substituted or unsubstituted lower cycloalkyl, and substituted or
unsubstituted lower heterocycloalkyl; [0452] R.sub.12 is H or lower
alkyl; or [0453] Y and R.sub.12 taken together form a 4-, 5-, or
6-membered heterocyclic ring; and [0454] pharmaceutically
acceptable active metabolites, pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof.
[0455] In further or alternative embodiments, G is selected from
among
##STR00044##
[0456] In further or alternative embodiments,
##STR00045##
is selected from among
##STR00046##
[0457] In further or alternative embodiment, the compound of
Formula (B2-B6) has the following structure of Formula (C2-C6):
##STR00047## ##STR00048## [0458] Y is alkylene or substituted
alkylene, or a 4-, 5-, or 6-membered cycloalkylene ring; [0459]
R.sub.12 is H or lower alkyl; or [0460] Y and R.sub.12 taken
together form a 4-, 5-, or 6-membered heterocyclic ring; [0461] G
is
##STR00049##
[0461] wherein, [0462] R.sub.6, R.sub.7 and R.sub.8 are
independently selected from among H, lower alkyl or substituted
lower alkyl, lower heteroalkyl or substituted lower heteroalkyl,
substituted or unsubstituted lower cycloalkyl, and substituted or
unsubstituted lower heterocycloalkyl; and [0463] pharmaceutically
acceptable active metabolites, pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof.
[0464] In a further or alternative embodiment, the "G" group of any
of Formula (A2-A6), Formula (B2-B6), or Formula (C2-C6) is any
group that is used to tailor the physical and biological properties
of the molecule. Such tailoring/modifications are achieved using
groups which modulate Michael acceptor chemical reactivity,
acidity, basicity, lipophilicity, solubility and other physical
properties of the molecule. The physical and biological properties
modulated by such modifications to G include, by way of example
only, enhancing chemical reactivity of Michael acceptor group,
solubility, in vivo absorption, and in vivo metabolism. In
addition, in vivo metabolism includes, by way of example only,
controlling in vivo PK properties, off-target activities, potential
toxicities associated with cypP450 interactions, drug-drug
interactions, and the like. Further, modifications to G allow for
the tailoring of the in vivo efficacy of the compound through the
modulation of, by way of example, specific and non-specific protein
binding to plasma proteins and lipids and tissue distribution in
vivo.
[0465] In one aspect are compounds (including irreversible
inhibitors of ACKs, including Btk and its cysteine homologs) having
the structure of Formula (D2-D6):
##STR00050## ##STR00051##
wherein [0466] L.sub.a is CH.sub.2, O, NH or S; [0467] Ar is an
optionally substituted aromatic carbocycle or an aromatic
heterocycle; [0468] Y is an optionally substituted alkylene,
heteroalkylene, carbocycloalkylene, heterocycloalkylene, or
combination thereof; [0469] Z is C(O), OC(O), NHC(O), C(S),
S(O).sub.x, OS(O).sub.x, NHS(O).sub.x, where x is 1 or 2; and
[0470] R.sub.6, R.sub.7, and R.sub.8 are independently selected
from H, alkyl, heteroalkyl, carbocycle, heterocycle, or
combinations thereof; and pharmaceutically active metabolites, or
pharmaceutically acceptable solvates, pharmaceutically acceptable
salts, or pharmaceutically acceptable prodrugs thereof.
[0471] In another embodiment are provided pharmaceutically
acceptable salts of compounds of Formula (D2-D6). By way of example
only, are salts of an amino group formed with inorganic acids such
as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid and perchloric acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid
or malonic acid. Further salts include those in which the
counterion is an anion, such as adipate, alginate, ascorbate,
aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, and valerate. Further salts
include those in which the counterion is an cation, such as sodium,
lithium, potassium, calcium, magnesium, ammonium, and quaternary
ammonium (substituted with at least one organic moiety)
cations.
[0472] In another embodiment are pharmaceutically acceptable esters
of compounds of Formula (D2-D6), including those in which the ester
group is selected from a formate, acetate, propionate, butyrate,
acrylate and ethylsuccinate.
[0473] In another embodiment are pharmaceutically acceptable
carbamates of compounds of Formula (D2-D6). In another embodiment
are pharmaceutically acceptable N-acyl derivatives of compounds of
Formula (D2-D6). Examples of N-acyl groups include N-acetyl and
N-ethoxycarbonyl groups.
[0474] In a further or alternative embodiment, L.sub.a is O.
[0475] In a further or alternative embodiment, Ar is phenyl.
[0476] In a further or alternative embodiment, Z is C(O).
[0477] In a further or alternative embodiment, each of R.sub.1,
R.sub.2, and R.sub.3 is H.
[0478] In one aspect are compounds (including irreversible
inhibitors of ACKs, including Btk and its cysteine homologs) having
the structure of Formula (D2-D6):
##STR00052## ##STR00053##
wherein: [0479] L.sub.a is CH.sub.2, O, NH or S; [0480] Ar is a
substituted or unsubstituted aryl, or a substituted or
unsubstituted heteroaryl; [0481] Y is an optionally substituted
group selected from among alkylene, heteroalkylene, cycloalkylene,
heterocycloalkylene, arylene, and heteroarylene; [0482] Z is
C(.dbd.O), OC(.dbd.O), NHC(.dbd.O), C(.dbd.S), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, NHS(.dbd.O).sub.x, where x is 1 or 2; [0483]
R.sub.7 and R.sub.8 are independently selected from among H,
unsubstituted C.sub.1-C.sub.4alkyl, substituted
C.sub.1-C.sub.4alkyl, unsubstituted C.sub.1-C.sub.4heteroalkyl,
substituted C.sub.1-C.sub.4heteroalkyl, unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted C.sub.3-C.sub.6cycloalkyl,
unsubstituted C.sub.2-C.sub.6heterocycloalkyl, and substituted
C.sub.2-C.sub.6heterocycloalkyl; or [0484] R.sub.7 and R.sub.8
taken together form a bond; [0485] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted C.sub.2-C.sub.8heterocycloalkyl,
substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and
pharmaceutically active metabolites, or pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof.
[0486] In another embodiment are provided pharmaceutically
acceptable salts of compounds of Formula (D2-D6). By way of example
only, are salts of an amino group formed with inorganic acids such
as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid and perchloric acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid
or malonic acid. Further salts include those in which the
counterion is an anion, such as adipate, alginate, ascorbate,
aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, and valerate. Further salts
include those in which the counterion is an cation, such as sodium,
lithium, potassium, calcium, magnesium, ammonium, and quaternary
ammonium (substituted with at least one organic moiety)
cations.
[0487] In another embodiment are pharmaceutically acceptable esters
of compounds of Formula (D2-D6), including those in which the ester
group is selected from a formate, acetate, propionate, butyrate,
acrylate and ethylsuccinate.
[0488] In another embodiment are pharmaceutically acceptable
carbamates of compounds of Formula (D2-D6). In another embodiment
are pharmaceutically acceptable N-acyl derivatives of compounds of
Formula (D2-D6).
[0489] For any and all of the embodiments, substituents can be
selected from among from a subset of the listed alternatives. For
example, in some embodiments, L.sub.a is CH.sub.2, O, or NH. In
other embodiments, L.sub.a is O or NH. In yet other embodiments,
L.sub.a is O.
[0490] In some embodiments, Ar is a substituted or unsubstituted
aryl. In yet other embodiments, Ar is a 6-membered aryl. In some
other embodiments, Ar is phenyl.
[0491] In some embodiments, x is 2. In yet other embodiments, Z is
C(.dbd.O), OC(.dbd.O), NHC(.dbd.O), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, or NHS(.dbd.O).sub.x. In some other embodiments,
Z is C(.dbd.O), NHC(.dbd.O), or S(.dbd.O).sub.2.
[0492] In some embodiments, R.sub.7 and R.sub.8 are independently
selected from among H, unsubstituted C.sub.1-C.sub.4 alkyl,
substituted C.sub.1-C.sub.4alkyl, unsubstituted
C.sub.1-C.sub.4heteroalkyl, and substituted
C.sub.1-C.sub.4heteroalkyl; or R.sub.7 and R.sub.8 taken together
form a bond. In yet other embodiments, each of R.sub.7 and R.sub.8
is H; or R.sub.7 and R.sub.8 taken together form a bond.
[0493] In some embodiments, R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.2alkyl-N(C.sub.1-C.sub.3alkyl).sub.2, substituted or
unsubstituted aryl, substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In some
other embodiments, R.sub.6 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.2alkyl-N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl). In yet other
embodiments, R.sub.6 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, --CH.sub.2--O--(C.sub.1-C.sub.3alkyl),
--CH.sub.2--N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(phenyl), or C.sub.1-C.sub.4alkyl(5- or
6-membered heteroaryl). In some embodiments, R.sub.6 is H,
substituted or unsubstituted C.sub.1-C.sub.4alkyl,
--CH.sub.2--O--(C.sub.1-C.sub.3alkyl),
--CH.sub.2--N(C.sub.1-C.sub.3alkyl).sub.2,
C.sub.1-C.sub.4alkyl(phenyl), or C.sub.1-C.sub.4alkyl(5- or
6-membered heteroaryl containing 1 or 2 N atoms), or
C.sub.1-C.sub.4alkyl(5- or 6-membered heterocycloalkyl containing 1
or 2 N atoms).
[0494] In some embodiments, Y is an optionally substituted group
selected from among alkylene, heteroalkylene, cycloalkylene, and
heterocycloalkylene. In other embodiments, Y is an optionally
substituted group selected from among C.sub.1-C.sub.6alkylene,
C.sub.1-C.sub.6heteroalkylene, 4-, 5-, 6- or 7-membered
cycloalkylene, and 4-, 5-, 6- or 7-membered heterocycloalkylene. In
yet other embodiments, Y is an optionally substituted group
selected from among C.sub.1-C.sub.6alkylene,
C.sub.1-C.sub.6heteroalkylene, 5-, or 6-membered cycloalkylene, and
5-, or 6-membered heterocycloalkylene containing 1 or 2 N atoms. In
some other embodiments, Y is a 5-, or 6-membered cycloalkylene, or
a 5-, or 6-membered heterocycloalkylene containing 1 or 2 N
atoms.
[0495] Any combination of the groups described above for the
various variables is contemplated herein.
[0496] In further aspects are compounds (including irreversible
inhibitors of ACKs, including Btk and its cysteine homologs) having
the structure of compounds of Formula (A1-A6), Formula (B1-B6),
Formula (C1-C6), Formula (D1-D6), including, but are not limited
to, compounds selected from the group consisting of:
##STR00054## ##STR00055## ##STR00056## ##STR00057## ##STR00058##
##STR00059## ##STR00060## ##STR00061## ##STR00062##
##STR00063##
[0497] In one aspect are compounds (including irreversible
inhibitors of ACKs, including Btk and its cysteine homologs)
selected from among:
(E)-4-(N-(2-hydroxyethyl)-N-methylamino)-1-(3-(4-phenoxyphenyl)-1H-pyrazo-
lo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)but-2-en-1-one (Compound 3);
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3-
-(1H-imidazol-4-yl)prop-2-en-1-one (Compound 4);
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-morpholinobut-2-en-1-one (Compound 5);
(E)-1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound 7);
(E)-N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-
-1-yl)cyclohexyl)-4-(dimethylamino)but-2-enamide (Compound 8);
N-((1r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)acrylamide (Compound 10);
(E)-1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)methyl)pyrolidin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound
11);
(E)-1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)methyl)pyrolidin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound
12);
1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 13);
1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 14);
1((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)me-
thyl)pyrrolidin-1-yl)but-2-yn-1-one (Compound 15);
1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)but-2-yn-1-one (Compound 16);
1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)but-2-yn-1-one (Compound 17);
(E)-N-((1,r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidi-
n-1-yl)cyclohexyl-4-(dimethylamino)but-2-enamide (Compound 18);
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
-N-methylacrylamide (Compound 19);
(E)-1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-4-
-morpholinobut-2-en-1-one (Compound 20);
(E)-1-((S-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)methyl)pyrrolidin-1-yl)-4-morpholinobut-2-en-1-one (Compound 21);
N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)but-2-ynamide (Compound 22);
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
acrylamide (Compound 23);
(E)-1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)piperidin-1-yl)-4-morpholinobut-2-en-1-one (Compound 24);
(E)-N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-
-1-yl)cyclohexyl)-4-morpholinobut-2-enamide (Compound 25).
[0498] The compounds of any of Formula (I), Formula (VII), Formula
(A1-A6), Formula (B1-B6), Formula (C1-C6), or Formula (D1-D6)
irreversibly inhibit Btk and are optionally used to treat patients
suffering from Bruton's tyrosine kinase-dependent or Bruton's
tyrosine kinase mediated conditions or diseases, including, but not
limited to, conditions or diseases characterized by the presence or
development of one or more solid tumors.
Preparation of Compounds
[0499] Compounds of any of Formula (A1-A6), Formula (B1-B6),
Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula (VII) are
optionally synthesized using standard synthetic techniques or using
such methods known in combination with methods described herein. In
additions, solvents, temperatures and other reaction conditions are
presented herein for illustration only, and not to limit the scope
of the methods and compositions described herein. As a further
guide the following synthetic methods may also be utilized.
[0500] The reactions are optionally employed in a linear sequence
to provide the compounds described herein or used to synthesize
fragments which are subsequently joined by the methods described
herein and/or documented elsewhere.
Formation of Covalent Linkages by Reaction of an Electrophile with
a Nucleophile
[0501] The compounds described herein can be modified using various
electrophiles or nucleophiles to form new functional groups or
substituents. Table 1 entitled "Examples of Covalent Linkages and
Precursors Thereof" lists selected examples of covalent linkages
and precursor functional groups which yield and can be used as
guidance toward the variety of electrophiles and nucleophiles
combinations available. Precursor functional groups are shown
TABLE-US-00001 TABLE 1 Examples of Covalent Linkages and Precursors
Thereof Covalent Linkage Product Electrophile Nucleophile
Carboxamides Activated esters amines/anilines Carboxamides acyl
azides amines/anilines Carboxamides acyl halides amines/anilines
Esters acyl halides alcohols/phenols Esters acyl nitriles
alcohols/phenols Carboxamides acyl nitriles amines/anilines Imines
Aldehydes amines/anilines Hydrazones aldehydes or ketones
Hydrazines Oximes aldehydes or ketones Hydroxylamines Alkyl amines
alkyl halides amines/anilines Esters alkyl halides carboxylic acids
Thioethers alkyl halides Thiols Ethers alkyl halides
alcohols/phenols Thioethers alkyl sulfonates Thiols Esters alkyl
sulfonates carboxylic acids Ethers alkyl sulfonates
alcohols/phenols Esters Anhydrides alcohols/phenols Carboxamides
Anhydrides amines/anilines Thiophenols aryl halides Thiols Aryl
amines aryl halides Amines Thioethers Azindines Thiols Boronate
esters Boronates Glycols Carboxamides carboxylic acids
amines/anilines Esters carboxylic acids Alcohols hydrazines
Hydrazides carboxylic acids N-acylureas or carbodiimides carboxylic
acids Anhydrides Esters diazoalkanes carboxylic acids Thioethers
Epoxides Thiols Thioethers haloacetamides Thiols Ammotriazines
halotriazines amines/anilines Triazinyl ethers halotriazines
alcohols/phenols Amidines imido esters amines/anilines Ureas
Isocyanates amines/anilines Urethanes Isocyanates alcohols/phenols
Thioureas isothiocyanates amines/anilines Thioethers Maleimides
Thiols Phosphite esters phosphoramidites Alcohols Silyl ethers
silyl halides Alcohols Alkyl amines sulfonate esters
amines/anilines Thioethers sulfonate esters Thiols Esters sulfonate
esters carboxylic acids Ethers sulfonate esters Alcohols
Sulfonamides sulfonyl halides amines/anilines Sulfonate esters
sulfonyl halides phenols/alcohols Alkyl thiol
.alpha.,.beta.-unsaturated ester thiols Alkyl ethers
.alpha.,.beta.-unsaturated ester alcohols Alkyl amines
.alpha.,.beta.-unsaturated ester amines Alkyl thiol Vinyl sulfone
thiols Alkyl ethers Vinyl sulfone alcohols Alkyl amines Vinyl
sulfone amines Vinyl sulfide Propargyl amide thiol
Use of Protecting Groups
[0502] In the reactions described, it may be necessary to protect
reactive functional groups, for example hydroxy, amino, imino, thio
or carboxy groups, where these are desired in the final product, to
avoid their unwanted participation in the reactions. Protecting
groups are used to block some or all reactive moieties and prevent
such groups from participating in chemical reactions until the
protective group is removed. In one embodiment, each protective
group be removable by a different means. Protective groups that are
cleaved under totally disparate reaction conditions fulfill the
requirement of differential removal. Protective groups can be
removed by acid, base, and hydrogenolysis. Groups such as trityl,
dimethoxytrityl, acetal and t-butyldimethylsilyl are acid labile
and may be used to protect carboxy and hydroxy reactive moieties in
the presence of amino groups protected with Cbz groups, which are
removable by hydrogenolysis, and Fmoc groups, which are base
labile. Carboxylic acid and hydroxy reactive moieties may be
blocked with base labile groups such as, but not limited to,
methyl, ethyl, and acetyl in the presence of amines blocked with
acid labile groups such as t-butyl carbamate or with carbamates
that are both acid and base stable but hydrolytically
removable.
[0503] Carboxylic acid and hydroxy reactive moieties may also be
blocked with hydrolytically removable protective groups such as the
benzyl group, while amine groups capable of hydrogen bonding with
acids may be blocked with base labile groups such as Fmoc.
Carboxylic acid reactive moieties may be protected by conversion to
simple ester compounds as exemplified herein, or they may be
blocked with oxidatively-removable protective groups such as
2,4-dimethoxybenzyl, while co-existing amino groups may be blocked
with fluoride labile silyl carbamates.
[0504] Allyl blocking groups are useful in then presence of acid-
and base-protecting groups since the former are stable and can be
subsequently removed by metal or pi-acid catalysts. For example, an
allyl-blocked carboxylic acid can be deprotected with a
Pd.sup.0-catalyzed reaction in the presence of acid labile t-butyl
carbamate or base-labile acetate amine protecting groups. Yet
another form of protecting group is a resin to which a compound or
intermediate may be attached. As long as the residue is attached to
the resin, that functional group is blocked and cannot react. Once
released from the resin, the functional group is available to
react.
[0505] Typically blocking/protecting groups may be selected
from:
##STR00064##
[0506] Other protecting groups, plus a detailed description of
techniques applicable to the creation of protecting groups and
their removal are described in Greene and Wuts, Protective Groups
in Organic Synthesis, 3rd Ed., John Wiley & Sons, New York,
N.Y., 1999, and Kocienski, Protective Groups, Thieme Verlag, New
York, N.Y., 1994, which are incorporated herein by reference for
such disclosure.
Synthesis of Compounds
[0507] In certain embodiments, provided herein are methods of
making and methods of using tyrosine kinase inhibitor compounds
described herein. In certain embodiments, compounds described
herein can be synthesized using the following synthetic schemes.
Compounds may be synthesized using methodologies analogous to those
described below by the use of appropriate alternative starting
materials.
[0508] Described herein are compounds that inhibit the activity of
tyrosine kinase(s), such as Btk, and processes for their
preparation. Also described herein are pharmaceutically acceptable
salts, pharmaceutically acceptable solvates, pharmaceutically
active metabolites and pharmaceutically acceptable prodrugs of such
compounds. Pharmaceutical compositions that include at least one
such compound or a pharmaceutically acceptable salt,
pharmaceutically acceptable solvate, pharmaceutically active
metabolite or pharmaceutically acceptable prodrug of such compound,
are provided.
[0509] The starting material used for the synthesis of the
compounds described herein is either synthesized or obtained from
commercial sources, such as, but not limited to, Aldrich Chemical
Co. (Milwaukee, Wis.), Bachem (Torrance, Calif.), or Sigma Chemical
Co. (St. Louis, Mo.). The compounds described herein, and other
related compounds having different substituents are optionally
synthesized using techniques and materials, such as described, for
example, in March, ADVANCED ORGANIC CHEMISTRY 4.sup.th Ed., (Wiley
1992); Carey and Sundberg, ADVANCED ORGANIC CHEMISTRY 4.sup.th Ed.,
Vols. A and B (Plenum 2000, 2001); Green and Wuts, PROTECTIVE
GROUPS IN ORGANIC SYNTHESIS 3.sup.rd Ed., (Wiley 1999); Fieser and
Fieser's Reagents for Organic Synthesis, Volumes 1-17 (John Wiley
and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5
and Supplementals (Elsevier Science Publishers, 1989); Organic
Reactions, Volumes 1-40 (John Wiley and Sons, 1991); and Larock's
Comprehensive Organic Transformations (VCH Publishers Inc., 1989).
Other methods for the synthesis of compounds described herein may
be found in International Patent Publication No. WO 01/01982901,
Arnold et al. Bioorganic & Medicinal Chemistry Letters 10
(2000) 2167-2170; Burchat et al. Bioorganic & Medicinal
Chemistry Letters 12 (2002) 1687-1690. As a guide the following
synthetic methods may be utilized.
[0510] The products of the reactions are optionally isolated and
purified, if desired, using conventional techniques, including, but
not limited to, filtration, distillation, crystallization,
chromatography and the like. Such materials are optionally
characterized using conventional means, including physical
constants and spectral data.
[0511] Compounds described herein are optionally prepared using the
synthetic methods described herein as a single isomer or a mixture
of isomers.
[0512] A non-limiting example of a synthetic approach towards the
preparation of compounds of any of Formula (A1-A6), Formula
(B1-B6), Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula
(VII) is shown in Scheme I.
##STR00065##
[0513] Halogenation of commercially available
1H-pyrazolo[3,4-d]pyrimidin-4-amine provides an entry into the
synthesis of compounds of Formula (A1-A6), (B1-B6), (C1-C6) and/or
(D1-D6). In one embodiment, 1H-pyrazolo[3,4-d]pyrimidin-4-amine is
treated with N-iodosuccinamide to give
3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine. Metal catalyzed cross
coupling reactions are then carried out on
3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine. In one embodiment,
palladium mediated cross-coupling of a suitably substituted phenyl
boronic acid under basic conditions constructs intermediate 2.
Intermediate 2 is coupled with N-Boc-3-hydroxypiperidine (as
non-limiting example) via Mitsunobu reaction to give the Boc
(tert-butyloxycarbonyl) protected intermediate 3. After
deprotection with acid, coupling with, but not limited to, an acid
chloride, such as, but not limited to, acryloyl chloride, completes
the synthesis to give Compound 13.
[0514] A non-limiting example of a synthetic approach towards the
preparation of compounds containing the imidazotriazine moiety,
##STR00066##
is shown in Scheme II.
##STR00067##
[0515] A non-limiting example of a synthetic approach towards the
preparation of compounds containing any imidazopyrazine moiety,
##STR00068##
is shown in Scheme III.
##STR00069##
[0516] A non-limiting example of a synthetic approach towards the
preparation of compounds containing the pyrrolopyrimidine
moiety,
##STR00070##
is shown in Scheme IV.
##STR00071##
[0517] A non-limiting example of a synthetic approach towards the
preparation of compounds containing the Azaindole moiety,
##STR00072##
is shown in Scheme V.
##STR00073##
[0518] A non-limiting example of a synthetic approach towards the
preparation of compounds containing the pyrrolopyrimidine
moiety,
##STR00074##
is shown in Scheme VI.
##STR00075##
[0519] Using the synthetic methods described herein, tyrosine
kinase inhibitors as disclosed herein are obtained in good yields
and purity. The compounds prepared by the methods disclosed herein
are purified by conventional means, such as, for example,
filtration, recrystallization, chromatography, distillation, and
combinations thereof.
[0520] Any combination of the groups described above for the
various variables is contemplated herein.
Further Forms of Compounds
[0521] Compounds disclosed herein have a structure of any of
Formula (A1-A6), Formula (B1-B6), Formula (C1-C6), Formula (D1-D6),
Formula (I), or Formula (VII). It is understood that when reference
is made to compounds described herein, it is meant to include
compounds of any of Formula (A1-A6), Formula (B1-B6), Formula
(C1-C6), Formula (D1-D6), Formula (I), or Formula (VII), as well as
to all of the specific compounds that fall within the scope of
these generic formulae, unless otherwise indicated.
[0522] The compounds described herein may possess one or more
stereocenters and each center may exist in the R or S
configuration. The compounds presented herein include all
diastereomeric, enantiomeric, and epimeric forms as well as the
appropriate mixtures thereof. Stereoisomers may be obtained, if
desired, by methods such as, for example, the separation of
stereoisomers by chiral chromatographic columns.
[0523] Diasteromeric mixtures can be separated into their
individual diastereomers on the basis of their physical chemical
differences by methods known, for example, by chromatography and/or
fractional crystallization. In one embodiment, enantiomers can be
separated by chiral chromatographic columns. In other embodiments,
enantiomers can be separated by converting the enantiomeric mixture
into a diastereomeric mixture by reaction with an appropriate
optically active compound (e.g., alcohol), separating the
diastereomers and converting (e.g., hydrolyzing) the individual
diastereomers to the corresponding pure enantiomers. All such
isomers, including diastereomers, enantiomers, and mixtures thereof
are considered as part of the compositions described herein.
[0524] The methods and formulations described herein include the
use of N-oxides, crystalline forms (also known as polymorphs), or
pharmaceutically acceptable salts of compounds described herein, as
well as active metabolites of these compounds having the same type
of activity. In some situations, compounds exist as tautomers. All
tautomers are included within the scope of the compounds presented
herein. In addition, the compounds described herein can exist in
unsolvated as well as solvated forms with pharmaceutically
acceptable solvents such as water, ethanol, and the like. The
solvated forms of the compounds presented herein are also
considered to be disclosed herein.
[0525] Compounds of any of Formula (A1-A6), Formula (B1-B6),
Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula (VII) in
unoxidized form can be prepared from N-oxides of compounds of any
of Formula (A1-A6), Formula (B1-B6), Formula (C1-C6), Formula
(D1-D6), Formula (I), or Formula (VII) by treating with a reducing
agent, such as, but not limited to, sulfur, sulfur dioxide,
triphenyl phosphine, lithium borohydride, sodium borohydride,
phosphorus trichloride, tribromide, or the like in a suitable inert
organic solvent, such as, but not limited to, acetonitrile,
ethanol, aqueous dioxane, or the like at 0 to 80.degree. C.
[0526] In some embodiments, compounds described herein are prepared
as prodrugs. A "prodrug" refers to an agent that is converted into
the parent drug in vivo. Prodrugs are often useful because, in some
situations, they may be easier to administer than the parent drug.
They may, for instance, be bioavailable by oral administration
whereas the parent is not. The prodrug may also have improved
solubility in pharmaceutical compositions over the parent drug. An
example, without limitation, of a prodrug is a compound described
herein, which is administered as an ester (the "prodrug") to
facilitate transmittal across a cell membrane where water
solubility is detrimental to mobility but which then is
metabolically hydrolyzed to the carboxylic acid, the active entity,
once inside the cell where water-solubility is beneficial. A
further example of a prodrug is a short peptide (polyaminoacid)
bonded to an acid group where the peptide is metabolized to reveal
the active moiety. In certain embodiments, upon in vivo
administration, a prodrug is chemically converted to the
biologically, pharmaceutically or therapeutically active form of
the compound. In certain embodiments, a prodrug is enzymatically
metabolized by one or more steps or processes to the biologically,
pharmaceutically or therapeutically active form of the compound. To
produce a prodrug, a pharmaceutically active compound is modified
such that the active compound will be regenerated upon in vivo
administration. The prodrug can be designed to alter the metabolic
stability or the transport characteristics of a drug, to mask side
effects or toxicity, to improve the flavor of a drug or to alter
other characteristics or properties of a drug. By virtue of
knowledge of pharmacodynamic processes and drug metabolism in vivo,
once a pharmaceutically active compound is known, prodrugs of
compounds can be designed (if desired) (for examples of this
procedure applied to other compounds, see, e.g., Nogrady (1985)
Medicinal Chemistry A Biochemical Approach, Oxford University
Press, New York, pages 388-392; Silverman (1992), The Organic
Chemistry of Drug Design and Drug Action, Academic Press, Inc., San
Diego, pages 352-401, Saulnier et al., (1994), Bioorganic and
Medicinal Chemistry Letters, Vol. 4, p. 1985).
[0527] Prodrug forms of the herein described compounds, wherein the
prodrug is metabolized in vivo to produce a derivative as set forth
herein are included within the scope of the claims. In some cases,
some of the compounds herein-described are prodrugs for another
derivative or active compound.
[0528] Prodrugs are often useful because, in some situations, they
are easier to administer than the parent drug. They are, for
instance, bioavailable by oral administration whereas the parent is
not. The prodrug optionally has improved solubility in
pharmaceutical compositions over the parent drug. Prodrugs may be
designed as reversible drug derivatives, for use as modifiers to
enhance drug transport to site-specific tissues. In some
embodiments, the design of a prodrug increases the effective water
solubility. See, e.g., Fedorak et al., Am. J. Physiol.,
269:G210-218 (1995); McLoed et al., Gastroenterol, 106:405-413
(1994); Hochhaus et al., Biomed. Chrom., 6:283-286 (1992); J.
Larsen and H. Bundgaard, Int. J. Pharmaceutics, 37, 87 (1987); J.
Larsen et al., Int. J Pharmaceutics, 47, 103 (1988); Sinkula et
al., J. Pharm. Sci., 64:181-210 (1975); T. Higuchi and V. Stella,
Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S.
Symposium Series; and Edward B. Roche, Bioreversible Carriers in
Drug Design, American Pharmaceutical Association and Pergamon
Press, 1987, all incorporated by reference for such disclosure.
[0529] Sites on the aromatic ring portion of compounds of any of
Formula (A1-A6), Formula (B1-B6), Formula (C1-C6), Formula (D1-D6),
Formula (I), or Formula (VII) can be susceptible to various
metabolic reactions, therefore incorporation of appropriate
substituents on the aromatic ring structures, such as, by way of
example only, halogens can reduce, minimize or eliminate this
metabolic pathway.
[0530] Compounds described herein include isotopically-labeled
compounds, which are identical to those recited in the various
formulas and structures presented herein, but for the fact that one
or more atoms are replaced by an atom having an atomic mass or mass
number different from the atomic mass or mass number usually found
in nature. Examples of isotopes that can be incorporated into the
present compounds include isotopes of hydrogen, carbon, nitrogen,
oxygen, fluorine and chlorine, such as .sup.2H, .sup.3H, .sup.13C,
.sup.14C, .sup.15N, .sup.18O, .sup.17O, .sup.35S, .sup.18F,
.sup.36Cl, respectively. Certain isotopically-labeled compounds
described herein, for example those into which radioactive isotopes
such as .sup.3H and .sup.14C are incorporated, are useful in drug
and/or substrate tissue distribution assays. Further, substitution
with isotopes such as deuterium, i.e., .sup.2H, can afford certain
therapeutic advantages resulting from greater metabolic stability,
for example increased in vivo half-life or reduced dosage
requirements.
[0531] In additional or further embodiments, the compounds
described herein are metabolized upon administration to an organism
in need to produce a metabolite that is then used to produce a
desired effect, including a desired therapeutic effect.
[0532] Compounds described herein (for example, compounds of any of
Formula (A1-A6), Formula (B1-B6), Formula (C1-C6), Formula (D1-D6),
Formula (I), or Formula (VII)) are optionally in the form of,
and/or used as, pharmaceutically acceptable salts. The type of
pharmaceutical acceptable salts, include, but are not limited to:
(1) acid addition salts, formed) by reacting the free base form of
the compound with a pharmaceutically acceptable: inorganic acid
such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric
acid, phosphoric acid, metaphosphoric acid, and the like; or with
an organic acid such as acetic acid, propionic acid, hexanoic acid,
cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic
acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric
acid, trifluoroacetic acid, tartaric acid, citric acid, benzoic
acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic
acid, methanesulfonic acid, ethanesulfonic acid,
1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid,
benzenesulfonic acid, toluenesulfonic acid, 2-naphthalenesulfonic
acid, 4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid,
glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-ene-1-carboxylic
acid), 3-phenylpropionic acid, trimethylacetic acid, tertiary
butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic
acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic
acid, and the like; (2) salts formed when an acidic proton present
in the parent compound either is replaced by a metal ion, e.g., an
alkali metal ion (e.g. lithium, sodium, potassium), an alkaline
earth ion (e.g. magnesium, or calcium), or an aluminum ion; or
coordinates with an organic base. Acceptable organic bases include
ethanolamine, diethanolamine, triethanolamine, tromethamine,
N-methylglucamine, and the like. Acceptable inorganic bases include
aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium
carbonate, sodium hydroxide, and the like.
[0533] The corresponding counterions of the pharmaceutically
acceptable salts are optionally analyzed and identified using
various methods including, but not limited to, ion exchange
chromatography, ion chromatography, capillary electrophoresis,
inductively coupled plasma, atomic absorption spectroscopy, mass
spectrometry, or any combination thereof.
[0534] The salts are recovered by using at least one of the
following techniques: filtration, precipitation with a non-solvent
followed by filtration, evaporation of the solvent, or, in the case
of aqueous solutions, lyophilization.
[0535] It should be understood that a reference to a
pharmaceutically acceptable salt includes the solvent addition
forms or crystal forms thereof, particularly solvates or
polymorphs. Solvates contain either stoichiometric or
non-stoichiometric amounts of a solvent, and are optionally formed
during the process of crystallization with pharmaceutically
acceptable solvents such as water, ethanol, and the like. Hydrates
are formed when the solvent is water, or alcoholates are formed
when the solvent is alcohol. Solvates of compounds described herein
can be conveniently prepared or formed during the processes
described herein. In addition, the compounds provided herein can
exist in unsolvated as well as solvated forms. In general, the
solvated forms are considered equivalent to the unsolvated forms
for the purposes of the compounds and methods provided herein.
[0536] It should be understood that a reference to a salt includes
the solvent addition forms or crystal forms thereof, particularly
solvates or polymorphs. Solvates contain either stoichiometric or
non-stoichiometric amounts of a solvent, and are often formed
during the process of crystallization with pharmaceutically
acceptable solvents such as water, ethanol, and the like. Hydrates
are formed when the solvent is water, or alcoholates are formed
when the solvent is alcohol. Polymorphs include the different
crystal packing arrangements of the same elemental composition of a
compound. Polymorphs usually have different X-ray diffraction
patterns, infrared spectra, melting points, density, hardness,
crystal shape, optical and electrical properties, stability, and
solubility. Various factors such as the recrystallization solvent,
rate of crystallization, and storage temperature may cause a single
crystal form to dominate.
[0537] Compounds described herein are optionally in various forms,
including but not limited to, amorphous forms, milled forms and
nano-particulate forms. In addition, compounds described herein
include crystalline forms, also known as polymorphs. Polymorphs
include the different crystal packing arrangements of the same
elemental composition of a compound. Polymorphs usually have
different X-ray diffraction patterns, infrared spectra, melting
points, density, hardness, crystal shape, optical and electrical
properties, stability, and solubility. Various factors such as the
recrystallization solvent, rate of crystallization, and storage
temperature may cause a single crystal form to dominate.
[0538] The screening and characterization of the pharmaceutically
acceptable salts, polymorphs and/or solvates may be accomplished
using a variety of techniques including, but not limited to,
thermal analysis, x-ray diffraction, spectroscopy, vapor sorption,
and microscopy. Thermal analysis methods address thermo chemical
degradation or thermo physical processes including, but not limited
to, polymorphic transitions, and such methods are used to analyze
the relationships between polymorphic forms, determine weight loss,
to find the glass transition temperature, or for excipient
compatibility studies. Such methods include, but are not limited
to, Differential scanning calorimetry (DSC), Modulated Differential
Scanning Calorimetry (MDCS), Thermogravimetric analysis (TGA), and
Thermogravi-metric and Infrared analysis (TG/IR). X-ray diffraction
methods include, but are not limited to, single crystal and powder
diffractometers and synchrotron sources. The various spectroscopic
techniques used include, but are not limited to, Raman, FTIR, UVIS,
and NMR (liquid and solid state). The various microscopy techniques
include, but are not limited to, polarized light microscopy,
Scanning Electron Microscopy (SEM) with Energy Dispersive X-Ray
Analysis (EDX), Environmental Scanning Electron Microscopy with EDX
(in gas or water vapor atmosphere), IR microscopy, and Raman
microscopy.
Cysteine-Targeted Kinase Inhibitor Discovery Platform
Kinases/Inhibitors SAR Approach
[0539] Protein kinases, which act on and modify the activity of
specific proteins, are used to transmit signals and control complex
processes in cells. Up to 518 different kinases have been
identified in humans. Many kinase inhibitor compounds
non-selectively bind and/or inhibit these kinases because the
active sites of some of these kinases are similar in structure.
Such cross-reactivity is not a desired feature of a kinase
inhibitor compound because of the potential for undesired side
effects when such a compound is being administered to treat a
disorder.
[0540] We have observed that small differences in the structure of
kinase inhibitor compounds have profound effects in the selectivity
of similarly-structured kinases (e.g., ACKs, including, Btk and the
Btk kinase cysteine homologs).
[0541] As a result, we have developed assays, methods, and systems
for converting a non-selective inhibitor compound into a
highly-selective inhibitor compound. In brief, the non-selective
inhibitor compound is provided with a Michael acceptor moiety and a
linker moiety that links the Michael acceptor moiety to the
remainder of the non-selective inhibitor compound. A series of
linker and Michael acceptor moieties provides a small library/panel
of test inhibitor compounds. The inhibitor library/panel is
contacted with a panel of structurally related kinases (e.g., Btk
and the Btk kinase cysteine homologs). Binding is determined by a
variety of means, included fluorescence detection (or via any other
detectable label), mass spectrometry, or a combination of
approaches. An Activity Probe is optionally used to detect binding
of members of the inhibitor library/panel to the kinase
library/panel. The binding data is then optionally collected and
analyzed to provide a structure-activity relationship (SAR) between
the structure of the members of the inhibitor panel/library (e.g.,
Michael acceptor and/or linker moieties) and the activity of
binding to and/or inhibiting members of the kinase panel. Based on
this information, further modifications are suggested if necessary.
We have successfully used this approach to improve the binding and
selectivity of Btk inhibitor compounds (see Examples herein,
including "Kinase Inhibitor Discovery Platform" example
section).
[0542] In some embodiments, a similar approach is used for
converting a selective inhibitor compound for a group of
similarly-structured ACKs (including, Btk and the Btk kinase
cysteine homologs) into a more highly-selective inhibitor compound
(e.g., more selective for a particular ACK over
structurally-similar ACKs), or for converting a selective inhibitor
compound for a particular ACK (e.g., Btk) into an even more
selective inhibitor of that particular ACK. For example, in brief,
the selective inhibitor compound (which, for example, contains an
active-site binding moiety, a linker moiety and a Michael acceptor
moiety) is modified. In one embodiment, a series of linker and
Michael acceptor moieties provides a small library/panel of test
inhibitor compounds. The inhibitor library/panel is contacted with
a panel of structurally related kinases (e.g., Btk and the Btk
kinase cysteine homologs). Binding is determined by a variety of
means, included fluorescence detection (or via any other detectable
label), mass spectrometry, or a combination of approaches. An
Activity Probe is optionally used to detect binding of members of
the inhibitor library/panel to the kinase library/panel. The
binding data is then optionally collected and analyzed to provide a
structure-activity relationship (SAR) between the structure of the
members of the inhibitor panel/library (e.g., Michael acceptor
and/or linker moieties) and the activity of binding to and/or
inhibiting members of the kinase panel. Based on this information,
further modifications are suggested if necessary. We have also
successfully used this approach to improve the binding and
selectivity of Btk inhibitor compounds (see Examples herein,
including "Kinase Inhibitor Discovery Platform" example
section).
[0543] Thus, for our highly selective BTK inhibitor Compound 1, we
engineered an electrophilic center capable of irreversibly
inactivating the target enzyme, BTK. That is, to an active site
binding moiety of a reversible inhibitor was added a linker moiety
and a Michael acceptor moiety that achieved a high degree of
potency and selectivity by (1) fitting the core scaffold into the
active site ATP binding pocket of kinase enzymes, and (2) forming a
covalent bond with Cysteine-481 located in BTK. The chemistry
required for covalent bond formation involves an electrophilic
moiety that acts as a Michael acceptor, which bonds with a
nucleophile (such as Cys-481) present in a precise location within
the active site.
[0544] In another example, the linker and Michael acceptor moiety
of Compound 1 was modified to provide Compound 9 which has a
different selectivity pattern. Table 1 is a table showing the
degree of inhibition of a panel of kinases for two example
compounds. IC.sub.50s were determined using the in vitro HotSpot
kinase assay (purified enzymes, 33P-ATP, an appropriate substrate
and 1 uM ATP.) Compared to Compound 1, Compound 9 has similar
potency toward Btk, but significantly less potency toward JAK-3,
ITK, and EGFR and significantly more potency toward the src-family
kinases lck, c-src, FGR, Fyn, Hck, and Lyn and Yes. Thus, subtle
modifications in the linker moiety and the Michael acceptor moiety
are important for the design of selective ACK inhibitors.
TABLE-US-00002 TABLE 1 Compound 1 Compound 9 Kinase IC50 (nM) IC50
(nM) BTK 0.5 1.0 ITK 11.7 909.9 Bmx/ETK 0.8 1.1 TEC 77.8 108.0 EFGR
0.5 20.6 HER4 9.4 1536.0 HER4 0.1 3.2 LCK 2.0 1.0 BLK 0.5 0.2 C-src
262.6 14.3 FGR 2.3 0.4 Fyn 95.6 7.1 HCK 3.7 1.0 Lyn 16.2 1.2 YES
6.5 0.8 ABL 86.1 32.3 Brk 3.3 3.3 CSK 2.2 2.4 FER 8,070.0 3,346.0
JAK3 10.4 8,278.0 SYK >10,000 >10,000
[0545] Table 2 of Example 1c in the "Kinase Discovery Platform and
Pulse Dosing" section of the examples section provides further
modifications of the linker moiety and/or the Michael acceptor
moiety and the impact of such changes of inhibitor selectivity.
[0546] Thus, in one aspect described herein are methods of
identifying an irreversible inhibitor of a kinase selected from
Btk, a Btk homolog, a Btk kinase cysteine homolog, an ACK, or HER4
(or indeed, any ACK) comprising: [0547] (1) contacting a
multiplicity of kinases selected from Btk, a Btk homolog, a Btk
kinase cysteine homolog, an ACK, or HER4 (or indeed any ACK) with a
compound that comprises a Michael acceptor moiety; [0548] (2)
contacting at least one non-kinase molecule having at least one
accessible SH group with the compound that comprises a Michael
acceptor moiety (this step allows for the selection of inhibitors
that have low selectivity for higher abundance biological molecules
that have moieties that irreversibly react with the inhibitor; thus
preventing the inhibitor from binding to the desire ACK when
administered as a drug to an individual); and [0549] (3)
determining the covalent binding of the compound that comprises a
Michael acceptor with the multiplicity of kinases and the at least
one non-kinase molecule; and repeating steps (1), (2), and (3) for
at least one other compound that comprises a Michael acceptor
moiety.
[0550] In a further aspect, the following steps are added: (4)
comparing the covalent binding of the compound that comprises a
Michael acceptor with the multiplicity of kinases and the at least
one non-kinase molecule; and repeating steps (1), (2), (3) and (4)
for at least one other compound that comprises a Michael acceptor
moiety.
[0551] In a further aspect the irreversible inhibitor compounds are
also contacted with at least one non-ACK kinase in order to
determine the selectivity of the irreversible inhibitor compound
for the ACK relative to the non-ACK.
[0552] By way of certain relevant examples of non-kinase molecules
with at least one accessible SH group are glutathione and/or
hemoglobin. Because of the high abundance of these molecules in
typical biological systems (e.g., in an individual), the desired
irreversible inhibitor compounds have low selectivity/reactivity
with these non-kinase molecules.
[0553] In certain embodiments of the Kinase Inhibitor Discovery
Platform, an Activity Probe (described in more detail herein) is
used as a rapid diagnostic method for determining whether a test
inhibitor compound has irreversibly inhibited an ACK. In one
embodiment, the Activity Probe is itself an irreversible inhibitor
of an ACK, an irreversible inhibitor of HER4, and further, has a
reporter moiety (e.g., a fluorescent moiety) as part of its
structure. When used in competition with a test irreversible
inhibitor, the absence of a `reporter` signal on an ACK is one
indication that the test irreversible inhibitor has prevented the
Activity Probe from binding to the ACK (and that the test
irreversible inhibitor has a higher binding affinity for the ACK
than the Activity Probe).
[0554] In certain embodiments, the Kinase Inhibitor Discovery
Platform, steps (1) and (2) are conducted in vivo and step (3) is
conducted in part using an Activity Probe. Further, in certain
embodiments, the determining step uses mass spectrometry,
fluorescence, or a combination thereof.
[0555] As described herein, in one embodiment, the inhibitor tested
with the Kinase Inhibitor Discovery Platform comprise an active
site binding moiety, a Michael acceptor moiety, and a linker moiety
that links the Michael acceptor moiety to the active site binding
moiety. For example, in such a scheme, the following information is
collected and analyzed: the structure-function activity
relationship between the structure of the linker moiety and/or the
Michael acceptor moiety of each compound, and the binding and/or
selectivity of each compound to at least one kinase. Further, in
certain embodiments, structure of the active site binding moiety of
each compound is not varied, whereas the structure of the linker
moiety and/or the Michael acceptor moiety is varied.
[0556] In one example, the inhibitors have the structure of Formula
(VII):
##STR00076##
wherein: [0557] wherein
##STR00077##
[0557] is a moiety that binds to the active site of a kinase,
including a tyrosine kinase, further including a Btk kinase
cysteine homolog; [0558] Y is an optionally substituted group
selected from among alkylene, heteroalkylene, arylene,
heteroarylene, heterocycloalkylene, cycloalkylene, alkylenearylene,
alkyleneheteroarylene, alkylenecycloalkylene, and
alkyleneheterocycloalkylene; [0559] Z is C(.dbd.O), OC(.dbd.O),
NHC(.dbd.O), NCH.sub.3C(.dbd.O), C(.dbd.S), S(.dbd.O).sub.x,
OS(.dbd.O).sub.x, NHS(.dbd.O).sub.x, where x is 1 or 2; [0560]
R.sub.7 and R.sub.8 are independently selected from among H,
unsubstituted C.sub.1-C.sub.4 alkyl, substituted
C.sub.1-C.sub.4alkyl, unsubstituted C.sub.1-C.sub.4heteroalkyl,
substituted C.sub.1-C.sub.4heteroalkyl, unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted C.sub.3-C.sub.6cycloalkyl,
unsubstituted C.sub.2-C.sub.6heterocycloalkyl, and substituted
C.sub.2-C.sub.6heterocycloalkyl; or [0561] R.sub.7 and R.sub.8
taken together form a bond; and [0562] R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted C.sub.2-C.sub.8heterocycloalkyl,
substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl).
[0563] In such a scheme, the following information is collected and
analyzed: the structure-function activity relationship between the
structure of Y--Z and/or
##STR00078##
of each compound, and the binding and/or selectivity of each
compound to at least one kinase. Further, the structure of
##STR00079##
of each compound is not varied, whereas the structure of the linker
moiety (Y--Z) and/or the Michael acceptor moiety
##STR00080##
is varied.
[0564] In certain embodiments of the Kinase Inhibitor Discovery
Platform, the resulting inhibitor is selective for one kinase
selected from Btk, a Btk homolog, an ACK, HER4, and a Btk kinase
cysteine homolog over at least one other kinase selected from Btk,
a Btk homolog, an ACK, HER4, and a Btk kinase cysteine homolog. In
some embodiments, this selectivity is at least 5.times., at least
10.times., at least 20.times., at least 50.times., or at least
100.times.. In further embodiments, the resulting inhibitor is
selective for at least one kinase selected from Btk, a Btk homolog,
an ACK, HER4, and a Btk kinase cysteine homolog over at least one
other non-kinase molecule having an accessible SH group. In some
embodiments, this selectivity is at least 5.times., at least
10.times., at least 20.times., at least 50.times., or at least
100.times..
[0565] In further embodiments, the resulting inhibitor is used in
the therapeutic methods described herein, or in the pharmaceutical
compositions described herein.
[0566] Activity Probe Compounds
[0567] Because of the Kinase Inhibitor Discovery Platform described
herein optionally utilizes an Activity Probe, the following section
describes the design, structure and use of non-limiting examples of
Activity Probes.
[0568] The Activity Probe compounds described herein are composed
of a moiety comprising an inhibitor of Btk, a Btk homolog, a Btk
kinase cysteine homolog, an ACK, or HER4 (hereinafter, a "Kinase
Inhibitor"), a linker moiety, and a reporter moiety. In one
embodiment, the Kinase Inhibitor is an irreversible inhibitor. In
another embodiment, the irreversible Kinase Inhibitor binds to a
non-catalytic residue in the ATP binding pocket of Btk, a Btk
homolog, a Btk kinase cysteine homolog, an ACK, or HER4
(hereinafter a "Kinase"); in further embodiments, the non-catalytic
residue is a cysteine residue. In some embodiments, the Activity
Probe forms a covalent bond with at least one non-catalytic residue
of a Kinase. In other embodiments, the Activity Probe forms a
non-covalent bond with at least one non-catalytic residue of a
Kinase. In a further embodiment, the Activity Probe forms hydrogen
bonding within the ATP binding pocket of a Kinase. In yet a further
embodiment, the Activity Probe has Van der Waals attractions with
the Kinase.
[0569] In some other embodiments, the Activity Probes described
herein are activity dependent such that the probe binds only an
active Kinase. In further embodiments, the Activity Probe binds a
Kinase that has been switched on by phosphorylation by upstream
kinases. In yet a further embodiment, the Activity Probes described
herein are activity independent such that the probe binds Kinases
that have not been switched on by phosphorylation by upstream
kinases. In some embodiments, the Activity Probe labels a
phosphorylated conformation of a Kinase. In other embodiments, the
Activity Probe labels a Kinase in a non-phosphorylated
conformation.
[0570] In some embodiments, the Activity Probe is permeable to
cells.
[0571] In further embodiments, the linker moiety is selected from a
bond, a substituted alkyl moiety, a substituted heterocycle moiety,
a substituted amide moiety, a ketone moiety, a substituted
carbamate moiety, an ester moiety, or any combination thereof. In
further embodiments, the reporter moiety is a moiety that is
detected using standard or modified laboratory equipment.
[0572] In one aspect is a Activity Probe of Formula (I)
comprising:
##STR00081##
wherein: [0573] A is a Kinase Inhibitor moiety; [0574] X and Y are
independently selected from the group consisting of: a bond,
--O(C.dbd.O)--, --NR.sup.a(C.dbd.O)--, --NR.sup.a--,
##STR00082##
[0574] --O--, --S--, --S--S--, --O--NR.sup.a--, --O(C.dbd.O)O--,
--O(C.dbd.)NR.sup.a, --NR.sup.a(C.dbd.O)NR.sup.a--,
--N.dbd.CR.sup.a--, --S(C.dbd.O)--, --S(O)--, and --S(O).sub.2--;
[0575] wherein
##STR00083##
[0575] forms a N-containing heterocycle; [0576] B is a linker
moiety; [0577] C is a reporter moiety; and [0578] R.sup.a is
hydrogen or alkyl.
[0579] In one embodiment, the moiety comprising an irreversible
Kinase Inhibitor is derived from an irreversible inhibitor of a
Kinase. In some embodiments, such irreversible Kinase Inhibitors
should possess at least one of the following characteristics:
potency, selectively and cell permeability. In further embodiments,
such irreversible Kinase Inhibitors possess at least two of the
aforementioned characteristics, and in further embodiments, at
least all of the aforementioned characteristics.
[0580] In another embodiment, the Kinase Inhibitor moiety is
derived from a Btk inhibitor having the structure of Formula
(II):
##STR00084##
wherein: [0581] L.sub.a is CH.sub.2, O, NH or S; [0582] Ar is a
substituted or unsubstituted aryl, or a substituted or
unsubstituted heteroaryl; and [0583] Y is an optionally substituted
group selected from among alkylene, heteroalkylene, arylene,
heteroarylene, heterocycloalkylene, cycloalkylene, alkylenearylene,
alkyleneheteroarylene, alkylenecycloalkylene, and
alkyleneheterocycloalkylene.
[0584] In some embodiments, L.sub.a is CH.sub.2, O, or NH. In other
embodiments, L.sub.a is O or NH. In yet other embodiments, L.sub.a
is O.
[0585] In other embodiments, Ar is a substituted or unsubstituted
aryl. In yet other embodiments, Ar is a 6-membered aryl. In some
other embodiments, Ar is phenyl.
[0586] In some embodiments, Y is an optionally substituted group
selected from among alkylene, heteroalkylene, arylene,
heteroarylene, heterocycloalkylene, cycloalkylene, alkylenearylene,
alkyleneheteroarylene, alkylenecycloalkylene, and
alkyleneheterocycloalkylene. In other embodiments, Y is an
optionally substituted group selected from among
C.sub.1-C.sub.6alkylene, C.sub.1-C.sub.6heteroalkylene, 4-, 5-, 6-,
or 7-membered cycloalkylene, and 4-, 5-, 6-, or 7-membered
heterocycloalkylene. In yet other embodiments, Y is an optionally
substituted group selected from among C.sub.1-C.sub.6alkylene,
C.sub.1-C.sub.6heteroalkylene 5- or 6-membered cycloalkylene, and
5- or 6-membered heterocycloalkylene containing 1 or 2 N atoms. In
some other embodiments, Y is a 5- or 6-membered cycloalkylene, or a
5- or 6-membered heterocycloalkylene containing 1 or 2 N atoms. In
some embodiments, Y is a 4-, 5-, 6-, or 7-membered cycloalkylene
ring; or Y is a 4-, 5-, 6-, or 7-membered heterocycloalkylene
ring.
[0587] In some embodiments, the Kinase Inhibitor moiety is derived
from a compound selected from among:
1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperi-
din-1-yl)prop-2-en-1-one;
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)but-2-en-1-one;
1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperi-
din-1-yl)sulfonylethene;
1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperi-
din-1-yl)prop-2-yn-1-one;
1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperi-
din-1-yl)prop-2-en-1-one; N-((1
s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyc-
lohexyl)acrylamide;
1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)py-
rrolidin-1-yl)prop-2-en-1-one;
1-((S)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)py-
rrolidin-1-yl)prop-2-en-1-one;
1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)prop-2-en-1-one;
1-((S)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)prop-2-en-1-one; and
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-(dimethylamino)but-2-en-1-one;
(E)-4-(N-(2-hydroxyethyl)-N-methylamino)-1-(3-(4-phenoxyphenyl)-1H-pyrazo-
lo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)but-2-en-1-one (Compound 3);
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3-
-(1H-imidazol-4-yl)prop-2-en-1-one (Compound 4);
(E)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-morpholinobut-2-en-1-one (Compound 5);
(E)-1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound 7);
(E)-N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-
-1-yl)cyclohexyl)-4-(dimethylamino)but-2-enamide (Compound 8);
N-((1r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)acrylamide (Compound 10);
(E)-1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)methyl)pyrolidin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound
11);
(E)-1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)methyl)pyrolidin-1-yl)-4-(dimethylamino)but-2-en-1-one (Compound
12);
1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 13);
1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 14);
1((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)me-
thyl)pyrrolidin-1-yl)but-2-yn-1-one (Compound 15);
1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)but-2-yn-1-one (Compound 16);
1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)but-2-yn-1-one (Compound 17);
(E)-N-((1,r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidi-
n-1-yl)cyclohexyl-4-(dimethylamino)but-2-enamide (Compound 18);
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
-N-methylacrylamide (Compound 19);
(E)-1-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-4-
-morpholinobut-2-en-1-one (Compound 20);
(E)-1-((S-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)methyl)pyrrolidin-1-yl)-4-morpholinobut-2-en-1-one (Compound 21);
N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)but-2-ynamide (Compound 22);
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
acrylamide (Compound 23);
(E)-1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)piperidin-1-yl)-4-morpholinobut-2-en-1-one (Compound 24);
(E)-N-((1s,4s)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-
-1-yl)cyclohexyl)-4-morpholinobut-2-enamide (Compound 25).
[0588] In another embodiment, the linker moiety is selected from a
bond, a polymer, a water soluble polymer, optionally substituted
alkyl, optionally substituted heteroalkyl, optionally substituted
heterocycloalkyl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkylalkyl, optionally substituted
heterocycloalkylalkenyl, optionally substituted aryl, optionally
substituted heteroaryl, and optionally substituted
heterocycloalkylalkenylalkyl. In some embodiments, the linker
moiety is an optionally substituted heterocycle. In other
embodiments, the heterocycle is selected from aziridine, oxirane,
episulfide, azetidine, oxetane, pyrroline, tetrahydrofuran,
tetrahydrothiophene, pyrrolidine, pyrazole, pyrrole, imidazole,
triazole, tetrazole, oxazole, isoxazole, oxirene, thiazole,
isothiazole, dithiolane, furan, thiophene, piperidine,
tetrahydropyran, thiane, pyridine, pyran, thiapyrane, pyridazine,
pyrimidine, pyrazine, piperazine, oxazine, thiazine, dithiane, and
dioxane. In some embodiments, the heterocycle is piperazine. In
further embodiments, the linker moiety is optionally substituted
with halogen, CN, OH, NO.sub.2, alkyl, S(O), and S(O).sub.2. In
other embodiments, the water soluble polymer is a PEG group.
[0589] In other embodiments, the linker moiety provides sufficient
spatial separation between the reporter moiety and the Kinase
Inhibitor moiety. In further embodiments, the linker moiety is
stable. In yet a further embodiment, the linker moiety does not
substantially affect the response of the reporter moiety. In other
embodiments the linker moiety provides chemical stability to the
Activity Probe. In further embodiments, the linker moiety provides
sufficient solubility to the Activity Probe.
[0590] In some embodiments, linkages such as water soluble polymers
are coupled at one end to a Kinase Inhibitor moiety and to a
reporter moiety at the other end. In other embodiments, the water
soluble polymers are coupled via a functional group or substituent
of the Kinase Inhibitor moiety. In further embodiments, the water
soluble polymers are coupled via a functional group or substituent
of the reporter moiety. In other embodiments, covalent attachment
of hydrophilic polymers to a Kinase Inhibitor moiety and a reporter
moiety represents one approach to increasing water solubility (such
as in a physiological environment), bioavailability, increasing
serum half-life, increasing pharmacodynamic parameters, or
extending the circulation time of the Activity Probe, including
proteins, peptides, and particularly hydrophobic molecules. In
further embodiments, additional important features of such
hydrophilic polymers include biocompatibility and lack of toxicity.
In other embodiments, for therapeutic use of the end-product
preparation, the polymer is pharmaceutically acceptable.
[0591] In some embodiments, examples of hydrophilic polymers
include, but are not limited to: polyalkyl ethers and alkoxy-capped
analogs thereof (e.g., polyoxyethylene glycol,
polyoxyethylene/propylene glycol, and methoxy or ethoxy-capped
analogs thereof, polyoxyethylene glycol, the latter is also known
as polyethylene glycol or PEG); polyvinylpyrrolidones;
polyvinylalkyl ethers; polyoxazolines, polyalkyl oxazolines and
polyhydroxyalkyl oxazolines; polyacrylamides, polyalkyl
acrylamides, and polyhydroxyalkyl acrylamides (e.g.,
polyhydroxypropylmethacrylamide and derivatives thereof);
polyhydroxyalkyl acrylates; polysialic acids and analogs thereof;
hydrophilic peptide sequences; polysaccharides and their
derivatives, including dextran and dextran derivatives, e.g.,
carboxymethyldextran, dextran sulfates, aminodextran; cellulose and
its derivatives, e.g., carboxymethyl cellulose, hydroxyalkyl
celluloses; chitin and its derivatives, e.g., chitosan, succinyl
chitosan, carboxymethylchitin, carboxymethylchitosan; hyaluronic
acid and its derivatives; starches; alginates; chondroitin sulfate;
albumin; pullulan and carboxymethyl pullulan; polyaminoacids and
derivatives thereof, e.g., polyglutamic acids, polylysines,
polyaspartic acids, polyaspartamides; maleic anhydride copolymers
such as: styrene maleic anhydride copolymer, divinylethyl ether
maleic anhydride copolymer; polyvinyl alcohols; copolymers thereof;
terpolymers thereof; mixtures thereof; and derivatives of the
foregoing. In other embodiments, the water soluble polymer is any
structural form including but not limited to linear, forked or
branched. In some embodiments, polymer backbones that are
water-soluble, with from 2 to about 300 termini, are particularly
useful. In further embodiments, multifunctional polymer derivatives
include, but are not limited to, linear polymers having two
termini, each terminus being bonded to a functional group which is
the same or different. In some embodiments, the water polymer
comprises a poly(ethylene glycol) moiety. In further embodiments,
the molecular weight of the polymer is of a wide range, including
but not limited to, between about 100 Da and about 100,000 Da or
more. In yet further embodiments, the molecular weight of the
polymer is between about 100 Da and about 100,000 Da, including but
not limited to, about 100,000 Da, about 95,000 Da, about 90,000 Da,
about 85,000 Da, about 80,000 Da, about 75,000 Da, about 70,000 Da,
about 65,000 Da, about 60,000 Da, about 55,000 Da, about 50,000 Da,
about 45,000 Da, about 40,000 Da, about 35,000 Da, 30,000 Da, about
25,000 Da, about 20,000 Da, about 15,000 Da, about 10,000 Da, about
9,000 Da, about 8,000 Da, about 7,000 Da, about 6,000 Da, about
5,000 Da, about 4,000 Da, about 3,000 Da, about 2,000 Da, about
1,000 Da, about 900 Da, about 800 Da, about 700 Da, about 600 Da,
about 500 Da, about 400 Da, about 300 Da, about 200 Da, and about
100 Da. In some embodiments, the molecular weight of the polymer is
between about 100 Da and 50,000 Da. In some embodiments, the
molecular weight of the polymer is between about 100 Da and 40,000
Da. In some embodiments, the molecular weight of the polymer is
between about 1,000 Da and 40,000 Da. In some embodiments, the
molecular weight of the polymer is between about 5,000 Da and
40,000 Da. In some embodiments, the molecular weight of the polymer
is between about 10,000 Da and 40,000 Da. In some embodiments, the
poly(ethylene glycol) molecule is a branched polymer. In further
embodiments, the molecular weight of the branched chain PEG is
between about 1,000 Da and about 100,000 Da, including but not
limited to, about 100,000 Da, about 95,000 Da, about 90,000 Da,
about 85,000 Da, about 80,000 Da, about 75,000 Da, about 70,000 Da,
about 65,000 Da, about 60,000 Da, about 55,000 Da, about 50,000 Da,
about 45,000 Da, about 40,000 Da, about 35,000 Da, about 30,000 Da,
about 25,000 Da, about 20,000 Da, about 15,000 Da, about 10,000 Da,
about 9,000 Da, about 8,000 Da, about 7,000 Da, about 6,000 Da,
about 5,000 Da, about 4,000 Da, about 3,000 Da, about 2,000 Da, and
about 1,000 Da. In some embodiments, the molecular weight of the
branched chain PEG is between about 1,000 Da and about 50,000 Da.
In some embodiments, the molecular weight of the branched chain PEG
is between about 1,000 Da and about 40,000 Da. In some embodiments,
the molecular weight of the branched chain PEG is between about
5,000 Da and about 40,000 Da. In some embodiments, the molecular
weight of the branched chain PEG is between about 5,000 Da and
about 20,000 Da. The foregoing list for substantially water soluble
backbones is by no means exhaustive and is merely illustrative, and
in some embodiments, the polymeric materials having the qualities
described above suitable for use in methods and compositions
described herein.
[0592] In further embodiments, the number of water soluble polymers
linked to a Kinase Inhibitor moiety and a reporter moiety described
herein is adjusted to provide an altered (including but not limited
to, increased or decreased) pharmacologic, pharmacokinetic or
pharmacodynamic characteristic such as in vivo half-life. In some
embodiments, the half-life of the Activity Probe is increased at
least about 10, about 20, about 30, about 40, about 50, about 60,
about 70, about 80, about 90 percent, about two fold, about
five-fold, about 10-fold, about 50-fold, or at least about 100-fold
over a Activity Probe without a water soluble linker.
[0593] In another embodiment, X is selected from the group
consisting of: a bond, --O(C.dbd.O)--, --NR.sup.a(C.dbd.O)--,
--NR.sup.a,
##STR00085##
--O--, --S--, --S--S--, --O--NR.sup.a--, --O(C.dbd.O)O--,
--O(C.dbd.O)NR.sup.a, --NR.sup.a(C.dbd.O)NR.sup.a--,
--N.dbd.CR.sup.a--, --S(C.dbd.O)--, --S(O)--, and --S(O).sub.2--;
wherein
##STR00086##
forms a N-containing heterocycle. In one embodiment, X is
NR.sup.a(C.dbd.O). In another embodiment, X is a bond. In another
embodiment, X is --O(C.dbd.O)--. In a further embodiment, Y is
selected from the group consisting of: a bond, --O(C.dbd.O)--,
--NR.sup.a(C.dbd.O)--, --NR.sup.a--,
##STR00087##
--O--, --S--, --S--S--, --O--NR.sup.a--, --O(C.dbd.O)O--,
--O(C.dbd.O)NR.sup.a, --NR.sup.a(C.dbd.O)NR.sup.a--,
--N.dbd.CR.sup.a--, --S(C.dbd.O)--, --S(O)--, and --S(O).sub.2--;
wherein
##STR00088##
forms a N-containing heterocycle. In yet a further embodiment, Y is
a bond. In one embodiment, Y is --NR.sup.a(C.dbd.O)--. In yet
another embodiment, R.sup.a is hydrogen. In yet a further
embodiment, R.sup.a is alkyl.
[0594] In a further embodiment, the reporter moiety is selected
from the group consisting of a label, a dye, a photocrosslinker, a
cytotoxic compound, a drug, an affinity label, a photoaffinity
label, a reactive compound, an antibody or antibody fragment, a
biomaterial, a nanoparticle, a spin label, a fluorophore, a
metal-containing moiety, a radioactive moiety, a novel functional
group, a group that covalently or noncovalently interacts with
other molecules, a photocaged moiety, an actinic radiation
excitable moiety, a ligand, a photoisomerizable moiety, biotin, a
biotin analog, a moiety incorporating a heavy atom, a chemically
cleavable group, a photocleavable group, a redox-active agent, an
isotopically labeled moiety, a biophysical probe, a phosphorescent
group, a chemiluminescent group, an electron dense group, a
magnetic group, an intercalating group, a chromophore, an energy
transfer agent, a biologically active agent, a detectable label, or
a combination thereof.
[0595] In another embodiment, the reporter moiety is a fluorophore.
In a further embodiment, the fluorophore is selected from the group
consisting of: BODIPY 493/503, BODIPY FL, BODIPY R6G, BODIPY
530/550, BODIPY TMR, BODIPY 558/568, BODIPY 564/570, BODIPY
576/589, BODIPY 581/591, BODIPY TR, Fluorescein,
5(6)-Carboxyfluorescein, 2,7-Dichlorofluorescein,
N,N-Bis(2,4,6-trimethylphenyl)-3,4:9,10-perylenebis(dicarboximide,
HPTS, Ethyl Eosin, DY-490XL MegaStokes, DY-485XL MegaStokes,
Adirondack Green 520, ATTO 465, ATTO 488, ATTO 495, YOYO-1, 5-FAM,
BCECF, BCECF, dichlorofluorescein, rhodamine 110, rhodamine 123,
Rhodamine Green, YO-PRO-1, SYTOX Green, Sodium Green, SYBR Green I,
Alexa Fluor 500, FITC, Fluo-3, Fluo-4, fluoro-emerald, YoYo-1
ssDNA, YoYo-1 dsDNA, YoYo-1, SYTO RNASelect, Diversa Green-FP,
Dragon Green, EvaGreen, Surf Green EX, Spectrum Green, Oregon Green
488, NeuroTrace 500525, NBD-X, MitoTracker Green FM, LysoTracker
Green DND-26, CBQCA, PA-GFP (post-activation), WEGFP
(post-activation), FlASH-CCXXCC, Azami Green monomeric, Azami
Green, EGFP (Campbell Tsien 2003), EGFP (Patterson 2001),
Fluorescein, Kaede Green,
7-Benzylamino-4-Nitrobenz-2-Oxa-1,3-Diazole, Bexl, Doxorubicin,
Lumio Green, and SuperGlo GFP.
[0596] In a further embodiment, the fluorophore is selected from
the group consisting of: BODIPY 493/503, BODIPY FL, BODIPY R6G,
BODIPY 530/550, BODIPY TMR, BODIPY 558/568, BODIPY 564/570, BODIPY
576/589, BODIPY 581/591, and BODIPY TR. In yet a further
embodiment, the fluorophore is BODIPY FL. In certain embodiments,
the fluorophore is not BODIPY 530. In some embodiments, the
fluorophore has an excitation maxima of between about 500 and about
600 nm. In some other embodiments, the fluorophore has an
excitation maxima of between about 500 and about 550 nm. In another
embodiments, the fluorophore has an excitation maxima of between
about 550 and about 600 nm. In yet a further embodiment, the
fluorophore has an excitation maxima of between about 525 and about
575 nm. In other embodiments, the fluorophore has an emission
maxima of between about 510 and about 670 nm. In another
embodiment, the fluorophore has an emission maxima of between about
510 and about 600 nm. In a further embodiment, the fluorophore has
an emission maxima of between about 600 and about 670 nm. In
another embodiment, the fluorophore has an emission maxima of
between about 575 and about 625 nm.
[0597] By way of example only and in some embodiments, the observed
potency, selectivity, and cell permeability of compounds such as
Compound 2 are appropriate to incorporate these molecules into a
Kinase-targeted, activity based probe that allows direct
visualization of Kinase activity in intact cells. In vitro
profiling against a panel of greater than 100 kinases showed
Compound 2 to be a highly potent and selective inhibitor of Tec
family kinases, including, Btk, as well as Src family kinases.
Without limiting the scope of the compositions and methods
described herein, it is postulated that the structural basis for
the selectivity is covalent modification of a non-catalytic
cysteine residue (Cys 481 in Btk) that is conserved in the ATP
binding pocket of the Tec family and several other kinases.
[0598] However, in other embodiments, any irreversible Kinase
Inhibitor that binds to the non-catalytic cysteine residue in the
ATP binding pocket of a Kinase is used in the compounds and methods
described herein.
General Synthesis and Characterization of an Illustrative Activity
Probe
[0599] Without limiting the scope of the compositions described
herein, an illustrative probe was synthesized by attaching a bodipy
FL fluorophore to an irreversible inhibitor via a piperazine
linker. The piperazine linker served to maintain probe solubility
and provided spatial separation between the fluorophore and the
pyrazolopyrimidine core.
##STR00089##
[0600] In some embodiments, the linkage formed is a stable linkage.
In other embodiments, in the case where the conjugate comprises two
components, the linker moiety forms a linkage, in some embodiments,
a stable linkage, between the Kinase Inhibitor moiety and the
reporter moiety. In some embodiments, the linker moiety is stable
and provides the means to control and determine the distance
between the Kinase Inhibitor moiety and the report moiety. Further,
in some embodiments, the linker moiety is selected such that the
probe's solubility is maintained. In some embodiments, the linker
moiety is a piperazinyl moiety. In further embodiments, a
piperazinyl-based linkage is formed by using a piperazine
containing compound. In other embodiments, the number and order of
units that comprise the linker moiety is selected such that the
length between the first component and the second component, as
well as the hydrophobic and hydrophilic characteristics of the
linker is controlled.
[0601] In the present context, spatial separation means a
thermochemically and photochemically non-active distance-making
group and in some embodiments is used to join two or more different
moieties of the types defined above. In other embodiments, spacers
are selected on the basis of a variety of characteristics including
their hydrophobicity, hydrophilicity, molecular flexibility and
length. The spacer, thus, in some embodiments, comprises a chain of
carbon atoms optionally interrupted or terminated with one or more
heteroatoms, such as oxygen atoms, nitrogen atoms, and/or sulphur
atoms. Thus, in some embodiments, the spacer comprises one or more
amide, ester, amino, ether, and/or thioether functionalities, and
optionally aromatic or mono/polyunsaturated hydrocarbons,
polyoxyethylene such as polyethylene glycol, oligo/polyamides such
as poly-.alpha.-alanine, polyglycine, polylysine, and peptides in
general, oligosaccharides, oligo/polyphosphates. Moreover, in other
embodiments, the spacer consists of combined units thereof. In
further embodiments, the length of the spacer varies, taking into
consideration the desired or necessary positioning and spatial
orientation of the active/functional part of the Activity
Probe.
[0602] Without limiting the scope of the compositions described
herein, in some embodiments the reporter moiety is Bodipy. In the
present context, the term reporter moiety means a group which is
detectable either by itself or as a part of a detection series.
[0603] In some embodiments, the labeled Activity Probes described
herein are purified by one or more procedures including, but are
not limited to, affinity chromatography; anion- or cation-exchange
chromatography (using, including but not limited to, DEAE
SEPHAROSE); chromatography on silica; reverse phase HPLC; gel
filtration (using, including but not limited to, SEPHADEX G-75);
hydrophobic interaction chromatography; size-exclusion
chromatography, metal-chelate chromatography;
ultrafiltration/diafiltration; ethanol precipitation; ammonium
sulfate precipitation; chromatofocusing; displacement
chromatography; electrophoretic procedures (including but not
limited to preparative isoelectric focusing), differential
solubility (including but not limited to ammonium sulfate
precipitation), or extraction. In other embodiments, apparent
molecular weight is estimated by GPC by comparison to globular
protein standards (PROTEIN PURIFICATION METHODS, A PRACTICAL
APPROACH (Harris & Angal, Eds.) IRL Press 1989, 293-306).
[0604] In one aspect, the in vitro inhibitory potency of a probe
against a panel of selected Kinases as a rapid means of confirming
accessibility of the reactive moiety to the Kinase active site is
tested. By way of example only, although less potent than the
parent Compound 2, the illustrative probe of Compound 3 retains
potency against Btk (IC.sub.50.about.90 nM). Thus, the piperazine
linker and bodipy fluorophore do not seriously compromise
accessibility of the illustrative probe to the enzyme active
site.
[0605] The Activity Probes described herein label kinases at the
non-catalytic Cys 481 (or a homologous cysteine) and that in some
embodiments, probe labeling does not require the catalytic
machinery per se. As such it differs from canonical activity-based
probes that target the enzyme catalytic machinery directly. In some
embodiments, the Kinase undergoes a phosphorylation dependent
conformational change that is tightly coupled to ATP binding and
kinase activation. In some embodiments, effective labeling by a
probe requires the Kinase to be in its active conformation in order
to directly detect Kinase activity in cells. In other embodiments,
effective labeling by an Activity Probe does not require the Kinase
to be in its active conformation in order to directly detect Kinase
activity in cells.
Therapeutic Uses of Irreversible Inhibitor Compounds
[0606] Described herein are methods, compositions, uses and
medicaments for the treatment of disorders characterized by the
presence of a solid tumor comprising administering to an individual
in need an irreversible inhibitor of an ACK. In some embodiments,
the disorder is a sarcoma, lymphoma, and/or carcinoma. In some
embodiments, the disorder is mammary ductal carcinoma, lobular
carcinoma, an adenocarcinoma (e.g. pancreatic cancer and colon
cancer), small cell lung carcinoma, non-small cell lung carcinoma,
and melanomas. In some embodiments, the disorder is mammary ductal
carcinoma, lobular carcinoma, or a combination thereof. In some
embodiments, the disorder is pancreatic cancer.
[0607] In some embodiments, the ACK is Btk or a Btk homolog. In yet
further embodiments, the ACK is tyrosine kinases that share
homology with Btk by having a cysteine residue (including a Cys 481
residue) that forms a covalent bond with the irreversible
inhibitor. See, e.g., protein kinases in FIG. 7. In some
embodiments, the ACK is HER4.
[0608] The methods described herein (which includes uses of a
pharmaceutical composition to treat a disorder, or uses of a
compound to form a medicament for treating a disorder) include
administering to an individual in need thereof a composition
containing a therapeutically effective amount of one or more
irreversible Btk inhibitor compounds described herein. In some
embodiments, the individual has been diagnosed with or is
predisposed to develop a sarcoma, lymphoma, and/or carcinoma. In
some embodiments, the individual has been diagnosed with or is
predisposed to develop mammary ductal carcinoma, lobular carcinoma,
an adenocarcinoma (e.g. pancreatic cancer and colon cancer), small
cell lung carcinoma, non-small cell lung carcinoma, and melanomas.
In some embodiments, the individual has been diagnosed with or is
predisposed to develop mammary ductal carcinoma, lobular carcinoma,
or a combination thereof. In some embodiments, the individual has
been diagnosed with or is predisposed to develop pancreatic
cancer.
[0609] Without being bound by theory, the diverse roles played by
Btk signaling in various hematopoietic cell functions show that
small molecule Btk inhibitors are useful for reducing the risk of
or treating a disorder characterized by the presence or development
of one or more solid tumors.
[0610] In some embodiments, are methods for treating a disorder
characterized by the presence of a solid tumor (e.g. lymphomas,
carcinomas, and/or sarcomas) comprising administering to an
individual in need a pharmaceutical formulation of any irreversible
inhibitor of Btk (or a Btk homolog) of Formula (A1-A6), Formula
(B1-B6), Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula
(VII). In some embodiments, the solid neoplasm is mammary ductal
carcinoma, lobular carcinoma, an adenocarcinoma (e.g. pancreatic
cancer and colon cancer), small cell lung carcinoma, non-small cell
lung carcinoma, and melanomas.
[0611] In still further embodiments are methods for treating a
disorder characterized by the presence of a solid tumor comprising
administering to an individual in need thereof a composition
containing a therapeutically effective amount of a compound that
forms a covalent bond with a cysteine sidechain of a Bruton's
tyrosine kinase or a Bruton's tyrosine kinase homolog. In some
embodiments, the individual has been diagnosed with or is
predisposed to develop a sarcoma, lymphoma, and/or carcinoma. In
some embodiments, the individual has been diagnosed with or is
predisposed to develop mammary ductal carcinoma, lobular carcinoma,
an adenocarcinoma (e.g. pancreatic cancer and colon cancer), small
cell lung carcinoma, non-small cell lung carcinoma, and melanomas.
In some embodiments, the individual has been diagnosed with or is
predisposed to develop mammary ductal carcinoma, lobular carcinoma,
or a combination thereof. In some embodiments, the individual has
been diagnosed with or is predisposed to develop pancreatic
cancer.
[0612] Further, in some embodiments, the irreversible Btk inhibitor
compounds described herein are used to inhibit a small subset of
other tyrosine kinases that share homology with Btk by having a
cysteine residue (including a Cys 481 residue) that is able to form
a covalent bond with the irreversible inhibitor. See, e.g., protein
kinases in FIG. 7. Thus, a subset of tyrosine kinases other than
Btk are also expected to be useful as therapeutic targets in a
number of health conditions, including lymphomas, carcinomas,
and/or sarcomas.
[0613] Symptoms, diagnostic tests, and prognostic tests for each of
the above-mentioned conditions include, e.g., Harrison's Principles
of Internal Medicine.COPYRGT.," 16th ed., 2004, The McGraw-Hill
Companies, Inc. Dey et al. (2006), Cytojournal 3(24), and the
"Revised European American Lymphoma" (REAL) classification system
(see, e.g., the website maintained by the National Cancer
Institute).
[0614] A number of animal models are useful for establishing a
range of therapeutically effective doses of irreversible
inhibitors, including irreversible Btk inhibitor compounds for
treating any of the foregoing diseases. For example, refer to
Examples 1-4 of the "Therapeutic Uses" section of the Examples
included herein. As an example, dosing of irreversible inhibitors
for the treatment of cancer can be examined in, e.g., a
human-to-mouse xenograft model in which human B-cell lymphoma cells
(e.g. Ramos cells) are implanted into immunodefficient mice (e.g.,
"nude" mice) as described in, e.g., Pagel et al. (2005), Clin
Cancer Res 11(13):4857-4866. Animal models for treatment of
thromboembolic disorders are also known.
[0615] In one embodiment, the therapeutic efficacy of the compound
for one of the foregoing diseases is optimized during a course of
treatment. For example, an individual being treated optionally
undergoes a diagnostic evaluation to correlate the relief of
disease symptoms or pathologies to inhibition of in vivo Btk
activity achieved by administering a given dose of an irreversible
Btk inhibitor. Cellular assays are used to determine in vivo
activity of Btk in the presence or absence of an irreversible Btk
inhibitor. For example, since activated Btk is phosphorylated at
tyrosine 223 (Y223) and tyrosine 551 (Y551), phospho-specific
immunocytochemical staining of P-Y223 or P-Y551-positive cells are
used to detect or quantify activation of Bkt in a population of
cells (e.g., by FACS analysis of stained vs unstained cells). See,
e.g., Nisitani et al. (1999), Proc. Natl. Acad. Sci, USA
96:2221-2226. Thus, the amount of the Btk inhibitor compound that
is administered to an individual is optionally increased or
decreased as needed so as to maintain a level of Btk inhibition
optimal for treating the subject's disease state.
[0616] In one embodiment are methods for identifying biomarkers
suitable for determining patient response to an irreversible ACK
inhibitor (including, e.g., a compound of Formula (I)) comprising
administering to a test subject a composition containing an amount
of the irreversible ACK inhibitor (including, e.g., a compound of
Formula (I)) sufficient to inhibit B cell receptor signaling and
correlating B cell receptor signaling with apoptosis. In another or
further embodiment are methods for selecting an individual for
treatment for lymphoma with an irreversible ACK inhibitor
(including, e.g., a compound of Formula (I)) comprising measuring
pErk or Erk transcriptional target levels in an individual sample,
and correlating a high level of transcriptional targets with a
positive response to the treatment. In another or further
embodiments are methods for measuring an individual's response to
treatment comprising administering to the patient an irreversible
ACK inhibitor (including, e.g., a compound of Formula (I)),
measuring pErk or Erk transcriptional target levels in an
individual sample, and correlating a reduced level of
transcriptional targets with a positive response to the
administration of the irreversible ACK inhibitor (including, e.g.,
a compound of Formula (I)).
Combination Treatments
[0617] In some embodiments, the irreversible Btk inhibitor
compositions described herein are used in combination with other
well known therapeutic reagents that are selected for their
therapeutic value for the condition to be treated. In general, the
compositions described herein and, in embodiments where
combinational therapy is employed, other agents do not have to be
administered in the same pharmaceutical composition, and are
optionally, because of different physical and chemical
characteristics, have to be administered by different routes. The
initial administration is made, for example, according to
established protocols, and then, based upon the observed effects,
the dosage, modes of administration and times of administration are
modified.
[0618] In certain instances, it is appropriate to administer at
least one irreversible Btk inhibitor compound described herein in
combination with another therapeutic agent. By way of example only,
if one of the side effects experienced by an individual upon
receiving one of the irreversible Btk inhibitor compounds described
herein is nausea, then it is appropriate to administer an
anti-nausea agent in combination with the initial therapeutic
agent. Or, by way of example only, the therapeutic effectiveness of
one of the compounds described herein is enhanced by administration
of an adjuvant (i.e., by itself the adjuvant has minimal
therapeutic benefit, but in combination with another therapeutic
agent, the overall therapeutic benefit to the patient is enhanced).
Or, by way of example only, the benefit experienced by an
individual is increased by administering one of the compounds
described herein with another therapeutic agent (which also
includes a therapeutic regimen) that also has therapeutic benefit.
In any case, regardless of the disease, disorder being treated, the
overall benefit experienced by the patient is in some embodiments
simply additive of the two therapeutic agents or in other
embodiments, the patient experiences a synergistic benefit.
[0619] The particular choice of compounds used will depend upon the
diagnosis of the attending physicians and their judgment of the
condition of the patient and the appropriate treatment protocol.
The compounds are optionally administered concurrently (e.g.,
simultaneously, essentially simultaneously or within the same
treatment protocol) or sequentially, depending upon the nature of
the disorder, the condition of the patient, and the actual choice
of compounds used. The determination of the order of
administration, and the number of repetitions of administration of
each therapeutic agent during a treatment protocol, is based on an
evaluation of the disease being treated and the condition of the
patient.
[0620] In some embodiments, therapeutically-effective dosages vary
when the drugs are used in treatment combinations. Methods for
experimentally determining therapeutically-effective dosages of
drugs and other agents for use in combination treatment regimens
are described in the literature. For example, the use of metronomic
dosing, i.e., providing more frequent, lower doses in order to
minimize toxic side effects, has been described extensively in the
literature Combination treatment further includes periodic
treatments that start and stop at various times to assist with the
clinical management of the patient.
[0621] For combination therapies described herein, dosages of the
co-administered compounds will of course vary depending on the type
of co-drug employed, on the specific drug employed, on the disorder
being treated and so forth. In addition, when co-administered with
one or more biologically active agents, the compound provided
herein is administered either simultaneously with the biologically
active agent(s), or sequentially. If administered sequentially, the
attending physician will decide on the appropriate sequence of
administering protein in combination with the biologically active
agent(s).
[0622] In any case, the multiple therapeutic agents (one of which
is a compound of Formula (A1-A6), (B1-B6), (C1-C6), or (D1-D6)
described herein) are optionally administered in any order or even
simultaneously. If simultaneously, the multiple therapeutic agents
are optionally provided in a single, unified form, or in multiple
forms (by way of example only, either as a single pill or as two
separate pills). In some embodiments, one of the therapeutic agents
is given in multiple doses, or both are given as multiple doses. If
not simultaneous, the timing between the multiple doses is from
about more than zero weeks to less than about four weeks. In
addition, the combination methods, compositions and formulations
are not to be limited to the use of only two agents; the use of
multiple therapeutic combinations are also envisioned.
[0623] It is understood that the dosage regimen to treat, prevent,
or ameliorate the condition(s) for which relief is sought, can be
modified in accordance with a variety of factors. These factors
include the disorder from which the subject suffers, as well as the
age, weight, sex, diet, and medical condition of the subject. Thus,
the dosage regimen actually employed can vary widely and therefore
can deviate from the dosage regimens set forth herein.
[0624] In some embodiments, the pharmaceutical agents which make up
the combination therapy disclosed herein are administered in a
combined dosage form, or in separate dosage forms intended for
substantially simultaneous administration. In some embodiments, the
pharmaceutical agents that make up the combination therapy are
administered sequentially, with either therapeutic compound being
administered by a regimen calling for two-step administration. In
some embodiments, the two-step administration regimen calls for
sequential administration of the active agents or spaced-apart
administration of the separate active agents. The time period
between the multiple administration steps ranges from a few minutes
to several hours, depending upon the properties of each
pharmaceutical agent, such as potency, solubility, bioavailability,
plasma half-life and kinetic profile of the pharmaceutical agent.
In some embodiments, circadian variation of the target molecule
concentration determines the optimal dose interval.
[0625] In addition, the compounds described herein also are
optionally used in combination with procedures that provide
additional or synergistic benefit to the patient. By way of example
only, patients are expected to find therapeutic and/or prophylactic
benefit in the methods described herein, wherein pharmaceutical
composition of a compound disclosed herein and/or combinations with
other therapeutics are combined with genetic testing to determine
whether that individual is a carrier of a mutant gene that is known
to be correlated with certain diseases or conditions.
[0626] The compounds described herein and combination therapies are
administered before, during or after the occurrence of a disorder,
and the timing of administering the composition containing a
compound is variable. In some embodiments, the compounds are used
as a prophylactic and are administered continuously to subjects
with a propensity to develop conditions or diseases in order to
prevent the occurrence of the disorder. In some embodiments, the
compounds and compositions are administered to an individual during
or as soon as possible after the onset of the symptoms. In some
embodiments, the administration of the compounds is initiated
within the first 48 hours of the onset of the symptoms, within the
first 6 hours of the onset of the symptoms, or within 3 hours of
the onset of the symptoms. In some embodiments, the initial
administration is via any route practical, such as, for example, an
intravenous injection, a bolus injection, infusion over 5 minutes
to about 5 hours, a pill, a capsule, transdermal patch, buccal
delivery, and the like, or combination thereof. A compound should
be administered as soon as is practicable after the onset of a
disorder is detected or suspected, and for a length of time
necessary for the treatment of the disease, such as, for example,
from about 1 month to about 3 months. The length of treatment can
vary for each subject, and the length can be determined using the
known criteria. In some embodiments, the compound or a formulation
containing the compound is administered for at least 2 weeks,
between about 1 month to about 5 years, or from about 1 month to
about 3 years.
[0627] Exemplary Therapeutic Agents for Use in Combination with an
Irreversible Inhibitor Compound
[0628] In some embodiments, where the subject is suffering from or
at risk of suffering from a disorder characterized by the presence
or development of one or more solid tumors, the subjected is
treated with an irreversible Btk inhibitor compound in any
combination with one or more other anti-cancer agents. In some
embodiments, one or more of the anti-cancer agents are proapoptotic
agents. Examples of anti-cancer agents include, but are not limited
to, any of the following: gossyphol, genasense, polyphenol E,
Chlorofusin, all trans-retinoic acid (ATRA), bryostatin, tumor
necrosis factor-related apoptosis-inducing ligand (TRAIL),
5-aza-2'-deoxycytidine, all trans retinoic acid, doxorubicin,
vincristine, etoposide, gemcitabine, imatinib (Gleevec.RTM.),
geldanamycin, 17-N-Allylamino-17-Demethoxygeldanamycin (17-AAG),
flavopiridol, LY294002, bortezomib, trastuzumab, BAY 11-7082,
PKC412, or PD184352, Taxol.TM., also referred to as "paclitaxel",
which is a well-known anti-cancer drug which acts by enhancing and
stabilizing microtubule formation, and analogs of Taxol.TM., such
as Taxotere.TM.. Compounds that have the basic taxane skeleton as a
common structure feature, have also been shown to have the ability
to arrest cells in the G2-M phases due to stabilized microtubules
and, in some embodiments, are useful for treating cancer in
combination with the compounds described herein.
[0629] Further examples of anti-cancer agents for use in
combination with an irreversible Btk inhibitor compound include
inhibitors of mitogen-activated protein kinase signaling, e.g.,
U0126, PD98059, PD184352, PD0325901, ARRY-142886, SB239063,
SP600125, BAY 43-9006, wortmannin, or LY294002; Syk inhibitors;
mTOR inhibitors; and antibodies (e.g., rituxan).
[0630] Other anti-cancer agents for use in combination with an
irreversible Btk inhibitor compound include Adriamycin,
Dactinomycin, Bleomycin, Vinblastine, Cisplatin, acivicin;
aclarubicin; acodazole hydrochloride; acronine; adozelesin;
aldesleukin; altretamine; ambomycin; ametantrone acetate;
aminoglutethimide; amsacrine; anastrozole; anthramycin;
asparaginase; asperlin; azacitidine; azetepa; azotomycin;
batimastat; benzodepa; bicalutamide; bisantrene hydrochloride;
bisnafide dimesylate; bizelesin; bleomycin sulfate; brequinar
sodium; bropirimine; busulfan; cactinomycin; calusterone;
caracemide; carbetimer; carboplatin; carmustine; carubicin
hydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin;
cladribine; crisnatol mesylate; cyclophosphamide; cytarabine;
dacarbazine; daunorubicin hydrochloride; decitabine; dexormaplatin;
dezaguanine; dezaguanine mesylate; diaziquone; doxorubicin;
doxorubicin hydrochloride; droloxifene; droloxifene citrate;
dromostanolone propionate; duazomycin; edatrexate; eflornithine
hydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine;
epirubicin hydrochloride; erbulozole; esorubicin hydrochloride;
estramustine; estramustine phosphate sodium; etanidazole;
etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride;
fazarabine; fenretinide; floxuridine; fludarabine phosphate;
fluorouracil; flurocitabine; fosquidone; fostriecin sodium;
gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicin
hydrochloride; ifosfamide; iimofosine; interleukin II (including
recombinant interleukin II, or rlL2), interferon alfa-2a;
interferon alfa-2b; interferon alfa-n1; interferon alfa-n3;
interferon beta-1 a; interferon gamma-1 b; iproplatin; irinotecan
hydrochloride; lanreotide acetate; letrozole; leuprolide acetate;
liarozole hydrochloride; lometrexol sodium; lomustine; losoxantrone
hydrochloride; masoprocol; maytansine; mechlorethamine
hydrochloride; megestrol acetate; melengestrol acetate; melphalan;
menogaril; mercaptopurine; methotrexate; methotrexate sodium;
metoprine; meturedepa; mitindomide; mitocarcin; mitocromin;
mitogillin; mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone
hydrochloride; mycophenolic acid; nocodazoie; nogalamycin;
ormaplatin; oxisuran; pegaspargase; peliomycin; pentamustine;
peplomycin sulfate; perfosfamide; pipobroman; piposulfan;
piroxantrone hydrochloride; plicamycin; plomestane; porfimer
sodium; porfiromycin; prednimustine; procarbazine hydrochloride;
puromycin; puromycin hydrochloride; pyrazofurin; riboprine;
rogletimide; safingol; safingol hydrochloride; semustine;
simtrazene; sparfosate sodium; sparsomycin; spirogermanium
hydrochloride; spiromustine; spiroplatin; streptonigrin;
streptozocin; sulofenur; talisomycin; tecogalan sodium; tegafur;
teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;
testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;
tirapazamine; toremifene citrate; trestolone acetate; triciribine
phosphate; trimetrexate; trimetrexate glucuronate; triptorelin;
tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;
verteporfin; vinblastine sulfate; vincristine sulfate; vindesine;
vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;
vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;
vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicin
hydrochloride.
[0631] Other anti-cancer agents for use in combination with an
irreversible Btk inhibitor compound include: 20-epi-1, 25
dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin;
acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK
antagonists; altretamine; ambamustine; amidox; amifostine;
aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole;
andrographolide; angiogenesis inhibitors; antagonist D; antagonist
G; antarelix; anti-dorsalizing morphogenetic protein-1;
antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston;
antisense oligonucleotides; aphidicolin glycinate; apoptosis gene
modulators; apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA;
arginine deaminase; asulacrine; atamestane; atrimustine;
axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin;
azatyrosine; baccatin III derivatives; balanol; batimastat; BCR/ABL
antagonists; benzochlorins; benzoylstaurosporine; beta lactam
derivatives; beta-alethine; betaclamycin B; betulinic acid; bFGF
inhibitor; bicalutamide; bisantrene; bisaziridinylspermine;
bisnafide; bistratene A; bizelesin; breflate; bropirimine;
budotitane; buthionine sulfoximine; calcipotriol; calphostin C;
camptothecin derivatives; canarypox IL-2; capecitabine;
carboxamide-amino-triazole; carboxyamidotriazole; CaRest M3; CARN
700; cartilage derived inhibitor; carzelesin; casein kinase
inhibitors (ICOS); castanospermine; cecropin B; cetrorelix;
chlorlns; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin;
cladribine; clomifene analogues; clotrimazole; collismycin A;
collismycin B; combretastatin A4; combretastatin analogue;
conagenin; crambescidin 816; crisnatol; cryptophycin 8;
cryptophycin A derivatives; curacin A; cyclopentanthraquinones;
cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;
cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;
dexamethasone; dexifosfamide; dexrazoxane; dexverapamil;
diaziquone; didemnin B; didox; diethylnorspermine;
dihydro-5-azacytidine; 9-dioxamycin; diphenyl spiromustine;
docosanol; dolasetron; doxifluridine; droloxifene; dronabinol;
duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab;
eflornithine; elemene; emitefur; epirubicin; epristeride;
estramustine analogue; estrogen agonists; estrogen antagonists;
etanidazole; etoposide phosphate; exemestane; fadrozole;
fazarabine; fenretinide; filgrastim; finasteride; flavopiridol;
flezelastine; fluasterone; fludarabine; fluorodaunorunicin
hydrochloride; forfenimex; formestane; fostriecin; fotemustine;
gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;
gelatinase inhibitors; gemcitabine; glutathione inhibitors;
hepsulfam; heregulin; hexamethylene bisacetamide; hypericin;
ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine;
ilomastat; imidazoacridones; imiquimod; immunostimulant peptides;
insulin-like growth factor-1 receptor inhibitor; interferon
agonists; interferons; interleukins; iobenguane; iododoxorubicin;
ipomeanol, 4-; iroplact; irsogladine; isobengazole;
isohomohalicondrin B; itasetron; jasplakinolide; kahalalide F;
lamellarin-N triacetate; lanreotide; leinamycin; lenograstim;
lentinan sulfate; leptolstatin; letrozole; leukemia inhibiting
factor; leukocyte alpha interferon;
leuprolide+estrogen+progesterone; leuprorelin; levamisole;
liarozole; linear polyamine analogue; lipophilic disaccharide
peptide; lipophilic platinum compounds; lissoclinamide 7;
lobaplatin; lombricine; lometrexol; lonidamine; losoxantrone;
lovastatin; loxoribine; lurtotecan; lutetium texaphyrin;
lysofylline; lytic peptides; maitansine; mannostatin A; marimastat;
masoprocol; maspin; matrilysin inhibitors; matrix metalloproteinase
inhibitors; menogaril; merbarone; meterelin; methioninase;
metoclopramide; MIF inhibitor; mifepristone; miltefosine;
mirimostim; mismatched double stranded RNA; mitoguazone;
mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast
growth factor-saporin; mitoxantrone; mofarotene; molgramostim;
monoclonal antibody, human chorionic gonadotrophin; monophosphoryl
lipid A+myobacterium cell wall sk; mopidamol; multiple drug
resistance gene inhibitor; multiple tumor suppressor 1-based
therapy; mustard anticancer agent; mycaperoxide B; mycobacterial
cell wall extract; myriaporone; N-acetyldinaline; N-substituted
benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin;
naphterpin; nartograstim; nedaplatin; nemorubicin; neridronic acid;
neutral endopeptidase; nilutamide; nisamycin; nitric oxide
modulators; nitroxide antioxidant; nitrullyn; O6-benzylguanine;
octreotide; okicenone; oligonucleotides; onapristone; ondansetron;
ondansetron; oracin; oral cytokine inducer; ormaplatin; osaterone;
oxaliplatin; oxaunomycin; palauamine; palmitoylrhizoxin; pamidronic
acid; panaxytriol; panomifene; parabactin; pazelliptine;
pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin;
pentrozole; perflubron; perfosfamide; perillyl alcohol;
phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil;
pilocarpine hydrochloride; pirarubicin; piritrexim; placetin A;
placetin B; plasminogen activator inhibitor; platinum complex;
platinum compounds; platinum-triamine complex; porfimer sodium;
porfiromycin; prednisone; propyl bis-acridone; prostaglandin J2;
proteasome inhibitors; protein A-based immune modulator; protein
kinase C inhibitor; protein kinase C inhibitors, microalgal;
protein tyrosine phosphatase inhibitors; purine nucleoside
phosphorylase inhibitors; purpurins; pyrazoloacridine;
pyridoxylated hemoglobin polyoxyethylerie conjugate; raf
antagonists; raltitrexed; ramosetron; ras farnesyl protein
transferase inhibitors; ras inhibitors; ras-GAP inhibitor;
retelliptine demethylated; rhenium Re 186 etidronate; rhizoxin;
ribozymes; RII retinamide; rogletimide; rohitukine; romurtide;
roquinimex; rubiginone B1; ruboxyl; safingol; saintopin; SarCNU;
sarcophytol A; sargramostim; Sdi 1 mimetics; semustine; senescence
derived inhibitor 1; sense oligonucleotides; signal transduction
inhibitors; signal transduction modulators; single chain
antigen-binding protein; sizofiran; sobuzoxane; sodium borocaptate;
sodium phenylacetate; solverol; somatomedin binding protein;
sonermin; sparfosic acid; spicamycin D; spiromustine; splenopentin;
spongistatin 1; squalamine; stem cell inhibitor; stem-cell division
inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;
superactive vasoactive intestinal peptide antagonist; suradista;
suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;
tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;
tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;
temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;
thaliblastine; thiocoraline; thrombopoietin; thrombopoietin
mimetic; thymalfasin; thymopoietin receptor agonist; thymotrinan;
thyroid stimulating hormone; tin ethyl etiopurpurin; tirapazamine;
titanocene bichloride; topsentin; toremifene; totipotent stem cell
factor; translation inhibitors; tretinoin; triacetyluridine;
triciribine; trimetrexate; triptorelin; tropisetron; turosteride;
tyrosine kinase inhibitors; tyrphostins; UBC inhibitors; ubenimex;
urogenital sinus-derived growth inhibitory factor; urokinase
receptor antagonists; vapreotide; variolin B; vector system,
erythrocyte gene therapy; velaresol; veramine; verdins;
verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole;
zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer.
[0632] Yet other anticancer agents for use in combination with an
irreversible Btk inhibitor compound include alkylating agents,
antimetabolites, natural products, or hormones, e.g., nitrogen
mustards (e.g., mechloroethamine, cyclophosphamide, chlorambucil,
etc.), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g.,
carmustine, lomusitne, etc.), or triazenes (decarbazine, etc.).
Examples of antimetabolites include but are not limited to folic
acid analog (e.g., methotrexate), or pyrimidine analogs (e.g.,
Cytarabine), purine analogs (e.g., mercaptopurine, thioguanine,
pentostatin).
[0633] Examples of natural products useful in combination with an
irreversible Btk inhibitor compound include but are not limited to
vinca alkaloids (e.g., vinblastin, vincristine),
epipodophyllotoxins (e.g., etoposide), antibiotics (e.g.,
daunorubicin, doxorubicin, bleomycin), enzymes (e.g.,
L-asparaginase), or biological response modifiers (e.g., interferon
alpha).
[0634] Examples of alkylating agents for use employed in
combination an irreversible Btk inhibitor compound include, but are
not limited to, nitrogen mustards (e.g., mechloroethamine,
cyclophosphamide, chlorambucil, meiphalan, etc.), ethylenimine and
methylmelamines (e.g., hexamethlymelamine, thiotepa), alkyl
sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine,
lomusitne, semustine, streptozocin, etc.), or triazenes
(decarbazine, etc.). Examples of antimetabolites include, but are
not limited to folic acid analog (e.g., methotrexate), or
pyrimidine analogs (e.g., fluorouracil, floxouridine, Cytarabine),
purine analogs (e.g., mercaptopurine, thioguanine, pentostatin.
[0635] Examples of hormones and antagonists useful in combination
with an irreversible Btk inhibitor compound include, but are not
limited to, adrenocorticosteroids (e.g., prednisone), progestins
(e.g., hydroxyprogesterone caproate, megestrol acetate,
medroxyprogesterone acetate), estrogens (e.g., diethlystilbestrol,
ethinyl estradiol), antiestrogen (e.g., tamoxifen), androgens
(e.g., testosterone propionate, fluoxymesterone), antiandrogen
(e.g., flutamide), gonadotropin releasing hormone analog (e.g.,
leuprolide). Other agents for use in the methods and compositions
described herein for the treatment or prevention of cancer include
platinum coordination complexes (e.g., cisplatin, carboblatin),
anthracenedione (e.g., mitoxantrone), substituted urea (e.g.,
hydroxyurea), methyl hydrazine derivative (e.g., procarbazine),
adrenocortical suppressant (e.g., mitotane, aminoglutethimide).
[0636] Examples of anti-cancer agents which act by arresting cells
in the G2-M phases due to stabilized microtubules and which can be
used in combination with an irreversible Btk inhibitor compound
include without limitation marketed drugs and drugs in
development.
[0637] Where the subject is suffering from or at risk of suffering
from a thromboembolic disorder (e.g., stroke), in some embodiments,
the individual is treated with an irreversible Btk inhibitor
compound in any combination with one or more other
anti-thromboembolic agents. Examples of anti-thromboembolic agents
include, but are not limited any of the following: thrombolytic
agents (e.g., alteplase anistreplase, streptokinase, urokinase, or
tissue plasminogen activator), heparin, tinzaparin, warfarin,
dabigatran (e.g., dabigatran etexilate), factor Xa inhibitors
(e.g., fondaparinux, draparinux, rivaroxaban, DX-9065a, otamixaban,
LY517717, or YM150), factor VIIa inhibitors, ticlopidine,
clopidogrel, CS-747 (prasugrel, LY640315), ximelagatran, or BIBR
1048.
Pharmaceutical Composition/Formulation
[0638] Pharmaceutical compositions are formulated in a conventional
manner using one or more physiologically acceptable carriers
including excipients and auxiliaries which facilitate processing of
the active compounds into preparations which can be used
pharmaceutically. Proper formulation is dependent upon the route of
administration chosen. A summary of pharmaceutical compositions
described herein is found, for example, in Remington: The Science
and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack
Publishing Company, 1995); Hoover, John E., Remington's
Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1975;
Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms,
Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage
Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams
& Wilkins 1999).
[0639] A pharmaceutical composition, as used herein, refers to a
mixture of a compound described herein, such as, for example,
compounds of any of Formula (A1-A6), Formula (B1-B6), Formula
(C1-C6), Formula (D1-D6), Formula (I), or Formula (VII), with other
chemical components, such as carriers, stabilizers, diluents,
dispersing agents, suspending agents, thickening agents, and/or
excipients. The pharmaceutical composition facilitates
administration of the compound to an organism. In practicing the
methods of treatment or use provided herein, therapeutically
effective amounts of compounds described herein are administered in
a pharmaceutical composition to a mammal having a disorder to be
treated. Preferably, the mammal is a human. The compounds can be
used singly or in combination with one or more therapeutic agents
as components of mixtures.
[0640] The pharmaceutical formulations described herein are
administered to an individual by any suitable administration route,
including but not limited to, oral, parenteral (e.g., intravenous,
subcutaneous, intramuscular), intranasal, buccal, topical, rectal,
or transdermal administration routes. The pharmaceutical
formulations described herein include, but are not limited to,
aqueous liquid dispersions, self-emulsifying dispersions, solid
solutions, liposomal dispersions, aerosols, solid dosage forms,
powders, immediate release formulations, controlled release
formulations, fast melt formulations, tablets, capsules, pills,
delayed release formulations, extended release formulations,
pulsatile release formulations, multiparticulate formulations, and
mixed immediate and controlled release formulations.
[0641] Pharmaceutical compositions including a compound described
herein are optionally manufactured in a conventional manner, such
as, by way of example only, by means of conventional mixing,
dissolving, granulating, dragee-making, levigating, emulsifying,
encapsulating, entrapping or compression processes.
[0642] The pharmaceutical compositions will include at least one
compound described herein, such as, for example, a compound of any
of Formula (A1-A6), Formula (B1-B6), Formula (C1-C6), Formula
(D1-D6), Formula (I), or Formula (VII), as an active ingredient in
free-acid or free-base form, or in a pharmaceutically acceptable
salt form. In addition, the methods and pharmaceutical compositions
described herein include the use of N-oxides, crystalline forms
(also known as polymorphs), as well as active metabolites of these
compounds having the same type of activity. In some situations,
compounds exist as tautomers. All tautomers are included within the
scope of the compounds presented herein. Additionally, in some
embodiments, the compounds described herein exist in unsolvated as
well as solvated forms with pharmaceutically acceptable solvents
such as water, ethanol, and the like. The solvated forms of the
compounds presented herein are also considered to be disclosed
herein.
[0643] A "carrier" or "carrier materials" includes excipients in
pharmaceutics and is selected on the basis of compatibility with
compounds disclosed herein, such as, compounds of any of Formula
(A1-A6), Formula (B1-B6), Formula (C1-C6), Formula (D1-D6), Formula
(I), or Formula (VII), and the release profile properties of the
desired dosage form. Exemplary carrier materials include, e.g.,
binders, suspending agents, disintegration agents, filling agents,
surfactants, solubilizers, stabilizers, lubricants, wetting agents,
diluents, and the like. See, e.g., Remington: The Science and
Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing
Company, 1995); Hoover, John E., Remington's Pharmaceutical
Sciences, Mack Publishing Co., Easton, Pa. 1975; Liberman, H. A.
and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker,
New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug
Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins
1999).
[0644] A "measurable serum concentration" or "measurable plasma
concentration" describes the blood serum or blood plasma
concentration, typically measured in mg, .mu.g, or ng of
therapeutic agent per ml, dl, or l of blood serum, absorbed into
the bloodstream after administration. As used herein, measurable
plasma concentrations are typically measured in ng/ml or
.mu.g/ml.
[0645] "Pharmacodynamics" refers to the factors which determine the
biologic response observed relative to the concentration of drug at
a site of action. "Pharmacokinetics" refers to the factors which
determine the attainment and maintenance of the appropriate
concentration of drug at a site of action.
[0646] "Steady state," as used herein, is when the amount of drug
administered is equal to the amount of drug eliminated within one
dosing interval resulting in a plateau or constant plasma drug
exposure.
[0647] Dosage Forms
[0648] Moreover, the pharmaceutical compositions described herein,
which include a compound of any of Formula (A1-A6), Formula
(B1-B6), Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula
(VII) are formulated into any suitable dosage form, including but
not limited to, aqueous oral dispersions, liquids, gels, syrups,
elixirs, slurries, suspensions and the like, for oral ingestion by
an individual to be treated, solid oral dosage forms, aerosols,
controlled release formulations, fast melt formulations,
effervescent formulations, lyophilized formulations, tablets,
powders, pills, dragees, capsules, delayed release formulations,
extended release formulations, pulsatile release formulations,
multiparticulate formulations, and mixed immediate release and
controlled release formulations.
[0649] The pharmaceutical solid dosage forms described herein
optionally include a compound described herein and one or more
pharmaceutically acceptable additives such as a compatible carrier,
binder, filling agent, suspending agent, flavoring agent,
sweetening agent, disintegrating agent, dispersing agent,
surfactant, lubricant, colorant, diluent, solubilizer, moistening
agent, plasticizer, stabilizer, penetration enhancer, wetting
agent, anti-foaming agent, antioxidant, preservative, or one or
more combination thereof. In still other aspects, using standard
coating procedures, such as those described in Remington's
Pharmaceutical Sciences, 20th Edition (2000), a film coating is
provided around the formulation of the compound of any of Formula
(A1-A6), Formula (B1-B6), Formula (C1-C6), Formula (D1-D6), Formula
(I), or Formula (VII). In one embodiment, some or all of the
particles of the compound of any of Formula (A1-A6), Formula
(B1-B6), Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula
(VII), are coated. In another embodiment, some or all of the
particles of the compound of any of Formula (A1-A6), Formula
(B1-B6), Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula
(VII), are microencapsulated. In still another embodiment, the
particles of the compound of any of Formula (A1-A6), Formula
(B1-B6), Formula (C1-C6), Formula (D1-D6), Formula (I), or Formula
(VII), are not microencapsulated and are uncoated.
[0650] Examples of Methods of Dosing and Treatment Regimens
[0651] In some embodiments, the compounds described herein are used
in the preparation of medicaments for the inhibition of Btk or a
homolog thereof, or for the treatment of diseases or conditions
that benefit, at least in part, from inhibition of Btk or a homolog
thereof. In some embodiments, the compounds described herein are
used in the preparation of medicaments for the inhibition of HER4
or a homolog thereof, or for the treatment of diseases or
conditions that benefit, at least in part, from inhibition of HER4
or a homolog thereof. In addition, a method for treating any of the
diseases or conditions described herein in an individual in need of
such treatment, involves administration of pharmaceutical
compositions containing at least one compound of any of Formula
(A1-A6), Formula (B1-B6), Formula (C1-C6), Formula (D1-D6), Formula
(I), or Formula (VII), described herein, or a pharmaceutically
acceptable salt, pharmaceutically acceptable N-oxide,
pharmaceutically active metabolite, pharmaceutically acceptable
prodrug, or pharmaceutically acceptable solvate thereof, in
therapeutically effective amounts to said subject.
[0652] In some embodiments, the compositions containing the
compound(s) described herein are administered for prophylactic
and/or therapeutic treatments. In therapeutic applications, the
compositions are administered to an individual already suffering
from a disorder, in an amount sufficient to cure or at least
partially arrest the symptoms of the disorder. Amounts effective
for this use will depend on the severity and course of the
disorder, previous therapy, the patient's health status, weight,
and response to the drugs, and the judgment of the treating
physician.
[0653] In prophylactic applications, compositions containing the
compounds described herein are administered to an individual
susceptible to or otherwise at risk of a particular disease,
disorder. Such an amount is defined to be a "prophylactically
effective amount or dose." In this use, the precise amounts also
depend on the patient's state of health, weight, and the like. When
used in an individual, effective amounts for this use will depend
on the severity and course of the disease, disorder, previous
therapy, the patient's health status and response to the drugs, and
the judgment of the treating physician.
[0654] In some embodiments, the irreversible kinase inhibitor is
administered to the patient on a regular basis, e.g., three times a
day, two times a day, once a day, every other day or every 3 days.
In other embodiments, the irreversible kinase inhibitor is
administered to the patient on an intermittent basis, e.g., twice a
day followed by once a day followed by three times a day; or the
first two days of every week; or the first, second and third day of
a week. In some embodiments, intermittent dosing is as effective as
regular dosing. In further or alternative embodiments, the
irreversible kinase inhibitor is administered only when the patient
exhibits a particular symptom, e.g., the onset of pain, or the
onset of a fever, or the onset of an inflammation, or the onset of
a skin disorder.
[0655] In the case wherein the patient's condition does not
improve, upon the doctor's discretion the administration of the
compounds may be administered chronically, that is, for an extended
period of time, including throughout the duration of the patient's
life in order to ameliorate or otherwise control or limit the
symptoms of the patient's disorder.
[0656] In the case wherein the patient's status does improve, upon
the doctor's discretion the administration of the compounds may be
given continuously; alternatively, the dose of drug being
administered may be temporarily reduced or temporarily suspended
for a certain length of time (i.e., a "drug holiday"). The length
of the drug holiday can vary between 2 days and 1 year, including
by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7
days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50
days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days,
250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. The
dose reduction during a drug holiday may be from 10%-100%,
including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%,
40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95%, or
100%.
[0657] Once improvement of the patient's conditions has occurred, a
maintenance dose is administered if necessary. Subsequently, the
dosage or the frequency of administration, or both, can be reduced,
as a function of the symptoms, to a level at which the improved
disease, disorder is retained. Patients can, however, require
intermittent treatment on a long-term basis upon any recurrence of
symptoms.
[0658] The amount of a given agent that will correspond to such an
amount will vary depending upon factors such as the particular
compound, disorder and its severity, the identity (e.g., weight) of
the subject or host in need of treatment, and is determined
according to the particular circumstances surrounding the case,
including, e.g., the specific agent being administered, the route
of administration, the condition being treated, and the subject or
host being treated. In general, however, doses employed for adult
human treatment will typically be in the range of 0.02-5000 mg per
day, or from about 1-1500 mg per day. The desired dose may
conveniently be presented in a single dose or as divided doses
administered simultaneously (or over a short period of time) or at
appropriate intervals, for example as two, three, four or more
sub-doses per day.
[0659] The pharmaceutical composition described herein may be in
unit dosage forms suitable for single administration of precise
dosages. In unit dosage form, the formulation is divided into unit
doses containing appropriate quantities of one or more compound.
The unit dosage may be in the form of a package containing discrete
quantities of the formulation. Non-limiting examples are packaged
tablets or capsules, and powders in vials or ampoules. Aqueous
suspension compositions can be packaged in single-dose
non-reclosable containers. Alternatively, multiple-dose reclosable
containers can be used, in which case it is typical to include a
preservative in the composition. By way of example only,
formulations for parenteral injection may be presented in unit
dosage form, which include, but are not limited to ampoules, or in
multi-dose containers, with an added preservative.
[0660] The foregoing ranges are merely suggestive, as the number of
variables in regard to an individual treatment regime is large, and
considerable excursions from these recommended values are not
uncommon. Such dosages may be altered depending on a number of
variables, not limited to the activity of the compound used, the
disorder to be treated, the mode of administration, the
requirements of the individual subject, the severity of the
disorder being treated, and the judgment of the practitioner.
[0661] Toxicity and therapeutic efficacy of such therapeutic
regimens can be determined by standard pharmaceutical procedures in
cell cultures or experimental animals, including, but not limited
to, the determination of the LD.sub.50 (the dose lethal to 50% of
the population) and the ED.sub.50 (the dose therapeutically
effective in 50% of the population). The dose ratio between the
toxic and therapeutic effects is the therapeutic index and it can
be expressed as the ratio between LD.sub.50 and ED.sub.50.
Compounds exhibiting high therapeutic indices are preferred. The
data obtained from cell culture assays and animal studies can be
used in formulating a range of dosage for use in human. The dosage
of such compounds lies preferably within a range of circulating
concentrations that include the ED.sub.50 with minimal toxicity.
The dosage may vary within this range depending upon the dosage
form employed and the route of administration utilized.
Dosing Strategies to Increase Selectivity
[0662] Described herein are irreversible kinase inhibitors that are
selective for one or more ACKs, including Btk, a Btk homolog, an
ACK, HER4, and a Btk kinase cysteine homolog. In some embodiments,
the irreversible inhibitors described herein also bind reversibly
to other kinases (some of which, in some embodiments, are also
ACKs). As a means of enhancing the selectivity profile, such
inhibitors are formulated (formulation includes chemical
modifications of the inhibitor, use of excipients in a
pharmaceutical composition, and combinations thereof) such that the
pharmacokinetic profile favors enhanced selectivity of the
inhibitors for an ACK over a non-ACK. By way of example only, an
ACK is formulated to have a short plasma half-life. In other
embodiments, an ACK is formulated to have an extended plasma
half-life.
[0663] For example, as shown in the Examples, Compound 1 and
Compound 12 have a short half-life in vivo. In contrast, Compound 7
and Compound 8 have a significantly longer in vivo half-life (FIG.
5). Compounds like 1 and 12 are predicted to have enhanced kinase
selectivity in vivo because inhibition will be sustained only for
those kinases that are irreversibly inhibited. Further, given that
the irreversible kinase inhibitors described herein have both
reversible (in general to non-ACKs) and irreversible (generally, to
ACKs) activities, in vivo properties of absorption, distribution,
metabolism and excretion (ADME) are selected in order to optimize
the therapeutic index. Specifically, in some embodiments, rapidly
cleared compounds cause only brief inhibition of reversibly
inhibited targets while maintaining sustained inhibition of
irreversibly inhibited targets. Depending on the degree to which
sustained inhibition of particular targets results in therapeutic
effects or toxicities, we identify compounds with an optimal
combination of in vitro selectivity profiles and in vivo ADME
properties.
[0664] In one embodiment are kinase inhibitors that selectively and
irreversibly binds to a protein tyrosine kinase selected from Btk,
a Btk homolog, an ACK, HER4, and a Btk kinase cysteine homolog, in
which the kinase inhibitor reversibly and non-selectively binds to
a multiplicity of protein tyrosine kinases, and further in which
the plasma half life of the kinase inhibitor is less than about 4
hours. In such an embodiment, the kinase inhibitor selectively and
irreversibly binds to at least one of Btk, Jak3, Blk, Bmx, Tec, and
Itk. In a further embodiment, the kinase inhibitor selectively and
irreversibly binds to Btk. In a further embodiment, the kinase
inhibitor selectively and irreversibly binds to Jak3. In a further
embodiment, the kinase inhibitor selectively and irreversibly binds
to Tec. In a further embodiment, the kinase inhibitor selectively
and irreversibly binds to Btk and Tec. In a further embodiment, the
kinase inhibitor selectively and irreversibly binds to Blk. In a
further embodiment, the kinase inhibitor reversibly and
non-selectively binds to a multiplicity of src-family protein
kinase inhibitors. In a further embodiment, the plasma half life of
the kinase inhibitor is less than about 3 hours. In a further
embodiment, the plasma half life of the kinase inhibitor is less
than about 2 hours.
[0665] In one embodiment are kinase inhibitors that selectively and
irreversibly binds to a protein tyrosine kinase selected from Btk,
a Btk homolog, an ACK, HER4, and a Btk kinase cysteine homolog, in
which the kinase inhibitor reversibly and non-selectively binds to
a multiplicity of protein tyrosine kinases, and further in which
the plasma half life of the kinase inhibitor is greater than about
12 hours. In such an embodiment, the kinase inhibitor selectively
and irreversibly binds to at least one of Btk, Jak3, Blk, Bmx, Tec,
and Itk. In a further embodiment, the kinase inhibitor selectively
and irreversibly binds to Btk. In a further embodiment, the kinase
inhibitor selectively and irreversibly binds to Jak3. In a further
embodiment, the kinase inhibitor selectively and irreversibly binds
to Tec. In a further embodiment, the kinase inhibitor selectively
and irreversibly binds to Btk and Tec. In a further embodiment, the
kinase inhibitor selectively and irreversibly binds to Blk. In a
further embodiment, the kinase inhibitor reversibly and
non-selectively binds to a multiplicity of src-family protein
kinase inhibitors In a further embodiment, the kinase inhibitor the
plasma half life of the kinase inhibitor is greater than about 16
hours.
[0666] In one particular embodiment of any of the aforementioned
kinase inhibitors, such kinase inhibitors have the structure of
Formula (VII):
##STR00090##
wherein: wherein
##STR00091##
is a moiety that binds to the active site of a kinase, including a
tyrosine kinase, further including a Btk kinase cysteine homolog; Y
is an optionally substituted group selected from among alkylene,
heteroalkylene, arylene, heteroarylene, heterocycloalkylene,
cycloalkylene, alkylenearylene, alkyleneheteroarylene,
alkylenecycloalkylene, and alkyleneheterocycloalkylene; Z is
C(.dbd.O), OC(.dbd.O), NHC(.dbd.O), NCH.sub.3C(.dbd.O), C(.dbd.S),
S(.dbd.O).sub.x, OS(.dbd.O).sub.x, NHS(.dbd.O).sub.x, where x is 1
or 2; R.sub.7 and R.sub.8 are independently selected from among H,
unsubstituted C.sub.1-C.sub.4 alkyl, substituted
C.sub.1-C.sub.4alkyl, unsubstituted C.sub.1-C.sub.4heteroalkyl,
substituted C.sub.1-C.sub.4heteroalkyl, unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted C.sub.3-C.sub.6cycloalkyl,
unsubstituted C.sub.2-C.sub.6heterocycloalkyl, and substituted
C.sub.2-C.sub.6heterocycloalkyl; or R.sub.7 and R.sub.8 taken
together form a bond; R.sub.6 is H, substituted or unsubstituted
C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted C.sub.2-C.sub.8heterocycloalkyl,
substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl); and
pharmaceutically active metabolites, or pharmaceutically acceptable
solvates, pharmaceutically acceptable salts, or pharmaceutically
acceptable prodrugs thereof.
[0667] In a further embodiment,
##STR00092##
on the kinase inhibitor is a substituted fused biaryl moiety
selected from
##STR00093##
[0668] In a further embodiment of such kinases:
Z is C(.dbd.O), NHC(.dbd.O), NCH.sub.3C(.dbd.O), or
S(.dbd.O).sub.2. The kinase inhibitor of claim 49, wherein: each of
R.sub.7 and R.sub.8 is H; or R.sub.7 and R.sub.8 taken together
form a bond.
[0669] In a further embodiment of such kinases:
R.sub.6 is H, substituted or unsubstituted C.sub.1-C.sub.4alkyl,
substituted or unsubstituted C.sub.1-C.sub.4heteroalkyl,
C.sub.1-C.sub.6alkoxyalkyl, C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, C.sub.1-C.sub.4alkyl(aryl),
C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl).
[0670] In a further embodiment of such kinases:
Y is a 4-, 5-, 6-, or 7-membered cycloalkylene ring; or Y is a 4-,
5-, 6-, or 7-membered heterocycloalkylene ring; or Y is a
C.sub.1-C.sub.4 alkylene, or 4-, 5-, 6-, or 7-membered
heterocycloalkylene ring.
[0671] In another aspect of such dosing methods are pharmaceutical
formulations comprising any of the aforementioned ACK inhibitors
and a pharmaceutically acceptable excipient. In some embodiments,
such pharmaceutical formulations are formulated for a route of
administration selected from oral administration, parenteral
administration, buccal administration, nasal administration,
topical administration, or rectal administration. In certain
embodiments, the pharmaceutical formulations are formulated for
oral administration.
[0672] In another aspect of such dosing methods are methods for
treating rheumatoid arthritis comprising administering to an
individual any of the aforementioned ACK inhibitors that
selectively and irreversibly binds to Btk and Tec.
[0673] In yet another aspect of such dosing strategies are methods
for increasing the selectivity of a test protein tyrosine kinase
inhibitor that irreversibly and selectively binds to at least one
protein kinase inhibitor selected from Btk, a Btk homolog, a Btk
kinase cysteine homolog, an ACK, or HER4, in which the test protein
tyrosine kinase inhibitor is chemically modified to decrease the
plasma half life to less than about 4 hours. In some embodiments,
the test protein tyrosine kinase inhibitor is chemically modified
to decrease the plasma half life to less than about 3 hours.
[0674] In further embodiments, the test protein tyrosine kinase
inhibitor has the structure of Formula (VII):
##STR00094##
wherein
##STR00095##
is a moiety that binds to the active site of a kinase, including a
tyrosine kinase, further including a Btk kinase cysteine homolog; Y
is an optionally substituted group selected from among alkylene,
heteroalkylene, arylene, heteroarylene, heterocycloalkylene,
cycloalkylene, alkylenearylene, alkyleneheteroarylene,
alkylenecycloalkylene, and alkyleneheterocycloalkylene; Z is
C(.dbd.O), OC(.dbd.O), NHC(.dbd.O), NCH.sub.3C(.dbd.O), C(.dbd.S),
S(.dbd.O).sub.x, OS(.dbd.O).sub.x, NHS(.dbd.O).sub.x, where x is 1
or 2; R.sub.7 and R.sub.8 are independently selected from among H,
unsubstituted C.sub.1-C.sub.4 alkyl, substituted
C.sub.1-C.sub.4alkyl, unsubstituted C.sub.1-C.sub.4heteroalkyl,
substituted C.sub.1-C.sub.4heteroalkyl, unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted C.sub.3-C.sub.6cycloalkyl,
unsubstituted C.sub.2-C.sub.6heterocycloalkyl, and substituted
C.sub.2-C.sub.6heterocycloalkyl; or R.sub.7 and R.sub.8 taken
together form a bond; and R.sub.6 is H, substituted or
unsubstituted C.sub.1-C.sub.4alkyl, substituted or unsubstituted
C.sub.1-C.sub.4heteroalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.8alkylaminoalkyl,
C.sub.1-C.sub.8hydroxyalkylaminoalkyl,
C.sub.1-C.sub.8alkoxyalkylaminoalkyl, substituted or unsubstituted
C.sub.3-C.sub.6cycloalkyl, substituted or unsubstituted aryl,
substituted or unsubstituted C.sub.2-C.sub.8heterocycloalkyl,
substituted or unsubstituted heteroaryl,
C.sub.1-C.sub.4alkyl(aryl), C.sub.1-C.sub.4alkyl(heteroaryl),
C.sub.1-C.sub.4alkyl(C.sub.3-C.sub.8cycloalkyl), or
C.sub.1-C.sub.4alkyl(C.sub.2-C.sub.8heterocycloalkyl).
[0675] In a further embodiment, the test protein tyrosine kinase
inhibitor non-selectively and reversibly binds to a multiplicity of
src-family protein tyrosine kinases.
[0676] In a further aspect of such dosing strategies are methods
for treating a disorder characterized by the presence or
development of one or more solid tumors comprising administering to
an individual in need a pharmaceutical composition of any of the
aforementioned ACK inhibitors. For example, as presented in the
Examples, brief exposure to Compound 1 in vitro is sufficient to
inhibit B cell activation in normal human B cells. This protocol
mimics the predicted exposure of cells to Compound 1 in vivo and
demonstrates that inhibition of B cells is sustained despite
washing out of Compound 1.
Kits/Articles of Manufacture
[0677] For use in the therapeutic applications described herein,
kits and articles of manufacture are also described herein. In some
embodiments, such kits include a carrier, package, or container
that is compartmentalized to receive one or more containers such as
vials, tubes, and the like, each of the container(s) including one
of the separate elements to be used in a method described herein.
Suitable containers include, for example, bottles, vials, syringes,
and test tubes. The containers can be formed from a variety of
materials such as glass or plastic.
[0678] The articles of manufacture provided herein contain
packaging materials. Packaging materials for use in packaging
pharmaceutical products include, e.g., U.S. Pat. Nos. 5,323,907,
5,052,558 and 5,033,252. Examples of pharmaceutical packaging
materials include, but are not limited to, blister packs, bottles,
tubes, inhalers, pumps, bags, vials, containers, syringes, bottles,
and any packaging material suitable for a selected formulation and
intended mode of administration and treatment. A wide array of
formulations of the compounds and compositions provided herein are
contemplated as are a variety of treatments for any disorder that
benefit by inhibition of Btk, or in which Btk is a mediator or
contributor to the symptoms or cause.
[0679] For example, the container(s) include one or more compounds
described herein, optionally in a composition or in combination
with another agent as disclosed herein. The container(s) optionally
have a sterile access port (for example the container is an
intravenous solution bag or a vial having a stopper pierceable by a
hypodermic injection needle). Such kits optionally comprising a
compound with an identifying description or label or instructions
relating to its use in the methods described herein.
[0680] A kit will typically include one or more additional
containers, each with one or more of various materials (such as
reagents, optionally in concentrated form, and/or devices)
desirable from a commercial and user standpoint for use of a
compound described herein. Non-limiting examples of such materials
include, but not limited to, buffers, diluents, filters, needles,
syringes; carrier, package, container, vial and/or tube labels
listing contents and/or instructions for use, and package inserts
with instructions for use. A set of instructions will also
typically be included.
[0681] In some embodiments, a label is on or associated with the
container. A label can be on a container when letters, numbers or
other characters forming the label are attached, molded or etched
into the container itself; a label can be associated with a
container when it is present within a receptacle or carrier that
also holds the container, e.g., as a package insert. A label can be
used to indicate that the contents are to be used for a specific
therapeutic application. The label can also indicate directions for
use of the contents, such as in the methods described herein.
[0682] In certain embodiments, the pharmaceutical compositions can
be presented in a pack or dispenser device which can contain one or
more unit dosage forms containing a compound provided herein. The
pack can for example contain metal or plastic foil, such as a
blister pack. The pack or dispenser device can be accompanied by
instructions for administration. The pack or dispenser can also be
accompanied with a notice associated with the container in form
prescribed by a governmental agency regulating the manufacture,
use, or sale of pharmaceuticals, which notice is reflective of
approval by the agency of the form of the drug for human or
veterinary administration. Such notice, for example, can be the
labeling approved by the U.S. Food and Drug Administration for
prescription drugs, or the approved product insert. Compositions
containing a compound provided herein formulated in a compatible
pharmaceutical carrier can also be prepared, placed in an
appropriate container, and labeled for treatment of an indicated
condition.
EXAMPLES
[0683] The following specific and non-limiting examples are to be
construed as merely illustrative, and do not limit the present
disclosure in any way whatsoever.
Synthesis of Compounds
Example 1: Preparation of
4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidine
(Intermediate 2)
[0684] 4-Amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidine
(Intermediate 2) is prepared as disclosed in International Patent
Publication No. WO 01/019829. Briefly, 4-phenoxybenzoic acid (48 g)
is added to thionyl chloride (100 mL) and heated under gentle
reflux for 1 hour. Thionyl chloride is removed by distillation, the
residual oil dissolved in toluene and volatile material removed at
80.degree. C./20 mbar. The resulting acid chloride is dissolved in
toluene (200 mL) and tetrahydrofuran (35 mL). Malononitrile (14.8
g) is added and the solution and stirred at -10.degree. C. while
adding diisopropylethylethylamine (57.9 g) in toluene (150 mL),
while maintaining the temperature below 0.degree. C. After 1 hour
at 0.degree. C., the mixture is stirred at 20.degree. C. overnight.
Amine hydrochloride is removed by filtration and the filtrate
evaporated in vacuo. The residue is taken up in ethyl acetate and
washed with 1.25 M sulphuric acid, then with brine and dried over
sodium sulfate. Evaporation of the solvents gives a semisolid
residue which is treated with a little ethyl acetate to give 4.1 g
of 1,1-dicyano-2-hydroxy-2-(4-phenoxyphenyl)ethene as a white solid
(m.p. 160-162.degree. C.). The filtrate on evaporation gives 56.58
(96%) of 1,1-dicyano-2-hydroxy-2-(4-phenoxyphenyl)ethene as a
grey-brown solid, which is sufficiently pure for further use.
[0685] 1,1-Dicyano-2-hydroxy-2-(4-phenoxyphenyl)ethene (56.5 g) in
acetonitrile (780 mL) and methanol (85 mL) is stirred under
nitrogen at 0.degree. C. while adding diisopropylethylamine (52.5
mL) followed by 2M trimethylsilyldiazomethane (150 mL) in THF. The
reaction is stirred for 2 days at 20.degree. C., and then 2 g of
silica is added (for chromatography). The brown-red solution is
evaporated in vacuo, the residue dissolved in ethyl acetate and
washed well with water then brine, dried and evaporated. The
residue is extracted with diethyl ether (3.times.250 mL), decanting
from insoluble oil. Evaporation of the ether extracts gives 22.5 g
of 1,1-dicyano-2-methoxy-2-(4-phenoxyphenyl)ethene as a pale orange
solid. The insoluble oil is purified by flash chromatography to
give 15.0 g of a red-orange oil.
1,1-Dicyano-2-methoxy-2-(4-phenoxyphenyl)ethene (22.5 g) and
1,1-dicyano-2-methoxy-2-(4-phenoxyphenyl)ethene oil (15 g) are
treated with a solution of hydrazine hydrate (18 mL) in ethanol (25
mL) and heated on the steambath for 1 hour. Ethanol (15 mL) is
added followed by water (10 mL). The precipitated solid is
collected and washed with ethanol:water (4:1) and then dried in air
to give 3-amino-4-cyano-5-(4-phenoxyphenyl)pyrazole as a pale
orange solid.
[0686] 3-Amino-4-cyano-5-(4-phenoxyphenyl)pyrazole (29.5 g) is
suspended in formamide (300 mL) and heated under nitrogen at
180.degree. C. for 4 hours. The reaction mixture is cooled to
30.degree. C. and water (300 mL) is added. The solid is collected,
washed well with water, then with methanol and dried in air to give
of 4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidine.
Example 2: Synthesis of
1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperi-
din-1-yl)prop-2-en-1-one (Compound 13)
##STR00096##
[0688] Synthesis of Compound 13; a) triphenylphosphine (TPP),
diisopropyl diazodicarboxylate (DIAD), tetrahydrofuran (THF); b)
TFA/CH.sub.2Cl.sub.2; then acryloyl chloride, diisopropylethylamine
(DIPEA), tetrahydrofuran (THF).
[0689] Compounds described herein were synthesized by following the
steps outlined in Scheme 1. A detailed illustrative example of the
reaction conditions shown in Scheme 1 is described for the
synthesis of
1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 13).
[0690] 0.5 g of
4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidine and 0.65 g
of triphenylphosphine(TPP) were mixed together with 15 mL of
tetrahydrofuran (THF). (R)-tert-butyl
2-(hydroxymethyl)pyrrolidine-1-carboxylate (0.5 g; 1.5 equivalents)
was added to the mixture followed by the addition of diisopropyl
diazodicarboxylate (0.5 mL). The reaction mixture was stirred at
room temperature for 4 hr. The reaction mixture was concentrated
and purified by flash chromatography (acetone/CH.sub.2Cl.sub.2=1/1)
to give intermediate 3 (1.49 g).
[0691] Intermediate 3 (1.49 g) was treated with 4 mL of TFA and 5
mL of CH.sub.2Cl.sub.2 and stirred overnight at room temperature
and then concentrated to dryness. The residue was dissolved in
ethyl acetate (100 mL) and then washed with dilute aq. NaHCO.sub.3
(100 mL). The ethyl acetate layer was dried (MgSO.sub.4), filtered
and concentrated to .about.20 mL and then 4.0 M HCl\dioxane (1 mL)
was added and a yellow precipitate formed. The solid was collected
by filtration and washed with ethyl acetate (20 mL). The solid was
suspended in ethyl acetate (100 mL) and again washed with dilute
aq. NaHCO.sub.3 (100 mL). The ethyl acetate was dried (MgSO.sub.4),
filtered and concentrated to provide 0.43 g of a light yellow
solid. The solid (0.14 g, 0.36 mmol) was stirred in THF (3 mL) and
TEA (015 mL, 1.1 mmol) was added, followed by cooling the reaction
with an ice bath for 30 min, then acryl chloride (30 .mu.L, 0.36
mmol) added and the reaction was stirred for 2 hr. The reaction
mixture was diluted with ethyl acetate (75 mL) and washed with
dilute aq. NaHCO.sub.3 (100 mL). The organic layer was dried
(MgSO.sub.4), filtered and concentrated. Flash chromatography (with
CH.sub.2Cl.sub.2/MeOH=20/1) gave 90 mg of compound 4 as a white
solid. EM (calc)=440.2; MS (M+1): 441.2.
Example 3: Synthesis of
1-((S)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)prop-2-en-1-one (Compound 14)
##STR00097##
[0693] The synthesis of Compound 14 was accomplished using a
procedure analogous to that described in Example 2. EM (calc.):
440.2; MS (M+1H): 441.2.
Example 4: Synthesis of
N-((1r,4r)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-y-
l)cyclohexyl)acrylamide
##STR00098##
[0695] The synthesis of this compound was accomplished using a
procedure analogous to that described for Example 2 EM (calc.):
454.21; MS (M+1): 455.2.
Example 5: Synthesis of
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
-N-methylacrylamide (Compound 19)
##STR00099##
[0697] The synthesis of this compound was accomplished using a
procedure analogous to that described for Example 2. EM (calc.):
414.18; MS (M+1H): 415.2.
Example 6: Synthesis of
N-(2-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-
acrylamide (Compound 23)
##STR00100##
[0699] The synthesis of this compound was accomplished using a
procedure analogous to that described for Example 2. EM (calc.):
400.16; MS (M+1H): 401.2.
Example 7: Synthesis of
1-((R)-3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pi-
peridin-1-yl)but-2-yn-1-one e (Compound 17)
##STR00101##
[0701] The synthesis of this compound was accomplished using a
procedure analogous to that described for Example 2. EM (calc.):
452.2; MS (M+1H): 453.2.
Example 8: Synthesis of
1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)m-
ethyl)pyrrolidin-1-yl)but-2-yn-1-one (Compound 15)
##STR00102##
[0703] The synthesis of this compound was accomplished using a
procedure analogous to that described for Example 2. EM (calc.):
452.2; MS (M+1H): 453.2.
Example 9: Synthesis of
(E)-1-((R)-2-((4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1--
yl)methyl)pyrrolidin-1-yl)-4-(dimethylamino)but-2-en-1-one
(Compound 11)
##STR00103##
[0704] The synthesis of this compound was accomplished using a
procedure analogous to that described for Example 2. EM (calc.):
497.25; MS (M+1H): 498.2.
Therapeutic Uses of Inhibitor Compounds
Example 1: Inhibition of Lymphoma Tumor Cell Growth
[0705] Compound 1 inhibits lymphoma tumor cell growth. A variety of
lymphoma cell lines were incubated with a range of concentrations
of Compound 1 to determine the GI50, the concentration that results
in 50% decrease in cell proliferation (FIG. 1A). Compound 1
inhibits tumor growth in DOHH2 and DLCL2 xenograft models (FIGS. 1B
and 1C).
[0706] For in vitro cell proliferation assays, cells were seeded in
96-well plates in standard growth media (in most cases RPMI+10%
fetal calf serum) and Compound 1 was added in a 9-point dilution
series ranging from 10 uM to 0.04 uM with DMSO at 0.1% final
concentration in all wells. After 72 hours, cell number was
measured using Alamar Blue using manufacturer's protocol. A
dilution series of untreated cells was run in parallel to verify
that the Alamar Blue assay reliably reflected cell number and that
growth conditions were not limiting. The GI50, the concentration
that results in a 50% decrease in cell number, was calculated using
Calcusyn to fit the dose-response curve. GI50 values were confirmed
in two or more separate experiments for each cell line.
[0707] For in vivo lymphoma xenograft studies, 5E6 DOHH2 or DLCL2
cells in 50% matrigel were implanted subcutaneously in SCID mice
and dosed orally with Compound 1 beginning when tumor size reached
100 mm2.
Example 2: Inhibition of Collagen-Induced Arthritis in a Mouse
[0708] Compound 1 inhibits collagen-induced arthritis in the mouse.
Male DBA/1OlaHsd mice were injected intradermally with 150
microliters of 2 mg/mL Type II collagen in Freund's complete
adjuvant with supplemental M. tuberculosis, 4 mg/mL and boosted
with the same injection 21 days later. After paw inflammation was
established, animals were randomized and Compound 1 or vehicle was
dosed orally once per day starting at day 1. Paw inflammation was
scored from 0-5 and averaged across all paws from all animals for
each group in the study. Compound 1 at 12.5 mg/kg and 50 mg/kg
regressed inflammation through the end of the study (day 11) while
3.125 mg/kg significantly reduced the increase in paw inflammation
(FIG. 2). Dexamethasone was included as a positive control.
[0709] In another study, Compound 1 was dosed at 12.5 mg/kg to such
mice over: (a) each day of an 11-day period; (b) days 1, 2, and 3
of an 11-day period; or (c) days 9, 10, and 11 of an 11-day period.
Intermittent dosing reduced the increase in paw inflammation. In
addition, Compound 9 was dosed to such mice at a level of 12.5
mg/kg or 50 mg/kg each day of an 11-day period. Compound 9 reduced
the increase in paw inflammation.
Example 3: Inhibition of Lupus in a Mouse Model
[0710] Compound 1 inhibits disease progression in the mouse MRL/lpr
model of lupus. Compound 1 at 3.125 mg/kg, 12.5 mg/kg, and 50 mg/kg
significantly reduced proteinuria, indicating amelioration of the
progressive autoimmune renal failure seen in this mouse strain
(FIG. 3). MRL/lpr mice (Jax strain 000485) were dosed orally once
per day from 12 weeks of age until 20 weeks of age and urine
protein levels were measured weekly using Clinitech Multistick
dipstick.
Example 4: Inhibition of Mast Cell Degranulation
[0711] Compound 1 inhibits mast cell degranulation in a mouse
passive cutaneous anaphylaxis model. Increasing doses of Compound 1
significantly decrease the amount of Evans Blue release, indicating
decreased mast cell activation and vascular permeabilization. (FIG.
4)
[0712] Mice were sensitized with an intradermal injection of
monoclonal anti-DNP-IgE in the back. 23 hours later they received a
single oral dose of Compound 1 or vehicle. After one hour, animals
were challenged with an intravenous injection of DNP-BSA and Evans
Blue dye. Mast cell degranulation leads to vascular permeability
and the distribution of the dye into the skin of the back. The area
of extravasation after 1 hour is measured.
Example 5: Pharmaceutical Compositions
[0713] The compositions described below are presented with a
compound of Formula (A1-A6) for illustrative purposes; any of the
compounds of any of Formulas (A1-A6), (B1-B6), (C1-C6), or (D1-D6)
are optionally used in such pharmaceutical compositions.
Example 5a: Parenteral Composition
[0714] To prepare a parenteral pharmaceutical composition suitable
for administration by injection, 100 mg of a water-soluble salt of
a compound of Formula (A1-A6) is dissolved in DMSO and then mixed
with 10 mL of 0.9% sterile saline. The mixture is incorporated into
a dosage unit form suitable for administration by injection.
Example 5b: Oral Composition
[0715] To prepare a pharmaceutical composition for oral delivery,
100 mg of a compound of Formula (A1-A6) is mixed with 750 mg of
starch. The mixture is incorporated into an oral dosage unit for,
such as a hard gelatin capsule, which is suitable for oral
administration.
Example 5c: Sublingual (Hard Lozenge) Composition
[0716] To prepare a pharmaceutical composition for buccal delivery,
such as a hard lozenge, mix 100 mg of a compound of Formula
(A1-A6), with 420 mg of powdered sugar mixed, with 1.6 mL of light
corn syrup, 2.4 mL distilled water, and 0.42 mL mint extract. The
mixture is gently blended and poured into a mold to form a lozenge
suitable for buccal administration.
Example 5d: Inhalation Composition
[0717] To prepare a pharmaceutical composition for inhalation
delivery, 20 mg of a compound of Formula (A1-A6) is mixed with 50
mg of anhydrous citric acid and 100 mL of 0.9% sodium chloride
solution. The mixture is incorporated into an inhalation delivery
unit, such as a nebulizer, which is suitable for inhalation
administration.
Example 5e: Rectal Gel Composition
[0718] To prepare a pharmaceutical composition for rectal delivery,
100 mg of a compound of Formula (A1-A6) is mixed with 2.5 g of
methylcellulose (1500 mPa), 100 mg of methylparaben, 5 g of
glycerin and 100 mL of purified water. The resulting gel mixture is
then incorporated into rectal delivery units, such as syringes,
which are suitable for rectal administration.
Example 5f: Topical Gel Composition
[0719] To prepare a pharmaceutical topical gel composition, 100 mg
of a compound of Formula (A1-A6) is mixed with 1.75 g of
hydroxypropyl cellulose, 10 mL of propylene glycol, 10 mL of
isopropyl myristate and 100 mL of purified alcohol USP. The
resulting gel mixture is then incorporated into containers, such as
tubes, which are suitable for topical administration.
Example 5g: Ophthalmic Solution Composition
[0720] To prepare a pharmaceutical ophthalmic solution composition,
100 mg of a compound of Formula (A1-A6) is mixed with 0.9 g of NaCl
in 100 mL of purified water and filtered using a 0.2 micron filter.
The resulting isotonic solution is then incorporated into
ophthalmic delivery units, such as eye drop containers, which are
suitable for ophthalmic administration.
Example 6: Levels of Tonic BCR Signaling Predict Response to
Compound 1
[0721] To identify biomarkers that correlate with response to
Compound 1, phosphorylation events in the BCR signal transduction
pathway were investigated. A panel of phospho-specific antibodies
that recognize activating phosphorylation sites on Syk, Btk, BLNK,
PLC-g1, PLC-g2, ERK, and AKT were used and tested the effects of
Compound 4 on both basal phosphorylation and phosphorylation
following BCR stimulation driven by anti-IgM or anti-IgG
cross-linking. We examined phosphorylation patterns in both a
Compound 1 sensitive cell line (DOHH2) and a Compound 1 resistant
cell line (Ramos).
[0722] Compound 1 inhibits most BCR-stimulus induced
phosphorylation events with similar potency in both cell lines.
However, when we examined basal phosphorylation levels, we found
higher basal phosphorylation in DOHH2 compared to Ramos, with
phospho-ERK in particular indicating higher levels of basal or
tonic signaling in DOHH2. Furthermore, Compound 4 significantly
decreased pERK levels in unstimulated DOHH2 cells (IC50<10 nM),
but not in Ramos cells.
[0723] A panel of nine Btk expressing B cell lymphoma cell lines
was screened for basal pERK levels. Seven lines expressed
significantly higher levels of basal pERK, and of these, 5 were
sensitive to Compound 1 (GI50<1.3 uM), while the two cell lines
with low pERK levels were resistant to Compound 1. This data shows
that tonic BCR signaling contributes to the survival of a subset of
lymphoma cell lines, and that inhibition of this signaling by
Compound 4 is correlated with induction of apoptosis.
[0724] Two additional experiments demonstrate that sensitivity to
Compound 1 is correlated with high levels of pERK. First 1 uM of
Compound 4 reduces expression of the known ERK transcriptional
target Egr-1 within 1 hr, with maximal downregulation (10-fold)
achieved by 4 hr. Second, in the lymphoma cell line WSU-DLCL2, BCR
cross-linking by anti-IgG (30 ug/ml) overcomes inhibition of pERK
by Compound 4, showing that strong BCR stimulus activates parallel
pathways to pERK that do not require Btk. BCR stimulus also rescues
WSU-DLCL2 from Compound 1 induced cytotoxicity, further confirming
that inhibition of pERK is correlated with apoptosis induction by
Compound 1. Taken together these data show high levels of pERK or
ERK transcriptional targets such as Egr-1 serve as useful markers
for lymphomas in which tonic BCR signaling is contributing to cell
survival and that these lymphomas are particularly sensitive to BCR
pathway inhibitors such as Compound 1.
Kinase Inhibitor Discovery Platform and Pulse Dosing
Example 1: Design of an Inhibitor
[0725] Because the ATP binding sites of the >500 kinases in the
human genome are highly conserved, it has proven difficult to
engineer selectivity for individual kinases using conventional
reversible binding inhibitors. For our highly selective BTK
inhibitor Compound 1, we engineered an electrophilic center capable
of irreversibly inactivating the target enzyme, BTK. The approach
employed structure based design to achieve a high degree of potency
and selectivity by (1) fitting the core scaffold into the active
site ATP binding pocket of kinase enzymes, and (2) forming a
covalent bond with Cysteine-481 located in BTK. The unique
chemistry required for covalent bond formation involves an
electrophilic moiety that acts as a Michael acceptor, which bonds
with a nucleophile (such as Cys-481) present in a precise location
within the active site.
Example 2: Inhibitor Screening Approach
[0726] By way of example only, a panel of 50-100 Cys-targeting
kinase inhibitors is generated. The molecular orientation and
positioning of the electrophilic group in these inhibitors in
relation to the Cysteine residue will affect the potency and
selectivity of a given inhibitor. Each inhibitor will then be
profiled for kinetics of kinase inhibition (K.sub.i) for each of
the ten Cys-containing kinases, effect on tumor cell proliferation
(GI.sub.50), effect on relevant off-targets (hERG, CYPs), drug-like
characteristics (solubility, c log P) and ability to block labeling
by the active site probe. This panel of diverse inhibitors are then
be used in cell assays (for example, inhibition of tumor growth) to
screen for a phenotype of interest. With the phenotype, the
identification of additional inhibited kinases is determined using
the active site probe and mass spectrometry.
Example 3: Inhibition of a Panel of Kinases for Compound 1 and
Compound 9
[0727] In another example, the linker and Michael acceptor moiety
of Compound 1 was modified to provide Compound 9 which has a
different selectivity pattern. Table 1 is a table showing the
degree of inhibition of a panel of kinases for two example
compounds. IC.sub.50s were determined using the in vitro HotSpot
kinase assay (purified enzymes, .sup.33P-ATP, an appropriate
substrate and 1 uM ATP.) Compared to Compound 1, Compound 9 has
similar potency toward Btk, but significantly less potency toward
JAK-3, ITK, and EGFR and significantly more potency toward the
src-family kinases lck, c-src, FGR, Fyn, Hck, and Lyn and Yes.
Thus, subtle modifications in the linker moiety and the Michael
acceptor moiety are important for the design of selective ACK
inhibitors.
TABLE-US-00003 TABLE 1 Compound 1 Compound 9 Kinase IC50 (nM) IC50
(nM) BTK 0.5 1.0 ITK 11.7 909.9 Bmx/ETK 0.8 1.1 TEC 77.8 108.0 EFGR
0.5 20.6 HER4 9.4 1536.0 HER4 0.1 3.2 LCK 2.0 1.0 BLK 0.5 0.2 C-src
262.6 14.3 FGR 2.3 0.4 Fyn 95.6 7.1 HCK 3.7 1.0 Lyn 16.2 1.2 YES
6.5 0.8 ABL 86.1 32.3 Brk 3.3 3.3 CSK 2.2 2.4 FER 8,070.0 3,346.0
JAK3 10.4 8,278.0 SYK >10,000 >10,000
Example 4: Modification of Linker and Michael Acceptor Moieties and
In Vitro Inhibitory Activity
[0728] In this example, compounds are selected based on in vitro
characteristics to optimize for potency of inhibition of particular
kinases and degree of covalent binding to off-target cysteines such
as glutathione. For example, in Table 2, Compound 9 and Compound 12
both inhibit Btk with a similar potency as Compound 1, but they are
both significantly less potent inhibitors of EGFR, ITK, and JAK-3.
As another example, Compound 11 is similar to Compound 1 for
inhibition of Btk but does not bind glutathione as readily.
[0729] A calculated value (e.g. (1/Btk IC.sub.50)/Glutathione
conjugation rate) as shown in the Table 2) is used to compare
compounds for their ratio between potency at inhibiting their
target and their non-specific binding to other SH groups, such as
those in glutathione. As shown in Table 2, this calculated value is
4.7 for Compound 1 and for 239.6 for Compound 11. Calculated ratios
such as these are used to quantitatively compare different
compounds and select compounds for further study.
Example 4a: Enzyme Inhibition
[0730] For enzyme inhibition assays, compounds were tested in range
of ten concentrations from 10 uM to 0.0005 uM using purified
enzymes and the Hotspot kinase assay. Reaction conditions were 1 uM
ATP, one hour incubation with inhibitor, and kinase activity
detected using 33-ATP phosphorylation of an appropriately selected
peptide substrate. Dose-response curves were fit using Prism, and
the IC.sub.50, the concentration at which enzyme inhibition is 50%
of maximal inhibition, was determined. See Table 2.
Example 4b: Glutathione Binding Assays
[0731] For the glutathione binding assays, 5 mM glutathione, 10
.mu.M Btk inhibitor in DMSO (10 .mu.L) and 6 equivalents of N'N'
Diisopropyl ethyl amine were combined in 1 mL potassium phosphate
buffer. The mixture was incubated for 0, 15, 60 minutes at room
temperature and the reaction was stopped with 10 equivalents of
formic acid. 50 .mu.L of each reaction mixture was injected on HPLC
(Mobil Phase A: 0.2% formic acid in water, Mobile Phase B: 0.2%
formic acid in acetonitrile, HPLC Column: Metasil Basic 3.mu.,
150.times.4.6 mm, 10% B, Gradient:10% to 90% B, Detection: UV/Vis
260 nM). Rate of reaction was reported as nmole GSH conjugate
conversion per minute from the normalized ratio for area under the
curve from HPLC chromatograms for both GSH conjugate and the
parent.
Example 4c: Cell Proliferation Assay
[0732] Analogs are generated that are Btk inhibitors and that are
cytotoxic to the lymphoma cell line DOHH2. See Table 2. For the
DOHH2 cell proliferation assay, cells were seeded in 96-well plates
in standard growth media (RPMI+10% fetal calf serum) and compounds
were added in a 9-point dilution series ranging from 10 uM to 0.04
uM with DMSO at 0.1% final concentration in all wells. After 72
hours, cell number was measured using Alamar Blue using
manufacturer's protocol. A dilution series of untreated cells was
run in parallel to verify that the Alamar Blue assay reliably
reflected cell number and that growth conditions were not limiting.
The GI.sub.50, the concentration that results in a 50% decrease in
cell number, was calculated using Calcusyn to fit the dose-response
curve.
TABLE-US-00004 TABLE 2 Glutathione (1/BTK DOH BTK ITK EGFR LCK JAK3
Conj IC.sub.50)/ H2 Compound IC.sub.50 IC.sub.50 IC.sub.50
IC.sub.50 IC.sub.50 Rate Glutathione GI.sub.50 # Structure (nM)
(nM) (nM) (nM) (nM) (nmol/min) Rate (.mu.M) 1 ##STR00104## 0.5 11.7
0.5 2.0 10.4 0.398 4.7 0.1 2 ##STR00105## 1.1 48.8 0.32 3
##STR00106## 21 74.5 4 ##STR00107## 22.2 487.6 5 ##STR00108## 5.6
326.0 0.004 44.5 6 ##STR00109## 3.1 60.9 0.39 0.8 7 ##STR00110##
6.3 6,123 268.7 2.6 >10,000 0.01 15.9 0.317 8 ##STR00111## 1.4
83.4 9 ##STR00112## 1.0 909.9 20.6 1.0 8278.0 0.011 10 ##STR00113##
1.31 1954 44.5 0.88 >10,000 <0.03 11 ##STR00114## 0.92 6891
18.85 2.43 >10,000 0.004525 239.6 >10 12 ##STR00115## 1.33
14290 698.3 5.97 >10,000 0.004361 172.2 >10 13 ##STR00116##
0.67 3013 18.75 1.56 12980 0.24 14 ##STR00117## 0.39 592.3 2.298
9.24 1456 0.37 15 ##STR00118## 4.16 21100 289.4 5.90 >10,000
0.59 16 ##STR00119## 3.14 >10,000 2807 3.82 >10,000 0.21 17
##STR00120## 2.00 2333 435.3 2.07 >10,000 0.0243 20.6 0.21 18
##STR00121## 1.38 2536 22.53 0.76 >10,000 <0.03 19
##STR00122## 1.58 534.6 28.22 6.62 5997 0.69 20 ##STR00123## 4.07
7993 303.60 98.59 >10,000 0.39 21 ##STR00124## 4.15 >10,000
6238.00 1346 >10,000 1.53 22 ##STR00125## 1.57 3691 156.30 22.12
>10,000 0.014 45.4 <0.04 23 ##STR00126## 0.32 830 70.49
208.00 3306.00 0.11 24 ##STR00127## 0.89 476 383.70 235.40 9077.00
0.44 25 ##STR00128## 3.48 >10,000 272.90 25.81 >10,000
0.05
Example 5: Kinase Inhibitor Selectivity Predicted by Dosing
[0733] Compound 1 and Compound 12 have a short half-life in vivo.
In contrast, Compound 7 and Compound 8 have a significantly longer
in vivo half-life (FIG. 5). Compounds like 1 and 12 are predicted
to have enhanced kinase selectivity in vivo because inhibition will
be sustained only for those kinases that are irreversibly
inhibited.
[0734] Male jugular vein cannulated rats were administered a single
dose of all test compounds at 8 mg/kg each, in combination by oral
gavage. Dose volumes were adjusted based on body weight data
collected immediately prior to dosing. Blood samples were collected
at 0.0833 (5 minutes), 0.333 (20 minutes), 1, 3, 6, 9, and 24 hours
post-dosing from orally dosed rats. The samples were collected into
plasma separator Microtainer tubes with anticoagulant (lithium
heparin). Plasma samples were prepared by centrifugation (5 min at
5000.times.g), and at least 100 .mu.L were transferred to storage
tubes and stored frozen at -80.degree. C. Plasma samples were
thawed and 75 uL aliquots were transferred to centrifuge tubes to
which 10 .mu.L aliquots of internal standard solution (1 .mu.g/mL)
were added. The samples were not diluted with blank plasma prior to
further processing. Soluble proteins were precipitated by the
addition of 200 L of acetonitrile, followed by centrifugation (20
min at 16,000.times.g). The samples were evaporated to dryness and
reconstituted in 200 .mu.L of water containing 0.2% formic acid and
10% methanol. All samples were loaded onto an autosampler
maintained at 6.degree. C. and evaluated for concentrations of test
compounds using LC-MS/MS.
Example 6: B Cell Inhibition
[0735] Brief exposure to Compound 1 in vitro is sufficient to
inhibit B cell activation in normal human B cells (FIG. 6). This
protocol mimics the predicted exposure of cells to Compound 1 in
vivo and demonstrates that inhibition of B cells is sustained
despite washing out of Compound 1.
[0736] B cells were purified from blood from healthy donors by
negative selecting using the RosetteSep Human B cell enrichment
cocktail. Cells were plated in growth media (10% RPMI+10% fetal
calf serum) and indicated concentrations of Compound 1 were added.
After incubation for 1 hour at 37.degree. C., cells were washed
three times using an 8-fold dilution in growth media for each wash.
Cells were then stimulated with 10 ug/ml of IgM F(ab')2 for 18
hours at 37.degree. C. Cells were then stained with anti-CD69-PE
antibody and analyzed by flow cytometry using standard
conditions.
Example 7: Optimizing the Therapeutic Index of Kinase
Inhibitors
[0737] Given that kinase inhibitors described above will have both
reversible and irreversible activities, we select their in vivo
properties of absorption, distribution, metabolism and excretion
(ADME) in order to optimize the therapeutic index. Specifically,
rapidly cleared compounds are expected to cause only brief
inhibition of reversibly inhibited targets while maintaining
sustained inhibition of irreversibly inhibited targets. Depending
on the degree to which sustained inhibition of particular targets
results in therapeutic effects or toxicities, we identify compounds
with an optimal combination of in vitro selectivity profiles and in
vivo ADME properties.
Example 8: Administration of Btk Inhibitor to a Mouse Model for
Colon Cancer
[0738] Under the skin of a mouse is implanted a colon cancer
xenograft. On a daily basis, administration of Compound 1 is
effected by intravenous administration at a level of 1 microgram
per gram of mouse weight. The size of the tumor xenograft is also
monitored daily. Success is determined by survival of the mouse for
a period of time statistically longer than survival of a mouse
administered with vehicle on the same dosing schedule.
Example 9: Pancreatic Cancer Clinical Trial
Length of Study
[0739] 8 months [length of time from FPV to LPV]
Objectives
[0740] The primary objective of this study will be to determine the
objective response rate (ORR) for Btk inhibitors when administered
every 2 weeks to patients with adenocarcinoma of the pancreas. The
secondary objectives of this study will be to measure time-to-event
variables including: time to objective tumor response for
responding patients (TtOR), duration of response for responding
patients, time to treatment failure (TtTF), time to progressive
disease (TtPD), progression-free survival (PFS), overall survival
(OS); the toxicities of therapy.
Study Design
[0741] The study will be a multi-center, double-blind, randomized,
placebo-controlled Phase 2 study. Tumor assessments will be
repeated every 4 cycles (approximately 8 weeks). Patients will
receive study therapy for 12 treatments, or until tumor progression
was documented, unacceptable toxicity was experienced, the patient
withdrew consent, or the patient is unable to fulfill the
responsibilities of study participation as determined by the
treating physician or the qualified investigator. After study
discontinuation, patients who have not progressed will have tumor
assessments performed approximately every 8 weeks until disease
progression. Once patients have disease progression, patients will
enter a post-study follow-up period, and will be followed every 12
weeks for 24 months for overall survival. Patients will also be
followed for ongoing or any new toxicities.
Diagnosis and Main Criteria for Inclusion.
[0742] Male and females .gtoreq.18 years of age will be eligible
for this study if they are diagnosed with adenocarcinoma of the
pancrease. Patients must have had as their initial presentation
pancreatic metastasis without evidence of pulmonary metastasis.
[0743] Main inclusion criteria will include: histologically proven
adenocarcinoma; performance Status of 0 or 1 on the Eastern
Cooperative Oncology Group (ECOG) scale; a complete history and
physical, chest x-ray, CT scan of abdomen and pelvis; barium enema,
or colonoscopy. Patients with pain will be requires to have had
their pain stabilized for 1 week prior to commencing therapy.
Patients requiring opioids for pain control will be required to
have been on a fixed analgesic regimen aimed to provide adequate
pain control with no more than three breakthrough (supplemental)
doses of analgesics per day to control pain. Patients will be
requires to demonstrate adequate bone marrow reserve (i.e.
Neutrophil count .gtoreq.1.5.times.109 cells/L; Platelets
.gtoreq.100.times.109 cells/L). Patients will be required to have
negative tumor markers for alpha-fetoprotein (AFP) and monoclonal
antichorionic gonadotropin (.beta.-subunit) (3HCG). Patients will
be required to demonstrate at least one unidimensionally measurable
lesion, meeting Response Evaluation Criteria in Solid Tumors
(RECIST). Patient will also be required to have an estimated life
expectancy of at least 12 weeks.
[0744] Main exclusion criteria will include: prior chemotherapy;
pregnancy or breastfeeding; inability or unwillingness to take
folic acid, vitamin B12 supplementation, or dexamethasone.
Study Drug, Dose, and Mode of Administration
[0745] Btk inhibitor of Formula VII dosage will be 500 mg/m.sup.2
and will be given as a 10-minute infusion on Day 1 of each 14-day
cycle. Folic acid and vitamin B12 supplementation, and
dexamethasone (or equivalent corticosteroid) prophylaxis will also
be administered.
Variables
[0746] Efficacy: Tumor response rate will be defined as the number
of patients with documented partial response (PR) or complete
response (CR) divided by the number of patients qualified for tumor
response analysis. Time-to-event analyses will be performed on the
observed distributions of time to objective progressive disease,
progression-free survival (PFS), time to treatment failure (TtTF),
and overall survival (OS) using the Kaplan-Meier (K-M) method. All
patients with best overall response of CR or PR will be analyzed
for response duration by using the K-M method.
[0747] Safety: Safety analyses will include adverse event (AE)
rates, serious AEs, vital signs, laboratory data, blood
transfusions required, and deaths. Toxicities using laboratory and
nonlaboratory adverse events will be evaluated using the common
terminology criteria for adverse events (CTCAE, version 3.0).
Evaluation Methods
[0748] Statistical: The primary analysis will be to estimate the
objective best overall response rate and its 95% confidence
interval (CI). Medians for each of the time-to-event endpoints, and
time-to-event variables will be estimated using the K-M method. All
estimates of treatment effects will be conducted at a two-sided
alpha level of 0.05, and CI for all parameters will be estimated
were to be constructed using a 95% level.
Example 10: Breast Cancer Clinical Trial
Length of Study
[0749] 6 months
Objectives
[0750] The primary objective of this study will be to determine the
objective response rate (ORR) for Btk inhibitors when administered
every 2 weeks to patients with breast cancer. The secondary
objectives of this study will be to measure time-to-event variables
including: time to objective tumor response for responding patients
(TtOR), duration of response for responding patients, time to
treatment failure (TtTF), time to progressive disease (TtPD),
progression-free survival (PFS), overall survival (OS); the
toxicities of therapy.
Study Design
[0751] The study will be a multi-center, double-blind, randomized,
placebo-controlled Phase 2 study. Tumor assessments will be
repeated every 4 cycles (approximately 6 weeks). Patients will
receive study therapy for 12 treatments, or until tumor progression
was documented, unacceptable toxicity was experienced, the patient
withdrew consent, or the patient is unable to fulfill the
responsibilities of study participation as determined by the
treating physician or the qualified investigator. After study
discontinuation, patients who have not progressed will have tumor
assessments performed approximately every 6 weeks until disease
progression. Once patients have disease progression, patients will
enter a post-study follow-up period, and will be followed every 12
weeks for 24 months for overall survival. Patients will also be
followed for ongoing or any new toxicities.
Diagnosis and Main Criteria for Inclusion.
[0752] Female patients aged 18 years or older
[0753] Histologically-confirmed ER-negative, progesterone receptor
(PgR)-positive or PgR-negative, metastatic breast cancer
[0754] Cancer not life-threatening
[0755] No previous endocrine or cytotoxic treatment for metastatic
breast cancer.
Study Drug, Dose, and Mode of Administration
[0756] Btk inhibitor of Formula VII dosage will be 1000 mg/m.sup.2
and will be given as a 60-minute infusion on Day 1 of each cycle.
Folic acid and vitamin B12 supplementation, and dexamethasone (or
equivalent corticosteroid) prophylaxis will also be
administered.
Variables
[0757] Efficacy: Tumor response rate will be defined as the number
of patients with documented partial response (PR) or complete
response (CR) divided by the number of patients qualified for tumor
response analysis. Time-to-event analyses will be performed on the
observed distributions of time to objective progressive disease,
progression-free survival (PFS), time to treatment failure (TtTF),
and overall survival (OS) using the Kaplan-Meier (K-M) method. All
patients with best overall response of CR or PR will be analyzed
for response duration by using the K-M method.
[0758] Safety: Safety analyses will include adverse event (AE)
rates, serious AEs, vital signs, laboratory data, blood
transfusions required, and deaths. Toxicities using laboratory and
nonlaboratory adverse events will be evaluated using the common
terminology criteria for adverse events (CTCAE, version 3.0).
Evaluation Methods
[0759] Statistical: The primary analysis will be to estimate the
objective best overall response rate and its 95% confidence
interval (CI). Medians for each of the time-to-event endpoints, and
time-to-event variables will be estimated using the K-M method. All
estimates of treatment effects will be conducted at a two-sided
alpha level of 0.05, and CI for all parameters will be estimated
were to be constructed using a 95% level.
Example 11: Breast Cancer Clinical Trial
[0760] An MDA-MB-453 breast cancer xenograft was implanted under
the skin of a nude mouse. On a daily basis, administration of
Compound 1 was effected by intravenous administration at a level of
(a) 50 mg/kg of mouse weight, or (b) 5 mg/kg of mouse weight. The
volume of the tumor xenograft was monitored daily.
Sequence CWU 1
1
14115PRTArtificialSynthetic peptide substrate 1Ala Val Leu Glu Ser
Glu Glu Glu Leu Tyr Ser Ser Ala Arg Gln1 5 10 15211PRTHomo sapiens
2Ile Thr Glu Tyr Met Ala Asn Gly Cys Leu Leu1 5 10311PRTHomo
sapiens 3Val Thr Glu Tyr Met Ala Arg Gly Cys Leu Leu1 5
10411PRTHomo sapiens 4Val Thr Glu Phe Met Glu Arg Gly Cys Leu Leu1
5 10511PRTHomo sapiens 5Val Thr Glu Phe Met Glu Asn Gly Cys Leu
Leu1 5 10611PRTHomo sapiens 6Val Phe Glu Phe Met Glu His Gly Cys
Leu Ser1 5 10711PRTHomo sapiens 7Ile Thr Gln Leu Met Pro Phe Gly
Cys Leu Leu1 5 10811PRTHomo sapiens 8Val Thr Gln Leu Met Pro Tyr
Gly Cys Leu Leu1 5 10911PRTHomo sapiens 9Val Thr Gln Leu Met Pro
His Gly Cys Leu Leu1 5 101011PRTHomo sapiens 10Val Met Glu Tyr Leu
Pro Ser Gly Cys Leu Arg1 5 101111PRTHomo sapiens 11Val Thr Glu Tyr
Leu Pro Ser Gly Cys Leu Leu1 5 101211PRTHomo sapiens 12Ile Thr Glu
Tyr Met Glu Asn Gly Ser Leu Val1 5 101311PRTHomo sapiens 13Ile Thr
Glu Tyr Met Ala Lys Gly Ser Leu Leu1 5 101411PRTHomo sapiens 14Val
Met Glu Met Ala Glu Leu Gly Pro Leu Asn1 5 10
* * * * *