U.S. patent application number 15/003537 was filed with the patent office on 2016-12-22 for bicyclic heteroaryl compounds.
The applicant listed for this patent is ARIAD Pharmaceuticals, Inc.. Invention is credited to David C. Dalgarno, Wei-Sheng Huang, Chester A. Metcalf, III, Jiwei Qi, Jan Antoinette C. Romero, Tomi K. Sawyer, William C. Shakespeare, Rajeswari Sundaramoorthi, Ranny M. Thomas, Yihan Wang, Xiaotian Zhu, Dong Zou.
Application Number | 20160368917 15/003537 |
Document ID | / |
Family ID | 38218600 |
Filed Date | 2016-12-22 |
United States Patent
Application |
20160368917 |
Kind Code |
A1 |
Zou; Dong ; et al. |
December 22, 2016 |
Bicyclic Heteroaryl Compounds
Abstract
This invention relates to compounds of the general formula:
##STR00001## in which the variable groups are as defined herein,
and to their preparation and use.
Inventors: |
Zou; Dong; (Concord, MA)
; Huang; Wei-Sheng; (Acton, MA) ; Thomas; Ranny
M.; (Sharon, MA) ; Romero; Jan Antoinette C.;
(Arlington, MA) ; Qi; Jiwei; (West Roxbury,
MA) ; Wang; Yihan; (Newton, MA) ; Zhu;
Xiaotian; (Newton, MA) ; Shakespeare; William C.;
(Southborough, MA) ; Sundaramoorthi; Rajeswari;
(Watertown, MA) ; Metcalf, III; Chester A.;
(Needham, MA) ; Dalgarno; David C.; (Brookline,
MA) ; Sawyer; Tomi K.; (Southborough, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
ARIAD Pharmaceuticals, Inc. |
Cambridge |
MA |
US |
|
|
Family ID: |
38218600 |
Appl. No.: |
15/003537 |
Filed: |
January 21, 2016 |
Related U.S. Patent Documents
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Application
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Patent Number |
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14296016 |
Jun 4, 2014 |
9278971 |
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15003537 |
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13801619 |
Mar 13, 2013 |
8778942 |
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14296016 |
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13357745 |
Jan 25, 2012 |
8470851 |
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13801619 |
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11644849 |
Dec 22, 2006 |
8114874 |
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13357745 |
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60754000 |
Dec 23, 2005 |
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60753962 |
Dec 23, 2005 |
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60756089 |
Jan 3, 2006 |
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60798472 |
May 8, 2006 |
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60833191 |
Jul 25, 2006 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 19/08 20180101;
A61P 35/00 20180101; A61P 3/00 20180101; A61P 19/00 20180101; A61P
29/00 20180101; C07D 471/04 20130101; A61P 19/02 20180101; A61P
19/10 20180101; C07D 473/34 20130101; A61P 43/00 20180101; C07D
487/04 20130101 |
International
Class: |
C07D 471/04 20060101
C07D471/04 |
Claims
1-22. (canceled)
23. A compound of the formula: ##STR00165## wherein: Ring A is a 5-
or 6-membered aryl or heteroaryl ring; Ring B is a 5- or 6-membered
aryl or heteroaryl ring; L.sup.1 is selected from NR.sup.1C(O),
C(O)NR.sup.1, NR.sup.1C(O)O, NR.sup.1C(O)NR.sup.1, and
OC(O)NR.sup.1; each occurrence of R.sup.a and R.sup.b is
independently halo, --CN, --NO.sub.2, --R.sup.4, --OR.sup.2,
--NR.sup.2R.sup.3, --C(O)YR.sup.2, --OC(O)YR.sup.2,
--NR.sup.2C(O)YR.sup.2, --SC(O)YR.sup.2,
--NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)YR.sup.2,
--YP(.dbd.O)(YR.sup.4)(YR.sup.4), --Si(R.sup.2).sub.3,
--NR.sup.2SO.sub.2R.sup.2, --S(O).sub.rR.sup.2,
--SO.sub.2NR.sup.2R.sup.3 or --NR.sup.2SO.sub.2NR.sup.2R.sup.3,
wherein each Y is independently a bond, --O--, --S-- or
--NR.sup.3--; R.sup.e, at each occurrence, is independently halo,
--CN, --NO.sub.2, --R.sup.4, --OR.sup.2, --NR.sup.2R.sup.3,
--C(O)YR.sup.2, --OC(O)YR.sup.2, --NR.sup.2C(O)YR.sup.2,
--SC(O)YR.sup.2, --NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)YR.sup.2,
--YP(.dbd.O)(YR.sup.4)(YR.sup.4), --Si(R.sup.2).sub.3,
--NR.sup.2SO.sub.2R.sup.2, --S(O).sub.rR.sup.2,
--SO.sub.2NR.sup.2R.sup.3 or --NR.sup.2SO.sub.2NR.sup.2R.sup.3,
wherein each Y is independently a bond, --O--, --S-- or
--NR.sup.3--; R.sup.1, R.sup.2 and R.sup.3 are independently H,
alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl,
aryl, heterocyclyl, or heteroaryl; or R.sup.2 and R.sup.3, taken
together with the atom to which they are attached, form a 5- or
6-membered saturated, partially saturated or unsaturated ring,
which contains 0-2 heteroatoms selected from N, O and S(O).sub.r;
each occurrence of R.sup.4 is independently alkyl, alkenyl,
alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl,
heterocyclyl, or heteroaryl; each of the alkyl, alkenyl, alkynyl,
cycloalkyl, cycloalkenyl, cycloalkynyl and heterocyclyl moieties is
optionally substituted with one or more groups selected from halo,
--CN, --R.sup.4, --OR.sup.2, --S(O).sub.rR.sup.2,
--SO.sub.2NR.sup.2R.sup.3, --NR.sup.2R.sup.3, --(CO)YR.sup.2,
--O(CO)YR.sup.2, --NR.sup.2(CO)YR.sup.2, --S(CO)YR.sup.2,
--NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)Y'R.sup.2,
--COCOR.sup.2,--COMCOR.sup.2, --YP(.dbd.O)(YR.sup.4)(YR.sup.4),
--Si(R.sup.2).sub.3, --NO.sub.2, --NR.sup.2SO.sub.2R.sup.2,
--NR.sup.2SO.sub.2NR.sup.2R.sup.3, .dbd.O, .dbd.S, .dbd.NH,
.dbd.NNR.sup.2R.sup.3, .dbd.NNHC(O)R.sup.2,
.dbd.NNHCO.sub.2R.sup.2, and .dbd.NNHSO.sub.2R.sup.2, wherein M is
a 1-6 carbon alkyl group; each of the aryl and heteroaryl moieties
is optionally substituted on an unsaturated carbon atom with one or
more groups selected from halo, --CN, --R.sup.4, --OR.sup.2,
--S(O).sub.rR.sup.2, --SO.sub.2NR.sup.2R.sup.3, --NR.sup.2R.sup.3,
--(CO)YR.sup.2, --O(CO)YR.sup.2, --NR.sup.2(CO)YR.sup.2,
--S(CO)YR.sup.2, --NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)Y'R.sup.2,
--COCOR.sup.2,--COMCOR.sup.2, --YP(.dbd.O)(YR.sup.4)(YR.sup.4),
--Si(R.sup.2).sub.3, --NO.sub.2, --NR.sup.2SO.sub.2R.sup.2, and
--NR.sup.2SO.sub.2NR.sup.2R.sup.3; m is 0, 1, 2, 3 or 4; p is 0, 1,
2, 3, 4 or 5; r is 0, 1 or 2; and s is 0, 1, 2, 3 or 4, or a
pharmaceutically acceptable salt-thereof.
24. The compound of claim 23 of the formula: ##STR00166## wherein:
Ring C is a 5- or 6-membered heterocyclyl, or heteroaryl ring,
comprising carbon atoms and 1-3 heteroatoms independently selected
from O, N and S(O).sub.r; R.sup.c, at each occurrence, is
independently selected from halo, .dbd.O, --CN, --NO.sub.2,
--R.sup.4, --OR.sup.2, --NR.sup.2R.sup.3, --C(O)YR.sup.2,
--OC(O)YR.sup.2, --NR.sup.2C(O)YR.sup.2, --Si(R.sup.2).sub.3,
--SC(O)YR.sup.2, --NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)YR.sup.2,
--YP(.dbd.O)(YR.sup.4)(YR.sup.4), --NR.sup.2SO.sub.2R.sup.2,
--S(O).sub.rR.sup.2, --SO.sub.2NR.sup.2R.sup.3 and
--NR.sup.2SO.sub.2NR.sup.2R.sup.3, wherein each Y is independently
a bond, --O--, --S-- or --NR.sup.3--; and, v is 0, 1, 2, 3, 4 or
5.
25. The compound of claim 24 wherein Ring A and Ring B are
independently a 5- or 6-membered aryl.
26. The compound of claim 25 wherein Ring C is imidazolyl.
27. The compound of claim 23 having the formula: ##STR00167##
wherein: Ring D represents a 5- or 6-membered heterocyclyl or
heteroaryl ring comprising carbon atoms and 1-3 heteroatoms
independently selected from O, N and S(O).sub.r; L.sup.2 is
(CH.sub.2).sub.z, O(CH.sub.2).sub.x, NR.sup.3(CH.sub.2).sub.x,
S(CH.sub.2).sub.x and (CH.sub.2).sub.xNR.sup.3C(O)(CH.sub.2).sub.x
in either direction; R.sup.d, at each occurrence, is H, halo,
.dbd.O, --CN, --NO.sub.2, --R.sup.4, --OR.sup.2, --NR.sup.2R.sup.3,
--C(O)YR.sup.2, --OC(O)YR.sup.2, --NR.sup.2C(O)YR.sup.2,
--SC(O)YR.sup.2, --NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)YR.sup.2,
--YP(.dbd.O)(YR.sup.4)(YR.sup.4), --Si(R.sup.2).sub.3,
--NR.sup.2SO.sub.2R.sup.2, --S(O).sub.rR.sup.2,
--SO.sub.2NR.sup.2R.sup.3 or --NR.sup.2SO.sub.2NR.sup.2R.sup.3,
wherein each Y is independently a bond, --O--, --S-- or
--NR.sup.3--; R.sup.2 and R.sup.3 are independently H, alkyl,
alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl,
heterocyclyl, or heteroaryl; or R.sup.2 and R.sup.3, taken together
with the atom to which they are attached, form a 5- or 6-membered
saturated, partially saturated or unsaturated ring, which contains
0-2 heteroatoms selected from N, O and S(O).sub.r; each occurrence
of R.sup.4 is independently alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkenyl, cycloalkynyl, aryl, heterocyclyl, or heteroaryl; each
of the alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,
cycloalkynyl, and heterocyclyl moieties is optionally substituted
with one or more groups selected from halo, --CN, --R.sup.4,
--OR.sup.2, --S(O).sub.rR.sup.2, --SO.sub.2NR.sup.2R.sup.3,
--NR.sup.2R.sup.3, --(CO)YR.sup.2, --O(CO)YR.sup.2,
--NR.sup.2(CO)YR.sup.2, --S(CO)YR.sup.2,
--NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)Y'R.sup.2,
--COCOR.sup.2,--COMCOR.sup.2, --YP(.dbd.O)(YR.sup.4)(YR.sup.4),
--Si(R.sup.2).sub.3, --NO.sub.2, --NR.sup.2SO.sub.2R.sup.2,
--NR.sup.2SO.sub.2NR.sup.2R.sup.3, .dbd.O, .dbd.S, .dbd.NH,
.dbd.NNR.sup.2R.sup.3, .dbd.NNHC(O)R.sup.2,
.dbd.NNHCO.sub.2R.sup.2, and .dbd.NNHSO.sub.2R.sup.2, wherein M is
a 1-6 carbon alkyl group; each of the aryl and heteroaryl moieties
is optionally substituted on an unsaturated carbon atom with one or
more groups selected from halo, --CN, --R.sup.4, --OR.sup.2,
--S(O).sub.rR.sup.2, --SO.sub.2NR.sup.2R.sup.3, --NR.sup.2R.sup.3,
--(CO)YR.sup.2, --O(CO)YR.sup.2, --NR.sup.2(CO)YR.sup.2,
--S(CO)YR.sup.2, --NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)Y'R.sup.2,
--COCOR.sup.2,--COMCOR.sup.2, --YP(.dbd.O)(YR.sup.4)(YR.sup.4),
--Si(R.sup.2).sub.3, --NO.sub.2, --NR.sup.2SO.sub.2R.sup.2, and
--NR.sup.2SO.sub.2NR.sup.2R.sup.3; w is 0, 1, 2, 3, 4 or 5; x is 0,
1, 2 or 3; and, z is 1, 2, 3 or 4.
28. The compound of claim 27, wherein Ring A and Ring B are
independently a 5- or 6-membered aryl.
29. The compound of claim 28, wherein Ring D is piperizinyl and
L.sup.2 is CH.sub.2.
30. The compound of claim 29 wherein s is 0, m is 1, p is 1,
R.sup.a is methyl, R.sup.b is CF.sub.3, and R.sup.d is --CH.sub.3
or --CH.sub.2CH.sub.2OH.
31. A composition comprising a compound of claim 23 or a
pharmaceutically acceptable salt thereof and a pharmaceutically
acceptable carrier, diluent or vehicle.
Description
BACKGROUND OF THE INVENTION
[0001] The protein kinases are a large family of proteins which
play a central role in the regulation of a wide variety of cellular
processes. A partial, non limiting, list of such kinases includes
abl, Akt, bcr-abl, Blk, Brk, c-kit, c-met, c-src, CDK1, CDK2, CDK3,
CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, cRaf1, CSK, EGFR, ErbB2,
ErbB3, ErbB4, Erk, Pak, fes, FGFR1, FGFR.sup.2, FGFR3, FGFR4,
FGFR5, Fgr, fit-1, Fps, Frk, Fyn, Hck, IGF-1R, INS-R, Jak, KDR,
Lck, Lyn, MEK, p38, PDGFR, PIK, PKC, PYK2, ros, tie, tie2, TRK and
Zap70. Abnormal protein kinase activity has been related to several
disorders, ranging from non-life threatening diseases such as
psoriasis to extremely serious diseases such as cancers.
[0002] In view of the large number of protein kinases and
associated diseases, there is an ever-existing need for new
inhibitors selective for various protein kinases which might be
useful in the treatment of related diseases.
[0003] This invention concerns a new family of acetylenic
heteroaryl compounds and their use in treating cancers, bone
disorders, metabolic disorders, inflammatory disorders and other
diseases.
DESCRIPTION OF THE INVENTION
1. General Description of Compounds of the Invention
[0004] The compounds of this invention have a broad range of useful
biological and pharmacological activities, permitting their use in
pharmaceutical compositions and methods for treating a variety of
diseases, including e.g., metabolic disorders, bone diseases (e.g.,
osteoporosis, Paget's Disease, etc.), inflammation (including
rheumatoid arthritis, among other inflammatory disorders) and
cancer (including solid tumors and leukemias, especially those
mediated by one or more kinases such as Src or kdr, or by
dysregulation of a kinase such as Abl and mutant variants thereof),
including, among others, advanced cases and cases which are
resistant or refractory to one or more other treatments.
[0005] Included are compounds of Formula I:
##STR00002##
[0006] or a tautomer or an individual isomer or a mixture of
isomers thereof in which:
[0007] Ring T is a 5-membered heteroaryl ring containing 1-2
nitrogens with the remaining ring atoms being carbon, substituted
on at least two ring atoms (each of which may be C or N) with
R.sup.t groups, at least two of which being located on adjacent
ring atoms, and, together with the atoms to which they are
attached, forming a saturated, partially saturated or unsaturated
5- or 6-membered ring (Ring E), containing 0-3 heteroatoms selected
from O, N, and S and being optionally substituted with 1-4 R.sup.e
groups;
[0008] Ring A represents a 5- or 6-membered aryl or heteroaryl ring
and is optionally substituted with 1-4 R.sup.a groups;
[0009] Ring B represents a 5- or 6-membered aryl or heteroaryl ring
and is optionally substituted with 1-5 R.sup.b groups;
[0010] L.sup.1 is selected from NR.sup.1C(O), C(O)NR.sup.1,
NR.sup.1C(O)O, NR.sup.1C(O)NR.sup.1, and OC(O)NR.sup.1;
[0011] each occurrence of R.sup.a, R.sup.b and R.sup.t is
independently selected from the group consisting of halo, --CN,
--NO.sub.2, --R.sup.4, --OR.sup.2, --NR.sup.2R.sup.3,
--C(O)YR.sup.2, --OC(O)YR.sup.2, --NR.sup.2C(O)YR.sup.2,
--SC(O)YR.sup.2, --NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)YR.sup.2,
--YP(.dbd.O)(YR.sup.4)(YR.sup.4), --Si(R.sup.2).sub.3,
--NR.sup.2SO.sub.2R.sup.2, --S(O).sub.rR.sup.2,
--SO.sub.2NR.sup.2R.sup.3 and --NR.sup.2SO.sub.2NR.sup.2R.sup.3,
wherein each Y is independently a bond, --O--, --S-- or
--NR.sup.3--;
[0012] R.sup.e, at each occurrence, is independently selected from
the group consisting of halo, .dbd.O, --CN, --NO.sub.2, --R.sup.4,
--OR.sup.2, --NR.sup.2R.sup.3, --C(O)YR.sup.2, --OC(O)YR.sup.2,
--NR.sup.2C(O)YR.sup.2, --SC(O)YR.sup.2,
--NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)YR.sup.2,
--YP(.dbd.O)(YR.sup.4)(YR.sup.4), --Si(R.sup.2).sub.3,
--NR.sup.2SO.sub.2R.sup.2, --S(O).sub.rR.sup.2,
--SO.sub.2NR.sup.2R.sup.3 and --NR.sup.2SO.sub.2NR.sup.2R.sup.3,
wherein each Y is independently a bond, --O--, --S-- or
--NR.sup.3--;
[0013] R.sup.1, R.sup.2 and R.sup.3 are independently selected from
H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl,
aryl, heterocyclic and heteroaryl;
[0014] alternatively, R.sup.2 and R.sup.3, taken together with the
atom to which they are attached, form a 5- or 6-membered saturated,
partially saturated or unsaturated ring, which can be optionally
substituted and which contains 0-2 heteroatoms selected from N, O
and S(O).sub.r;
[0015] each occurrence of R.sup.4 is independently selected from
alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl,
aryl, heterocyclic and heteroaryl;
[0016] each of the alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkenyl, cycloalkynyl, aryl, heterocyclic and heteroaryl
moieties in this Section 1 is optionally substituted;
[0017] m is 0, 1, 2, 3 or 4;
[0018] n is 2 or 3;
[0019] p is 0, 1, 2, 3, 4 or 5; and,
[0020] r is 0, 1 or 2;
[0021] or a pharmaceutically acceptable salt, solvate or hydrate
thereof.
[0022] The foregoing definitions are further elaborated upon and
exemplified below and apply to all subsequent occurrences except to
the extent otherwise specified.
2. Featured Classes of Compounds and their Use, Generally
[0023] Compounds of this invention include those in which Ring T
has the following structure:
##STR00003##
[0024] where Ring E is a 5- or 6-membered unsaturated ring (formed
by two R.sup.t groups together with the Ring T atoms to which they
are attached, as described above) and s is 0, 1, 2, 3 or 4. These
are illustrated by the compounds of formula I in which the fused
Ring T ring system is one of the following (in which one of the
optional R.sup.e substituents is depicted):
##STR00004##
[0025] Other classes of particular interest are compounds of
Formula I, as described in Part 1, in which Ring E is a 6-membered
ring, otherwise as described above. Illustrative examples of such
compounds include compounds of Formula I in which Ring T (with its
attached Ring E) is a fused bicyclic heteroaryl of the following
types:
##STR00005##
[0026] For the previously described class and subclasses of
compounds, as in all compounds of this invention, Ring A and Ring B
are as previously defined in Part 1.
[0027] Illustrative examples of substituted Ring A groups are:
##STR00006##
[0028] Ring B represents a 5 or 6-membered aryl or heteroaryl ring
as defined above in Part 1.
[0029] Illustrative examples of substituted Ring B groups
include:
##STR00007## ##STR00008##
[0030] Of special interest is the class of compounds of Formula I
as described above in Part 1, in which one of the R.sup.b
substituents is a 5- or 6-membered ring (Ring C), which may be
heteroaryl or heterocyclic, comprising carbon atoms and 1-3
heteroatoms independently selected from O, N and S(O).sub.r, and
Ring C being optionally substituted on carbon or heteroatom(s) with
1 to 5 substituents R.sup.c.
[0031] This class is represented by Formula II:
##STR00009##
[0032] in which the previously defined variables, e.g., n, m, p, A,
B, T, L.sup.1, R.sup.1, R.sup.t, R.sup.a and R.sup.b, are as
defined above in part 1, and
[0033] R.sup.c, at each occurrence, is independently selected from
halo, .dbd.O, --CN, --NO.sub.2, --R.sup.4, --OR.sup.2,
--NR.sup.2R.sup.3, --C(O)YR.sup.2, --OC(O)YR.sup.2,
--NR.sup.2C(O)YR.sup.2, --Si(R.sup.2).sub.3, --SC(O)YR.sup.2,
--NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)YR.sup.2,
--YP(.dbd.O)(YR.sup.4)(YR.sup.4), --NR.sup.2SO.sub.2R.sup.2,
--S(O).sub.rR.sup.2, --SO.sub.2NR.sup.2R.sup.3 and
--NR.sup.2SO.sub.2NR.sup.2R.sup.3, wherein each Y is independently
a bond, --O--, --S-- or --NR.sup.3-- and r, R.sup.2, R.sup.3 and
R.sup.4, are as defined previously in Part 1; and,
[0034] v is 0, 1, 2, 3, 4 or 5.
[0035] Illustrative examples of Ring C systems include but are not
limited to the following types:
##STR00010##
[0036] in which R.sup.c and v are as defined above.
[0037] Of special interest is the class of compounds of formula II
in which Ring T has the following structure:
##STR00011##
[0038] in which the indicated variables, e.g., R.sup.e, s and Ring
E, are as defined previously.
[0039] Illustrative subsets of such compounds include those having
the following structures:
##STR00012##
[0040] as embodied by the following non-limiting illustrative
examples:
##STR00013## ##STR00014##
[0041] in which several illustrative -[Ring A]-[L.sup.1]-[Ring
B]-[Ring C]- portions are depicted.
[0042] Compounds of interest include among others, compounds of
Formula II in which Ring C is an imidazole ring, optionally
substituted with one or more R.sup.c groups. Of particular
interest, are compounds of this subclass in which Ring C bears a
single lower alkyl (e.g., methyl) R.sup.c group.
[0043] A further feature of the invention relates to compounds of
Formula I as described in Part 1, in which one R.sup.b substituent
is -[L.sup.2]-[Ring D]. This class is represented by Formula
III:
##STR00015##
[0044] in which the previously defined variables, e.g., n, m, p,
Ring T, Ring A, Ring B, L.sup.1, R.sup.1, R.sup.t, R.sup.a and
R.sup.b, are defined above in part 1, and
[0045] L.sup.2 is selected from (CH.sub.2).sub.z,
O(CH.sub.2).sub.x, NR.sup.3(CH.sub.2).sub.x, S(CH.sub.2).sub.x, and
(CH.sub.2).sub.xNR.sup.3C(O)(CH.sub.2).sub.x, and the linker moiety
L.sup.2 can be included in either direction;
[0046] Ring D represents a 5- or 6-membered heterocyclic or
heteroaryl ring comprising carbon atoms and 1-3 heteroatoms
independently selected from O, N and S(O).sub.r, and Ring D is
optionally substituted on carbon or heteroatom(s) with 1-5 R.sup.d
groups;
[0047] R.sup.d, at each occurrence, is independently selected from
halo, .dbd.O, --CN, --NO.sub.2,--R.sup.4,--OR.sup.2,
--NR.sup.2R.sup.3,--Si(R.sup.2).sub.3, --C(O)YR.sup.2,
--OC(O)YR.sup.2, --NR.sup.2C(O)YR.sup.2, --SC(O)YR.sup.2,
--NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, --YC(.dbd.NR.sup.3)YR.sup.2,
--YP(.dbd.O)(YR.sup.4)(YR.sup.4), --NR.sup.2SO.sub.2R.sup.2,
--S(O).sub.rR.sup.2, --SO.sub.2NR.sup.2R.sup.3 and
--NR.sup.2SO.sub.2NR.sup.2R.sup.3, wherein each Y is independently
a bond, --O--, --S-- or --NR.sup.3-- and r, R.sup.2, R.sup.3 and
R.sup.4 are as previously defined in Part 1;
[0048] w is 0, 1, 2, 3, 4 or 5;
[0049] x is 0, 1, 2 or 3; and,
[0050] z is 1, 2, 3 or 4.
[0051] Non-limiting, illustrative examples of -[Ring
B]-[L.sup.2]-[Ring D] moieties in compounds of Formula III include
among others:
##STR00016## ##STR00017##
[0052] Of special interest is the class of compounds of formula III
in which Ring T has the following structure:
##STR00018##
[0053] in which the previously defined variables, e.g., R.sup.e, s,
and Ring E, are as defined previously.
[0054] Non-limiting examples of such compounds include those having
the following structures:
##STR00019##
[0055] as illustrated by the following examples:
##STR00020##
[0056] Compounds of interest include among others, compounds of
Formula III in which Ring D is a piperazine ring, substituted on
nitrogen with R.sup.d. Of particular current interest, are
compounds of this subclass in which R.sup.d is a substituted or
unsubstituted lower (i.e., 1-6 carbon) alkyl as illustrated by
N-methylpiperazine moieties in some of the foregoing examples.
[0057] Of special interest are compounds of formula II and formula
III in which Ring T is an optionally substituted
imidazo[1,2-a]pyridine, imidazo[1,2-b]pyridazine,
imidazo[1,2-a]pyrazine, pyrazolo[1,5-a]pyrimidine,
pyrazolo[1,5-a]pyridine, pyrazolo[1,5-c]pyrimidine, and
pyrazolo[1,5-a][1,3,5]triazine.
[0058] Also of interest are compounds of formula II and formula III
in which Rings A and B are aryl.
[0059] Another subclass of interest are compounds of Formulas II
and III, in which Ring T is any 6/5 fused heteroaryl ring system,
optionally substituted with up to three R.sup.e groups. Of
particular interest are compounds in which s is 0. Also of interest
are those in which s is 1-3 and at least one R.sup.e is halo, lower
alkyl, alkoxy, amino, --NH-alkyl, --C(O)NH-alkyl, --NHC(O)-alkyl,
--NHC(O)NH-alkyl, --NHC(NH)-alkyl, --NHC(NH)NH.sub.2,
--NH(CH.sub.2).sub.x-heteroaryl, --NH(CH.sub.2).sub.x-heterocycle,
--NH(CH.sub.2).sub.x-aryl or --(CH.sub.2).sub.xC(O)NH.sub.2, in
which x is 0, 1, 2 or 3 and "alkyl" includes straight (i.e.,
unbranched and acyclic), branched and cyclic alkyl groups and in
which aryl, heteroaryl, heterocyclyl rings are optionally
substituted. Illustrative, non limiting, examples of the foregoing
include compounds of formulas II and III in which Ring T is one of
the following:
##STR00021##
[0060] Illustrative, non limiting examples of this subclass include
compounds of formulas IIa, IIb, IIc, IIIa, IIIb and IIIc:
##STR00022## ##STR00023##
[0061] in which the previously defined variables, e.g., R.sup.a,
R.sup.b, R.sup.c, R.sup.d, R.sup.e, m and p, are as previously
defined, e.g., in part 1, and s is an integer from 0 to 4.
[0062] One subset of interest includes compounds of Formulas IIa,
IIb and IIc in which s is 0; m, p and v are 1; and, R.sup.a is
CH.sub.3, R.sup.b is CF.sub.3 and R.sup.C is methyl.
[0063] Another includes compounds of Formulas IIIa, IIIb, IIIc in
which s is 0; m and p are 1; and, R.sup.a is CH.sub.3, R.sup.b is
CF.sub.3 and R.sup.d is CH.sub.3 or CH.sub.2CH.sub.2OH.
[0064] Compounds of this invention of particular interest include
those with one or more of the following characteristics: [0065] a
molecular weight of less than 1000, preferably less than 750 and
more preferably less than 600 mass units (not including the weight
of any solvating or co-crystallizing species, of any counter-ion in
the case of a salt); or [0066] inhibitory activity against a wild
type or mutant (especially a clinically relevant mutant) kinase,
especially a Src family kinase such as Src, Yes, Lyn or Lck; a
VEGF-R such as VEGF-R1 (Fit-1), VEGF-R2 (kdr), or VEGF-R3; a
PDGF-R; an Abl kinase or another kinase of interest with an IC50
value of 1 .mu.M or less (as determined using any scientifically
acceptable kinase inhibition assay), preferably with an IC50 of 500
nM or better, and optimally with an IC50 value of 250 nM or better;
or [0067] inhibitory activity against a given kinase with an IC50
value at least 100-fold lower than their IC50 values for other
kinases of interest; or [0068] inhibitory activity against both Src
and kdr with a 1 .mu.M or better IC50 value against each; or [0069]
a cytotoxic or growth inhibitory effect on cancer cell lines
maintained in vitro, or in animal studies using a scientifically
acceptable cancer cell xenograft model, (especially preferred are
compounds of the invention which inhibit proliferation of cultured
K562 cells with a potency at least as great as Gleevec, preferably
with a potency at least twice that of Gleevec, and more preferably
with a potency at least 10 times that of Gleevec as determined by
comparative studies.).
[0070] Also provided is a composition comprising at least one
compound of the invention or a salt, hydrate or other solvate
thereof, and at least one pharmaceutically acceptable excipient or
additive. Such compositions can be administered to a subject in
need thereof to inhibit the growth, development and/or metastasis
of cancers, including solid tumors (e.g., breast, colon,
pancreatic, CNS and head and neck cancers, among others) and
various forms of leukemia, including leukemias and other cancers
which are resistant to other treatment, including those which are
resistant to treatment with Gleevec or another kinase inhibitor,
and generally for the treatment and prophylaxis of diseases or
undesirable conditions mediated by one or more kinases which are
inhibited by a compound of this invention.
[0071] The cancer treatment method of this invention involves
administering (as a monotherapy or in combination with one or more
other anti-cancer agents, one or more agents for ameliorating side
effects, radiation, etc) a therapeutically effective amount of a
compound of the invention to a human or animal in need of it in
order to inhibit, slow or reverse the growth, development or spread
of cancer, including solid tumors or other forms of cancer such as
leukemias, in the recipient. Such administration constitutes a
method for the treatment or prophylaxis of diseases mediated by one
or more kinases inhibited by one of the disclosed compounds or a
pharmaceutically acceptable derivative thereof. "Administration" of
a compound of this invention encompasses the delivery to a
recipient of a compound of the sort described herein, or a prodrug
or other pharmaceutically acceptable derivative thereof, using any
suitable formulation or route of administration, as discussed
herein. Typically the compound is administered one or more times
per month, often one or more times per week, e.g. daily, every
other day, 5 days/week, etc. Oral and intravenous administrations
are of particular current interest.
[0072] The phrase, "pharmaceutically acceptable derivative", as
used herein, denotes any pharmaceutically acceptable salt, ester,
or salt of such ester, of such compound, or any other adduct or
derivative which, upon administration to a patient, is capable of
providing (directly or indirectly) a compound as otherwise
described herein, or a metabolite or residue (MW>300) thereof.
Pharmaceutically acceptable derivatives thus include among others
pro-drugs. A pro-drug is a derivative of a compound, usually with
significantly reduced pharmacological activity, which contains an
additional moiety which is susceptible to removal in vivo yielding
the parent molecule as the pharmacologically active species. An
example of a pro-drug is an ester which is cleaved in vivo to yield
a compound of interest. Pro-drugs of a variety of compounds, and
materials and methods for derivatizing the parent compounds to
create the pro-drugs, are known and may be adapted to the present
invention.
[0073] Particularly favored derivatives and prodrugs of a parent
compound are those derivatives and prodrugs that increase the
bioavailability of the compound when administered to a mammal
(e.g., by permitting enhanced absorption into the blood following
oral administration) or which enhance delivery to a biological
compartment of interest (e.g., the brain or lymphatic system)
relative to the parent compound. Preferred prodrugs include
derivatives of a compound of this invention with enhanced aqueous
solubility or active transport through the gut membrane, relative
to the parent compound.
[0074] One important aspect of this invention is a method for
treating cancer in a subject in need thereof, which comprises
administering to the subject a treatment effective amount of a
composition containing a compound of this invention. Various
cancers which may be thus treated are noted elsewhere herein and
include, among others, cancers which are or have become resistant
to another anticancer agent such as Gleevec, Iressa, Tarceva or one
of the other agents noted herein. Treatment may be provided in
combination with one or more other cancer therapies, include
surgery, radiotherapy (e.g., gamma-radiation, neutron beam
radiotherapy, electron beam radiotherapy, proton therapy,
brachytherapy, and systemic radioactive isotopes, etc.), endocrine
therapy, biologic response modifiers (e.g., interferons,
interleukins, and tumor necrosis factor (TNF) to name a few),
hyperthermia, cryotherapy, agents to attenuate any adverse effects
(e.g., antiemetics), and other cancer chemotherapeutic drugs. The
other agent(s) may be administered using a formulation, route of
administration and dosing schedule the same or different from that
used with the compound of this invention.
[0075] Such other drugs include but not limited to one or more of
the following: an anti-cancer alkylating or intercalating agent
(e.g., mechlorethamine, chlorambucil, Cyclophosphamide, Melphalan,
and Ifosfamide); antimetabolite (e.g., Methotrexate); purine
antagonist or pyrimidine antagonist (e.g., 6-Mercaptopurine,
5-Fluorouracil, Cytarabile, and Gemcitabine); spindle poison (e.g.,
Vinblastine, Vincristine, Vinorelbine and Paclitaxel);
podophyllotoxin (e.g., Etoposide, Irinotecan, Topotecan);
antibiotic (e.g., Doxorubicin, Bleomycin and Mitomycin);
nitrosourea (e.g., Carmustine, Lomustine); inorganic ion (e.g.,
Cisplatin, Carboplatin, Oxaliplatin or oxiplatin); enzyme (e.g.,
Asparaginase); hormone (e.g., Tamoxifen, Leuprolide, Flutamide and
Megestrol); mTOR inhibitor (e.g., Sirolimus (rapamycin),
Temsirolimus (CCI779), Everolimus (RAD001), AP23573 or other
compounds disclosed in U.S. Pat. No. 7,091,213); proteasome
inhibitor (such as Velcade, another proteasome inhibitor (see e.g.,
WO 02/096933) or another NF-kB inhibitor, including, e.g., an IkK
inhibitor); other kinase inhibitors (e.g., an inhibitor of Src,
BRC/Abl, kdr, flt3, aurora-2, glycogen synthase kinase 3 ("GSK-3"),
EGF-R kinase (e.g., Iressa, Tarceva, etc.), VEGF-R kinase, PDGF-R
kinase, etc); an antibody, soluble receptor or other receptor
antagonist against a receptor or hormone implicated in a cancer
(including receptors such as EGFR, ErbB2, VEGFR, PDGFR, and IGF-R;
and agents such as Herceptin, Avastin, Erbitux, etc.); etc. For a
more comprehensive discussion of updated cancer therapies see,
http://www.nci.nih.gov/, a list of the FDA approved oncology drugs
at http://www.fda.gov/cder/cancer/druglistframe.htm, and The Merck
Manual, Seventeenth Ed. 1999, the entire contents of which are
hereby incorporated by reference. Examples of other therapeutic
agents are noted elsewhere herein and include among others,
Zyloprim, alemtuzmab, altretamine, amifostine, nastrozole,
antibodies against prostate-specific membrane antigen (such as
MLN-591, MLN591RL and MLN2704), arsenic trioxide, bexarotene,
bleomycin, busulfan, capecitabine, Gliadel Wafer, celecoxib,
chlorambucil, cisplatin-epinephrine gel, cladribine, cytarabine
liposomal, daunorubicin liposomal, daunorubicin, daunomycin,
dexrazoxane, docetaxel, doxorubicin, Elliott's B Solution,
epirubicin, estramustine, etoposide phosphate, etoposide,
exemestane, fludarabine, 5-FU, fulvestrant, gemcitabine,
gemtuzumab-ozogamicin, goserelin acetate, hydroxyurea, idarubicin,
idarubicin, Idamycin, ifosfamide, imatinib mesylate, irinotecan (or
other topoisomerase inhibitor, including antibodies such as MLN576
(XR11576)), letrozole, leucovorin, leucovorin levamisole, liposomal
daunorubicin, melphalan, L-PAM, mesna, methotrexate, methoxsalen,
mitomycin C, mitoxantrone, MLN518 or MLN608 (or other inhibitors of
the fit-3 receptor tyrosine kinase, PDFG-R or c-kit), itoxantrone,
paclitaxel, Pegademase, pentostatin, porfimer sodium, Rituximab
(RITUXAN.RTM.), talc, tamoxifen, temozolamide, teniposide, VM-26,
topotecan, toremifene, 2C4 (or other antibody which interferes with
HER.sup.2-mediated signaling), tretinoin, ATRA, valrubicin,
vinorelbine, or pamidronate, zoledronate or another
bisphosphonate.
[0076] This invention further comprises the preparation of a
compound of any of Formulas I, II, III, IIa, IIb, IIc, IIIa, IIIb,
IIIc or of any other of the compounds of this invention.
[0077] The invention also comprises the use of a compound of the
invention, or a pharmaceutically acceptable derivative thereof, in
the manufacture of a medicament for the treatment either acutely or
chronically of cancer (including leukemias and solid tumors,
primary or metastatic, including cancers such as noted elsewhere
herein and including cancers which are resistant or refractory to
one or more other therapies). The compounds of this invention are
useful in the manufacture of an anti-cancer medicament. The
compounds of the present invention are also useful in the
manufacture of a medicament to attenuate or prevent disorders
through inhibition of one or more kinases such as Src, kdr, abl.
etc.
[0078] Other disorders which may be treated with a compound of this
invention include metabolic disorders, inflammatory disorders and
osteoporosis and other bone disorders. In such cases the compound
of this invention may be used as a monotherapy or may be
administered in conjunction with administration of another drug for
the disorder, e.g., a bisphosphonate in the case of osteoporosis or
other bone-related illnesses.
[0079] This invention further encompasses a composition comprising
a compound of the invention, including a compound of any of the
described classes or subclasses, including those of any of the
formulas noted above, among others, preferably in a
therapeutically-effective amount, in association with a least one
pharmaceutically acceptable carrier, adjuvant or diluent.
[0080] Compounds of this invention are also useful as standards and
reagents for characterizing various kinases, especially but not
limited to kdr and Src family kinases, as well as for studying the
role of such kinases in biological and pathological phenomena; for
studying intracellular signal transduction pathways mediated by
such kinases, for the comparative evaluation of new kinase
inhibitors; and for studying various cancers in cell lines and
animal models.
3. Definitions
[0081] In reading this document, the following information and
definitions apply unless otherwise indicated. In addition, unless
otherwise indicated, all occurrences of a functional group are
independently chosen, as the reader is in some cases reminded by
the use of a slash mark or prime to indicate simply that the two
occurrences may be the same or different (e.g., R, R', R'', or Y,
Y', Y'' etc.).
[0082] The term "Alkyl" is intended to include linear (i.e.,
unbranched or acyclic), branched, cyclic, or polycyclic non
aromatic hydrocarbon groups, which are optionally substituted with
one or more functional groups. Unless otherwise specified, "alkyl"
groups contain one to eight, and preferably one to six carbon
atoms. C.sub.1-6 alkyl, is intended to include C.sub.1, C.sub.2,
C.sub.3, C.sub.4, C.sub.5, and C.sub.6 alkyl groups. Lower alkyl
refers to alkyl groups containing 1 to 6 carbon atoms. Examples of
Alkyl include, but are not limited to, methyl, ethyl, n-propyl,
isopropyl, cyclopropyl, butyl, isobutyl, sec-butyl, tert-butyl,
cyclobutyl, pentyl, isopentyl tert-pentyl, cyclopentyl, hexyl,
isohexyl, cyclohexyl, etc. Alkyl may be substituted or
unsubstituted. Illustrative substituted alkyl groups include, but
are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl,
2-fluoroethyl, 3-fluoropropyl, hydroxymethyl, 2-hydroxyethyl,
3-hydroxypropyl, benzyl, substituted benzyl, phenethyl, substituted
phenethyl, etc.
[0083] The term "Alkoxy" represent a subset of alkyl in which an
alkyl group as defined above with the indicated number of carbons
attached through an oxygen bridge. For example, "alkoxy" refers to
groups --O-alkyl, wherein the alkyl group contains 1 to 8 carbons
atoms of a linear, branched, cyclic configuration. Examples of
"alkoxy" include, but are not limited to, methoxy, ethoxy,
n-propoxy, i-propoxy, t-butoxy, n-butoxy, s-pentoxy and the
like.
[0084] "Haloalkyl" is intended to include both branched and linear
chain saturated hydrocarbon having one or more carbon substituted
with a Halogen. Examples of haloalkyl, include, but are not limited
to, trifluoromethyl, trichloromethyl, pentafluoroethyl and the
like.
[0085] The term "alkenyl" is intended to include hydrocarbon chains
of linear, branched, or cyclic configuration having one or more
unsaturated Carbon-carbon bonds that may occur in any stable point
along the chain or cycle. Unless otherwise specified, "alkenyl"
refers to groups usually having two to eight, often two to six
carbon atoms. For example, "alkenyl" may refer to prop-2-enyl,
but-2-enyl, but-3-enyl, 2-methylprop-2-enyl, hex-2-enyl,
hex-5-enyl, 2,3-dimethylbut-2-enyl, and the like. Furthermore,
alkenyl groups may be substituted or unsubstituted.
[0086] The term "alkynyl" is intended to include hydrocarbon chains
of either linear or branched configuration, having one or more
carbon-carbon triple bond that may occur in any stable point along
the chain. Unless otherwise specified, "alkynyl" groups refer
refers to groups having two to eight, preferably two to six
carbons. Examples of "alkynyl" include, but are not limited to
prop-2-ynyl, but-2-ynyl, but-3-ynyl, pent-2-ynyl,
3-methylpent-4-ynyl, hex-2-ynyl, hex-5-ynyl, etc. Furthermore,
alkynyl groups may be substituted or unsubstituted.
[0087] Cycloalkyl is a subset of alkyl and includes any stable
cyclic or polycyclic hydrocarbon groups of from 3 to 13 carbon
atoms, any of which is saturated. Examples of such cycloalkyl
include, but are not limited to cyclopropyl, norbornyl,
[2.2.2]bicyclooctane, [4.4.0]bicyclodecane, and the like, which, as
in the case of other alkyl moieties, may optionally be substituted.
The term "cycloalkyl" may be used interchangeably with the term
"carbocycle".
[0088] Cycloalkenyl is a subset of alkenyl and includes any stable
cyclic or polycyclic hydrocarbon groups of from 3 to 13 carbon
atoms, preferably from 5 to 8 carbon atoms, which contains one or
more unsaturated carbon-carbon double bonds that may occur in any
point along the cycle. Examples of such cycloalkenyl include, but
are not limited to cyclopentenyl, cyclohexenyl and the like.
[0089] Cycloalkynyl is a subset of alkynyl and includes any stable
cyclic or polycyclic hydrocarbon groups of from 5 to 13 carbon
atoms, which contains one or more unsaturated carbon-carbon triple
bonds that may occur in any point along the cycle. As in the case
of other alkenyl and alkynyl moieties, cycloalkenyl and
cycloalkynyl may optionally be substituted.
[0090] "Heterocycle", "heterocyclyl", or "heterocyclic" as used
herein refers to non-aromatic ring systems having five to fourteen
ring atoms, preferably five to ten, in which one or more ring
carbons, preferably one to four, are each replaced by a heteroatom
such as N, O, or S. Non-limiting examples of heterocyclic rings
include 3-1H-benzimidazol-2-one,
(1-substituted)-2-oxo-benzimidazol-3-yl, 2-tetrahydrofuranyl,
3-tetrahydrofuranyl, 2-tetrahydrothiophenyl,
3-tetrahydrothiophenyl, 2-morpholinyl, 3-morpholinyl,
4-morpholinyl, 2-thiomorpholinyl, 3-thiomorpholinyl,
4-thiomorpholinyl, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl,
1-piperazinyl, 2-piperazinyl, 1-piperidinyl, 2-piperidinyl,
3-piperidinyl, 4-piperidinyl, 4-thiazolidinyl, diazolonyl,
N-substituted diazolonyl, 1-phthalimidinyl, benzoxanyl,
benzopyrrolidinyl, benzopiperidinyl, benzoxolanyl, benzothiolanyl,
and benzothianyl. Also included within the scope of the term
"heterocyclyl" or "heterocyclic", as it is used herein, is a group
in which a non-aromatic heteroatom-containing ring is fused to one
or more aromatic or non-aromatic rings, such as in an indolinyl,
chromanyl, phenanthridinyl, or tetrahydroquinolinyl, where the
radical or point of attachment is on the non-aromatic
heteroatom-containing ring. The term "heterocycle", "heterocyclyl",
or "heterocyclic" whether saturated or partially unsaturated, also
refers to rings that are optionally substituted.
[0091] The term "aryl" used alone or as part of a larger moiety as
in "aralkyl", "aralkoxy", or "aryloxy-alkyl", refers to aromatic
ring groups having six to fourteen ring atoms, such as phenyl,
1-naphthyl, 2-naphthyl, 1-anthracyl and 2-anthracyl. An "aryl" ring
may contain one or more substituents. The term "aryl" may be used
interchangeably with the term "aryl ring". "Aryl" also includes
fused polycyclic aromatic ring systems in which an aromatic ring is
fused to one or more rings. Non-limiting examples of useful aryl
ring groups include phenyl, hydroxyphenyl, halophenyl,
alkoxyphenyl, dialkoxyphenyl, trialkoxyphenyl, alkylenedioxyphenyl,
naphthyl, phenanthryl, anthryl, phenanthro and the like, as well as
1-naphthyl, 2-naphthyl, 1-anthracyl and 2-anthracyl. Also included
within the scope of the term "aryl", as it is used herein, is a
group in which an aromatic ring is fused to one or more
non-aromatic rings, such as in a indanyl, phenanthridinyl, or
tetrahydronaphthyl, where the radical or point of attachment is on
the aromatic ring.
[0092] The term "heteroaryl" as used herein refers to stable
heterocyclic, and polyheterocyclic aromatic moieties having 5-14
ring atoms. Heteroaryl groups may be substituted or unsubstituted
and may comprise one or more rings. Examples of typical heteroaryl
rings include 5-membered monocyclic ring groups such as thienyl,
pyrrolyl, imidazolyl, pyrazolyl, furyl, isothiazolyl, furazanyl,
isoxazolyl, thiazolyl and the like; 6-membered monocyclic groups
such as pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl and
the like; and polycyclic heterocyclic ring groups such as
benzo[b]thienyl, naphtho[2,3-b]thienyl, thianthrenyl,
isobenzofuranyl, chromenyl, xanthenyl, phenoxathienyl, indolizinyl,
isoindolyl, indolyl, indazolyl, purinyl, isoquinolyl, quinolyl,
phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl,
benzothiazole, benzimidazole, tetrahydroquinoline cinnolinyl,
pteridinyl, carbazolyl, beta-carbolinyl, phenanthridinyl,
acridinyl, perimidinyl, phenanthrolinyl, phenazinyl, isothiazolyl,
phenothiazinyl, phenoxazinyl, and the like (see e.g. Katritzky,
Handbook of Heterocyclic Chemistry). Further specific examples of
heteroaryl rings include 2-furanyl, 3-furanyl, N-imidazolyl,
2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl,
4-isoxazolyl, 5-isoxazolyl, 2-oxadiazolyl, 5-oxadiazolyl,
2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 1-pyrrolyl, 2-pyrrolyl,
3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl,
4-pyrimidyl, 5-pyrimidyl, 3-pyridazinyl, 2-thiazolyl, 4-thiazolyl,
5-thiazolyl, 5-tetrazolyl, 2-triazolyl, 5-triazolyl, 2-thienyl,
3-thienyl, carbazolyl, benzimidazolyl, benzothienyl, benzofuranyl,
indolyl, quinolinyl, benzotriazolyl, benzothiazolyl, benzooxazolyl,
benzimidazolyl, isoquinolinyl, indolyl, isoindolyl, acridinyl, or
benzoisoxazolyl. Heteroaryl groups further include a group in which
a heteroaromatic ring is fused to one or more aromatic or
nonaromatic rings where the radical or point of attachment is on
the heteroaromatic ring. Examples include tetrahydroquinoline,
tetrahydroisoquinoline, and pyrido[3,4-d]pyrimidinyl,
imidazo[1,2-a]pyrimidyl, imidazo[1,2-a]pyrazinyl,
imidazo[1,2-a]pyiridinyl, imidazo[1,2-c]pyrimidyl,
pyrazolo[1,5-a][1,3,5]triazinyl, pyrazolo[1,5-c]pyrimidyl,
imidazo[1,2-b]pyridazinyl, imidazo[1,5-a]pyrimidyl,
pyrazolo[1,5-b][1,2,4]triazine, quinolyl, isoquinolyl, quinoxalyl,
imidazotriazinyl, pyrrolo[2,3-d]pyrimidyl, triazolopyrimidyl,
pyridopyrazinyl. The term "heteroaryl" also refers to rings that
are optionally substituted. The term "heteroaryl" may be used
interchangeably with the term "heteroaryl ring" or the term
"heteroaromatic".
[0093] An aryl group (including the aryl portion of an aralkyl,
aralkoxy, or aryloxyalkyl moiety and the like) or heteroaryl group
(including the heteroaryl portion of a heteroaralkyl or
heteroarylalkoxy moiety and the like) may contain one or more
substituents. Examples of suitable substituents on the unsaturated
carbon atom of an aryl or heteroaryl group include halogen (F, Cl,
Br or I), --CN, --R.sup.4, --OR.sup.2, --S(O).sub.rR.sup.2,
(wherein r is an integer of 0, 1 or 2), --SO.sub.2NR.sup.2R.sup.3,
--NR.sup.2R.sup.3, --(CO)YR.sup.2, --O(CO)YR.sup.2,
--NR.sup.2(CO)YR.sup.2, --S(CO)YR.sup.2,
--NR.sup.2C(.dbd.S)YR.sup.2, --OC(.dbd.S)YR.sup.2,
--C(.dbd.S)YR.sup.2, wherein each occurrence of Y is independently
--O--, --S--, --NR.sup.3--, or a chemical bond; --(CO)YR.sup.2 thus
encompasses --C(.dbd.O)R.sup.2, --C(.dbd.O)OR.sup.2 and
--C(.dbd.O)NR.sup.2R.sup.3. Additional substituents include
--YC(.dbd.NR.sup.3)Y'R.sup.2, --COCOR.sup.2,--COMCOR.sup.2 (where M
is a 1-6 carbon alkyl group),
--YP(.dbd.O)(YR.sup.4)(YR.sup.4)(including among others
--P(.dbd.O)(R.sup.4).sub.2), --Si(R.sup.2).sub.3, --NO.sub.2,
--NR.sup.2SO.sub.2R.sup.2 and --NR.sup.2SO.sub.2NR.sup.2R.sup.3. To
illustrate further, substituents in which Y is --NR.sup.3 thus
include among others, --NR.sup.3C(.dbd.O)R.sup.2,
--NR.sup.3C(.dbd.O)NR.sup.2R.sup.3, --NR.sup.3C(.dbd.O)OR.sup.2 and
--NR.sup.3C(.dbd.NH)NR.sup.2R.sup.3. R.sup.2 and R.sup.3
substituents at each occurrence are independently selected from
hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,
cycloalkynyl, aryl, heteroaryl, heterocyclyl, and R.sup.2 and
R.sup.3 (and R.sup.4) substituents may themselves be substituted or
unsubstituted. Examples of substituents allowed on R.sup.2, R.sup.3
and R.sup.4 include, among others amino, alkylamino, dialkylamino,
aminocarbonyl, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl,
carbocycle, heterocycle, alkylaminocarbonyl, dialkylaminocar-bonyl,
alkylaminocarbonyloxy, dialkylaminocarbonyloxy, nitro, cyano,
carboxy, alkoxycar-bonyl, alkylcarbonyl, hydroxy, alkoxy,
haloalkoxy groups. Additional illustrative examples include
protected OH (such as acyloxy), phenyl, substituted phenyl,
--O-phenyl, --O-(substituted) phenyl, -benzyl, substituted benzyl,
--O-phenethyl (i.e., --OCH.sub.2CH.sub.2C.sub.6H.sub.5),
--O-(substituted)phenethyl. Non-limiting illustrations of a
substituted R.sup.2, R.sup.3 or R.sup.4 moiety include haloalkyl
and trihaloalkyl, alkoxyalkyl, halophenyl, -M-heteroaryl,
-M-heterocycle, -M-aryl, -M-OR.sup.2, -M-SR.sup.2,
-M-NR.sup.2R.sup.3, -M-OC(O)NR.sup.2R.sup.3,
-M-C(.dbd.NR.sup.2)NR.sup.2R.sup.3, -M-C(.dbd.NR.sup.2)OR.sup.3,
-M-P(O)R.sup.2R.sup.3, Si(R.sup.2).sub.3, -M-NR.sup.2C(O)R.sup.3,
-M-NR.sup.2C(O)OR.sup.2, -M-C(O)R.sup.2, -M-C(.dbd.S)R.sup.2,
-M-C(.dbd.S)NR.sup.2R.sup.3, -M-C(O)NR.sup.2R.sup.3,
-M-C(O)NR.sup.2-M-NR.sup.2R.sup.3,
-M-NR.sup.2C(NR.sup.3)NR.sup.2R.sup.3,
-M-NR.sup.2C(S)NR.sup.2R.sup.3, -M-S(O).sub.2R3, -M-C(O)R.sup.3,
-M-OC(O)R.sup.3, -MC(O)SR.sup.2, -M-S(O).sub.2NR.sup.2R.sup.3,
--C(O)-M-C(O)R.sup.2, -MCO.sub.2R.sup.2,
-MC(.dbd.O)NR.sup.2R.sup.3, -M-C(.dbd.NH)NR.sup.2R.sup.3 and
-M-OC(.dbd.NH)NR.sup.2R.sup.3 (wherein M is a 1-6 carbon alkyl
group).
[0094] Some more specific examples include but are not limited to
chloromethyl, trichloromethyl, trifluoromethyl, methoxyethyl,
alkoxyphenyl, halophenyl, --CH.sub.2-aryl, --CH.sub.2-heterocycle,
--CH.sub.2C(O)NH.sub.2, --C(O)CH.sub.2N(CH.sub.3).sub.2,
--CH.sub.2CH.sub.2OH, --CH.sub.2OC(O)NH.sub.2,
--CH.sub.2CH.sub.2NH.sub.2, --CH.sub.2CH.sub.2CH.sub.2N Et.sub.2,
--CH.sub.2OCH.sub.3, --C(O)NH.sub.2,
--CH.sub.2CH.sub.2-heterocycle, --C(.dbd.S)CH.sub.3,
--C(.dbd.S)NH.sub.2, --C(.dbd.NH)NH.sub.2, --C(.dbd.NH)OEt,
--C(O)NH-cyclopropyl, C(O)NHCH.sub.2CH.sub.2-heterocycle,
--C(O)NHCH.sub.2CH.sub.2OCH.sub.3,
--C(O)CH.sub.2CH.sub.2NHCH.sub.3, --CH.sub.2CH.sub.2F,
--C(O)CH.sub.2-heterocycle, --CH.sub.2C(O)NHCH.sub.3,
--CH.sub.2CH.sub.2P(O)(CH.sub.3).sub.2, Si(CH.sub.3).sub.3 and the
like.
[0095] An aliphatic, i.e., alkyl, alkenyl, alkynyl, alkoxy,
haloalkyl, cycloalkyl, cycloalkenyl, cycloalkynyl or non-aromatic
heterocyclic, group may thus also contain one or more substituents.
Examples of suitable substituents on such groups include, but are
not limited to those listed above for the carbon atoms of an aryl
or heteroaryl group and in addition include the following
substituents for a saturated carbon atom: .dbd.O, .dbd.S, .dbd.NH,
.dbd.NNR.sup.2R.sup.3, .dbd.NNHC(O)R.sup.2,
.dbd.NNHCO.sub.2R.sup.2, or .dbd.NNHSO.sub.2R.sup.2, wherein
R.sup.2 at each occurrence is independently H, alkyl, alkenyl,
alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl,
heterocyclyl.
[0096] Illustrative examples of substituents on an aliphatic,
heteroaliphatic or heterocyclic group include amino, alkylamino,
dialkylamino, aminocarbonyl, halogen, alkyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylaminocarbonyloxy,
dialkylaminocarbonyloxy, alkoxy, nitro, --CN, carboxy,
alkoxycarbonyl, alkylcarbonyl, --OH, haloalkoxy or haloalkyl
groups.
[0097] Illustrative substituents on a nitrogen, e.g., in an aryl,
heteroaryl or non-aromatic heterocyclic ring, include R.sup.4,
--NR.sup.2R.sup.3, --C(.dbd.O)R.sup.2, --C(.dbd.O)OR.sup.2,
--C(.dbd.O)SR.sup.2, --C(.dbd.O)NR.sup.2R3,
--C(.dbd.NR.sup.2)NR.sup.2R3, --C(.dbd.NR.sup.2)OR.sup.2,
--C(.dbd.NR.sup.2)R.sup.3, --COCOR.sup.2,--COMCOR.sup.2, --CN,
--SO.sub.2R.sup.3, S(O)R.sup.3, --P(.dbd.O)(YR.sup.2)(YR.sup.2),
--NR.sup.2SO.sub.2R.sup.3 and --NR.sup.2SO.sub.2NR.sup.2R3, wherein
each occurrence of R.sup.2 and R.sup.3 is independently hydrogen,
alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl,
aryl, heteroary, heterocyclyl.
[0098] This invention encompasses only those combinations of
substituents and variables that result in a stable or chemically
feasible compound. A stable compound or chemically feasible
compound is one that has stability sufficient to permit its
preparation and detection. Preferred compounds of this invention
are sufficiently stable that they are not substantially altered
when kept at a temperature of 40.degree. C. or less, in the absence
of moisture or other chemically reactive conditions, for at least a
week.
[0099] Certain compounds of this invention may exist in tautomeric
forms, and this invention includes all such tautomeric forms of
those compounds unless otherwise specified.
[0100] Unless otherwise stated, structures depicted herein are also
meant to include all stereochemical forms of the structure; i.e.,
the R and S configurations for each asymmetric center. Thus, single
stereochemical isomers as well as enantiomeric and diastereomeric
mixtures of the present compounds are within the scope of the
invention. Thus, this invention encompasses each diasteriomer or
enantiomer substantially free of other isomers (>90%, and
preferably >95%, free from other stereoisomers on a molar basis)
as well as a mixture of such isomers.
[0101] Particular optical isomers can be obtained by resolution of
the racemic mixtures according to conventional processes, e.g., by
formation of diastereoisomeric salts, by treatment with an
optically active acid or base. Examples of appropriate acids are
tartaric, diacetyltartaric, dibenzoyltartaric, ditoluoyltartaric,
and camphorsulfonic acid and then separation of the mixture of
diastereoisomers by crystallization followed by liberation of the
optically active bases from these salts. A different process for
separation of optical isomers involves the use of a chiral
chromatography column optimally chosen to maximize the separation
of the enantiomers. Still another method involves synthesis of
covalent diastereoisomeric molecules by reacting compounds of the
invention with an optically pure acid in an activated form or an
optically pure isocyanate. The synthesized diastereoisomers can be
separated by conventional means such as chromatography,
distillation, crystallization or sublimation, and then hydrolyzed
to deliver the enantiomerically pure compound.
[0102] Optically active compounds of the invention can be obtained
by using active starting materials. These isomers may be in the
form of a free acid, a free base, an ester or a salt.
[0103] The compounds of this invention can exist in radiolabelled
form, i.e., said compounds may contain one or more atoms containing
an atomic mass or mass number different from the atomic mass or
mass number: ordinarily found in nature. Radioisotopes of hydrogen,
carbon, phosphorous, fluorine and chlorine include .sup.3H,
.sup.14C, .sup.32P, .sup.35S, .sup.43F and .sup.36Cl, respectively.
Compounds of this invention which contain those radioisotopes
and/or other radioisotopes of other atoms are within the scope of
this invention. Tritiated, i.e., .sup.3H, and carbon-14, i.e.,
.sup.14C, radioisotopes are particularly preferred for their ease
of preparation and detectability.
[0104] Radiolabelled compounds of this invention can generally be
prepared by methods well known to those skilled in the art.
Conveniently, such radiolabelled compounds can be prepared by
carrying out the procedures disclosed herein except substituting a
readily available radiolabelled reagent for a non-radiolabelled
reagent.
4. Synthetic Overview
[0105] The practitioner has a well-established literature of
heterocyclic and other relevant chemical transformations, recovery
and purification technologies to draw upon, in combination with the
information contained in the examples which follow, for guidance on
synthetic strategies, protecting groups, and other materials and
methods useful for the synthesis, recovery and characterization of
the compounds of this invention, including compounds containing the
various choices for the R.sup.t, R.sup.a, R.sup.b, R.sup.c,
R.sup.d, R.sup.e and Rings T, A, B, C and D. The following
references, and the references cited therein, may be of particular
interest: WO 01/27109, WO 02/066478, WO 02/30428, WO 02/080911, WO
02/080914, WO 2004/033453, WO 2004/035578, WO 2004/23972, WO
2005/105798, US 2003/0119842, US 2004/0023972, US 2004/0122044, US
2004/0142961, US 2005/0239822, U.S. Pat. No. 6,420,365 and U.S.
Pat. No. 6,703,404 are referring to the preparation of
imidazo[1,2-a]pyridines; WO 05/030218, WO 03/022850 are referring
to imidazo[1,2-a]pyrimidines; WO 05/047290, WO 03/089434, U.S. Pat.
No. 6,589,952 are referring to imidazopyrazines, WO 04/011466 and
U.S. Pat. No. 5,145,850 are referring to the preparation of
imidazo[1,2-b]pyridazines; and WO 05/070431, WO 96/35690, WO
04/089471 are referring to pyrazolo[1,5-a]pyrimidines.
[0106] Various synthetic approaches may be used to produce the
compounds described herein, including those approaches depicted
schematically below. The practitioner will appreciate that
protecting groups may be used in these approaches. "Protecting
groups", are moieties that are used to temporarily block chemical
reaction at a potentially reactive site (e.g., an amine, hydroxy,
thiol, aldehyde, etc.) so that a reaction can be carried out
selectively at another site in a multifunctional compound. In
preferred embodiments, a protecting group reacts selectively in
good yield to give a protected substrate that is suitable for the
planned reactions; the protecting group should be selectively
removable in good yield by readily available, preferably nontoxic
reagents that do not unduly attack the other functional groups
present; the protecting group preferably forms an readily separable
derivative (more preferably without the generation of new
stereogenic centers); and the protecting group preferably has a
minimum of additional functionality to avoid the complication of
further sites of reaction. A wide variety of protecting groups and
strategies, reagents and conditions for deploying and removing them
are known in the art. See, e.g., "Protective Groups in Organic
Synthesis" Third Ed. Greene, T. W. and Wuts, P. G., Eds., John
Wiley & Sons, New York: 1999. For additional background
information on protecting group methodologies (materials, methods
and strategies for protection and deprotection) and other synthetic
chemistry transformations useful in producing the compounds
described herein, see in R. Larock, Comprehensive organic
Transformations, VCH Publishers (1989); T. W. Greene and P. G. M.
Wuts, Protective Groups in Organic Synthesis, 3rd. Ed., John Wiley
and Sons (1999); L. Fieser and M. Fieser, Fieser and Fieser's
Reagents for Organic Synthesis, John Wiley and Sons (1994); and L.
Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John
Wiley and Sons (1995). The entire contents of these references are
hereby incorporated by reference.
[0107] Also, one may chose reagents enriched for a desired isotope,
e.g. deuterium in place of hydrogen, to create compounds of this
invention containing such isotope(s). Compounds containing
deuterium in place of hydrogen in one or more locations, or
containing various isotopes of C, N, P and O, are encompassed by
this invention and may be used, for instance, for studying
metabolism and/or tissue distribution of the compounds or to alter
the rate or path of metabolism or other aspects of biological
functioning.
[0108] The compounds of the this invention can be synthesized using
the methods described below, together with synthetic methods known
in the art of synthetic organic chemistry, or by a variation
thereon as appreciated by those skilled in the art. Preferred
methods include, but are not limited to those described below. The
reactions are preformed in a solvent appropriate to the reagents
and materials employed and suitable for the transformation being
effected. It will be understood by those skilled in the art of
organic synthesis that the functionality present on the molecule
should be consistent the transformations proposed. This will
sometimes required some judgment to modify the order of the
synthetic steps or to select one particular process scheme over
anotherin order to obtain a desired compound of the invention.
[0109] A compound of the present invention could be prepared as
outlined in Scheme I to Scheme XIX and via standard methods known
to those skilled in the art.
[0110] A palladium catalyzed Sonogashira coupling reaction is used
to link the `top` Ring T to the `bottom` [Ring A]-[L.sup.1]-[Ring
B] moiety as illustrated in Scheme I and II. In Scheme I the
Sonogashira coupling reaction is performed with an acetylenic `top`
Ring T and a `[Ring A]-[L.sup.1]-[Ring B] moiety which has been
activated by the presence of a reactive group, W, which is an I, a
Br or another reactive group permitting the desired coupling
reaction. The variables in the W-[Ring A]-[L.sup.1]-[Ring B] are as
defined previously, Rings A and B being substituted with permitted
R.sup.a and R.sup.b groups, respectively.
##STR00024##
[0111] An alternative coupling reaction is described in Scheme II,
in which Ring T is "activated" by the presence of a reactive group
W (such as I or Br) and is coupled to the `bottom` acetylenic
[RingA]-L.sup.1-[RingB] under similar Palladium catalyzed coupling
conditions.
##STR00025##
[0112] The Sonogashira coupling conditions described in Scheme I
and II are applicable to all bicyclic heteroaryl Ring T's and
useful to synthesize all compounds of this invention.
[0113] Several illustrative overall synthetic approaches to the
preparation of the acetylenic Ring T moieties, based on known
transformations, are illustrated below in Schemes III to VIII:
##STR00026##
##STR00027##
##STR00028##
##STR00029##
##STR00030##
##STR00031##
[0114] For the coupling step, see Malleron, J-L., Fiaud, J-C.,
Legros, J-Y. Handbook of Palladium Catalyzed Organic Reactions. San
Diego: academic Press, 1997.
[0115] As one of ordinary skill in the art would recognize, these
methods for the preparation of various substituted acetylenic Ring
T groups, are widely applicable to various other fused bicyclic
ring systems not shown.
[0116] Schemes IX to XIII below depict the synthesis of compounds
of the formula W-[Ring A]-[L.sup.1]-[Ring B] which are useful as
intermediates in the coupling reaction described in Schemes I and
II.
[0117] It should be apparent that intermediates of the formula:
##STR00032##
are of particular interest as their coupling reaction with the
`top` heteroaryl rings produces compounds of the present invention.
The variable groups A, L.sup.1 and B are as previously defined and
are optionally substituted as described herein, and W is I or an
alternative reactive group permitting the desired coupling
reaction.
[0118] Illustrative such intermediates include among others those
of those following structures:
##STR00033## ##STR00034##
[0119] wherein the previously defined variables, e.g., R.sup.a,
R.sup.b, R.sup.c and R.sup.d, are as previously defined. For
instance, R.sup.a in some embodiments is chosen from F or alkyl,
e.g., Me, among others, and Rb in some embodiments is chosen from
Cl, F, Me, t-butyl, --CF.sub.3 or --OCF.sub.3 among others. Those
and other compounds of the formula W-[Ring A]-[L.sup.1]-[Ring B]
with the various permitted substituents are useful for preparing
the corresponding compounds of the invention as are defined in the
various formulae, classes and subclasses disclosed herein.
[0120] Some illustrative synthetic routes for the preparation of
reagents and representative intermediates are presented below:
[0121] Scheme IX describes an illustrative synthesis of W-[Ring
A]-[L.sup.1]-[Ring B] in which Rings A and B are phenyl and L.sup.1
is NHC(O).
##STR00035##
[0122] Scheme X depicts the synthesis of a variant of the foregoing
in which Ring B is a 2-pyridine and L.sup.1 is C(O)NH (i.e., in the
other orientation).
##STR00036##
[0123] Schemes XI and XII, below, illustrate the synthesis of
W-[Ring A]-[L.sup.1]-[Ring B] in which Rings A and B are phenyl and
Ring C is a heteroaryl ring. These intermediates are useful for
making compounds of formula II.
[0124] More specifically, Scheme XI describes the preparation of
intermediates in which Ring C is an imidazole ring.
##STR00037##
[0125] Scheme XII describes the preparation of intermediates in
which Ring C is a pyrrole or an oxazole ring.
##STR00038##
[0126] Scheme XIII illustrates the synthesis of W-[Ring
A]-[L.sup.1]-[Ring B] in which Rings A and B are phenyl and an
R.sup.b substituent is -L.sup.2-[Ring D]. These intermediates are
useful for making compounds of formula III in which Ring D is a 5
or 6-membered heterocycle, containing one or two heteroatoms.
##STR00039##
[0127] In this scheme, non limiting examples of substituents
R.sup.b on Ring B are halo, e.g., Cl; lower alkyl groups, e.g.,
isopropyl; and substituted lower alkyl groups, e.g. --CF.sub.3; and
non limiting examples of Ring D are N,N-dimethylpyrrolidine,
N-(2-hydroxyethyl)piperazine, and N-methylpiperazine.
[0128] Intermediates W-[Ring A]-[L.sup.1]-[Ring B], such as those
presented in the various synthetic schemes above, can be reacted
with an acetylenic Ring T using the Sonogashira coupling conditions
described in the general Scheme I.
[0129] An example is depicted below in Scheme XIV, in which Ring T
moiety can be further derivatized after the Sonogashira coupling
step, to generate various interesting substituted analogs of this
invention.
##STR00040##
[0130] Alternatively, the W-[Ring A]-[L.sup.1]-[Ring B] can be
reacted under Sonogashira conditions with trimethylsilylacetylene,
prior to the coupling with an iodo- or a bromo-activated Ring T as
otherwise described in the general Scheme II.
[0131] An example is depicted in Scheme XV:
##STR00041##
[0132] In other embodiments, the steps can be carried out in a
different order. For example, the Sonogashira Coupling reaction can
be used to Ring T to Ring A prior to linking that portion to Ring B
and/or [Ring B]-[L.sup.2]-[Ring D] and/or [Ring B]-[Ring C] as
shown in Scheme XVI.
##STR00042##
[0133] In a non-limiting example in which Ring A and Ring B are
phenyl and L.sup.1 is CONH, Scheme XVII describes Sonogashira
Coupling of an acetylenic Ring T with 3-iodo-4-methylbenzoic acid
(a Ring A moiety) to generate a [Ring T]-[Ring A] intermediate
which then undergoes an amide coupling with an optionally
substituted Ring B moiety:
##STR00043##
[0134] This approach is illustrated in Scheme XVIII which depicts
the coupling of an acetylenic Ring T (i.e.,
3-ethynylimidazo[1,2-b]pyridazine) with a substituted W-[Ring A]
(i.e., 3-iodo-4-methylbenzoic acid), followed by an amide coupling
of the resultant [Ring T]-[Ring A]-COOH intermediate with a
H.sub.2N-[Ring B]-L2-[Ring C] moiety (i.e.,
4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethylaniline):
##STR00044##
[0135] Alternatively, as another illustration of the practitioner's
range of assembly options, the 3-iodo-4-methylbenzoic acid Ring A
intermediate can be reacted in a Sonogashira reaction with
trimethylsilylacetylene, which after silyl deprotection, can a
second Sonogashira coupling reaction with an activated Ring T as
illustrated in Scheme XIX.
##STR00045##
[0136] With synthetic approaches such as the foregoing, combined
with the examples which follow, additional information provided
herein and conventional methods and materials, the practitioner can
prepare the full range of compounds disclosed herein.
5. Uses, Formulations, Administration
[0137] Pharmaceutical Uses; indications
[0138] This invention provides compounds having biological
properties which make them of interest for treating or
amerliorating diseases in which kinases may be involved, symptoms
of such disease, or the effect of other physiological events
mediated by kinases. For instance, a number of compounds of this
invention have been shown to inhibit tyrosine kinase activity of
Src and abl, among other tyrosine kinases which are believed to
mediate the growth, development and/or metastasis of cancer. A
number of compounds of the invention have also been found to
possess potent in vitro activity against cancer cell lines,
including among others K-562 leukemia cells. Observed potencies
have been as much as 10-fold more powerful than Gleevec in
conventional antiproliferation assays with K562 cells.
[0139] Such compounds are thus of interest for the treatment of
cancers, including both primary and metastatic cancers, including
solid tumors as well as lymphomas and leukemias (including CML, AML
and ALL), and including cancers which are resistant to other
therapies, including other therapies involving the administration
of kinase inhibitors such as Gleevec, Tarceva or Iressa.
[0140] Such cancers include, among others, cancers of the breast,
cervix, colon and rectum, lung, ovaries, pancreas, prostate, head
and neck, gastrointestinal stroma, as well as diseases such as
melanoma, multiple myeloma, non-Hodgkin's lymphoma, melanoma,
gastric cancers and leukemias (e.g., myeloid, lymphocytic,
myelocytic and lymphoblastic leukemias) including cases which are
resistant to one or more other therapies, including among others,
Gleevec, Tarceva or Iressa.
[0141] Resistance to various anticancer agents can arise from one
or more mutations in a mediator or effector of the cancer (e.g.,
mutation in a kinase such as Src or Abl) which correlate with
alteration in the protein's drug binding properties, phosphate
binding properties, protein binding properties, autoregulation or
other characteristics. For example, in the case of BCR-Abl, the
kinase associated with chronic myeloid leukemia, resistance to
Gleevec has been mapped to a variety of BCR/Abl mutations which are
linked to a variety of functional consequences, including among
others, steric hindrance of drug occupancy at the kinase's active
site, alteration in deformability of the phosphate binding P loop,
effects on the conformation of the activation loop surrounding the
active site, and others. See e.g. Shah et al, 2002, Cancer Cell 2,
117-125 and Azam et al, 2003, Cell 112, 831-843 and references
cited therein for representative examples of such mutations in
Bcr/Abl which correlate with drug resistance. See also the
following references for additional background information on
BCR/Abl, its mechanistic role in CML and drug-resistance-conferring
mechanisms and mutations: Kurzrock et al., Philadelphia
chromosome-positive leukemias: from basic mechanisms to molecular
therapeutics, Ann Intern Med. 2003 May 20; 138(10):819-30; O'Dwyer
et al., Demonstration of Philadelphia chromosome negative abnormal
clones in patients with chronic myelogenous leukemia during major
cytogenetic responses induced by imatinib mesylate. Leukemia. 2003
March; 17(3):481-7; Hochhaus et al., Molecular and chromosomal
mechanisms of resistance to imatinib (ST1571) therapy, Leukemia.
2002 November; 16(11):2190-6; O'Dwyer et al., The impact of clonal
evolution on response to imatinib mesylate (ST1571) in accelerated
phase CML. Blood. 2002 Sep. 1; 100(5):1628-33; Braziel et al.,
Hematopathologic and cytogenetic findings in imatinib
mesylate-treated chronic myelogenous leukemia patients: 14 months'
experience. Blood. 2002 Jul. 15; 100(2):435-41; Corbin et al.,
Analysis of the structural basis of specificity of inhibition of
the Abl kinase by ST1571. J Biol Chem. 2002 Aug. 30;
277(35):32214-9; Wertheim et al., BCR-ABL-induced adhesion defects
are tyrosine kinase-independent. Blood. 2002 Jun. 1;
99(11):4122-30; Kantarjian et al., Hematologic and cytogenetic
responses to imatinib mesylate in chronic myelogenous leukemia, N
Engl J Med. 2002 Feb. 28; 346(9):645-52. Erratum in: N Engl J Med
2002 Jun. 13; 346(24):1923; Hochhaus et al., Roots of clinical
resistance to STI-571 cancer therapy. Science. 2001 Sep. 21;
293(5538):2163; Druker et al., Activity of a specific inhibitor of
the BCR-ABL tyrosine kinase in the blast crisis of chronic myeloid
leukemia and acute lymphoblastic leukemia with the Philadelphia
chromosome. N Engl J Med. 2001 Apr. 5; 344(14):1038-42. Erratum in:
N Engl J Med 2001 Jul. 19; 345(3):232; Mauro et al., Chronic
myelogenous leukemia. Curr Opin Oncol. 2001 January; 13(1):3-7.
Review; Kolibaba et al., CRKL binding to BCR-ABL and BCR-ABL
transformation. Leuk Lymphoma. 1999 March; 33(1-2):119-26; Bhat et
al., Interactions of p62(dok) with p210(bcr-abl) and
Bcr-Abl-associated proteins. J Biol Chem. 1998 Nov. 27;
273(48):32360-8; Senechal et al., Structural requirements for
function of the Crkl adapter protein in fibroblasts and
hematopoietic cells. Mol Cell Biol. 1998 September; 18(9):5082-90;
Kolibaba et al., Protein tyrosine kinases and cancer. Biochim
Biophys Acta. 1997 Dec. 9; 1333(3):F217-48. Review; Heaney et al.,
Direct binding of CRKL to BCR-ABL is not required for BCR-ABL
transformation. Blood. 1997 Jan. 1; 89(1):297-306; Hallek et al.,
Interaction of the receptor tyrosine kinase p145c-kit with the
p210bcr/abl kinase in myeloid cells. Br J Haematol. 1996 July;
94(1):5-16; Oda et al., The SH2 domain of ABL is not required for
factor-independent growth induced by BCR-ABL in a murine myeloid
cell line. Leukemia. 1995 February; 9(2):295-301; Carlesso et al.,
Use of a temperature-sensitive mutant to define the biological
effects of the p210BCR-ABL tyrosine kinase on proliferation of a
factor-dependent murine myeloid cell line. Oncogene. 1994 January;
9(1):149-56.
[0142] Again, we contemplate that compounds of this invention, both
as monotherapies and in combination therapies, will be useful
against leukemias and other cancers, including those which are
resistant in whole or part to other anticancer agents, specifically
including Gleevec and other kinase inhibitors, and specifically
including leukemias involving one or more mutations in BCR/Abl,
within or outside the kinase domain, including but not limited to
those noted in any of the foregoing publications. See in particular
Azam et al. and references cited therein for examples of such
mutations in BCR/Abl, including, among others, mutations in the
drug binding cleft, the phosphate binding P loop, the activation
loop, the conserved VAVK of the kinase beta-3 sheet, the catalytic
alpha-1 helix of the small N lobe, the long alpha-3 helix within
the large C lobe, and the region within the C lobe downstream of
the activation loop.
[0143] Pharmaceutical Methods
[0144] The method of the invention comprises administering to a
subject in need thereof a therapeutically effective amount of a
compound of the invention.
[0145] A "therapeutically effective amount" is that amount
effective for detectable killing or inhibition of the growth or
spread of cancer cells; the size or number of tumors; or other
measure of the level, stage, progression or severity of the cancer.
The exact amount required will vary from subject to subject,
depending on the species, age, and general condition of the
subject, the severity of the disease, the particular anticancer
agent, its mode of administration, combination treatment with other
therapies, and the like.
[0146] The compound, or a composition containing the compound, may
be administered using any amount and any route of administration
effective for killing or inhibiting the growth of tumors or other
forms of cancer.
[0147] The anticancer compounds of the invention are preferably
formulated in dosage unit form for ease of administration and
uniformity of dosage. The expression "dosage unit form" as used
herein refers to a physically discrete unit of anticancer agent
appropriate for the patient to be treated. As is normally the case,
the total daily usage of the compounds and compositions of the
present invention will be decided by the attending physician using
routine reliance upon sound medical judgment. The specific
therapeutically effective dose level for any particular patient or
organism will depend upon a variety of factors including the
disorder being treated; the severity of the disorder; the potency
of the specific compound employed; the specific composition
employed; the age, body weight, general health, sex and diet of the
patient; the route and schedule of administration; the rate of
metabolism and/or excretion of the compound; the duration of the
treatment; drugs used in combination or coincident with
administration of the compound of this invention; and like factors
well known in the medical arts.
[0148] Furthermore, after formulation with an appropriate
pharmaceutically acceptable carrier in a desired dosage, the
compositions of this invention can be administered to humans and
other animals orally, rectally, parenterally, intracisternally,
intravaginally, intraperitoneally, topically (as by transdermal
patch, powders, ointments, or drops), sublingually, bucally, as an
oral or nasal spray, or the like.
[0149] The effective systemic dose of the compound will typically
be in the range of 0.01 to 500 mg of compound per kg of patient
body weight, preferably 0.1 to 125 mg/kg, and in some cases 1 to 25
mg/kg, administered in single or multiple doses. Generally, the
compound may be administered to patients in need of such treatment
in a daily dose range of about 50 to about 2000 mg per patient.
Administration may be once or multiple times daily, weekly (or at
some other multiple-day interval) or on an intermittent schedule.
For example, the compound may be administered one or more times per
day on a weekly basis (e.g. every Monday) indefinitely or for a
period of weeks, e.g. 4-10 weeks. Alternatively, it may be
administered daily for a period of days (e.g. 2-10 days) followed
by a period of days (e.g. 1-30 days) without administration of the
compound, with that cycle repeated indefinitely or for a given
number of repetitions, e.g. 4-10 cycles. As an example, a compound
of the invention may be administered daily for 5 days, then
discontinued for 9 days, then administered daily for another 5 day
period, then discontinued for 9 days, and so on, repeating the
cycle indefinitely, or for a total of 4-10 times.
[0150] The amount of compound which will be effective in the
treatment or prevention of a particular disorder or condition will
depend in part on well known factors affecting drug dosage. In
addition, in vitro or in vivo assays may optionally be employed to
help identify optimal dosage ranges. A rough guide to effective
doses may be extrapolated from dose-response curves derived from in
vitro or animal model test systems. The precise dosage level should
be determined by the attending physician or other health care
provider and will depend upon well known factors, including route
of administration, and the age, body weight, sex and general health
of the individual; the nature, severity and clinical stage of the
disease; the use (or not) of concomitant therapies; and the nature
and extent of genetic engineering of cells in the patient.
[0151] When administered for the treatment or inhibition of a
particular disease state or disorder, the effective dosage of the
compound of this invention may vary depending upon the particular
compound utilized, the mode of administration, the condition, and
severity thereof, of the condition being treated, as well as the
various physical factors related to the individual being treated.
In many cases, satisfactory results may be obtained when the
compound is administered in a daily dosage of from about 0.01
mg/kg-500 mg/kg, preferably between 0.1 and 125 mg/kg, and more
preferably between 1 and 25 mg/kg. The projected daily dosages are
expected to vary with route of administration. Thus, parenteral
dosing will often be at levels of roughly 10% to 20% of oral dosing
levels.
[0152] When the compound of this invention is used as part of a
combination regimen, dosages of each of the components of the
combination are administered during a desired treatment period. The
components of the combination may administered at the same time;
either as a unitary dosage form containing both components, or as
separate dosage units; the components of the combination can also
be administered at different times during a treatment period, or
one may be administered as a pretreatment for the other.
[0153] Regarding the Compounds
[0154] Compounds of present invention can exist in free form for
treatment, or where appropriate, as a pharmaceutically acceptable
salt or other derivative. As used herein, the term
"pharmaceutically acceptable salt" refers to those salts which are,
within the scope of sound medical judgment, suitable for use in
contact with the tissues of humans and lower animals without undue
toxicity, irritation, allergic response and the like, and are
commensurate with a reasonable benefit/risk ratio. Pharmaceutically
acceptable salts of amines, carboxylic acids, phosphonates and
other types of compounds, are well known in the art. For example,
S. M. Berge, et al. describe pharmaceutically acceptable salts in
detail in J. Pharmaceutical Sciences, 66: 1-19 (1977), incorporated
herein by reference. The salts can be prepared in situ during the
isolation and purification of the compounds of the invention, or
separately by reacting the free base or free acid of a compound of
the invention with a suitable base or acid, respectively. Examples
of pharmaceutically acceptable, nontoxic acid addition salts are
salts of an amino group formed with inorganic acids such as
hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid
and perchloric acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid
or malonic acid or by using other methods used in the art such as
ion exchange. Other pharmaceutically acceptable salts include
adipate, alginate, ascorbate, aspartate, benzenesulfonate,
benzoate, bisulfate, borate, butyrate, camphorate,
camphorsulfonate, citrate, cyclopentanepropionate, digluconate,
dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate,
glycerophosphate, gluconate, hernisulfate, heptanoate, hexanoate,
hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate,
laurate, lauryl sulfate, malate, maleate, malonate,
methane-sulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,
oleate, oxalate, palmitate, pamoate, pectinate, persulfate,
3-phenylpropionate, phosphate, picrate, pivalate, propionate,
stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like.
Representative alkali or alkaline earth metal salts include sodium,
lithium, potassium, calcium, magnesium, and the like. Further
pharmaceutically acceptable salts include, when appropriate,
nontoxic ammonium, quaternary ammonium, and amine cations formed
using counterions such as halide, hydroxide, carboxylate, sulfate,
phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate.
[0155] Additionally, as used herein, the term "pharmaceutically
acceptable ester" refers preferably to esters which hydrolyze in
vivo and include those that break down readily in the human body to
leave the parent compound or a salt thereof. Suitable ester groups
include, for example, those derived from pharmaceutically
acceptable aliphatic carboxylic acids, particularly alkanoic,
alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl
or alkenyl moiety advantageously has not more than 6 carbon atoms.
Examples of particular esters include formates, acetates,
propionates, butyrates, acrylates and ethylsuccinates. Obviously,
esters can be formed with a hydroxyl or carboxylic acid group of
the compound of the invention.
[0156] Furthermore, the term "pharmaceutically acceptable prodrugs"
as used herein refers to those prodrugs of the compounds of the
present invention which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of humans
and lower animals with undue toxicity, irritation, allergic
response, and the like, commensurate with a reasonable benefit/risk
ratio, and effective for their intended use, as well as the
zwitterionic forms, where possible, of the compounds of the
invention. The term "prodrug" refers to compounds that are
transformed in vivo to yield the parent compound of the above
formula, for example by hydrolysis in blood. See, e.g., T. Higuchi
and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the
A.C.S. Symposium Series, and Edward B. Roche, ed., Bioreversible
Carriers in Drug Design, American Pharmaceutical Assocn. and
Pergamon Press, 1987, both of which are incorporated herein by
reference.
[0157] Compositions
[0158] Compositions are provided which comprise any one of the
compounds described herein (or a prodrug, pharmaceutically
acceptable salt or other pharmaceutically acceptable derivative
thereof), and one or more pharmaceutically acceptable carriers or
excipients. These compositions optionally further comprise one or
more additional therapeutic agents. Alternatively, a compound of
this invention may be administered to a patient in need thereof in
combination with the administration of one or more other
therapeutic regimens (e.g. Gleevec or other kinase inhibitors,
interferon, bone marrow transplant, farnesyl transferase
inhibitors, bisphosphonates, thalidomide, cancer vaccines, hormonal
therapy, antibodies, radiation, etc). For example, additional
therapeutic agents for conjoint administration or inclusion in a
pharmaceutical composition with a compound of this invention may be
another one or more anticancer agents.
[0159] As described herein, the compositions of the present
invention comprise a compound of the invention together with a
pharmaceutically acceptable carrier, which, as used herein,
includes any and all solvents, diluents, or other vehicle,
dispersion or suspension aids, surface active agents, isotonic
agents, thickening or emulsifying agents, preservatives, solid
binders, lubricants and the like, as suited to the particular
dosage form desired. Remington's Pharmaceutical Sciences, Fifteenth
Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1975)
discloses various carriers used in formulating pharmaceutical
compositions and known techniques for the preparation thereof.
Except insofar as any conventional carrier medium is incompatible
with the compounds of the invention, such as by producing any
undesirable biological effect or otherwise interacting in a
deleterious manner with any other component(s) of the
pharmaceutical composition, its use is contemplated to be within
the scope of this invention. Some examples of materials which can
serve as pharmaceutically acceptable carriers include, but are not
limited to, sugars such as lactose, glucose and sucrose; starches
such as corn starch and potato starch; cellulose and its
derivatives such as sodium carboxymethyl cellulose, ethyl cellulose
and cellulose acetate; powdered tragacanth; malt; gelatin; talc;
excipients such as cocoa butter and suppository waxes; oils such as
peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil;
corn oil and soybean oil; glycols; such a propylene glycol; esters
such as ethyl oleate and ethyl laurate; agar; buffering agents such
as magnesium hydroxide and aluminum hydroxide; alginic acid;
pyrogen-free water; isotonic saline; Ringer's solution; ethyl
alcohol, and phosphate buffer solutions, as well as other non-toxic
compatible lubricants such as sodium lauryl sulfate and magnesium
stearate, as well as coloring agents, releasing agents, coating
agents, sweetening, flavoring and perfuming agents, preservatives
and antioxidants can also be present in the composition.
[0160] Formulations
[0161] This invention also encompasses a class of compositions
comprising the active compounds of this invention in association
with one or more pharmaceutically-acceptable carriers and/or
diluents and/or adjuvants (collectively referred to herein as
"carrier" materials) and, if desired, other active ingredients. The
active compounds of the present invention may be administered by
any suitable route, preferably in the form of a pharmaceutical
composition adapted to such a route, and in a dose effective for
the treatment intended. The compounds and compositions of the
present invention may, for example, be administered orally,
mucosally, topically, rectally, pulmonarily such as by inhalation
spray, or parentally including intravascularly, intravenously,
intraperitoneally, subcutaneously, intramuscularly, intrasternally
and infusion techniques, in dosage unit formulations containing
conventional pharmaceutically acceptable carriers, adjuvants, and
vehicles.
[0162] The pharmaceutically active compounds of this invention can
be processed in accordance with conventional methods of pharmacy to
produce medicinal agents for administration to patients, including
humans and other mammals.
[0163] For oral administration, the pharmaceutical composition may
be in the form of, for example, a tablet, capsule, suspension or
liquid. The pharmaceutical composition is preferably made in the
form of a dosage unit containing a particular amount of the active
ingredient.
[0164] Examples of such dosage units are tablets or capsules. For
example, these may contain an amount of active ingredient from
about I to 2000 mg, preferably from about I to 500 mg, more
commonly from about 5 to 200 mg. A suitable daily dose for a human
or other mammal may vary depending on the condition of the patient
and other factors, but, once again, can be determined using routine
methods.
[0165] The amount of compounds which are administered and the
dosage regimen for treating a disease condition with the compounds
and/or compositions of this invention depends on a variety of
factors, including the age, weight, sex and medical condition of
the subject, the type of disease, the severity of the disease, the
route and frequency of administration, and the particular compound
employed. Thus, the dosage regimen may vary widely, but can be
determined routinely using standard methods. A typical daily dose
is in the range of 0.01 to 500 mg of compound per kg body weight,
preferably between 0.1 and 125 mg/kg body weight and in some cases
between 1 and 25 mg/kg body weight. As mentioned previously, the
daily dose can be given in one administration or may be divided
between 2, 3, 4 or more administrations.
[0166] For therapeutic purposes, the active compounds of this
invention are ordinarily combined with one or more adjuvants,
excipients or carriers appropriate to the indicated route of
administration. If administered per os, the compounds may be
admixed with lactose, sucrose, starch powder, cellulose esters of
alkanoic acids, cellulose alkyl esters, talc, stearic acid,
magnesium stearate, magnesium oxide, sodium and calcium salts of
phosphoric and sulfuric acids, gelatin, acacia gum, sodium
alginate, polyvinylpyrrolidone, and/or polyvinyl alcohol, and then
tableted or encapsulated for convenient administration. Such
capsules or tablets may contain a controlled-release formulation as
may be provided in a dispersion of active compound in
hydroxypropylmethyl cellulose. In the case of skin conditions, it
may be preferable to apply a topical preparation of compounds of
this invention to the affected area two to four times a day.
Formulations suitable for topical administration include liquid or
semi-liquid preparations suitable for penetration through the skin
(e.g., liniments, lotions, ointments, creams, or pastes) and drops
suitable for administration to the eye, ear, or nose. A suitable
topical dose of active ingredient of a compound of the invention is
0.1 mg to 150 mg administered one to four, preferably one or two
times daily. For topical administration, the active ingredient may
comprise from 0.001% to 10% w/w, e.g., from 1% to 2% by weight of
the formulation, although it may comprise as much as 10% w/w, but
preferably not more than 5% w/w, and more preferably from 0.1% to
1% of the formulation.
[0167] When formulated in an ointment, the active ingredients may
be employed with either paraffinic or a water-miscible ointment
base. Alternatively, the active ingredients may be formulated in a
cream with an oil-in-water cream base. If desired, the aqueous
phase of the cream base may include, for example at Least 30% w/w
of a polyhydric alcohol such as propylene glycol, butane-1,3-diol,
mannitol, sorbitol, glycerol, polyethylene glycol and mixtures
thereof. The topical formulation may desirably include a compound
which enhances absorption or penetration of the active ingredient
through the skin or other affected areas. Examples of such dermal
penetration enhancers include dimethylsulfoxide and related
analogs.
[0168] The compounds of this invention can also be administered by
a transdermal device. Preferably transdermal administration will be
accomplished using a patch either of the reservoir and porous
membrane type or of a solid matrix variety. In either case, the
active agent is delivered--continuously from the reservoir or
microcapsules through a membrane into the active agent permeable
adhesive, which is in contact with the skin or mucosa of the
recipient. If the active agent is absorbed through the skin, a
controlled and predetermined flow of the active agent is
administered to the recipient. In the case of microcapsules, the
encapsulating agent may also function as the membrane. The oily
phase of the emulsions of this invention may be constituted from
known ingredients in a known manner.
[0169] While the phase may comprise merely an emulsifier, it may
comprise a mixture of at least one emulsifier with a fat or an oil
or with both a fat and an oil. Preferably, a hydrophilic emulsifier
is included together with a lipophilic emulsifier which acts as a
stabilizer. It is also preferred to include both an oil and a fat.
Together, the emulsifier(s) with or without stabilizer(s) make-up
the socalled emulsifying wax, and the wax together with the oil and
fat make up the so-called emulsifying ointment base which forms the
oily dispersed phase of the cream formulations. Emulsifiers and
emulsion stabilizers suitable for use in the formulation of the
present invention include Tween 60, Span 80, cetostearyl alcohol,
myristyl alcohol, glyceryl monostearate, sodium lauryl sulfate,
glyceryl distearate alone or with a wax, or other materials well
known in the art.
[0170] The choice of suitable oils or fats for the formulation is
based on achieving the desired cosmetic properties, since the
solubility of the active compound in most oils likely to be used in
pharmaceutical emulsion formulations is very low. Thus, the cream
should preferably be a non-greasy, non-staining and washable
product with suitable consistency to avoid leakage from tubes or
other containers. Straight or branched chain, mono- or dibasic
alkyl esters such as di-isoadipate, isocetyl stearate, propylene
glycol diester of coconut fatty acids, isopropyl myristate, decyl
oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate
or a blend of branched chain esters may be used. These may be used
alone or in combination depending on the properties required.
[0171] Alternatively, high melting point lipids such as white soft
paraffin and/or liquid paraffin or other mineral oils can be
used.
[0172] Formulations suitable for topical administration to the eye
also include eye drops wherein the active ingredients are dissolved
or suspended in suitable carrier, especially an aqueous solvent for
the active ingredients.
[0173] The active ingredients are preferably present in such
formulations in a concentration of 0.5 to 20%, advantageously 0.5
to 10% and particularly about 1.5% w/w. Formulations for parenteral
administration may be in the form of aqueous or non-aqueous
isotonic sterile injection solutions or suspensions. These
solutions and suspensions may be prepared from sterile powders or
granules using one or more of the carriers or diluents mentioned
for use in the formulations for oral administration or by using
other suitable dispersing or wetting agents and suspending agents.
The compounds may be dissolved in water, polyethylene glycol,
propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil,
sesame oil, benzyl alcohol, sodium chloride, tragacanth gum, and/or
various buffers. Other adjuvants and modes of administration are
well and widely known in the pharmaceutical art. The active
ingredient may also be administered by injection as a composition
with suitable carriers including saline, dextrose, or water, or
with cyclodextrin (i.e. Captisol), cosolvent solubilization (i.e.
propylene glycol) or micellar solubilization (i.e. Tween 80).
[0174] The sterile injectable preparation may also be a sterile
injectable solution or suspension in a non-toxic parenterally
acceptable diluent or solvent, for example as a solution in
1,3-butanediol. Among the acceptable vehicles and solvents that may
be employed are water, Ringer's solution, and isotonic sodium
chloride solution. In addition, sterile, fixed oils are
conventionally employed as a solvent or suspending medium. For this
purpose any bland fixed oil may be employed, including synthetic
mono- or diglycerides. In addition, fatty acids such as oleic acid
find use in the preparation of injectables.
[0175] For pulmonary administration, the pharmaceutical composition
may be administered in the form of an aerosol or with an inhaler
including dry powder aerosol.
[0176] Suppositories for rectal administration of the drug can be
prepared by mixing the drug with a suitable nonirritating excipient
such as cocoa butter and polyethylene glycols that are solid at
ordinary temperatures but liquid at the rectal temperature and will
therefore melt in the rectum and release the drug.
[0177] The pharmaceutical compositions may be subjected to
conventional pharmaceutical operations such as sterilization and/or
may contain conventional adjuvants, such as preservatives,
stabilizers, wetting agents, emulsifiers, buffers etc. Tablets and
pills can additionally be prepared with enteric coatings. Such
compositions may also comprise adjuvants, such as wetting,
sweetening, flavoring, and perfuming agents. Pharmaceutical
compositions of this invention comprise a compound of the formulas
described herein or a pharmaceutically acceptable salt thereof; an
additional agent selected from a kinase inhibitory agent (small
molecule, polypeptide, antibody, etc.), an immunosuppressant, an
anticancer agent, an anti-viral agent, antiinflammatory agent,
antifungal agent, antibiotic, or an anti-vascular
hyperproliferation compound; and any pharmaceutically acceptable
carrier, adjuvant or vehicle.
[0178] Alternate compositions of this invention comprise a compound
of the formulae described herein or a pharmaceutically acceptable
salt thereof; and a pharmaceutically acceptable carrier, adjuvant
or vehicle. Such compositions may optionally comprise one or more
additional therapeutic agents, including, for example, kinase
inhibitory agents (small molecule, polypeptide, antibody, etc.),
immunosuppressants, anti-cancer agents, anti-viral agents,
antiinflammatory agents, antifungal agents, antibiotics, or
anti-vascular hyperproliferation compounds.
[0179] The term "pharmaceutically acceptable carrier or adjuvant"
refers to a carrier or adjuvant that may be administered to a
patient, together with a compound of this invention, and which does
not destroy the pharmacological activity thereof and is nontoxic
when administered in doses sufficient to deliver a therapeutic
amount of the compound. Pharmaceutically acceptable carriers,
adjuvants and vehicles that may be used in the pharmaceutical
compositions of this invention include, but are not limited to, ion
exchangers, alumina, aluminum stearate, lecithin, selfemulsifying
drug delivery systems (SEDDS) such as d-atocopherol
polyethyleneglycol 1000 succinate, surfactants used in
pharmaceutical dosage forms such as Tweens or other similar
polymeric delivery matrices, serum proteins, such as human serum
albumin, buffer substances such as phosphates, glycine, sorbic
acid, potassium sorbate, partial glyceride mixtures of saturated
vegetable fatty acids, water, salts or electrolytes, such as
protamine sulfate, disodium hydrogen phosphate, potassium hydrogen
phosphate, sodium chloride, zinc salts, colloidal silica, magnesium
trisilicate, polyvinyl pyrrolidone, cellulose-based substances,
polyethylene glycol, sodium carboxymethylcellulose, polyacrylates,
waxes, polyethylene-polyoxypropylene-block polymers, polyethylene
glycol and wool fat. Cyclodextrins such as u-, P-, and
y-cyclodextrin, or chemically modified derivatives such as
hydroxyalkylcyclodextrins, including 2 and
3-hydroxypropyl-cyclodextrins, or other solubilized derivatives may
also be advantageously used to enhance delivery of compounds of the
formulae described herein.
[0180] The pharmaceutical compositions may be orally administered
in any orally acceptable dosage form including, but not limited to,
capsules, tablets, emulsions and aqueous suspensions, dispersions
and solutions. In the case of tablets for oral use, carriers which
are commonly used include lactose and corn starch. Lubricating
agents, such as magnesium stearate, are also typically added. For
oral administration in a capsule form, useful diluents include
lactose and dried corn starch. When aqueous suspensions and/or
emulsions are administered orally, the active ingredient may be
suspended or dissolved in an oily phase is combined with
emulsifying and/or suspending agents.
[0181] If desired, certain sweetening, flavoring and/or coloring
agents may be added. The pharmaceutical compositions may comprise
formulations utilizing liposome or microencapsulation techniques,
various examples of which are known in the art.
[0182] The pharmaceutical compositions may be administered by nasal
aerosol or inhalation. Such compositions are prepared according to
techniques well known in the art of pharmaceutical formulation and
may be prepared as solutions in saline, employing benzyl alcohol or
other suitable preservatives, absorption promoters to enhance
bioavailability, fluorocarbons, and/or other solubilizing or
dispersing agents, examples of which are also well known in the
art.
[0183] Combinations
[0184] While the compounds of the invention can be administered as
the sole active pharmaceutical agent, they can also be used in
combination with one or more other compounds of the invention or
with one or more other agents. When administered as a combination,
the therapeutic agents can be formulated as separate compositions
that are administered at the same time or sequentially at different
times, or the therapeutic agents can be given as a single
composition.
[0185] The phrase "combination therapy", in referring to the use of
a compound of this invention together with another pharmaceutical
agent, means the coadministration of each agent in a substantially
simultaneous manner as well as the administration of each agent in
a sequential manner, in either case, in a regimen that will provide
beneficial effects of the drug combination. Coadministration
includes inter alia the simultaneous delivery, e.g., in a single
tablet, capsule, injection or other dosage form having a fixed
ratio of these active agents, as well as the simultaneous delivery
in multiple, separate dosage forms for each agent respectively.
[0186] Thus, the administration of compounds of the present
invention may be in conjunction with additional therapies known to
those skilled in the art in the prevention or treatment of cancer,
such as radiation therapy or cytostatic agents, cytotoxic agents,
other anti-cancer agents and other drugs to amerliorate symptoms of
the cancer or side effects of any of the drugs.
[0187] If formulated as a fixed dose, such combination products
employ the compounds of this invention within the accepted dosage
ranges. Compounds of this invention may also be administered
sequentially with other anticancer or cytotoxic agents when a
combination formulation is inappropriate. The invention is not
limited in the sequence of administration; compounds of this
invention may be administered prior to, simulateously with, or
after administration of the other anticancer or cytotoxic
agent.
[0188] Currently, standard treatment of primary tumors consists of
surgical excision, when appropriate, followed by either radiation
or chemotherapy, and typically administered intravenously (IV). The
typical chemotherapy regime consists of either DNA alkylating
agents, DNA intercalating agents, CDK inhibitors, or microtubule
poisons. The chemotherapy doses used are just below the maximal
tolerated dose and therefore dose limiting toxicities typically
include, nausea, vomiting, diarrhea, hair loss, neutropenia and the
like.
[0189] There are large numbers of antineoplastic agents available
in commercial use, in clinical evaluation and in pre-clinical
development, which would be selected for treatment of cancer by
combination drug chemotherapy. And there are several major
categories of such antineoplastic agents, namely, antibiotic-type
agents, alkylating agents, antimetabolite agents, hormonal agents,
immunological agents, interferon-type agents and a category of
miscellaneous agents.
[0190] A first family of antineoplastic agents which may be used in
combination with compounds of the present invention includes
antimetabolite-type/thymidilate synthase inhibitor antineoplastic
agents. Suitable antimetabolite antineoplastic agents may be
selected from but not limited to the group consisting of
5-FU-fibrinogen, acanthifolic acid, aminothiadiazole, brequinar
sodium, carmofur, CibaGeigy CGP-30694, cyclopentyl cytosine,
cytarabine phosphate stearate, cytarabine conjugates, Lilly DATHF,
Merrel Dow DDFC, dezaguanine, dideoxycytidine, dideoxyguanosine,
didox, Yoshitomi DMDC, doxifluridine, Wellcome EHNA, Merck &
Co. EX-015, fazarabine, floxuridine, fludarabine phosphate,
5fluorouracil, N-(21-furanidyl) fluorouracil, Daiichi Seiyaku
FO-152, isopropyl pyrrolizine, Lilly LY-188011, Lilly LY-264618,
methobenzaprim, methotrexate, Wellcome MZPES, norspermidine, NCI
NSC-127716, NCI NSC-264880, NCI NSC-39661, NCI NSC-612567,
Warner-Lambert PALA, pentostatin, piritrexim, plicamycin, Asahi
Chemical PL-AC, Takeda TAC788, thioguanine, tiazofurin, Erbamont
TIF, trimetrexate, tyrosine kinase inhibitors, Taiho UFT and
uricytin.
[0191] A second family of antineoplastic agents which may be used
in combination with compounds of the present invention consists of
alkylating-type antineoplastic agents. Suitable alkylating-type
antineoplastic agents may be selected from but not limited to the
group consisting of Shionogi 254-S, aldo-phosphamide analogues,
altretamine, anaxirone, Boehringer Mannheim BBR-2207, bestrabucil,
budotitane, Wakunaga CA-102, carboplatin, carmustine, Chinoin-139,
Chinoin-153, chlorambucil, cisplatin, cyclophosphamide, American
Cyanamid CL-286558, Sanofi CY-233, cyplatate, Degussa D 384,
Sumimoto DACHP(Myr)2, diphenylspiromustine, diplatinum cytostatic,
Erba distamycin derivatives, Chugai DWA-2114R, ITI E09, elmustine,
Erbamont FCE-24517, estramustine phosphate sodium, fotemustine,
Unimed G M, Chinoin GYKI-17230, hepsulfam, ifosfamide, iproplatin,
lomustine, mafosfamide, mitolactolf Nippon Kayaku NK-121, NCI
NSC-264395, NCI NSC-342215, oxaliplatin, Upjohn PCNU,
prednimustine, Proter PTT-119, ranimustine, semustine, SmithKline
SK&F-101772, Yakult Honsha SN-22, spiromus-tine, Tanabe Seiyaku
TA-077, tauromustine, temozolomide, teroxirone, tetraplatin and
trimelamol.
[0192] A third family of antineoplastic agents which may be used in
combination with compounds of the present invention consists of
antibiotic-type antineoplastic agents. Suitable antibiotic-type
antineoplastic agents may be selected from but not limited to the
group consisting of Taiho 4181-A, aclarubicin, actinomycin D,
actinoplanone, Erbamont ADR-456, aeroplysinin derivative, Ajinomoto
AN II, Ajinomoto AN3, Nippon Soda anisomycins, anthracycline,
azino-mycin-A, bisucaberin, Bristol-Myers BL-6859, Bristol-Myers
BMY-25067, Bristol-Myers BNY-25551, Bristol-Myers BNY-26605
IBristolMyers BNY-27557, Bristol-Myers BMY-28438, bleomycin
sulfate, bryostatin-1, Taiho C-1027, calichemycin, chromoximycin,
dactinomycin, daunorubicin, Kyowa Hakko DC-102, Kyowa Hakko DC-79,
Kyowa Hakko DC-88A, Kyowa Hakko, DC89-AI, Kyowa Hakko DC92-B,
ditrisarubicin B, Shionogi DOB-41, doxorubicin,
doxorubicin-fibrinogen, elsamicin-A, epirubicin, erbstatin,
esorubicin, esperamicin-AI, esperamicin-Alb, Erbamont FCE21954,
Fujisawa FK-973, fostriecin, Fujisawa FR-900482, glidobactin,
gregatin-A, grincamycin, herbimycin, idarubicin, illudins,
kazusamycin, kesarirhodins, Kyowa Hakko KM-5539, Kirin Brewery
KRN-8602, Kyowa Hakko KT-5432, Kyowa Hakko KT-5594, Kyowa Hakko
KT-6149, American Cyanamid LL-D49194, Meiji Seika ME 2303,
menogaril, mitomycin, mitoxantrone, SmithKline M-TAG, neoenactin,
Nippon Kayaku NK-313, Nippon Kayaku NKT-01, SRI International
NSC-357704, oxalysine, oxaunomycin, peplomycin, pilatin,
pirarubicin, porothramycin, pyrindanycin A, Tobishi RA-I,
rapamycin, rhizoxin, rodorubicin, sibanomicin, siwenmycin, Sumitomo
SM5887, Snow Brand SN-706, Snow Brand SN-07, sorangicin-A,
sparsomycin, SS Pharmaceutical SS-21020, SS Pharmaceutical
SS-7313B, SS Pharmaceutical SS-9816B, steffimycin B, Taiho 4181-2,
talisomycin, Takeda TAN-868A, terpentecin, thrazine, tricrozarin A,
Upjohn U-73975, Kyowa Hakko UCN-10028A, Fujisawa WF-3405, Yoshitomi
Y-25024 and zorubicin.
[0193] A fourth family of antineoplastic agents which may be used
in combination with compounds of the present invention consists of
a miscellaneous family of antineoplastic agents, including tubulin
interacting agents, topoisomerase II inhibitors, topoisomerase I
inhibitors and hormonal agents, selected from but not limited to
the group consisting of (xcarotene, (X-difluoromethyl-arginine,
acitretin, Biotec AD-5, Kyorin AHC-52, alstonine, amonafide,
amphethinile, amsacrine, Angiostat, ankinomycin, anti-neoplaston
A10, antineoplaston A2, antineoplaston A3, antineoplaston A5.
antineoplaston AS2-1F Henkel APD, aphidicolin glycinate,
asparaginase, Avarol, baccharin, batracylin, benfluron, benzotript,
Ipsen-Beaufour BIM-23015, bisantrene, BristoMyers BNY-40481, Vestar
boron-10, bromofosfamide, Wellcome BW-502, Wellcome BW-773,
caracemide, carmethizole hydrochloride, Ajinomoto CDAF,
chlorsulfaquinoxalone, Chemes CHX-2053, Chemex CHX-100,
Warner-Lambert CI-921, WarnerLambert CI-937, Warner-Lambert CI-941,
Warner-Lambert C1958, clanfenur, claviridenone, ICN compound 1259,
ICN compound 4711, Contracan, Yakult Honsha CPT-11, crisnatol,
curaderm, cytochalasin B. cytarabine, cytocytin, Merz D-609, DABIS
maleate, dacarbazine, datelliptinium, didemnin-B,
dihaematoporphyrin ether, dihydrolenperone, dinaline, distamycin,
Toyo Pharmar DM-341, Toyo Pharmar DM-75, Daiichi Seiyaku DN-9693,
docetaxel elliprabin, elliptinium acetate, Tsumura EPMTC, the
epothilones, ergotamine, etoposide, etretinate, fenretinide,
Fujisawa FR-57704t gallium nitrate, genkwadaphnin, Chugai GLA-43,
Glaxo GR-63178, grifolan NMF5N, hexadecylphosphocholine, Green
Cross HO-221, homoharringtonine, hydroxyurea, BTG ICRF-187,
ilmofosine, isoglutamine, isotretinoin, Otsuka JI-36, Ramot K-477,
Otsuak K-76COONa, Kureha Chemical K-AM, MECT Corp KI-8110, American
Cyanamid L-623, leukoregulin, lonidamine, Lundbeck LU 1121 Lilly
LY-186641, NCI (US) MAP, marycin, Merrel Dow MDL-27048, Medco
MEDR-340, merbarone, merocyanlne derivatives,
methylanilinoacridine, Molecular Genetics MGI136, minactivin,
mitonafide, mitoquidone mopidamol, motretinide, Zenyaku Kogyo
MST-16, N-(retinoyl)amino acids, Nisshin Flour Milling N-021,
N-acylated-dehydroalanines, nafazatrom, Taisho NCU-190, nocodazole
derivative, Normosang, NCI NSC-145813, NCI NSC-361456, NCI
NSC-604782, NCI NSC-95580, ocreotide, Ono ONO-112, oquizanocine,
Akzo Org-10172, paclitaxel, pancratistatin, pazelliptine,
WarnerLambert PD-111707, Warner-Lambert PD-115934, Warner-Lambert
PD-131141, Pierre Fabre PE-1001, ICRT peptide D, piroxantrone,
polyhaematoporphyrin, polypreic acid, Efamol porphyrin, probimane,
procarbazine, proglumide, Invitron protease nexin I, Tobishi
RA-700, razoxane, Sapporo Breweries RBS, restrictin-P,
retelliptine, retinoic acid, Rhone-Poulenc RP-49532, Rhone-Poulenc
RP-56976, SmithKline SK&F-104864, Sumitomo SM-108, Kuraray
SMANCS, SeaPharm SP10094, spatol, spirocyclopropane derivatives,
spirogermanium, Unimed, SS Pharmaceutical SS-554, strypoldinone,
Stypoldione, Suntory SUN 0237, Suntory SUN 2071, superoxide
dismutase, Toyama T-506, Toyama T-680, taxol, Teijin TEI-0303,
teniposide, thaliblastine, Eastman Kodak TJB-29, tocotrienol,
topotecan, Topostin, Teijin TT82, Kyowa Hakko UCN-01, Kyowa Hakko
UCN-1028, ukrain, Eastman Kodak USB-006, vinblastine sulfate,
vincristine, vindesine, vinestramide, vinorelbine, vintriptol,
vinzolidine, withanolides and Yamanouchi YM Alternatively, the
present compounds may also be used in co-therapies with other
anti-neoplastic agents, such as acemannan, aclarubicin,
aldesleukin, alemtuzumab, alitretinoin, altretamine, amifostine,
aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole,
ANCER, ancestim, ARGLABIN, arsenic trioxide, BAM 002 (Novelos),
bexarotene, bicalutamide, broxuridine, capecitabine, celmoleukin,
cetrorelix, cladribine, clotrimazole, cytarabine ocfosfate, DA 3030
(Dong-A), daclizumab, denileukin diftitox, deslorelin, dexrazoxane,
dilazep, docetaxel, docosanol, doxercalciferol, doxifluridine,
doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil,
HIT diclofenac, interferon alfa, daunorubicin, doxorubicin,
tretinoin, edelfosine, edrecolomab eflornithine, emitefur,
epirubicin, epoetin beta, etoposide phosphate, exemestane,
exisulind, fadrozole, filgrastim, finasteride, fludarabine
phosphate, formestane, fotemustine, gallium nitrate, gemcitabine,
gemtuzumab zogamicin, gimeracil/oteracil/tegafur combination,
glycopine, goserelin, heptaplatin, human chorionic gonadotropin,
human fetal alpha fetoprotein, ibandronic acid, idarubicin,
(imiquimod, interferon alfa, interferon alfa, natural, interferon
alfa-2, interferon alfa-2a, interferon alfa-2b, interferon alfa-NI,
interferon alfa-n3, interferon alfacon1, interferon alpha, natural,
interferon beta, interferon beta-la, interferon beta-lb, interferon
gamma, natural interferon gamma-la, interferon gamma-lb,
interleukin-I beta, iobenguane, irinotecan, irsogladine,
lanreotide, LC 9018 (Yakult), leflunomide, lenograstim, lentinan
sulfate, letrozole, leukocyte alpha interferon, leuprorelin,
levamisole+fluorouracil, liarozole, lobaplatin, Ionidamine,
lovastatin, masoprocol, melarsoprol, metoclopramide, mifepristone,
miltefosine, mirimostim, mismatched double stranded RNA,
mitoguazone, mitolactol, mitoxantrone, molgramostim, nafarelin,
naloxone+pentazocine, nartograstim, nedaplatin, nilutamide,
noscapine, novel erythropoiesis stimulating protein, NSC 631570
octreotide, oprelvekin, osaterone, oxaliplatin, paclitaxel,
pamidronic acid, pegaspargase, peginterferon alfa-2b, pentosan
polysulfate sodium, pentostatin, picibanil, pirarubicin, rabbit
antithymocyte polyclonal antibody, polyethylene glycol interferon
alfa-2a, porfimer sodium, raloxifene, raltitrexed, rasburicase,
rhenium Re 186 etidronate, RII retinamide, rituximab, romurtide,
samarium (153 Sm) lexidronam, sargramostim, sizofiran, sobuzoxane,
sonermin, strontium-89 chloride, suramin, tasonermin, tazarotene,
tegafur, temoporfin, temozolomide, teniposide,
tetrachlorodecaoxide, thalidomide, thymalfasin, thyrotropin alfa,
topotecan, toremifene, tositumomab-iodine 131, trastuzumab,
treosulfan, tretinoin, trilostane, trimetrexate, triptorelin, tumor
necrosis factor alpha, natural, ubenimex, bladder cancer vaccine,
Maruyama. vaccine, melanoma lysate vaccine, valrubicin,
verteporfin, vinorelbine, VIRULIZIN, zinostatin stimalamer, or
zoledronic acid; abarelix; AE 941 (Aeterna), ambamustine, antisense
oligonucleotide, bcl-2 (Genta), APC 8015 (Dendreon), cetuximab,
decitabine, dexaminoglutethimide, diaziquone, EL 532 (Elan), EM 800
(Endorecherche), eniluracil, etanidazole, fenretinidel filgrastim
SDO1 (Amgen), fulvestrant, galocitabine, gastrin 17 immunogen,
HLA-B7 gene therapy (Vical), granulocyte macrophage colony
stimulating factor, histamine dihydrochloride, ibritumomab
tiuxetan, ilomastat, IM 862 (Cytran), interleukin iproxifene, LDI
200 (Milkhaus), leridistim, lintuzumab, CA 125 MAb (Biomira),
cancer MAb (Japan Pharmaceutical Development), HER-2 and Fc MAb
(Medarex), idiotypic 105AD7 MAb (CRC Technology), idiotypic CEA MAb
(Trilex), LYM iodine 131 MAb (Techniclone), polymorphic epithelial
mucin-yttrium 90 MAb (Antisoma), marimastat, menogaril, mitumomab,
motexafin, gadolinium, MX 6 (Galderma), nelarabine, nolatrexed, P
30 protein, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL
0903 (Shire), rubitecan, satraplatin, sodium phenylacetate,
sparfosic acid, SRL 172 (SR Pharma), SU 5416 (SUGEN)y SU 6668
(SUGEN), TA 077 (Tanabe), tetrathiomolybdate, thaliblastine,
thrombopoietin, tin ethyl etiopurpurin, tirapazamine, cancer
vaccine (Biomira), melanoma vaccine (New York University), melanoma
vaccine (Sloan Kettering Institute), melanoma oncolysate vaccine
(New York Medical College), viral melanoma cell lysates vaccine
(Royal Newcastle Hospital), or valspodar.
[0194] Treatment Kits
[0195] In other embodiments, the present invention relates to a kit
for conveniently and effectively carrying out the methods in
accordance with the present invention. In general, the
pharmaceutical pack or kit comprises one or more containers filled
with one or more of the ingredients of the pharmaceutical
compositions of the invention. Such kits are especially suited for
the delivery of solid oral forms such as tablets or capsules. Such
a kit preferably includes a number of unit dosages, and may also
include a card having the dosages oriented in the order of their
intended use. If desired, a memory aid can be provided, for example
in the form of numbers, letters, or other markings or with a
calendar insert, designating the days in the treatment schedule in
which the dosages can be administered. Optionally associated with
such container(s) can be a notice in the form prescribed by a
governmental agency regulating the manufacture, use or sale of
pharmaceutical products, which notice reflects approval by the
agency of manufacture, use or sale for human administration.
[0196] The following representative examples contain important
additional information, exemplification and guidance which can be
adapted to the practice of this invention in its various
embodiments and the equivalents thereof. These examples are
intended to help illustrate the invention, and are not intended to,
nor should they be construed to, limit its scope. Indeed, various
modifications of the invention, and many further embodiments
thereof, in addition to those shown and described herein, will
become apparent to those skilled in the art upon review of this
document, including the examples which follow and the references to
the scientific and patent literature cited herein. The contents of
those cited references are incorporated herein by reference to help
illustrate the state of the art.
[0197] In addition, for purposes of this invention, the chemical
elements are identified in accordance with the Periodic Table of
the Elements, CAS version, Handbook of Chemistry and Physics,
75.sup.th Ed., inside cover. Additionally, general principles of
organic chemistry, as well as specific functional moieties and
reactivity, are described in "Organic Chemistry", Thomas Sorrell,
University Science Books, Sausalito: 1999, and "Organic Chemistry",
Morrison & Boyd (3d Ed), the entire contents of both of which
are incorporated herein by reference.
EXAMPLES
[0198] Some of the compounds described in the following examples
have been converted into an HCl salt. The general procedure for
generating HCl salts is described below: To the final product was
added just enough MeOH saturated with HCl (g) to dissolve, cooled
to 0.degree. C. for 0.5-1 h, filtered, washed solid with ice cold
MeOH then Et.sub.2O, and the resulting solid dried in a vacuum
desiccator to provide in most cases the tris HCl salt.
Example 1
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-(imidazo[1,2-a]pyrazi-
n-3-ylethynyl)-4-methylbenzamide
##STR00046##
[0199] Imidazo[1,2-a]pyrazine
[0200] A solution of aminopyrazine (1 g, 10.5 mmol) and
chloroacetaldehyde (50% wt in H.sub.2O; 1.98 g, 12.6 mmol) in 1.6
mL of EtOH was heated at 90.degree. C. in a sealed tube for 5 h.
Upon cooling to ambient temperature, the reaction mixture was
concentrated and diluted with dichloromethane (DCM). The organic
layer washed with saturated aqueous NaHCO.sub.3 then dried over
MgSO.sub.4 and concentrated. The crude product was purified by
silica gel flash chromatography (eluted with 10% MeOH/DCM) to
provide 0.8 g of product.
3-((Trimethylsilyl)ethynyl)imidazo[1,2-a]pyrazine
[0201] A mixture of 3-bromoimidazo[1,2-a]pyrazine (0.15 g, 0.76
mmol; prepared according to J. Bradac, et al. J. Org. Chem. (1977),
42, 4197-4201), 0.09 g (0.91 mmol) of ethynyltrimethylsilane, 0.044
g (0.038 mmol) of Pd(PPh.sub.3).sub.4, 0.014 g (0.076 mmol) of CuI,
and 0.26 mL (1.52 mmol) of diisopropylethylamine in 3.8 mL of DMF
was heated at 50.degree. C. overnight under an atmosphere of
N.sub.2. Upon cooling to ambient temperature, the reaction mixture
was concentrated and the crude product was purified by silica gel
flash chromatography (eluted with 50% EtOAc/hexanes) to provide
0.15 g of product: 216 m/z (M+H).
3-Ethynylimidazo[1,2-a]pyrazine
[0202] To a solution of 3-((Trimethylsilyl)ethynyl)imidazo
[1,2-a]pyrazine (0.15 g, 0.7 mmol) in 3.5 mL of THF was added 1.05
mL (1.05 mmol) of tetrabutylammonium fluoride (1.0M in THF) at
ambient temperature. The solution was stirred for 15 min,
concentrated, and the crude product purified by silica gel flash
chromatography (eluted with 50% EtOAc/hexanes) to provide 0.078 g
of product.
3-(1H-imidazol-1-yl)-5-(trifluoromethyl)aniline
[0203] A mixture of 3-Amino-5-bromobenzotrifluoride (4.0 g, 0.0167
mol), 8-hydroxy quinoline (0.362 g, 0.0025 mol), CuI (0.476 g,
0.025 mol), imidazole (1.36 g, 0.0199 mol), and potassium carbonate
(2.52 g, 0.0183 mol) in 17 mL of DMSO (degassed with argon for
.about.10 min) was heated at 120.degree. C. under an atmosphere of
argon for 15 h; the HPLC indicated no starting material. A 14%
aqueous solution of ammonium hydroxide was added to the cooled
mixture and this was stirred for 1 h at ambient temperature. Water
(50 mL) and EtOAc (200 mL) were added and the aqueous layer was
extracted with EtOAc (3.times.30 mL). The combined organic layers
were dried over Na.sub.2SO.sub.4 and concentrated. The crude
product was purified by silica gel flash chromatography (eluted
with EtOAc/hexanes) to provide 2.51 g of product.
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-iodo-4-methylbenzamid-
e
[0204] To 3-Iodo-4-methylbenzoic acid (3.07 g, 0.0117 mol) was
added thionyl chloride (10 mL) and refluxed for 2 h. The excess
thionyl chloride was carefully removed and the resulting acid
chloride was dried in vacuo for 2 h. The residue was then dissolved
in DCM (anhydrous, 25 mL) and cooled on ice. To the cooled solution
was added 3-(1H-imidazol-1-yl)-5-(trifluoromethyl)aniline 5 (3.46
g, 0.0152 mol) in DCM followed by the dropwise addition of
diisopropylethylamine (8.2 mL, 0.047 mol). This was stirred at
ambient temperature for 21 h. The white solid that separated was
filtered and washed with water and dried to provide 4.65 g of
product. Additional product could be obtained from the filtrate
following concentration and purification by silica gel flash
chromatography in EtOAc/hexanes.
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-(imidazo[1,2-a]pyrazi-
n-3-ylethynyl)-4-methylbenzamide
[0205] A mixture of 3-Ethynylimidazo[1,2-a]pyrazine (0.075 g, 0.52
mmol), 0.245 g (0.52 mmol) of
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-iodo-4-methylbenzami-
de, 0.030 g (0.026 mmol) of Pd(PPh.sub.3).sub.4, 0.007 g (0.039
mmol) of CuI, and 0.14 mL (0.78 mmol) of diisopropylethylamine in
3.0 mL of DMF was stirred at ambient temperature overnight under an
atmosphere of N.sub.2. The reaction mixture was concentrated and
the crude product was purified by silica gel flash chromatography
(eluted with 10% EtOAc/hexanes, then 100% EtOAc, then 10%
MeOH/EtOAc) to provide 0.090 g of product as a solid: 487 m/z
(M+H).
Alternative Synthesis of
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-(imidazo[1,2-a]pyraz-
in-3-ylethynyl)-4-methylbenzamide
[0206] 3-((Trimethylsilyl)ethynyl)imidazo[1,2-a]pyrazine can be
prepared as described previously. In one variation, the reaction
can also be carried out in THF instead of DMF. The crude product
can also be purified by silica gel pad chromatography (eluted with
ethyl acetate/hexane) and a brief treatment with activated charcoal
(Darco) can be carried out to help further reduce contamination
with the homo coupling product.
3-Ethynylimidazo[1,2-a]pyrazine
[0207] To a solution of 3-((trimethylsilyl)ethynyl)
imidazo[1,2-a]pyrazine (1.39 mol) in 10.times. volume of Ethyl
acetate and 1.5.times. volume of Methanol is added two and a half
equivalents of potassium carbonate at ambient temperature and the
solution stirred for 1 hour. Potassium carbonate is filtered off
and the organic stream is washed with water and with saturated
sodium chloride solution (two or more times). Aqueous phases can be
combined and re-extracted with ethyl acetate. Organic streams can
then be combined and concentrated under vacuum to about 0.5 L.
Solids can be allowed to precipitate out upon concentration. Slurry
is cooled, e.g. to about -5.degree. C., stored overnight, filtered,
and washed with about 0.3 L of cold ethyl acetate. The solids can
then be dried under vacuum.
[0208] 3-(imidazo[1,2-a]pyrazin-3-ylethynyl)-4-methylbenzoic acid
can be prepared in a manner similar to that described above for the
Sonogashira reaction. 3-Ethynylimidazo[1,2-a]pyrazine and
3-iodo-4-methylbenzoic acid are used as coupling partners.
Alternatively, the solvent (DMF) can be replaced by ethyl acetate
and the base (Hunig base) can be replaced by triethylamine. The
product can be isolated by filtration of the crude reaction
mixture. The filter cake is washed sequentially with a solvent such
as ethyl acetate and then water, then dried in a vacuum oven.
Further purification can be achieved by slurrying the solids in
water adjusted to pH 3 with the addition of concentrated HCl. After
filtration and water wash, the product can be dried in a vacuum
oven.
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-(imidazo[1,2-a]pyrazi-
n-3-ylethynyl)-4-methylbenzamide
[0209] 3-(imidazo[1,2-a]pyrazin-3-ylethynyl)-4-methylbenzoic acid
(18 mmol) is dissolved in methylene chloride (100 mL). To this
solution is added 3 equivalents of 4-methylmorpholine (NMM)
followed by 1.05 equivalents of oxalyl chloride. After stirring at
ambient temperature for 30 minutes, 0.8 equivalents of
3-(1H-imidazol-1-yl)-5-(trifluoromethyl)aniline (prepared as above)
is added along with 5 mole % of DMAP. After initially stirring at
ambient temperature, the mixture is brought to reflux and stirred
overnight. After 16 h an additional 0.2 equivalents of the aniline
is added, bringing the total charge to 1 equivalent. The mixture
can then be stirred for an additional 2 h, quenched with water, and
the layers separated. The aqueous layer can be extracted with
methylene chloride (2.times.50 mL) and the combined extracts can be
washed with water. The combined methylene chloride layers can then
be evaporated and the residue dissolved in 100 mL of ethyl acetate
(20 mL). After standing for 1 h, the product is allowed to
crystallize. The mixture is cooled, e.g. to 0.degree. C., filtered,
and the solid product is washed with cold ethyl acetate.
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-(imidazo[1,2-a]pyrazi-
n-3-ylethynyl)-4-methylbenzamide mono hydrochloride salt
[0210]
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-(imidazo[1,2-a-
]pyrazin-3-ylethynyl)-4-methylbenzamide (0.94 mmol) can be
suspended in MeCN (10 ml) and heated with stirring to a temperature
of 45 to 55.degree. C. (hot plate temperature). Hydrochloric acid
(1.1 eq 1M solution in EtOH) is added to obtain dissolution. Within
a few minutes, a precipitate is allowed to form. The suspension can
be cooled to ambient temperature and then filtered and washed with
MeCN (1.times.1.5 ml liquors+1.times.1.5 ml fresh). The solid can
be dried at 50.degree. C. under vacuum to constant weight.
Example 2
3-(Imidazo[1,2-a]pyrazin-3-ylethynyl)-4-methyl-N-(4-((4-methylpiperazin-1--
yl)methyl)-3-(trifluoromethyl)phenyl)benzamide
##STR00047##
[0212] The title compound was synthesized from
3-ethynylimidazo[1,2-a]pyrazine and
3-iodo-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)-
phenyl)benzamide in a manner similar to that described for Example
1. The product was obtained as a solid: 533 m/z (M+H).
1-(Bromomethyl)-4-nitro-2-(trifluoromethyl)benzene
[0213] A suspension of 2-methyl-5-nitrobenzotrifluoride (3.90 g, 19
mmol), N-bromosuccinimide (NBS, 3.56 g, 20 mmol),
2,2'-azobis(2-methylpropionitrile) (AIBN, 94 mg, 0.6 mmol) in
CCl.sub.4 (40 mL) was refluxed under N.sub.2 for 16 h. HPLC
indicated ca. 50% conversion. More NBS (10 mmol) and AIBN (0.6
mmol) was added, and the mixture was refluxed for another 14 h.
HPLC indicated ca. 80% conversion. The reaction mixture was cooled
down, and the solid was filtered off and washed with EtOAc. The
combined filtrate was washed with aq. NaHCO.sub.3, dried over
Na.sub.2SO.sub.4, filtered, concentrated on rotovap and further
dried under vacuum. 1H NMR shows the ratio of desired product to
unreacted 2-methyl-5-nitrobenzotrifluoride is 75:25. This material
was not purified but used directly in the next step.
1-Methyl-4-(4-nitro-2-(trifluoromethyl)benzyl)piperazine
[0214] To a solution of crude
1-(bromomethyl)-4-nitro-2-(trifluoromethyl)benzene (13.33 mmol, 75%
pure) in DCM (10 mL) was added Et.sub.3N (1.4 mL, 10 mmol) and
1-methylpiperazine (1.1 mL, 10 mmol). After stirring for 3 h at rt,
aq. NaHCO.sub.3 was added, and the mixture was extracted with DCM.
The combined organic layer was dried over Na.sub.2SO.sub.4,
filtered, concentrated, and the resulting residue was purified by
silica gel chromatography (eluted with 10% MeOH/DCM) to provide
2.21 g of product as a pale yellow oil.
4-((4-Methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline
[0215] A suspension of
1-methyl-4-(4-nitro-2-(trifluoromethyl)benzyl)piperazine (1.23 g, 4
mmol) and sodium hydrosulfite (7.0 g, 85% pure from Aldrich, 40
mmol) in acetone and water (1:1, 20 mL) was refluxed for 3 h. Upon
cooling, the volatile components (mainly acetone) were removed on
rotavap, and the resulting mixture was subjected to filtration. The
solid was thoroughly washed with EtOAc. The combined filtrate was
extracted with n-BuOH (4.times.), and the combined organic layer
was washed with saturated aq. NaHCO.sub.3, dried
(Na.sub.2SO.sub.4), filtered, concentrated, and the resulting
residue was purified by silica gel chromatography (eluted with 5%
MeOH/DCM, MeOH was pre-saturated with ammonia gas) to provide 0.71
g of product as a pale yellow solid.
3-Iodo-4-meth
yl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)
Benzamide
[0216] 3-Iodo-4-methylbenzoyl chloride (0.48 g, 1.7 mmol), prepared
from the reaction of 3-iodo-4-methylbenzoic acid and SOCl.sub.2 (as
previously described), was added to a solution of
4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (0.47
g, 1.7 mmol), N,N-diisopropylethylamine (0.26 g, 2.0 mmol), and a
catalytic amount of DMAP in THF (10 mL). After stirring at rt for 2
h, the reaction was quenched with water. EtOAc was added and the
layers separated. The combined organic layers were concentrated to
dryness and purified by silica gel chromatography (eluted with 5%
MeOH/DCM, MeOH was pre-saturated with ammonia gas), to provide 0.51
g of product as an off-white solid.
Alternative synthesis of
3-(Imidazo[1,2-a]pyrazin-3-ylethynyl)-4-methyl-N-(4-((4-methylpiperazin-1-
-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide
[0217]
3-(Imidazo[1,2-a]pyrazin-3-ylethynyl)-4-methyl-N-(4-((4-methylpiper-
azin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide and its mono
hydrochloride salt can be prepared in an alternative synthesis
similar to that described in Example 1 from
3-(imidazo[1,2-a]pyrazin-3-ylethynyl)-4-methylbenzoic acid and
4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (as
prepared above).
Example 3
N-(3-(2-((dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethyl)pheny-
l)-3-(imidazo[1,2-a]pyrazin-3-ylethynyl)-4-methylbenzamide
##STR00048##
[0219] The title compound was synthesized from
3-ethynylimidazo[1,2-a]pyrazine and
N-(3-(2-((dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethyl)phen-
yl)-3-iodo-4-methylbenzamide in a manner similar to that described
for Example 1. The product was obtained as a solid: 544 m/z
(M+H).
1-(1H-imidazol-2-yl)-N,N-dimethylmethanamine
[0220] To a two-necked round-bottomed flask equipped with a reflux
condenser and a pressure-equalizing addition funnel, was added
2-imidazolecarboxaldehyde (6 g, 62.5 mmol) in MeOH (60 mL). To this
suspension (ambient temperature) was added a solution of
dimethylamine (40% aqueous, 60 mL) at a fast dropping rate (20
min). After the addition was complete, solid sodium borohydride (7
g, 186.8 mmol) was CAUTIOUSLY added portionwise over 45 min.
Foaming occurred after each portion, and the internal temperature
was allowed to maintain .about.50.degree. C. without external
cooling. The reaction mixture was then heated to 65.degree. C. for
3 h and allowed to cool to ambient temperature for overnight. The
reaction contents were concentrated in vacuo and the resultant
residue was taken up in EtOAc (2.times.30 mL) washed with brine and
with CHCl.sub.3 (4.times.100 mL). The EtOAc extract was discarded.
The CHCl.sub.3 extract was dried over (NaSO.sub.4), filtered, and
concentrated in vacuo to give 3.7 g of the desired product as a
waxy solid.
3-(2-((Dimethylamino)methyl)-H-imidazol-1-yl)-5-(trifluoromethyl)aniline
[0221] 3-Amino-5-bromobenzotrifluoride (6 g, 25 mmol) and
1-(1H-imidazol-2-yl)-N,N-dimethylmethanamine (3.7 g, 29.6 mmol)
were dissolved in anhydrous DMSO (25 mL). To this was added CuI
(0.95 g, 7.5 mmol), 8-hydroxy quinoline (0.72 g, 7.5 mmol) and
K.sub.2CO.sub.3 (6.9 g, 50 mmol). The mixture was stirred
vigorously and degassed with N.sub.2 for 15 minutes. The flask was
then equipped with a condenser and heated at 120.degree. C. for 18
h. The resultant heterogeneous mixture was cooled to rt, poured
into 14% aq. NH.sub.4OH (100 mL) and extracted with EtOAc
(3.times.300 ml). The combined extracts were dried over NaSO.sub.4
and concentrated in vacuo. The residue was chromatographed over
silica gel eluting with MeOH/DCM (5:95) to furnish 3.5 g of the
desired product as a tan colored material: 285 m/z (M+H).
N-(3-(2-((dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethyl)pheny-
l)-3-iodo-4-methylbenzamide
[0222] 3-Iodo-4-methylbenzoyl chloride (2.2 g, 7.88 mmol),
dissolved in anhydrous THF (13 mL), was added dropwise to a
solution of
3-(2-((dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethyl)aniline
(1.5 g, 5.5 mmol), DIPEA (2.1 mL, 11.8 mmol) in THF (30 mL) at
.about.5.degree. C. The resultant solution was stirred at ambient
temperature overnight. The solvent was removed in vacuo and the
crude residue was redissolved in CH.sub.2Cl.sub.2 and washed with
1N NaOH. The organic layer was then washed with water, and brine
then dried over NaSO.sub.4 before being concentrated in vacuo. The
brown colored residue was then triturated in a mixture of
hexanes/DCM to precipitate 1.4 g of the desired product as an
off-white powder: 529 m/z (M+H).
Alternative Synthesis of
N-(3-(2-((dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethyl)phen-
yl)-3-(imidazo[1,2-a]pyrazin-3-ylethynyl)-4-methylbenzamide
[0223]
N-(3-(2-((dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethy-
l)phenyl)-3-(imidazo[1,2-a]pyrazin-3-ylethynyl)-4-methylbenzamide
and its mono hydrochloride salt can be prepared in an alternative
synthesis similar to that described in Example 1 from
3-(imidazo[1,2-a]pyrazin-3-ylethynyl)-4-methylbenzoic acid and
3-(2-((Dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethyl)aniline
(as prepared above).
Example 4
3-(Imidazo[1,2-a]pyridin-3-ylethynyl)-4-methyl-N-(3-(4-methyl-1H-imidazol--
1-yl)-5-(trifluoromethyl)phenyl)benzamide
##STR00049##
[0224] 3-Ethynylimidazo[1,2-a]pyridine
[0225] To 3-bromoimidazo[1,2-a]pyridine (5 g, 0.0254 mol) in
acetonitrile (50 mL) in a sealed tube was added
bis(triphenylphosphine) palladium(II) dichloride (0.445 g, 0.634
mmol), CuI (0.17 g, 0.89 mmol), dicyclohexylamine (5.6 mL, 0.028
mol) and ethynyltrimethylsilane (7.2 mL, 0.051 mol). The solution
was purged with argon for 15 minutes, sealed and heated at
80.degree. C. for 3 h. At this point the HPLC did not show any
starting bromide. The solvents were concentrated and to the residue
was added water and dichloromethane (25 mL each). The organic layer
was separated and the aqueous layer was repeatedly extracted with
dichloromethane (3.times.20 mL). The combined extracts were dried
(Na.sub.2SO.sub.4), and concentrated (Rf, 0.47 in 1/1 hexanes/ethyl
acetate). The resulting residue was dissolved in THF (100 mL) and
treated with tetrabutyl ammonium fluoride monohydrate (8.3 g, 0.032
mol) in water (5 mL) and the mixture was stirred at rt for 2 h. The
solvents were concentrated and the resulting residue was
partitioned between water (25 mL) and dichloromethane (150 mL). The
aqueous layer was extracted with dichloromethane (2.times.30 mL).
The combined extracts were dried (Na.sub.2SO.sub.4), and
concentrated. The resulting residue was purified by combiflash on
silica gel using hexanes/ethyl acetate. The desired product was
eluted with 50/50 hexane/ethyl acetate and isolated as an off-white
solid: MS (M+H).sup.+ 200.
3-(4-Methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)aniline
[0226] A suspension of 3-bromo-5-(trifluoromethyl)aniline (4.8 g,
20 mmol), 4-methylimidazole (1.97 g, 24 mmol), potassium carbonate
(3.04 g, 22 mmol), CuI (0.57 g, 3 mmol), and 8-hydroxyquinoline
(0.44 g, 3 mmol) in dry DMSO (20 mL) in a pressure tube was
degassed by bubbling N.sub.2 into the suspension for 10 minutes
while stirring. The tube was sealed tightly. The mixture was heated
at 120.degree. C. (oil bath temperature) for 15 h. The mixture was
cooled down to 45-50.degree. C. and 14% aq. NH.sub.4OH (20 mL) was
added. The mixture was maintained at this temperature for 1 h.
After cooling to rt, water and ethyl acetate were added. The
aqueous layer was extracted with ethyl acetate and the combined
organic layers were passed through a short silica gel column to
remove most of green/blue Cu salts. The filtrate was dried over
sodium sulfate and concentrated on a rotavap. The crude product was
recrystallized from EtOAc/hexanes, giving pure pale yellow needles.
The mother liquor was concentrated and the residue was purified on
silica gel column (5% methanol/methylene chloride), yielding a
second crop as pale yellow needles.
3-Iodo-4-methyl-N-(3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl-
) Benzamide
[0227] 3-Iodo-4-methylbenzoic acid (2.62 g, 10 mmol) was refluxed
in SOCl.sub.2 (10 mL) for 1 h. The volatile components were removed
on a rotavap and the residue was dissolved in benzene (10 mL),
concentrated to dryness on a rotavap and further dried under
vacuum. The resulting acyl chloride was added to a solution
3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)benzenamine (2.46
g, 10.2 mmol), N,N-diisopropylethylamine (1.56 g, 12 mmol), and a
catalytic amount of DMAP in THF (20 mL). After stirring at rt for 2
h, the reaction was quenched with water. EtOAc was added and the
layers separated. The combined organic layers were concentrated to
dryness and used without purification in next step.
3-(Imidazo[1,2-a]pyridin-3-ylethynyl)-4-methyl-N-(3-(4-methyl-1H-imidazol--
1-yl)-5-(trifluoromethyl)phenyl)benzamide
[0228] To a solution of
3-iodo-4-methyl-N-(3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)pheny-
l)benzamide (0.11 g, 0.22 mmol) in DMF (1 mL) in a sealed tube was
added Pd[(PPh.sub.3).sub.4] (0.013 g, 0.011 mmol), CuI (3 mg, 0.016
mmol), diethylisopropylamine (0.057 mL, 0.33 mmol), followed by
3-ethynylimidazo[1,2-a]pyridine (0.040 g, 0.28 mmol.). The mixture
was purged with argon for 15 minutes, sealed and stirred at rt for
28 h. The solvent was concentrated and the residue was taken up in
methylene chloride (50 mL). The organic layer was washed with
water, dried (Na.sub.2SO.sub.4) and evaporated to leave a brown
residue which was purified by combiflash (hexane/ethyl
acetate/methanol) to yield the desired material: MS
(M+H).sup.+500.
Alternative Synthesis of
3-(Imidazo[1,2-a]pyridin-3-ylethynyl)-4-methyl-N-(3-(4-methyl-1H-imidazol-
-1-yl)-5-(trifluoromethyl)phenyl)benzamide
[0229]
3-(Imidazo[1,2-a]pyridin-3-ylethynyl)-4-methyl-N-(3-(4-methyl-1H-im-
idazol-1-yl)-5-(trifluoromethyl)phenyl)benzamide and its mono
hydrochloride salt can be prepared in an alternative synthesis
similar to that described in Example 1 from
3-(imidazo[1,2-a]pyridin-3-ylethynyl)-4-methylbenzoic acid and
3-(4-Methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)aniline (as
prepared above). The
3-(imidazo[1,2-a]pyridin-3-ylethynyl)-4-methylbenzoic acid is
prepared in a manner similar to that described in Example 1 using
3-Ethynylimidazo[1,2-a]pyridine and 3-iodo-4-methylbenzoic acid as
Sonogashira coupling partners.
Example 5
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-(imidazo[1,2-a]pyridi-
n-3-ylethynyl)-4-methylbenzamide
##STR00050##
[0231] The titled compound was made as for example 1 using
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-iodo-4-methylbenzami-
de and 3-ethynylimidazo[1,2-a]pyridine: MS (M+H).sup.+ 486. The
titled compound can also be prepared according to the alternative
synthesis described in example 1 from
3-(imidazo[1,2-a]pyridin-3-ylethynyl)-4-methylbenzoic acid and
3-(1H-imidazol-1-yl)-5-(trifluoromethyl)aniline (as prepared in
Example 1). The
3-(imidazo[1,2-a]pyridin-3-ylethynyl)-4-methylbenzoic acid is
prepared in a manner similar to that described in Example 1 using
3-Ethynylimidazo[1,2-a]pyridine and 3-iodo-4-methylbenzoic acid as
Sonogashira coupling partners.
Example 6
3-(Imidazo[1,2-a]pyridin-3-ylethynyl)-4-methyl-N-(4-(trifluoromethyl)pyrid-
in-2-yl)benzamide
##STR00051##
[0233] The titled compound was made as for example 1 using
3-iodo-4-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide and
3-ethynylimidazo[1,2-a]pyridine: MS (M+H).sup.+ 421.39.
Example 7
N-(5-tert-butylisoxazol-3-yl)-3-(imidazo[1,2-a]
pyridin-3-ylethynyl)-4-methylbenzamide
##STR00052##
[0235] The titled compound was made as for example 1 using
N-(5-tert-butylisoxazol-3-yl)-3-iodo-4-methylbenzamide and
3-ethynylimidazo[1,2-a]pyridine: MS (M+H).sup.+ 399.
Example 8
3-(Imidazo[1,2-a]pyridin-3-ylethynyl)-4-methyl-N-(4-((4-methylpiperazin-1--
yl)methyl)-3-(trifluoromethyl)phenyl)benzamide
##STR00053##
[0237] 3-Ethynylimidazo[1,2-a]pyridine (37 mg, 0.26 mmol),
3-iodo-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)-
phenyl)benzamide (103.4 mg, 0.2 mmol), (prepared as in Example 2),
Pd[(PPh.sub.3).sub.4] (11.6 mg, 5 mol %), and CuI (2.9 mg, 7.5 mmol
%) was placed in a vial with rubber septum. The mixture underwent 3
cycles of vacuum/filling with N.sub.2, and DMF (1.5 ml) and N,
N-diisopropylethylamine (53 mL, 0.3 mmol) was added. The mixture
was stirred at rt for 16 h, and the reaction was quenched with
H.sub.2O. EtOAc and more water were added for extraction. The
combined organic layer was dried (Na.sub.2SO.sub.4), filtered,
concentrated, and the resulting residue was purified by silica gel
chromatography (eluent: 5% MeOH in methylene chloride, MeOH was
pre-saturated with ammonia gas), giving the titled compound as an
off-white solid (53%, 56 mg): MS (M+H).sup.+ 532.
Alternative Synthesis of
3-(Imidazo[1,2-a]pyridin-3-ylethynyl)-4-methyl-N-(4-((4-methylpiperazin-1-
-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide
[0238]
3-(Imidazo[1,2-a]pyridin-3-ylethynyl)-4-methyl-N-(4-((4-methylpiper-
azin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide and its mono
hydrochloride salt can be prepared in an alternative synthesis
similar to that described in Example 1 from
3-(imidazo[1,2-a]pyridin-3-ylethynyl)-4-methylbenzoic acid and
4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (as
prepared in example 2). The
3-(imidazo[1,2-a]pyridin-3-ylethynyl)-4-methylbenzoic acid is
prepared in a manner similar to that described in Example 1 using
3-Ethynylimidazo[1,2-a]pyridine and 3-iodo-4-methylbenzoic acid as
Sonogashira coupling partners.
Example 9
N-(3-(2-((dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethyl)pheny-
l)-3-(imidazo[1,2-a]pyridin-3-ylethynyl)-4-methylbenzamide
##STR00054##
[0240] To 3-ethynylimidazo[1,2-a]pyridine (0.032 g, 0.22 mmol) in
anhydrous DMF (1.26 mL) was added
N-(3-(2-((dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethyl)phen-
yl)-3-iodo-4-methylbenzamide (prepared as in Example 3),
Pd(PPh.sub.3).sub.4(0.013 g, 0.011 mmol), CuI (0.0032 mg, 0.0165
mmol) and DIPEA (0.064 mL, 0.44 mmol). The solution was degassed
with argon for 15 minutes then stirred overnight at rt. The solvent
was removed and the resultant residue was chromatographed over
silica gel eluting initially with EtOAc and then with
methanol/methylene chloride (5:95) to furnish the desired product:
(0.07 g, 59%) MS (M+H).sup.+ 542.
Alternative Synthesis of
N-(3-(2-((dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethyl)phen-
yl)-3-(imidazo[1,2-a]pyridin-3-ylethynyl)-4-methylbenzamide
[0241]
N-(3-(2-((dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethy-
l)phenyl)-3-(imidazo[1,2-a]pyridin-3-ylethynyl)-4-methylbenzamide
and its mono hydrochloride salt can be prepared in an alternative
synthesis similar to that described in Example 1 from
3-(imidazo[1,2-a]pyridin-3-ylethynyl)-4-methylbenzoic acid and
3-(2-((Dimethylamino)methyl)-1H-imidazol-1-yl)-5-(trifluoromethyl)aniline
(as prepared in Example 3). The
3-(imidazo[1,2-a]pyridin-3-ylethynyl)-4-methylbenzoic acid is
prepared in a manner similar to that described in Example 1 using
3-Ethynylimidazo[1,2-a]pyridine and 3-iodo-4-methylbenzoic acid as
Sonogashira coupling partners.
Example 10
3-((8-Acetamidoimidazo[1,2-a]pyridin-3-yl)ethynyl)-4-methyl-N-(4-(trifluor-
omethyl)pyridin-2-yl)benzamide
##STR00055##
[0242] N-(3-Ethynylimidazo[1,2-a]pyridin-8-yl)acetamide
[0243] N-(3-Ethynylimidazo[1,2-a]pyridin-8-yl)acetamide was
synthesized as for example 1A from
N-(3-bromoimidazo[1,2-a]pyridin-8-yl)acetamide (E. Smakula Hand and
William W. Paudler, J. Org. Chem., 1978, 43, 2900-2906). The titled
compound was isolated as an off-white solid, Rf, 0.6
(hexane/ethylacetate 50/50): MS (M+H).sup.+200.
3-((8-Acetamidoimidazo[1,2-a]pyridin-3-yl)
ethynyl)-4-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide
[0244] The titled compound was made as for example 1 using
3-iodo-4-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide and
N-(3-ethynylimidazo[1,2-a]pyridin-8-yl)acetamide: MS (M+H).sup.+
478.4.
Example 11
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-((8-acetamidoimidazo[-
1,2-a]pyridin-3-yl)ethynyl)-4-methylbenzamide
##STR00056##
[0246] The titled compound was made as for example 10 using
N-(3-(1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-iodo-4-methylbenzami-
de and N-(3-ethynylimidazo[1,2-a]pyridin-8-yl)acetamide: MS (M+H)
543.
Example 12
4-Methyl-3-((8-(4-(methylsulfonyl)phenylamino)imidazo[1,2-a]pyridin-3-yl)e-
thynyl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide
##STR00057##
[0247] 8-(Benzyloxy)-3-bromoimidazo[1,2-a]pyridine
[0248] To a solution of 2-amino-3-benzyloxypyridine (25.0 g, 124.9
mmol) and chloroacetaldehyde (50% wt in H.sub.2O; 16.7 mL, 131.2
mmol) in 250 mL of EtOH was heated at reflux in a sealed tube for
19 h. Upon cooling to ambient temperature, the reaction mixture was
concentrated and the resulting brown oil added 125 mL 1N NaOH then
extracted with dichloromethane (DCM). The combined organic layers
were washed with H.sub.2O, dried over Na.sub.2SO.sub.4 and
concentrated. Upon concentrating the solution, a tan solid formed
which was filtered and dried to provide 25.8 g of crude
product.
[0249] To a solution of crude 8-(benzyloxy)imidazo[1,2-a]pyridine
(8.73 g, 38.9 mmol) in 100 mL of EtOH was added, dropwise, 4.8 mL
(46.7 mmol) of a solution of 1:1 Br.sub.2/H.sub.2O at ambient
temperature under an atmosphere of N.sub.2. The resulting dark
orange suspension was stirred at ambient temperature for 30 min,
added 60 mL 1N NaOH, and the reaction mixture extracted with DCM.
The combined organic layers were dried over Na.sub.2SO.sub.4 and
concentrated. The crude product was purified by silica gel flash
chromatography (eluted with 30% EtOAc/hexanes) to provide 7.04 g of
product.
8-(Benzyloxy)-3-((trimethylsilyl)ethynyl)imidazo[1,2-a]pyridine
[0250] A mixture of 8-(benzyloxy)-3-bromoimidazo[1,2-a]pyridine
(10.0 g, 33.0 mmol), 9.39 mL (66.0 mmol) of ethynyltrimethylsilane,
0.580 g (0.825 mmol) of Pd(PPh.sub.3).sub.2Cl.sub.2, 0.230 g (1.19
mmol) of CuI, and 5.09 mL (36.3 mmol) of diisopropylamine in 100 mL
of acetonitrile was heated at reflux for 3 h under an atmosphere of
N.sub.2. Upon cooling to ambient temperature, the reaction mixture
was concentrated and the crude product was purified by silica gel
flash chromatography (eluted with 20-50% EtOAc/hexanes) to provide
6.74 g of product: 321 m/z (M+H).
3-((Trimethylsilyl)ethynyl)imidazo[1,2-a]pyridin-8-yl
trifluoromethanesulfonate
[0251] To a cooled (0.degree. C.) solution of
8-(benzyloxy)-3-((trimethylsilyl)ethynyl)imidazo[1,2-a]pyridine
(3.44 g, 10.7 mmol) in 400 mL of DCM, under an atmosphere of
N.sub.2, was added via cannulation 100 mL (100 mmol) of boron
trichloride (1.0M solution in hexanes). The reaction solution was
stirred at 0.degree. C./N.sub.2 for 30 min, to which was added
(0.degree. C.) 200 mL H.sub.2O followed by extraction with DCM. The
combined organic layers were washed with brine, dried over
Na.sub.2SO.sub.4 and concentrated. The crude product was purified
by silica gel flash chromatography (eluted with 30% EtOAc/hexanes
then 10% MeOH/DCM) to provide 2.32 g of deprotected product: 231
m/z (M+H).
[0252] To a cooled (-78.degree. C.) solution of
8-(hydroxy)-3-((trimethylsilyl)ethynyl)imidazo[1,2-a]pyridine (2.32
g, 10.1 mmol) and 1.63 mL (20.1 mmol) of pyridine in 50 mL of DCM,
under an atmosphere of N.sub.2, was added 2.03 mL (12.1 mmol) of
trifluoromethanesulfonic anhydride via syringe. Upon removing the
cooling bath, the reaction solution was stirred at ambient
temperature (N.sub.2) for 2 h. The reaction mixture was poured into
a stirring solution of 100 mL 1.0N HCl, the layers separated, and
the organic layer washed successively with 1.0N HCl, H.sub.2O,
saturated aqueous NaHCO.sub.3, and brine. The organic layer was
dried over Na.sub.2SO.sub.4 and concentrated. The crude product was
filtered through a small plug of silica gel (eluted with 30%
EtOAc/hexanes), concentrated, and further dried in vacuo to provide
3.63 g of product: 363 m/z (M+H).
N-(4-(Methylsulfonyl)phenyl)-3-((trimethylsilyl)ethynyl)imidazo[1,2-a]pyri-
din-8-amine
[0253] A mixture of
3-((trimethylsilyl)ethynyl)imidazo[1,2-a]pyridin-8-yl
trifluoromethanesulfonate (0.329 g, 0.91 mmol), 0.186 (1.09 mmol)
of 4-(methylsulfonyl)aniline, 0.083 g (0.091 mmol) of
Pd.sub.2(dba).sub.2, 0.087 g (0.181 mmol) of
2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl, and 0.385 g
(1.81 mmol) of potassium phosphate in 8 mL of DME was heated at
80.degree. C. in a sealed tube overnight under an atmosphere of
N.sub.2. Upon cooling to ambient temperature, the reaction mixture
was concentrated and the crude product was purified by silica gel
flash chromatography (triethylamine-treated silica gel; eluted with
0-80% EtOAc/hexanes) to provide 0.058 g of product: 384 m/z
(M+H).
3-Ethynyl-N-(4-(methylsulfonyl)phenyl)imidazo[1,2-a]pyridin-8-amine
[0254] To a solution of
N-(4-(methylsulfonyl)phenyl)-3-((trimethylsilyl)ethynyl)imidazo[1,2-a]pyr-
idin-8-amine (0.058 g, 0.15 mmol) in 1.5 mL of THF was added 0.23
mL (0.23 mmol) of tetrabutylammonium fluoride (1.0M in THF) at
ambient temperature. The solution was stirred for 15 min,
concentrated, and the crude product purified by silica gel flash
chromatography (triethylamine-treated silica gel; eluted with 100%
DCM then 5% MeOH/DCM) to provide a quantitative yield (0.047 g) of
product: 312 m/z (M+H).
4-Methyl-3-((8-(4-(methylsulfonyl)phenylamino)imidazo[1,2-a]pyridin-3-yl)e-
thynyl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide
[0255] A mixture of 3-ethynyl-N-(4-(methylsulfonyl)
phenyl)imidazo[1,2-a]pyridin-8-amine 5 (0.048 g, 0.154 mmol), 0.069
g (0.170 mmol) of
3-iodo-4-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide, 0.009
g (0.008 mmol) of Pd(PPh.sub.3).sub.4, 0.002 g (0.012 mmol) of CuI,
and 0.04 mL (0.23 mmol) of diisopropylethylamine in 0.8 mL of DMF
was stirred at ambient temperature overnight under an atmosphere of
N.sub.2. The reaction mixture was concentrated and the crude
product was purified by silica gel flash chromatography
(triethylamine-treated silica gel; eluted with 10% EtOAc/hexanes to
100% EtOAc) to provide 0.047 g of product as a solid: 590 m/z
(M+H).
Example 13
4-methyl-3-((8-(4-sulfamoylphenylamino)imidazo[1,2-a]pyridin-3-yl)ethynyl)-
-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide
##STR00058##
[0257] The title compound was synthesized from
3-ethynyl-N-(4-sulfamoylphenyl)imidazo[1,2-a]pyridin-8-amine and
3-iodo-4-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide in a
manner similar to that described for Example 12. The product was
obtained as a solid: 591 m/z (M+H).
Example 14
(R)--N-(4-((3-(Dimethylamino)pyrrolidin-1-yl)methyl)-3-(trifluoromethyl)ph-
enyl)-3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzamide
##STR00059##
[0258] 3-((Trimethylsilyl)ethynyl)imidazo[1,2-b]pyridazine
[0259] A mixture of 3-bromoimidazo[1,2-b]pyridazine (36.78 g, 0.186
mol; prepared according to Stanovnik, B. et al. Synthesis (1981),
12, 987-989), ethynyltrimethylsilane (21.89 g, 0.223 mol),
Pd(PPh.sub.3).sub.4 (10.73 g, 9.29 mmol), CuI (5.30 g, 0.028 mol),
and diisopropylethylamine (32.4 mL, 0.279 mol) in 150 mL of DMF was
stirred at ambient temperature, under an atmosphere of N.sub.2, for
1 h. The reaction mixture was concentrated and the crude product
was purified by silica gel flash chromatography (eluted with 0-5%
MeOH/DCM) to provide 28.46 g of product.
3-Ethynylimidazo[1,2-b]pyridazine
[0260] To a solution of 3-((trimethylsilyl)ethynyl)
imidazo[1,2-b]pyridazine (28.46 g, 0.132 mol) in 200 mL of THF was
added 145 mL (0.145 mol) of tetrabutylammonium fluoride (1.0M in
THF) at ambient temperature. The solution was stirred for 15 min,
concentrated, and the crude product purified by silica gel flash
chromatography (eluted with 0-5% MeOH/DCM) to provide 17.84 g of
product.
1-(Bromomethyl)-4-nitro-2-(trifluoromethyl)benzene
[0261] A suspension of 2-methyl-5-nitrobenzotrifluoride (3.90 g, 19
mmol), N-bromosuccinimide (NBS, 3.56 g, 20 mmol), and
2,2'-azobis(2-methylpropionitrile) (AIBN, 0.094 g, 0.6 mmol) in 40
mL of CCl.sub.4 was heated at reflux under N.sub.2 for 16 h. HPLC
indicated ca. 50% conversion. Additional NBS (10 mmol) and AIBN
(0.6 mmol) were added and the mixture was heated at reflux for
another 14 h. HPLC indicated ca. 80% conversion. The reaction
mixture was cooled to ambient temperature, and the solid was
filtered and washed with EtOAc. The combined filtrate was washed
with aq. NaHCO.sub.3, dried over Na.sub.2SO.sub.4, filtered,
concentrated on rotovap, and further dried under vacuum. .sup.1H
NMR indicated the ratio of desired product to unreacted
2-methyl-5-nitrobenzotrifluoride to be 75:25. This material was
used directly in the next step.
(R)--N,N-Dimethyl-1-(4-nitro-2-(trifluoromethyl)benzyl)pyrrolidin-3-amine
[0262] To a solution of crude
1-(bromomethyl)-4-nitro-2-(trifluoromethyl)benzene (17.5 mmol, 75%
pure) in 40 mL of DCM was added Et.sub.3N (2.69 mL, 19.3 mmol) and
(R)-(+)-3-(dimethylamino)pyrrolidine (2.0 g, 17.5 mmol). After
stirring overnight at ambient temperature under an atmosphere of
N.sub.2, the reaction solution was concentrated, added aq.
NaHCO.sub.3 (100 mL), and the resulting mixture extracted with DCM
(4.times.50 mL). The combined organic layer was dried over
Na.sub.2SO.sub.4, filtered, concentrated, and the resulting residue
was purified by silica gel chromatography (eluted with 0-10%
MeOH/DCM) to provide 3.35 g of product as a yellow oil.
(R)-1-(4-Amino-2-(trifluoromethyl)benzyl)-N,N-dimethylpyrrolidin-3-amine
[0263] To a solution of
(R)--N,N-dimethyl-1-(4-nitro-2-(trifluoromethyl)benzyl)pyrrolidin-3-amine
(1.20 g, 3.79 mmol) in 20 mL of wet EtOH was added 0.26 g of Pd/C
(10% Pd on C) and the mixture shaken in a Parr apparatus (pressure
reaction vessel purged thoroughly with H.sub.2 and pressure
regulated at 45 psi throughout) for 2-3 h. The reaction mixture was
filtered through a small pad of celite, washed with EtOAc, and the
combined organics concentrated to provide a quantitative yield of a
light yellow oil. This material was used directly in the next
step.
(R)--N-(4-((3-(Dimethylamino)pyrrolidin-1-yl)methyl)-3-(trifluoromethyl)ph-
enyl)-3-iodo-4-methylbenzamide
[0264] To a cooled (0.degree. C.) solution of
(R)-1-(4-amino-2-(trifluoromethyl)benzyl)-N,N-dimethylpyrrolidin-3-amine
(3.79 mmol) in 14 mL DCM, under an atmosphere of N.sub.2, was added
3-Iodo-4-methylbenzoyl chloride (1.17 g, 4.17 mmol; CAS#52107-98-9,
prepared from the reaction of 3-iodo-4-methylbenzoic acid and
SOCl.sub.2) followed by dropwise addition of
N,N-diisopropylethylamine (2.64 mL, 15.2 mmol). After stirring to
ambient temperature over 1.5 h, the reaction mixture was
concentrated and the crude product was purified by silica gel
chromatography (eluted with 0-8% MeOH/DCM; MeOH was pre-saturated
with ammonia gas), to provide 0.71 g of product as a thick yellow
oil.
(R)--N-(4-((3-(dimethylamino)pyrrolidin-1-yl)methyl)-3-(trifluoromethyl)ph-
enyl)-3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzamide
[0265] A mixture of 3-ethynylimidazo[1,2-b]pyridazine (0.051 g,
0.34 mmol), 0.150 g (0.28 mmol) of
(R)--N-(4-((3-(dimethylamino)pyrrolidin-1-yl)methyl)-3-(trifluoromethyl)p-
henyl)-3-iodo-4-methylbenzamide, 0.016 g (0.014 mmol) of
Pd(PPh.sub.3).sub.4, 0.004 g (0.021 mmol) of CuI, and 0.09 mL (0.51
mmol) of N,N-diisopropylethylamine in 3.5 mL of DMF was stirred at
ambient temperature, under an atmosphere of N.sub.2, for 3 days
(reaction pushed to completion with additional equivalents of
reagents and heating to 80.degree. C.). The reaction mixture was
concentrated and the crude product was purified by silica gel
chromatography (eluted with 0-10% MeOH/DCM; MeOH was pre-saturated
with ammonia gas) to provide 0.020 g of product as a solid: 547 m/z
(M+H).
Alternative Synthesis of
(R)--N-(4-((3-(Dimethylamino)pyrrolidin-1-yl)methyl)-3-(trifluoromethyl)p-
henyl)-3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzamide
[0266]
(R)--N-(4-((3-(Dimethylamino)pyrrolidin-1-yl)methyl)-3-(trifluorome-
thyl)phenyl)-3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzamide
and its mono hydrochloride salt can be prepared in an alternative
synthesis similar to that described in Example 1 from
3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzoic acid and
(R)-1-(4-Amino-2-(trifluoromethyl)benzyl)-N,N-dimethylpyrrolidin-3-amine
(as prepared above). The
3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzoic acid is
prepared in a manner similar to that described in Example 1 using
3-Ethynylimidazo[1,2-b]pyridazine and 3-iodo-4-methylbenzoic acid
as Sonogashira coupling partners.
Example 15
N-(3-(Imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylphenyl)-4-((4-methylpip-
erazin-1-yl)methyl)-3-(trifluoromethyl)benzamide
##STR00060##
[0268] The title compound was synthesized from
3-ethynylimidazo[1,2-b]pyridazine and
N-(3-iodo-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoro-
methyl)benzamide in a manner similar to that described for Example
14. The product was obtained as a solid: 533 m/z (M+H).
N-(3-Iodo-4-methylphenyl)-4-((4-methylpiperazin-1-yl)methyl)-3-(trifluorom-
ethyl)benzamide
[0269] To a flask containing 1.0 g (2.67 mmol) of
4-[(4-methyl-1-piperazinyl)methyl]-3-(trifluoromethyl)-benzoic acid
(CAS#859027-02-4; prepared according to Asaki, T. et al. Bioorg.
Med. Chem. Lett. (2006), 16, 1421-1425), 0.62 g (2.67 mmol) of
3-lodo-4-methylaniline, 0.77 g (4.0 mmol) of
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(EDAC), and 0.43 g (3.2 mmol) of N-hydroxybenzotriazole monohydrate
(HOBt.H.sub.2O) was added 5 mL of DCM and 5 mL of triethylamine.
The solution was stirred at ambient temperature under an atmosphere
of N.sub.2 for 3 days, concentrated, and the crude product purified
by silica gel chromatography (eluted with 100% EtOAc then 10%
MeOH/EtOAc), to provide 0.69 g of product as a white solid.
Example 16
3-(Imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-(4-((4-methylpiperazin--
1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide
##STR00061##
[0271] The title compound was synthesized in a manner similar to
that described for Example 14, from
3-ethynylimidazo[1,2-b]pyridazine and
3-iodo-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)-
phenyl)benzamide (Prepared as described in Example 2). The product
was obtained as a solid: 533 m/z (M+H).
Alternative Synthesis of
3-(Imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-(4-((4-methylpiperazin-
-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide
[0272]
3-(Imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-(4-((4-methylpip-
erazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide and its
mono hydrochloride salt can be prepared in an alternative synthesis
similar to that described in Example 1 from
3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzoic acid and
4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl) aniline (as
prepared in example 2). The
3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzoic acid is
prepared in a manner similar to that described in Example 1 using
3-Ethynylimidazo[1,2-b]pyridazine and 3-iodo-4-methylbenzoic acid
as Sonogashira coupling partners.
Example 17
N-(3-Chloro-4-((4-methylpiperazin-1-yl)methyl)phenyl)-3-(imidazo[1,2-b]pyr-
idazin-3-ylethynyl)-4-methylbenzamide
##STR00062##
[0274] The title compound was synthesized according to Example 14,
from 3-ethynylimidazo[1,2-b]pyridazine and
N-(3-chloro-4-((4-methylpiperazin-1-yl)methyl)phenyl)-3-iodo-4-methylbenz-
amide. The product was obtained as a solid: 499 m/z (M+H).
1-(Bromomethyl)-2-chloro-4-nitro-benzene
[0275] A suspension of 2-chloro-4-nitrotoluene (10.0 g, 58.3 mmol),
N-bromosuccinimide (NBS, 10.9 g, 61.2 mmol), and
2,2'-azobis(2-methylpropionitrile) (AIBN, 0.29 g, 1.75 mmol) in 120
mL of CCl.sub.4 was heated at reflux under an atmosphere of N.sub.2
for 12 h. The reaction mixture was cooled to ambient temperature,
and the solid was filtered and washed with EtOAc. The combined
filtrate was washed with aq. NaHCO.sub.3, dried over
Na.sub.2SO.sub.4, filtered, concentrated on rotovap, and further
dried under vacuum. .sup.1H NMR indicated the ratio of desired
product to unreacted 2-chloro-4-nitrotoluene to be 50:50. This
material was used directly in the next step.
1-(2-Chloro-4-nitrobenzyl)-4-methylpiperazine
[0276] To a solution of crude
1-(bromomethyl)-2-chloro-4-nitro-benzene (29.1 mmol; 50% pure) in
30 mL of DCM was added Et.sub.3N (4.2 mL, 30 mmol) and
1-methylpiperazine (3.4 mL, 30 mmol). After stirring for 3 h at
ambient temperature, aq. NaHCO.sub.3 was added and the mixture was
extracted with DCM. The combined organic layer was dried over
Na.sub.2SO.sub.4, filtered, concentrated, and the resulting residue
was purified by silica gel chromatography (eluted with 5% MeOH/DCM)
to provide 6.80 g of product as a dark yellow oil.
3-Chloro-4-((4-methylpiperazin-1-yl)methyl)aniline
[0277] To a solution of
1-(2-chloro-4-nitrobenzyl)-4-methylpiperazine (0.96 g, 3.6 mmol) in
MeOH/water (4:1, 50 mL) was added 1.80 g (33.7 mmol) of NH.sub.4Cl
and 1.47 g (26.3 mmol) of Fe dust and the mixture heated at reflux
under an atmosphere of N.sub.2 for 2 h (HPLC indicated no
progress). To this was added 4 mL of glacial acetic acid and the
mixture heated at reflux for an additional 2 h. The reaction
mixture was cooled to ambient temperature, filtered, and the
filtrate concentrated. The residue was partitioned between EtOAc
and saturated aq. NaHCO.sub.3, the separated aqueous layer was
extracted with EtOAc, and the combined organics washed with brine
and dried over Na.sub.2SO.sub.4. Upon concentration, the crude
product was purified by silica gel chromatography (eluted with 5-7%
MeOH/DCM; silica gel deactivated with 1% triethylamine/DCM) to
provide 0.53 g of product.
Alternative Synthesis of
N-(3-Chloro-4-((4-methylpiperazin-1-yl)methyl)phenyl)-3-(imidazo[1,2-b]py-
ridazin-3-ylethynyl)-4-methylbenzamide
[0278] N-(3-Chloro-4-((4-methyl
piperazin-1-yl)methyl)phenyl)-3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-m-
ethylbenzamide and its mono hydrochloride salt can be prepared in
an alternative synthesis similar to that described in Example 1
from 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzoic acid
and 3-Chloro-4-((4-methylpiperazin-1-yl)methyl)aniline (as prepared
above). The 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzoic
acid is prepared in a manner similar to that described in Example 1
using 3-Ethynylimidazo[1,2-b]pyridazine and 3-iodo-4-methylbenzoic
acid as Sonogashira coupling partners.
Example 18
N-(3-Cyclopropyl-4-((4-methylpiperazin-1-yl)methyl)phenyl)-3-(imidazo[1,2--
b]pyridazin-3-ylethynyl)-4-methylbenzamide
##STR00063##
[0280] The title compound was synthesized from
3-ethynylimidazo[1,2-b]pyridazine and
N-(3-cyclopropyl-4-((4-methylpiperazin-1-yl)methyl)phenyl)-3-iodo-4-methy-
lbenzamide in a manner similar to that described for Example 14
(nitro reduction performed in a manner similar to that described
for Example 17; 0.25M in MeOH/10% AcOH). The product was obtained
as a solid: 505 m/z (M+H).
1-(2-Cyclopropyl-4-nitrobenzyl)-4-methylpiperazine
[0281] A mixture of 1-(2-bromo-4-nitrobenzyl)-4-methylpiperazine
(0.94 g, 3.0 mmol), 0.77 g (9.0 mmol) of cyclopropylboronic acid,
0.067 g (0.30 mmol) of Pd(OAc).sub.2, 2.87 g (13.5 mmol) of
K.sub.3PO.sub.4, and 0.168 g (0.60 mmol) of tricyclohexylphosphine
in 18 mL of toluene/water (5:1) was heated at reflux under an
atmosphere of N.sub.2 for 19 h. The reaction mixture was
concentrated and the crude product was purified by silica gel
chromatography (eluted with 5% MeOH/DCM; MeOH was pre-saturated
with ammonia gas) to provide 0.80 g of product.
Example 19
3-(Imidazo[1,2-b]pyridazin-3-ylethynyl)-N-(4-((4-methylpiperazin-1-yl)meth-
yl)-3-(trifluoromethyl)phenyl)benzamide
##STR00064##
[0283] The title compound was synthesized from
3-ethynylimidazo[1,2-b]pyridazine and
3-iodo-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)be-
nzamide in a manner similar to that described for Example 14. The
product was obtained as a solid: 519 m/z (M+H).
[0284] The titled compound can also be prepared according to the
alternative synthesis described in. example 1 from
3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzoic acid and
4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)aniline (as
prepared in example 2). The
3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzoic acid is
prepared in a manner similar to that described in Example 1 using
3-Ethynylimidazo[1,2-b]pyridazine and 3-iodo-4-methylbenzoic acid
as Sonogashira coupling partners.
Example 20
N-(4-((4-(2-Hydroxyethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)-
-3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylbenzamide
##STR00065##
[0286] The title compound was synthesized from
3-ethynylimidazo[1,2-b]pyridazine and
N-(4-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl-
)-3-iodo-4-methylbenzamide in a manner similar to that described
for Example 14. The product was obtained as a solid: 563 m/z
(M+H).
Example 21
3-(Imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-(4-(piperazin-1-ylmethy-
l)-3-(trifluoromethyl)phenyl)benzamide
##STR00066##
[0288] The title compound was synthesized from
3-ethynylimidazo[1,2-b]pyridazine and tert-butyl
4-(4-(3-iodo-4-methylbenzamido)-2-(trifluoromethyl)benzyl)piperazine-1-ca-
rboxylate in a manner similar to that described for Example 14.
Following deprotection using saturated MeOH/HCl (g), the product
was obtained as a tris HCl salt: 519 m/z (M+H).
Example 22
Biological Evaluation of Compounds
[0289] Compounds of this invention are evaluated in a variety of
assays to determine their biological activities. For example, the
compounds of the invention can be tested for their ability to
inhibit various protein kinases of interest. Some of the compounds
tested displayed potent nanomolar activity against the following
kinases: Abl, Abl T315I, Src and FGFR. Furthermore, several of
these compounds were screened for antiproliferative activity in
BaF3 cells transfected with either wild-type Bcr-Abl or the Bcr-Abl
T315I mutant and demonstrated activity in the range of 1-100
nM.
[0290] The compounds can also be evaluated for their cytotoxic or
growth inhibitory effects on tumor cells of interest, e.g., as
described in more detail below and as shown above for some
representative compounds. See e.g., WO 03/000188, pages 115-136,
the full contents of which are incorporated herein by
reference.
[0291] Some representative compounds are depicted below.
TABLE-US-00001 T315I cell proliferation Compounds of the Invention
(nM) ##STR00067## <1000 ##STR00068## <1000 ##STR00069##
<1000 ##STR00070## <1000 ##STR00071## <1000 ##STR00072##
<1000 ##STR00073## <1000 ##STR00074## <1000 ##STR00075##
<1000 ##STR00076## <1000 ##STR00077## <1000 ##STR00078##
<1000 ##STR00079## <1000 ##STR00080## <1000 ##STR00081##
<1000 ##STR00082## <1000 ##STR00083## <1000 ##STR00084##
<1000 ##STR00085## <1000 ##STR00086## <1000 ##STR00087##
<1000 ##STR00088## <1000 ##STR00089## <1000 ##STR00090##
<1000 ##STR00091## <1000 ##STR00092## <1000 ##STR00093##
<1000 ##STR00094## <1000 ##STR00095## <1000 ##STR00096##
<1000 ##STR00097## <1000 ##STR00098## <1000 ##STR00099##
<1000 ##STR00100## <1000 ##STR00101## <1000 ##STR00102##
<1000 ##STR00103## <1000 ##STR00104## <1000 ##STR00105##
<1000 ##STR00106## <1000 ##STR00107## <1000 ##STR00108##
<1000 ##STR00109## <1000 ##STR00110## <1000 ##STR00111##
<1000 ##STR00112## <1000 ##STR00113## <1000 ##STR00114##
<1000 ##STR00115## <1000 ##STR00116## <1000 ##STR00117##
<1000 ##STR00118## <1000 ##STR00119## <1000 ##STR00120##
<1000 ##STR00121## <1000 ##STR00122## <1000 ##STR00123##
<1000 ##STR00124## <1000 ##STR00125## <1000 ##STR00126##
<1000 ##STR00127## <1000 ##STR00128## <1000 ##STR00129##
<1000 ##STR00130## <1000 ##STR00131## <1000 ##STR00132##
<1000 ##STR00133## <1000 ##STR00134## <1000 ##STR00135##
<1000 ##STR00136## <1000 ##STR00137## <1000 ##STR00138##
<1000 ##STR00139## <1000 ##STR00140## <1000 ##STR00141##
<1000 ##STR00142## <1000 ##STR00143## <1000 ##STR00144##
<1000 ##STR00145## <1000 ##STR00146## <1000 ##STR00147##
<1000 ##STR00148## <1000 ##STR00149## <1000 ##STR00150##
<1000 ##STR00151## <1000 ##STR00152## <1000 ##STR00153##
<1000 ##STR00154## <1000 ##STR00155## <1000 ##STR00156##
<1000 ##STR00157## <1000 ##STR00158## <1000 ##STR00159##
<1000
[0292] The compounds listed in the table below also showed
inhibitory activity against various protein kinase of interest.
##STR00160## ##STR00161## ##STR00162## ##STR00163##
##STR00164##
Kinase Inhibition
[0293] More specifically, the compounds described herein are
screened for kinase inhibition activity as follows. Kinases
suitable for use in the following protocol include, but are not
limited to: Abl, Lck, Lyn, Src, Fyn, Syk, Zap-70, Itk, Tec, Btk,
EGFR, ErbB2, Kdr, FIt1, Flt-3, Tek, c-Met, InsR, and AKT.
[0294] Kinases are expressed as either kinase domains or full
length constructs fused to glutathione S-transferase (GST) or
polyHistidine tagged fusion proteins in either E. coli or
Baculovirus-High Five expression systems. They are purified to near
homogeneity by affinity chromatography as previously described
(Lehr et al., 1996; Gish et al., 1995). In some instances, kinases
are co-expressed or mixed with purified or partially purified
regulatory polypeptides prior to measurement of activity.
[0295] Kinase activity and inhibition can be measured by
established protocols (see e.g., Braunwalder et al., 1996). In such
cases, the transfer of .sup.33P0.sub.4 from ATP to the synthetic
substrates poly(Glu, Tyr) 4:1 or poly(Arg, Ser) 3:1 attached to the
bioactive surface of microtiter plates is taken as a measure of
enzyme activity. After an incubation period, the amount of
phosphate transferred is measured by first washing the plate with
0.5% phosphoric acid, adding liquid scintillant, and then counting
in a liquid scintillation detector. The IC50 is determined by the
concentration of compound that causes a 50% reduction in the amount
of .sup.33P incorporated onto the substrate bound to the plate.
[0296] In one method, the activated kinase is incubated with a
biotinylated substrate peptide (containing tyr) with or without the
presence of a compound of the invention. After the kinase assay
incubation period, excess kinase inhibitor is added to kill the
kinase reaction along with Europium-labeled anti-phosphotyrosine
antibody (Eu-Ab) and Allophycocyanin-Streptavidin (SA-APC). The
biotinylated substrate peptide (with or without phosphorylated
Tyrosine) in solution binds to the SA-APC via Biotin-Avidin
binding. The Eu-Ab binds only to substrate with phosphorylated
tryrosine. When the solution is excited at 615 nm, there is an
energy transfer from the Europium to the APC when they are in close
proximity (i.e. attached to the same molecule of biotinylated and
phosphorylated substrate peptide). The APC then fluoresces at a
wavelength of 665 nm. Excitation and emission take place in a
Wallac Victor.sup.2 V plate reader where the plate is read
fluorometrically and absorbances at 615 and 665 nm are recorded.
These data are then processed by an Excel plate processor which
calculates IC50s of test compounds by converting the fluorescence
into amounts of phosphorylated substrate made and determining the
concentration of test compound that would be required to inhibit
the development of phosphorylated substrate by 50% (IC50).
[0297] Other methods relying upon the transfer of phosphate to
peptide or polypeptide substrate containing tyrosine, serine,
threonine or histidine, alone, in combination with each other, or
in combination with other amino acids, in solution or immobilized
(i.e., solid phase) are also useful.
[0298] For example, transfer of phosphate to a peptide or
polypeptide can also be detected using scintillation proximity,
Fluorescence Polarization or homogeneous time-resolved
fluorescence. Alternatively, kinase activity can be measured using
antibody-based methods in which an antibody or polypeptide is used
as a reagent to detect phosphorylated target polypeptide.
[0299] For additional background information on such assay
methodologies, see e.g., Braunwalder et al., 1996, Anal. Biochem.
234(l):23; Cleaveland et al., 1990, Anal Biochem. 190(2):249 Gish
et al. (1995). Protein Eng. 8(6):609 Kolb et al. (1998). Drug
Discov. Toda V. 3:333 Lehr et al. (1996). Gene 169(2):27527-87
Seethala et al. (1998). Anal Biochem. 255(2):257 Wu et al.
(2000).
[0300] IC50 values in the low nanomolar range have been observed
for compounds of this invention against various kinases, including
Src, Abl and kdr.
Cell-Based Assays
[0301] Certain compounds of this invention have also been
demonstrated cytotoxic or growth inhibitory effects on tumor and
other cancer cell lines and thus may be useful in the treatment of
cancer and other cell proliferative diseases. Compounds are assayed
for anti-tumor activity using in vivo and in vitro assays which are
well known to those skilled in the art. Generally, initial screens
of compounds to identify candidate anti-cancer drugs are performed
in cellular assays. Compounds identified as having
anti-proliferative activity in such cell-based assays can then be
subsequently assayed in whole organisms for anti-tumor activity and
toxicity. Generally speaking, cell-based screens can be performed
more rapidly and cost-effectively relative to assays that use whole
organisms. For purposes of this invention, the terms "anti-tumor"
and "anti-cancer" activity are used interchangeably.
[0302] Cell-based methods for measuring antiproliferative activity
are well known and can be used for comparative characterization of
compounds of this invention. In general, cell proliferation and
cell viability assays are designed to provide a detectable signal
when cells are metabolically active. Compounds may be tested for
antiproliferative activity by measuring any observed decrease in
metabolic activity of the cells after exposure of the cells to
compound. Commonly used methods include, for example, measurement
of membrane integrity (as a measure of cell viability)(e.g. using
trypan blue exclusion) or measurement of DNA synthesis (e.g. by
measuring incorporation of BrdU or 3H-thymidine).
[0303] Some methods for assaying cell proliferation use a reagent
that is converted into a detectable compound during cell
proliferation. Particularly preferred compounds are tetrazolium
salts and include without limitation MTT (3-(4,
5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide;
Sigma-Aldrich, St. Louis, Mo.), MTS
(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl-
)-2H-tetrazolium), XTT
(2,3-bis(2-Methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide)-
, INT, NBT, and NTV (Bernas et al. Biochim Biophys Acta
1451(1):73-81, 1999). Preferred assays utilizing tetrazolium salts
detect cell proliferation by detecting the product of the enzymatic
conversion of the tetrazolium salts into blue formazan derivatives,
which are readily detected by spectroscopic methods (Mosman. J.
Immunol. Methods. 65:55-63, 1983).
[0304] Generally, preferred methods for assaying cell proliferation
involve incubating cells in a desired growth medium with and
without the compounds to be tested. Growth conditions for various
prokaryotic and eukaryotic cells are well-known to those of
ordinary skill in the art (Ausubel et al. Current Protocols in
Molecular Biology. Wiley and Sons. 1999; Bonifacino et al. Current
Protocols in Cell Biology. Wiley and Sons. 1999 both incorporated
herein by reference). To detect cell proliferation, the tetrazolium
salts are added to the incubated cultured cells to allow enzymatic
conversion to the detectable product by active cells. Cells are
processed, and the optical density of the cells is determined to
measure the amount of formazan derivatives. Furthermore,
commercially available kits, including reagents and protocols, are
availabe for examples, from Promega Corporation (Madison, Wis.),
Sigma-Aldrich (St. Louis, Mo.), and Trevigen (Gaithersburg,
Md.).
[0305] More specifically, the cell proliferation assay we currently
perform is using CellTiter 96 AQueous One Solution Cell
Proliferation assay kit (Promaga, Cat#G3581). This assay is a
colorimetric method for determining the number of alive cells in
proliferation or cytotoxicity assays. The assay utilizing
terazolium salts detect cell proliferation by detecting the product
of the enzymatic conversion of the tetrazolium salts into blue
formazan derivatives, which can be measured by the absorbance at
490 nm in a plate reader, Wallac Victor.sup.2V (PerkinElmer).
[0306] An example of cell-based assay is shown as below. The cell
lines used in the assay are Ba/F3, a murine pro-B cell line, which
have been stably transfected with full-length wild-type Bcr-Abl or
Bcr-Abl with various kinase domain point mutations (including
T351I, Y253F, E255K, H396P, M351T etc) constructs. Parental Ba/F3
cell line is used as control. These cell lines were obtained from
Brian J. Druker (Howard Hughes Medical Institute, Oregon Health and
Science University, Portland, Oreg., USA). Ba/F3 cell expressing
Bcr-Abl or Bcr-Abl mutants were maintained in PRMI 1640 growth
medium with 200 .mu.M L-gultamine, 10% FCS, penicillin (200 U/ml),
and streptomycin (200 .mu.g/ml). Parental Ba/F3 cells were culture
in the same medium supplemented with 10 ng/ml IL-3.
[0307] Parental Ba/F3 cells (supplemented with IL-3) or Ba/F3 cells
expressing WT or mutant Bcr-Abl are plated in duplicate at
1.times.10.sup.4 cells/well in 96-well plates with the compounds in
different concentrations in the media. The compounds are first
dissolved and diluted in DMSO by preparation of 4-fold dilution;
next equal volumes of compounds with DMSO are transferred to medium
and then transferred to cell plates. The final compound
concentrations start from 10 .mu.M to 6 nM. DMSO at same percentage
is used as control. After compound was incubated with cells for 3
days, the numbers of active cells are measured using CellTiter 96
AQueous One Solution Cell Proliferation assay kit following the kit
instruction. Basically, the tetrazolium salts are added to the
incubated cultured cells to allow enzymatic conversion to the
detectable product by active cells. Cells are processed, and the
optical density of the cells is determined to measure the amount of
formazan derivatives. Mean+/-SD are generated from duplicated wells
and reported as the percentage absorbance of control. IC50s are
calculated in best-fit curves using Microsoft Excel-fit
software.
[0308] In addition, a wide variety of cell types may be used to
screen compounds for antiproliferative activity, including the
following cell lines, among others: COLO 205 (colon cancer), DLD-1
(colon cancer), HCT-15 (colon cancer), HT29 (colon cancer), HEP G2
(Hepatoma), K-562 (Leukemia), A549 (Lung), NCI-H249 (Lung), MCF7
(Mammary), MDA-MB-231 (Mammary), SAOS-2 (Osteosarcoma), OVCAR-3
(Ovarian), PANC-1 (Pancreas), DU-145 (Prostate), PC-3 (Prostate),
ACHN (Renal), CAKI-1 (Renal), MG-63 (Sarcoma).
[0309] While the cell line is preferably mammalian, lower order
eukaryotic cells such as yeast may also be used to screen
compounds. Preferred mammalian cell lines are derived from humans,
rats, mice, rabbits, monkeys, hamsters, and guinea pigs since cells
lines from these organisms are well-studied and characterized.
However, others may be used as well.
[0310] Suitable mammalian cell lines are often derived from tumors.
For example, the following tumor cell-types may be sources of cells
for culturing cells: melanoma, myeloid leukemia, carcinomas of the
lung, breast, ovaries, colon, kidney, prostate, pancreas and
testes), cardiomyocytes, endothelial cells, epithelial cells,
lymphocytes (T-cell and B cell), mast cells, eosinophils, vascular
intimal cells, hepatocytes, leukocytes including mononuclear
leukocytes, stem cells such as haemopoetic, neural, skin, lung,
kidney, liver and myocyte stem cells (for use in screening for
differentiation and de-differentiation factors), osteoclasts,
chondrocytes and other connective tissue cells, keratinocytes,
melanocytes, liver cells, kidney cells, and adipocytes.
Non-limiting examples of mammalian cells lines that have been
widely used by researchers include HeLa, NIH/3T3, HT1080, CHO,
COS-1, 293T, WI-38 and CV1/EBNA-1.
[0311] Other cellular assays may be used which rely upon a reporter
gene to detect metabolically active cells. Non-limiting examples of
reporter gene expression systems include green fluorescent protein
(GFP), and luciferase. As an example of the use of GFP to screen
for potential antitumor drugs, Sandman et al. (Chem Biol. 6:541-51;
incorporated herein by reference) used HeLa cells containing an
inducible variant of GFP to detect compounds that inhibited
expression of the GFP, and thus inhibited cell proliferation.
[0312] Compounds identified by such cellular assays as having
anti-cell proliferation activity are then tested for anti-tumor
activity in whole organisms. Preferably, the organisms are
mammalian. Well-characterized mammalians systems for studying
cancer include rodents such as rats and mice. Typically, a tumor of
interest is transplanted into a mouse having a reduced ability to
mount an immune response to the tumor to reduce the likelihood of
rejection. Such mice include for example, nude mice (athymic) and
SCID (severe combined immunodeficiency) mice. Other transgenic mice
such as oncogene containing mice may be used in the present assays
(see for example U.S. Pat. No. 4,736,866 and U.S. Pat. No.
5,175,383). For a review and discussion on the use of rodent models
for antitumor drug testing see Kerbel (Cancer Metastasis Rev.
17:301-304, 1998-99).
[0313] In general, the tumors of interest are implanted in a test
organism preferably subcutaneously. The organism containing the
tumor is treated with doses of candidate anti-tumor compounds. The
size of the tumor is periodically measured to determine the effects
of the test compound on the tumor. Some tumor types are implanted
at sites other than subcutaneous sites (e.g. intraperitoneal sites)
and survival is measured as the endpoint. Parameters to be assayed
with routine screening include different tumor models, various
tumor and drug routes, and dose amounts and schedule. For a review
of the use of mice in detecting antitumor compounds see Corbett et
al. (Invest New Drugs. 15:207-218, 1997; incorporated herein by
reference).
Example 23
Pharmaceutical Compositions
[0314] Representative pharmaceutical dosage forms of the compounds
of this invention (the active ingredient being referred to as
"Compound"), are provided for therapeutic or prophylactic use in
humans:
TABLE-US-00002 (a) Tablet I mg/tablet Compound 100 Lactose Ph. Eur
182.75 Croscarmellose sodium 12.0 Maize starch paste (5% w/v paste)
2.25 Magnesium stearate 3.0 (b) Tablet II mg/tablet Compound 50
Lactose Ph. Eur 223.75 Croscarmellose sodium 6.0 Maize starch 15.0
Polyvinylpyffolidone (5% w/v paste) 2.25 Magnesium stearate 3.0 (c)
Tablet III mg/tablet Compound 1.0 Lactose Ph. Eur 93.25
Croscarmellose sodium 4.0 Maize starch paste (5% w/v paste) 0.75
Magnesium stearate 1.0-76 (d) Capsule mg/capsule Compound 10
Lactose Ph. Eur 488.5 Magnesium 1.5 (e) Injection I (50 mg/ml)
Compound 5.0% w/v 1M Sodium hydroxide solution 15.0% v/v 0. lM
Hydrochloric acid (to adjust pH to 7.6) Polyethylene glycol 400
4.5% w/v Water for injection to 100% (f) Injection II (10 mg/ml)
Compound 1.0% W/v Sodium phosphate BP 3.6% w/v O. 1M Sodium
hydroxide solution 15.0% v/v Water for injection to 100% (g)
Injection III (1 mg/ml, buffered to pH 6) Compound 0. l % w/v
Sodium phosphate BP 2.26% w/v Citric acid 0.38% w/v Polyethylene
glycol 400 3.5% w/v Water for injection to 100% (h) Aerosol l mg/ml
Compound 10.0 Sorbitan trioleate 13.5 Trichlorofluoromethane 910.0
Dichlorodifluoromethane 490.0 (i) Aerosol II mg/ml Compound 0.2
Sorbitan trioleate 0.27 Trichlorofluoromethane 70.0
Dichlorodifluoromethane 280.0 Dichlorotetrafluoroethane 1094.0 (j)
Aerosol III mg/ml Compound 2.5 Sorbitan trioleate 3.38
Trichlorofluoromethane 67.5 Dichlorodifluoromethane 1086.0
Dichlorotetrafluoroethane 191.6 (k) Aerosol IV mg/ml Compound 2.5
Soya lecithin 2.7 Trichlorofluoromethane 67.5
Dichlorodifluoromethane 1086.0 Dichlorotetrafluoroethane 191.6 (1)
Ointment ml Compound 40 mg Ethanol 300 .mu.l Water 300 .mu.l
1-Dodecylazacycloheptan one 50 .mu.l Propylene glycol to 1 ml
[0315] Note: These formulations may be prepared using conventional
procedures well known in the pharmaceutical art. The tablets
(a)-(c) may be enteric coated by conventional means, if desired to
provide a coating of cellulose acetate phthalate, for example. The
aerosol formulations (h)-(k) may be used in conjunction with
standard, metered dose aerosol dispensers, and the suspending
agents sorbitan trioleate and soya lecithin may be replaced by an
alternative suspending agent such as sorbitan monooleate, sorbitan
sesquioleate, polysorbate 80, polyglycerol oleate or oleic
acid.
* * * * *
References