U.S. patent application number 11/567717 was filed with the patent office on 2007-06-07 for pyrimidine-2,4-diamines and their uses.
Invention is credited to Somasekhar Phamidipati, Rajinder Singh.
Application Number | 20070129360 11/567717 |
Document ID | / |
Family ID | 38119605 |
Filed Date | 2007-06-07 |
United States Patent
Application |
20070129360 |
Kind Code |
A1 |
Phamidipati; Somasekhar ; et
al. |
June 7, 2007 |
Pyrimidine-2,4-diamines and Their Uses
Abstract
The present invention relates to diaryl pyrimidine-2,4-diamines,
pharmaceutical compositions thereof, and the use of the compounds
and compositions for the inhibition of kinases. The compounds,
analogs, and pharmaceutically acceptable salts thereof, and
pharmaceutical compositions can be used in the treatment and
prevention of cancer.
Inventors: |
Phamidipati; Somasekhar;
(Foster City, CA) ; Singh; Rajinder; (Belmont,
CA) |
Correspondence
Address: |
FENWICK & WEST LLP
SILICON VALLEY CENTER
801 CALIFORNIA STREET
MOUNTAIN VIEW
CA
94041
US
|
Family ID: |
38119605 |
Appl. No.: |
11/567717 |
Filed: |
December 6, 2006 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
11295752 |
Dec 6, 2005 |
|
|
|
11567717 |
Dec 6, 2006 |
|
|
|
Current U.S.
Class: |
514/230.5 ;
514/232.8; 544/105; 544/122; 544/295; 544/324 |
Current CPC
Class: |
A61P 19/02 20180101;
C07D 498/04 20130101; A61P 43/00 20180101; A61P 35/00 20180101 |
Class at
Publication: |
514/230.5 ;
544/122; 544/295; 544/324; 514/232.8; 544/105 |
International
Class: |
A61K 31/538 20060101
A61K031/538; C07D 491/02 20060101 C07D491/02 |
Claims
1. A compound of formula (I) ##STR23## wherein R.sub.3 is aryl or
heteroaryl, each optionally substituted; X.sub.3 and X.sub.4 are
independently selected from CH or N; X.sub.5 is selected from the
group consisting of CR.sub.12R.sub.13, O, S, SO, SO.sub.2, and
NR.sub.14 wherein R.sub.12 and R.sub.13 are independently selected
from H, OH, lower alkyl, or together form an oxo group; R.sub.4 is
an electronegative group; R', R.sub.5, R.sub.6 and R.sub.14 are
independently selected from H, lower alkyl, a progroup, cycloalkyl
or aryl, and wherein at least one of R', R.sub.5, R.sub.6 or
R.sub.14 is said progroup which is linked via a carbamate,
thiocarbamate, dithiocarbamate, urea, or thiourea; R.sub.7 and
R.sub.8 are independently selected from the group consisting of H,
halogen, lower alkyl, cycloalkyl, aryl, and heteroaryl; and R.sub.9
and R.sub.10 are independently selected from the group consisting
of H, halogen, --OH, -alkoxy, lower alkyl, cycloalkyl, aryl, and
heteroaryl wherein R.sub.7 and R.sub.8, or R.sub.9 and R.sub.10
together form an oxo group, and wherein R.sub.9 or R.sub.10 are not
--OH or alkoxy when X.sub.5 is NR.sub.14.
2. A compound of formula (II) ##STR24## wherein X.sub.1 is selected
from the group consisting of O, S, and NR.sub.11 wherein R.sub.11
is H or lower alkyl; X.sub.2 is O or S; X.sub.3 and X.sub.4 are
independently selected from CH or N; X.sub.5 is selected from the
group consisting of CR.sub.12R.sub.13, O, S, SO, SO.sub.2, and
NR.sub.14 wherein R.sub.12 and R.sub.13 are independently selected
from H, OH, lower alkyl or together form an oxo group, and R.sub.14
is H or lower alkyl; R is selected from the group consisting of
straight or branched, saturated or unsaturated alkyl, allyl,
cycloalkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl,
prenylalkaryl and heteroarylalkyl; R.sub.1 and R.sub.2 are each
independently selected from the group consisting of H, OH,
--OR.sub.11, NR.sub.15R.sub.15, halo, lower alkyl, --C(O)O-alkyl,
--C(O)OH, --OP(.dbd.O)(OR.sub.11).sub.2, --OC(.dbd.O)OR.sub.11,
--OC(.dbd.O)R.sub.11, cycloalkyl, aryl, and heteroaryl or together
form an oxo, wherein each R.sub.15 is independently selected from
the group consisting of H, lower alkyl, prenyl, allyl,
--C(O)O-alkyl, cycloalkyl, aryl, heteroaryl, alkaryl and
alkheteroaryl, or two of R.sub.15 combine to form an optionally
substituted cycloheteroalkyl; R.sub.3 is aryl or heteroaryl, each
optionally substituted; R.sub.4 is an electronegative group;
R.sub.5 and R.sub.6 are independently selected from H, lower alkyl,
cycloalkyl or aryl; R.sub.7, R.sub.8, R.sub.9, and R.sub.10 are
independently selected from the group consisting of H, OH, halogen,
lower alkyl, cycloalkyl, aryl, and heteroaryl, or wherein R.sub.7
and R.sub.8 or R.sub.9 and R.sub.10 together form an oxo group; and
n is an integer from 0 to 10.
3. The compound of claim 2, wherein X.sub.1 is O.
4. The compound of claim 3, wherein X.sub.2 is O.
5. The compound of claim 2, wherein R.sub.3 is optionally
substituted aryl.
6. The compound of claim 5, wherein aryl is alkoxyphenyl,
dialkoxyphenyl, or trialkoxyphenyl.
7. The compound of claim 6, wherein trialkoxyphenyl is
trimethoxyphenyl.
8. The compound of claim 2, wherein R.sub.4 is H, OH, halogen,
cyano, nitro, trifluoromethyl, difluoromethyl, fluoromethyl,
trifluoromethoxy, difluoromethoxy, or fluoromethoxy.
9. The compound of claim 8, wherein R.sub.4 is F.
10. The compound of claim 2, wherein R.sub.5 and R.sub.6 are H.
11. The compound of claim 2, wherein X.sub.4 is CH.
12. The compound of claim 11, wherein X.sub.4 is N.
13. The compound of claim 2, wherein X.sub.5 is CH.sub.2 or O.
14. The compound of claim 2, wherein R.sub.7 and R.sub.8 are H or
together form the oxo group.
15. The compound of claim 2, wherein R.sub.9 and R.sub.10 are
methyl.
16. The compound of claim 2, wherein R is cycloheteroalkyl.
17. The compound of claim 16, wherein R is substituted or
unsubstituted morpholine, or substituted or unsubstituted
pyrrolidine.
18. The compound of claim 2, wherein R heteroaryl.
19. The compound of claim 18, wherein heteroaryl is substituted or
unsubstituted indole.
20. The compound of claim 2, wherein R is selected from the group
consisting of acetate, amino, and dialkylamino.
21. The compound of claim 2, wherein n is 1, 2, or 3.
22. The compound of claim 1, wherein X.sub.1 is NR.sub.11.
23. The compound of claim 22, wherein X.sub.2 is O.
24. The compound of claim 22, wherein X.sub.2 is S.
25. The compound of claim 1, wherein X.sub.1 is S.
26. The compound of claim 25, wherein X.sub.2 is O.
27. A compound of formula (III): ##STR25## wherein R is selected
from the group consisting of straight or branched, saturated or
unsaturated alkyl, allyl, cycloalkyl, cycloheteroalkyl, aryl, and
heteroaryl; R.sub.11 is H or lower alkyl; and n is and integer
between 0 and 10.
28. The compound of claim 27, wherein R is morpholine.
29. The compound of claim 27, wherein R is 1-methylpiperidine.
30. The compound of claim 27, wherein R is piperazine or
3-piperazinepropane sulfonate.
31. The compound of claim 27, wherein R is dimethylamine.
32. The compound of claim 27, wherein R is tryptamine or
N-tert-butylaceyltryptamine.
33. The compound of claim 27, wherein n is 0, 1, 2, or 3.
34. A method of inhibiting a kinase, the method comprising
contacting the kinase with a compound of formula (II) ##STR26##
wherein X.sub.1 is selected from the group consisting of O, S, and
NR.sub.11 wherein R.sub.11 is H or lower alkyl; X.sub.2 is O or S;
X.sub.3 and X.sub.4 are independently selected from CH or N;
X.sub.5 is selected from the group consisting of CR.sub.12R.sub.13,
O, S, SO, SO.sub.2, and NR.sub.14 wherein R.sub.12 and R.sub.13 are
independently selected from H, OH, lower alkyl together form an oxo
group, and R.sub.14 is H or lower alkyl; R is selected from the
group consisting of straight or branched, saturated or unsaturated
alkyl, allyl, cycloalkyl, cycloalkyl, cycloheteroalkyl, aryl,
heteroaryl, prenylalkaryl and heteroarylalkyl; R.sub.1 and R.sub.2
are each independently selected from the group consisting of H, OH,
--OR.sub.11, NR.sub.15R.sub.15, halo, lower alkyl, --C(O)O-alkyl,
--C(O)OH, --OP(.dbd.O)(OR.sub.11).sub.2, --OC(.dbd.O)OR.sub.11,
--OC(.dbd.O)R.sub.11, cycloalkyl, aryl, and heteroaryl or together
form an oxo, wherein each R.sub.15 is independently selected from
the group consisting of H, lower alkyl, prenyl, allyl,
--C(O)O-alkyl, cycloalkyl, aryl, heteroaryl, alkaryl and
alkheteroaryl, or two of R.sub.15 combine to form an optionally
substituted cycloheteroalkyl; R.sub.3 is aryl or heteroaryl, each
optionally substituted; R.sub.4 is an electronegative group;
R.sub.5 and R.sub.6 are independently selected from H, lower alkyl,
cycloalkyl or aryl; R.sub.7, R.sub.8, R.sub.9, and R.sub.10 are
independently selected from the group consisting of H, OH, halogen,
lower alkyl, cycloalkyl, aryl, and heteroaryl, or wherein R.sub.7
and R.sub.8 or R.sub.9 and R.sub.10 together form an oxo group; and
n is an integer from 0 to 10.
35. The method of claim 34, wherein the compound is according to
the following formula: ##STR27## and salts thereof.
36. The method of claim 34, wherein the compound is according to
the following formula: ##STR28## and salts thereof.
37. The method of claim 34, wherein the compound is according to
the following formula: ##STR29## and salts thereof.
38. The method of claim 34, wherein the compound is according to
the following formula: ##STR30## and salts thereof.
39. The method of claim 34, wherein the compound is according to
the following formula: ##STR31## and salts thereof.
40. The method of claim 34, wherein the compound is according to
the following formula: ##STR32## and salts thereof.
41. The method of claim 34, wherein the compound is according to
the following formula: ##STR33## and salts thereof.
42. The method of claim 34, wherein the kinase is a tyrosine
kinase.
43. The method of claim 42, wherein the tyrosine kinase is Syk
kinase.
44. A method for treating or preventing a disorder associated with
a kinase, the method comprising: administering to a subject an
effective amount of a compound of claim 1 or acceptable salts,
N-oxides, hydrates, or solvates thereof, and
pharmaceutically-acceptable carrier or diluent.
45. The method of claim 44, wherein the disorder is cancer.
46. The method of claim 45, wherein the cancer is breast cancer,
colon cancer, lung cancer, prostate cancer, or hematopoietic tumors
of lymphoid lineage.
47. The method of claim 44, wherein the subject is a domestic
animal.
48. The method of claim 44, wherein the subject is a human.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation of U.S. patent
application Ser. No. 11/295,752 filed Dec. 6, 2005, from which
priority is claimed and which application is incorporated herein by
reference in its entirety.
FIELD OF INVENTION
[0002] The invention relates to compounds containing the
pyrimidine-2,4-diamine moiety, particularly diaryl
pyrimidine-2,4-diamines, compositions comprising the compounds, and
methods of using the compounds and compositions for the inhibition
of kinases. The compounds and compositions are useful for treating
or modulating disease in which kinases may be involved, symptoms of
such disease, or the effect of other physiological events mediated
by kinases.
BACKGROUND OF THE INVENTION
[0003] The protein kinases constitute a large family of
structurally related enzymes that are responsible for the control
of a wide variety of signal transduction processes within the cell.
(Hardie and Hanks (1995) The Protein Kinase Facts Book, I and II,
Academic Press, San Diego, Calif.). Protein kinases are thought to
have evolved from a common ancestral gene due to the conservation
of their structure and catalytic function. Almost all kinases
contain a similar 250-300 amino acid catalytic domain. Protein
kinases catalyze phosphorylation of the hydroxyl moiety of serine,
threonine or tyrosine. Thus, the kinases may be categorized into
families by the substrates they phosphorylate (e.g.,
protein-tyrosine, protein-serine/threonine, lipids, etc.), and
sequence motifs generally corresponding to each of the kinase
families have been identified.
[0004] The phosphorylation and dephosphorylation is an important
post-translational control element in eukaryotic signal
transduction. The phosphorylation state of a given protein can
govern its enzyme activity, protein-protein binding interactions,
and cellular distribution. Thus, protein kinases represent a large
family of proteins which play a central role in the regulation of a
wide variety of cellular processes, maintaining control over
cellular function. A partial list of such kinases includes abl,
AKT, bcr-abl, Blk, Brk, Btk, c-kit, c-met, c-src, CDK1, CDK2, CDK3,
CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, cRafl, CSFir, CSK, EGFR,
ErbB2, ErbB3, ErbB4, Erk, Fak, fes, FGFR1, FGFR2, FGFR3, FGFR4,
FGFR5, Fgr, flt-1, Fps, Frk, Fyn, Hck, IGF-1R, INS-R, Jak, KDR,
Lck, Lyn, MEK, p38, PDGFR, PIK, PKC, PYK2, ron, Syk, Src, tie,
tie2, TRK, Yes, and Zap70. Inhibition of the kinases has become an
important therapeutic target.
[0005] Kinases regulate many different cell processes including,
but not limited to, proliferation, differentiation, apoptosis,
motility, transcription, translation and other signaling processes,
by adding phosphate groups to target proteins. Phosphorylation of
target proteins occurs in response to a variety of extracellular
signals (hormones, neurotransmitters, growth and differentiation
factors, etc.), cell cycle events, environmental or nutritional
stresses, etc. The appropriate protein kinase functions in
signaling pathways to activate or inactivate, for example, a
metabolic enzyme, regulatory protein, receptor, cytoskeletal
protein, ion channel or pump, or transcription factor. Uncontrolled
signaling due to defective control of protein phosphorylation has
been implicated in a number of diseases, including, for example,
inflammation, cancer, allergy/asthma, disease and conditions of the
immune system, disease and conditions of the central nervous
system, and angiogenesis.
[0006] For example, the Src family is composed of ten highly
homologous cytosolic kinases which are critical components in an
array of cell signaling pathways ranging from lymphocyte activation
to cell growth and proliferation. Constitutive activation of these
enzymes can lead to oncogenic cell transformation, making them
putative drug targets for cancer therapies. Because of their
importance in the regulation of these fundamental cellular
processes, many studies have focused on developing inhibitors for
the Src family kinase. However, the potent inhibitors that have
been discovered lack the high selectivity that would be required
for probing the cellular inhibition of an individual target kinase.
Conventional inhibitor screens have produced few if any molecules
which can discriminate between the active sites of the various Src
family kinases.
[0007] WO01/00213 describes substituted pyrimidines as Src kinase
inhibitors. WO01/40218 describes arylamine derivatives for use as
anti-telomerase agents. WO00/39101 describes substituted
pyrimidines as anti-cancer agents. WO01/29009 describes substituted
pyrimidines as kinase inhibitors, while WO00/39101, WO00/59892, and
WO01/47921 describe amino substituted pyrimidines as kinase
inhibitors. U.S. Pat. No. 6,080,858 describes a process for
preparing substituted pyrimidines. WO01/19825 describes amino
substituted pyrimidines as synthetic intermediates. WO01/72745
describes 4-heteroaryl-substituted pyrimidines as inhibitors of
CDK's. WO01/72717 describes 4-amino-5-cyanopyrimidines as
inhibitors of CDK's. WO02/22601 describes
4-(pyrazol-5-ylamino)-pyrimidines as kinase inhibitors. WO02/46184
describes 4-(4-pyrazolyl)-pyrimidines as kinase inhibitors.
WO02/46170 and WO02/46171 describes 2-anilino-pyrimidines as
inhibitors of JNK and IKK, respectively. WO02/47690 describes
4-arylamino-pyrimidines as kinase inhibitors.
[0008] Many of the active 2,4-pyrimidinediamine compounds are also
potent inhibitors of the tyrosine kinase Syk kinase. Examples of
such 2,4-pyrimidinediamine are described, for example, in U.S.
application Ser. No. 10/355,543 filed Jan. 31, 2003
(US2004/0029902A1), international application Serial No.
PCT/US03/03022 filed Jan. 31, 2003 (WO 03/063794), U.S. application
Ser. No. 10/631,029 filed Jul. 29, 2003, international application
Serial No. PCT/US03/24087 (WO2004/014382), U.S. application Ser.
No. 10/903,263 filed Jul. 30, 2004 (US2005/0234049), and
international application Serial No. PCT/US2004/24716
(WO2005/016893).
[0009] The development of selective protein kinase inhibitors that
can block the disease pathologies and/or symptoms resulting from
aberrant protein kinase activity has generated much interest.
However, additional compounds for inhibition of kinases and
treatment and prevention of diseases associated with them are
needed.
SUMMARY OF THE INVENTION
[0010] The present invention provides prodrugs of
pyrimidine-2,4-diamine compounds, compositions comprising the
prodrugs, methods and intermediates useful for synthesizing the
prodrugs and methods of using the prodrugs, including in the
treatment and/or prevention of diseases mediated by kinases.
[0011] The compounds of the invention generally comprise a
biologically active pyrimidine-2,4-diamine compound that is
substituted at the nitrogen atom of one or more primary or
secondary amine groups with a progroup RP. The progroup generally
includes a group or moiety that is metabolized under the conditions
of use to yield the active pyrimidine-2,4-diamine drug, and is
covalently attached to the drug via a carbamate, a thiocarbamate, a
dithiocarbamate, a urea, or a thiourea linkage.
[0012] Virtually any known pyrimidine-2,4-diamine compound that has
biological, and hence therapeutic, activity can be protected at an
available primary or secondary amine of the parent drug molecule
with one or more progroups R.sup.P as described herein. Suitable
active pyrimidine-2,4-diamine compounds are described, for example,
in U.S. application Ser. No. 10/355,543 filed Jan. 31, 2003
(US2004/0029902A1), international application Serial No.
PCT/US03/03022 filed Jan. 31, 2003 (WO 03/063794), U.S. application
Ser. No. 10/631,029 filed Jul. 29, 2003, international application
Serial No. PCT/US03/24087 (WO2004/014382), U.S. application Ser.
No. 10/903,263 filed July 30, (US2005/0234049), and international
application Serial No. PCT/US2004/24716 (WO2005/016893). In such
pyrimidine-2,4-diamine compounds, the progroup(s) can be attached
to any available primary or secondary amine, including, for
example, the N2 nitrogen atom of the 2,4-pyrimidinediamine moiety,
the N4 nitrogen atom of the pyrimidine-2,4-diamine moiety, and/or a
primary or secondary nitrogen atom included in a substituent on the
pyrimidine-2,4-diamine compound.
[0013] The compounds of the invention are potent inhibitors of
kinases. Accordingly, in still another aspect, the present
invention provides methods of inhibiting kinases comprising
contacting a kinase with an effective amount of a compound or
composition of the invention effective for inhibition. The methods
can be practiced either in vitro or in vivo, and can be used as a
therapeutic approach towards the treatment and/or prevention of
diseases such as treatment of neoplasia including cancer and
metastasis, promoting apoptosis, and in the treatment and
prevention of other diseases associated with protein kinases.
[0014] Compounds of the present invention are useful for, but not
limited to, the prevention or treatment of cancer and related
diseases. The compounds of the invention have kinase inhibitory
activity, therefore, the compounds of the invention can be useful
in therapy as antineoplasia agents. Compounds of the invention can
be useful for the treatment of carcinomas, hematopoietic tumors,
solid tumors, sarcomas, retinoblastoma, hematopoietic malignancies,
including leukemias and lymphomas, tumor-induced pleural or
pericardial effusions, and are also useful for promoting
apoptosis.
[0015] The compounds of this invention can act as inhibitors of
protein kinases, such as Syk, Src, ErbB, KDR, CDK-2, LCK, CDK-5,
IKK, JNK3, and thus be effective in the treatment of diseases
associated with these protein kinases.
[0016] In one aspect, the present invention provides compounds
containing the pyrimidine-2,4-diamine moiety, particularly diaryl
pyrimidine-2,4-diamine moiety, and compositions comprising the
compounds. The compounds have the general structure shown below:
##STR1## where X and Y are independently selected from oxygen,
amino or substituted amino, S(O).sub.0-2, or substituted or
unsubstituted carbon; R', R'', and R''' are optional substituents;
and R'''' is H or a progroup, R.sup.P. The progroup R.sup.P is
covalently attached via a carbamate, a thiocarbamate, a
dithiocarbamate, a urea, or a thiourea linkage to any one or more
of the 2'-N, the 4'-N, or to X or Y when they are amino. The
compounds and compositions can be used in methods for the
inhibition of kinases.
[0017] In one aspect, the compounds of the invention have the
formula (I): ##STR2## wherein R.sub.3 is aryl or heteroaryl that is
optionally substituted; X.sub.3 and X.sub.4 are independently
selected from CH or N; X.sub.5 is selected from the group
consisting of CR.sub.12R.sub.13, O, S, SO, SO.sub.2, and NR.sub.14,
wherein R.sub.12 and R.sub.13 are independently selected from H,
OH, or lower alkyl; R.sub.4 is an electronegative group; R',
R.sub.5, R.sub.6 and R.sub.14 are independently selected from H,
lower alkyl, a progroup, cycloalkyl or aryl, and wherein at least
one of R', R.sub.5, R.sub.6 or R.sub.14 is the aforementioned
progroup linked via a constitutent carbamate, a thiocarbamate, a
dithiocarbamate, a urea, or a thiourea linkage; R.sub.7 and R.sub.8
are independently selected from the group consisting of H, halogen,
lower alkyl, cycloalkyl, aryl, and heteroaryl; R.sub.9, and
R.sub.10 are independently selected from the group consisting of H,
halogen, --OH, -alkoxy, lower alkyl, cycloalkyl, aryl, and
heteroaryl; wherein R.sub.7 and R.sub.8, or R.sub.9 and R.sub.10
together form an oxo group, and provided R.sub.9 or R.sub.10 are
not --OH or alkoxy when X.sub.5 is NR.sub.14.
[0018] In another aspect, the present invention provides compounds
of formula (II): ##STR3## wherein X.sub.1 is O, S, or NR.sub.11;
X.sub.2 is selected from the group consisting of O and S; X.sub.3
and X.sub.4 are independently selected from CH or N; X.sub.5 is
selected from the group consisting of CR.sub.12R.sub.13, O, S, SO,
SO.sub.2, and NR.sub.14 wherein R.sub.12 and R.sub.13 are
independently selected from H, OH, lower alkyl, or together form an
oxo; and R.sub.14 is H or lower alkyl; R is selected from the group
consisting of straight or branched, saturated or unsaturated alkyl,
allyl, cycloalkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl,
prenylalkaryl and heteroarylalkyl, each of which is optionally
substituted; R.sub.1 and R.sub.2 are each independently selected
from the group consisting of H, OH, --OR.sub.11, NR.sub.15R.sub.15,
halo, lower alkyl, --C(O)O-alkyl, --C(O)OH,
--OP(.dbd.O)(OR.sub.11).sub.2, --OC(.dbd.O)OR.sub.11,
--OC(.dbd.O)R.sub.11, cycloalkyl, aryl, heteroaryl or together form
an oxo, wherein each R.sub.15 is independently selected from H,
lower alkyl, prenyl, allyl, --C(O)O-alkyl, cycloalkyl, aryl,
heteroaryl, alkaryl and alkheteroaryl, or two of R.sub.15 combine
to form an optionally substituted cycloheteroalkyl; R.sub.3 is aryl
or heteroaryl, each optionally substituted; each R.sub.11 is
independently H or lower alkyl; R.sub.4 is an electronegative group
such as NO.sub.2, fluorine, halogen, CN, haloalkyl, alkoxy,
carboxylate, CF.sub.3, CHF.sub.2, CH.sub.2F, CF.sub.3O--, and the
like; R.sub.5 and R.sub.6 are independently selected from H, lower
alkyl, cycloalkyl or aryl; R.sub.7, R.sub.8, R.sub.9, and R.sub.10
are independently selected from the group consisting of H, OH,
halogen, lower alkyl, cycloalkyl, aryl, and heteroaryl, or wherein
R.sub.7 and R.sub.8 or R.sub.9 and R.sub.10 together form an oxo
group; and n is an integer from 0 to 10.
[0019] In another aspect, the present invention provides methods of
treating and/or preventing cancer. The methods generally involve
administering to a subject that has cancer or that is at risk of
developing cancer an amount of a compound or composition of the
invention effective to treat or prevent the disease. The method may
be practiced in animals or in humans.
[0020] Many of the active 2,4-pyrimidinediamine compounds are also
potent inhibitors of the tyrosine kinase Syk kinase. Examples of
such 2,4-pyrimidinediamine compounds are described, for example, in
U.S. application Ser. No. 10/355,543 filed Jan. 31, 2003
(US2004/0029902A1), international application Serial No.
PCT/US03/03022 filed Jan. 31, 2003 (WO 03/063794), U.S. application
Ser. No. 10/631,029 filed Jul. 29, 2003, international application
Serial No. PCT/US03/24087 (WO2004/014382), U.S. application Ser.
No. 10/903,263 filed Jul. 30, 2004 (US2005/0234049), and
international application Serial No. PCT/US2004/24716
(WO2005/016893). Thus, in still another aspect, the present
disclosure provides methods of regulating, and in particular
inhibiting, Syk kinase activity. The method generally involves
contacting a Syk kinase or a cell comprising a Syk kinase with an
amount of a suitable prodrug, or an acceptable salt, hydrate,
solvate, N-oxide and/or composition thereof, effective to regulate
or inhibit Syk kinase activity. In one embodiment, the Syk kinase
is an isolated or recombinant Syk kinase. In another embodiment,
the Syk kinase is an endogenous or recombinant Syk kinase expressed
by a cell, for example a mast cell or a basophil cell. The method
can be practiced in in vitro wherein the contacting is performed
under conditions in which the progroup(s) metabolize to yield the
active 2,4-pyrimidinediamine compound, or in in vivo for the
treatment or prevention of diseases characterized by, caused by or
associated with Syk kinase activity.
[0021] In another aspect, the present disclosure provides methods
of regulating, and in particular inhibiting, signal transduction
cascades in which Syk plays a role. The method generally involves
contacting a Syk-dependent receptor or a cell expressing a
Syk-dependent receptor with an amount of a suitable compound of the
invention described herein, or an acceptable salt, hydrate,
solvate, N-oxide and/or composition thereof, effective to regulate
or inhibit the signal transduction cascade. The methods can also be
used to regulate, and in particular inhibit, downstream processes
or cellular responses elicited by activation of the particular
Syk-dependent signal transduction cascade. The methods can be
practiced to regulate any signal transduction cascade involving
Syk.
BRIEF DESCRIPTION OF DRAWINGS
[0022] FIG. 1 illustrates carbamate-derived prodrugs of the
pyrimidine-2,4-diamines.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
Definitions
[0023] Unless otherwise stated, the following terms used in this
application, including the specification and claims, have the
definitions given below. It must be noted that, as used in the
specification and the appended claims, the singular forms "a," "an"
and "the" include plural referents unless the context clearly
dictates otherwise. Definition of standard chemistry terms may be
found in reference works, including Carey and Sundberg (1992)
"Advanced Organic Chemistry 3.sup.rd Ed." Vols. A and B, Plenum
Press, New York. The practice of the present invention will employ,
unless otherwise indicated, conventional methods of mass
spectroscopy, protein chemistry, biochemistry, recombinant DNA
techniques and pharmacology, within the skill of the art.
[0024] As used herein, the following terms are intended to have the
following meanings:
[0025] "Alkyl," by itself or as part of another substituent, refers
to a saturated or unsaturated, branched, straight-chain or cyclic
monovalent hydrocarbon radical derived by the removal of one
hydrogen atom from a single carbon atom of a parent alkane, alkene
or alkyne. Typical alkyl groups include, but are not limited to,
methyl; ethyls such as ethanyl, ethenyl, ethynyl; propyls such as
propan-1-yl, propan-2-yl, cyclopropan-1-yl, prop-1-en-1-yl,
prop-1-en-2-yl, prop-2-en-1-yl(allyl), cycloprop-1-en-1-yl;
cycloprop-2-en-1-yl, prop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butyls
such as butan-1-yl, butan-2-yl, 2-methyl-propan-1-yl,
2-methyl-propan-2-yl, cyclobutan-1-yl, but-1-en-1-yl,
but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl,
but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl,
cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl,
but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the
like.
[0026] The term "alkyl" is specifically intended to include groups
having any degree or level of saturation, i.e., groups having
exclusively single carbon-carbon bonds, groups having one or more
double carbon-carbon bonds, groups having one or more triple
carbon-carbon bonds and groups having mixtures of single, double
and triple carbon-carbon bonds. Where a specific level of
saturation is intended, the expressions "alkanyl," "alkenyl," and
"alkynyl" are used. Preferably, an alkyl group comprises from 1 to
15 carbon atoms (C.sub.1-C.sub.15 alkyl), more preferably from 1 to
10 carbon atoms (C.sub.1-C.sub.10 alkyl) and even more preferably
from 1 to 6 carbon atoms (C.sub.1-C.sub.6 alkyl or lower
alkyl).
[0027] "Alkanyl," by itself or as part of another substituent,
refers to a saturated branched, straight-chain or cyclic alkyl
radical derived by the removal of one hydrogen atom from a single
carbon atom of a parent alkane. Typical alkanyl groups include, but
are not limited to, methanyl; ethanyl; propanyls such as
propan-1-yl, propan-2-yl(isopropyl), cyclopropan-1-yl, etc.;
butanyls such as butan-1-yl, butan-2-yl(sec-butyl),
2-methyl-propan-1-yl(isobutyl), 2-methyl-propan-2-yl(t-butyl),
cyclobutan-1-yl; pentanyls, such as pent-1-yl, pent-2-yl,
pent-3-yl, cyclopent-1-yl; hexanyls, such as hexan-1-yl,
hexan-3-yl, cyclohexan-1-yl, etc.; heptanyls, such as heptan-1-yl,
heptan-2-yl, cycloheptan-1-yl, etc.; and the like.
[0028] "Alkenyl," by itself or as part of another substituent,
refers to an unsaturated branched, straight-chain or cyclic alkyl
radical having at least one carbon-carbon double bond derived by
the removal of one hydrogen atom from a single carbon atom of a
parent alkene. The group may be in either the cis or trans
conformation about the double bond(s). Typical alkenyl groups
include, but are not limited to, ethenyl; propenyls such as
prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl(allyl),
prop-2-en-2-yl, cycloprop-1-en-1-yl; cycloprop-2-en-1-yl; butenyls
such as but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl,
but-2-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, buta-1,3-dien-1-yl,
buta-1,3-dien-2-yl, cyclobut-1-en-1-yl, cyclobut-1-en-3-yl,
cyclobuta-1,3-dien-1-yl, etc.; and the like.
[0029] "Alkynyl," by itself or as part of another substituent
refers to an unsaturated branched, straight-chain or cyclic alkyl
radical having at least one carbon-carbon triple bond derived by
the removal of one hydrogen atom from a single carbon atom of a
parent alkyne. Typical alkynyl groups include, but are not limited
to, ethynyl; propynyls such as prop-1-yn-1-yl, prop-2-yn-1-yl,
etc.; butynyls such as but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl,
etc.; and the like.
[0030] "Alkyldiyl" by itself or as part of another substituent
refers to a saturated or unsaturated, branched, straight-chain or
cyclic divalent hydrocarbon group derived by the removal of one
hydrogen atom from each of two different carbon atoms of a parent
alkane, alkene or alkyne, or by the removal of two hydrogen atoms
from a single carbon atom of a parent alkane, alkene or alkyne. The
two monovalent radical centers or each valency of the divalent
radical center can form bonds with the same or different atoms.
Typical alkyldiyl groups include, but are not limited to,
methandiyl; ethyldiyls such as ethan-1,1-diyl, ethan-1,2-diyl,
ethen-1,1-diyl, ethen-1,2-diyl; propyldiyls such as
propan-1,1-diyl, propan-1,2-diyl, propan-2,2-diyl, propan-1,3-diyl,
cyclopropan-1,1-diyl, cyclopropan-1,2-diyl, prop-1-en-1,1-diyl,
prop-1-en-1,2-diyl, prop-2-en-1,2-diyl, prop-1-en-1,3-diyl,
cycloprop-1-en-1,2-diyl, cycloprop-2-en-1,2-diyl,
cycloprop-2-en-1,1-diyl, prop-1-yn-1,3-diyl, etc.; butyldiyls such
as, butan-1,1-diyl, butan-1,2-diyl, butan-1,3-diyl, butan-1,4-diyl,
butan-2,2-diyl, 2-methyl-propan-1,1-diyl, 2-methyl-propan-1,2-diyl,
cyclobutan-1,1-diyl; cyclobutan-1,2-diyl, cyclobutan-1,3-diyl,
but-1-en-1,1-diyl, but-1-en-1,2-diyl, but-1-en-1,3-diyl,
but-1-en-1,4-diyl, 2-methyl-prop-1-en-1,1-diyl,
2-methanylidene-propan-1,1-diyl, buta-1,3-dien-1,1-diyl,
buta-1,3-dien-1,2-diyl, buta-1,3-dien-1,3-diyl,
buta-1,3-dien-1,4-diyl, cyclobut-1-en-1,2-diyl,
cyclobut-1-en-1,3-diyl, cyclobut-2-en-1,2-diyl,
cyclobuta-1,3-dien-1,2-diyl, cyclobuta-1,3-dien-1,3-diyl,
but-1-yn-1,3-diyl, but-1-yn-1,4-diyl, buta-1,3-diyn-1,4-diyl, etc.;
and the like. Where specific levels of saturation are intended, the
nomenclature alkanyldiyl, alkenyldiyl and/or alkynyldiyl is used.
Where it is specifically intended that the two valencies are on the
same carbon atom, the nomenclature "alkylidene" is used. In
preferred embodiments, the alkyldiyl group comprises from 1 to 6
carbon atoms (C1-C6 alkyldiyl). Also preferred are saturated
acyclic alkanyldiyl groups in which the radical centers are at the
terminal carbons, e.g., methandiyl(methano);
ethan-1,2-diyl(ethano); propan-1,3-diyl(propano);
butan-1,4-diyl(butano); and the like (also referred to as
alkylenos, defined infra).
[0031] "Alkoxy," by itself or as part of another substituent,
refers to a radical of the formula --OR, where R is an alkyl or
cycloalkyl group as defined herein. Representative examples alkoxy
groups include, but are not limited to, methoxy, ethoxy, propoxy,
isopropoxy, butoxy, tert-butoxy, cyclopropyloxy, cyclopentyloxy,
cyclohexyloxy and the like.
[0032] "Alkoxycarbonyl," by itself or as part of another
substituent, refers to a radical of the formula --C(O)-alkoxy,
where alkoxy is as defined herein.
[0033] "Alkylthio," by itself or as part of another substituent,
refers to a radical of the formula --SR, where R is an alkyl or
cycloalkyl group as defined herein. Representative examples of
Alkylthio groups include, but are not limited to, methylthio,
ethylthio, propylthio, isopropylthio, butylthio tert-butylthio,
cyclopropylthio, cyclopentylthio, cyclohexylthio, and the like.
[0034] "Aryl," by itself or as part of another substituent, refers
to a monovalent aromatic hydrocarbon group derived by the removal
of one hydrogen atom from a single carbon atom of a parent aromatic
ring system, as defined herein. Typical aryl groups include, but
are not limited to, groups derived from aceanthrylene,
acenaphthylene, acephenanthrylene, anthracene, azulene, benzene,
chrysene, coronene, fluoranthene, fluorene, hexacene, hexaphene,
hexalene, as-indacene, s-indacene, indane, indene, naphthalene,
octacene, octaphene, octalene, ovalene, penta-2,4-diene, pentacene,
pentalene, pentaphene, perylene, phenalene, phenanthrene, picene,
pleiadene, pyrene, pyranthrene, rubicene, triphenylene,
trinaphthalene and the like. Preferably, an aryl group comprises
from 6 to 20 carbon atoms (C.sub.6-C.sub.20 aryl), more preferably
from 6 to 15 carbon atoms (C.sub.6-C.sub.15 aryl) and even more
preferably from 6 to 10 carbon atoms (C.sub.6-C.sub.10 aryl).
[0035] "Arylalkyl," by itself or as part of another substituent,
refers to an acyclic alkyl group in which one of the hydrogen atoms
bonded to a carbon atom, typically a terminal or sp.sup.3 carbon
atom, is replaced with an aryl group as, as defined herein. Typical
arylalkyl groups include, but are not limited to, benzyl,
2-phenylethan-1-yl, 2-phenylethen-1-yl, naphthylmethyl,
2-naphthylethan-1-yl, 2-naphthylethen-1-yl, naphthobenzyl,
2-naphthophenylethan-1-yl and the like. Where specific alkyl
moieties are intended, the nomenclature arylalkanyl, arylalkenyl
and/or arylalkynyl is used. Preferably, an arylalkyl group is
(C.sub.6-C.sub.30) arylalkyl, e.g., the alkanyl, alkenyl or alkynyl
moiety of the arylalkyl group is (C.sub.1-C.sub.10) alkyl and the
aryl moiety is (C.sub.6-C.sub.20) aryl, more preferably, an
arylalkyl group is (C.sub.6-C.sub.20) arylalkyl, e.g., the alkanyl,
alkenyl or alkynyl moiety of the arylalkyl group is
(C.sub.1-C.sub.8) alkyl and the aryl moiety is (C.sub.6-C.sub.12)
aryl, and even more preferably, an arylalkyl group is
(C.sub.6-C.sub.15) arylalkyl, e.g., the alkanyl, alkenyl or alkynyl
moiety of the arylalkyl group is (C.sub.1-C.sub.5) alkyl and the
aryl moiety is (C.sub.6-C.sub.10) aryl.
[0036] "Aryloxy," by itself or as part of another substituent,
refers to a radical of the formula --O-aryl, where aryl is as
defined herein.
[0037] "Arylalkyloxy, by itself or as part of another substituent,
refers to a radical of the formula --O-arylalkyl, where arylalkyl
is as defined herein.
[0038] "Aryloxycarbonyl," by itself or as part of another
substituent, refers to a radical of the formula --C(O)--O-aryl,
where aryl is as defined herein.
[0039] "Atropisomers" are stereoisomers resulting from hindered
rotation about single bonds where the barrier to rotation is high
enough to allow for the isolation of the conformers (Eliel, E. L.;
Wilen, S. H. Stereochemistry of Organic Compounds; Wiley &
Sons: New York, 1994; Chapter 14). Atropisomerism is significant
because it introduces an element of chirality in the absence of
stereogenic atoms. The invention is meant to encompass
atropisomers.
[0040] "Carbamoyl," by itself or as part of another substituent,
refers to a radical of the formula --C(O)NR'R'', where R' and R''
are each, independently of one another, selected from the group
consisting of hydrogen, alkyl and cycloalkyl as defined herein, or
alternatively, R' and R'', taken together with the nitrogen atom to
which they are bonded, form a 5-, 6- or 7-membered cycloheteroalkyl
ring as defined herein, which may optionally include from 1 to 4 of
the same or different additional heteroatoms selected from the
group consisting of O, S and N.
[0041] "Compounds of the invention" refers to compounds encompassed
by the various descriptions and structural formulae disclosed
herein. The compounds of the invention may be identified by either
their chemical structure and/or chemical name. When the chemical
structure and chemical name conflict, the chemical structure is
determinative of the identity of the compound. The compounds of the
invention may contain one or more chiral centers and/or double
bonds and therefore may exist as stereoisomers, such as double-bond
isomers (i.e., geometric isomers), rotamers, enantiomers or
diastereomers. Accordingly, when stereochemistry at chiral centers
is not specified, the chemical structures depicted herein encompass
all possible configurations at those chiral centers including the
stereoisomerically pure form (e.g., geometrically pure,
enantiomerically pure or diastereomerically pure) and enantiomeric
and stereoisomeric mixtures. Enantiomeric and stereoisomeric
mixtures can be resolved into their component enantiomers or
stereoisomers using separation techniques or chiral synthesis
techniques well known to the skilled artisan. The compounds of the
invention may also exist in several tautomeric forms including the
enol form, the keto form and mixtures thereof. Accordingly, the
chemical structures depicted herein encompass all possible
tautomeric forms of the illustrated compounds. The compounds of the
invention may also include isotopically labeled compounds where one
or more atoms have an atomic mass different from the atomic mass
conventionally found in nature. Examples of isotopes that may be
incorporated into the compounds of the invention include, but are
not limited to, .sup.2H, .sup.3H, .sup.11C, .sup.13C, .sup.14C,
.sup.15N, .sup.13O, .sup.17O, .sup.31P, .sup.32P, .sup.35S,
.sup.18F and .sup.36Cl. Compounds of the invention may exist in
unsolvated forms as well as solvated forms, including hydrated
forms and as N-oxides. In general, the hydrated, solvated and
N-oxide forms are within the scope of the present invention.
Certain compounds of the present invention may exist in multiple
crystalline or amorphous forms. In general, all physical forms are
equivalent for the uses contemplated by the present invention and
are intended to be within the scope of the present invention.
[0042] "Cycloalkyl," by itself or as part of another substituent,
refers to a saturated or unsaturated cyclic alkyl radical, as
defined herein. Where a specific level of saturation is intended,
the nomenclature "cycloalkanyl" or "cycloalkenyl" is used. Typical
cycloalkyl groups include, but are not limited to, groups derived
from cyclopropane, cyclobutane, cyclopentane, cyclohexane,
cycloheptane, and the like. Preferably, the cycloalkyl group
comprises from 3 to 10 ring atoms (C.sub.3-C.sub.10 cycloalkyl) and
more preferably from 3 to 7 ring atoms (C.sub.3-C.sub.7
cycloalkyl).
[0043] "Cycloheteroalkyl," by itself or as part of another
substituent, refers to a saturated or unsaturated cyclic alkyl
radical in which one or more carbon atoms (and optionally any
associated hydrogen atoms) are independently replaced with the same
or different heteroatom. Typical heteroatoms to replace the carbon
atom(s) include, but are not limited to, N, P, O, S, Si, etc. Where
a specific level of saturation is intended, the nomenclature
"cycloheteroalkanyl" or "cycloheteroalkenyl" is used. Typical
cycloheteroalkyl groups include, but are not limited to, groups
derived from epoxides, azirines, thiiranes, imidazolidine,
morpholine, piperazine, piperidine, pyrazolidine, pyrrolidone,
quinuclidine, and the like. Preferably, the cycloheteroalkyl group
comprises from 3 to 10 ring atoms (3-10 membered cycloheteroalkyl)
and more preferably from 5 to 7 ring atoms (5-7 membered
cycloheteroalkyl).
[0044] A cycloheteroalkyl group may be substituted at a heteroatom,
for example, a nitrogen atom, with a lower alkyl group. As specific
examples, N-methyl-imidazolidinyl, N-methyl-morpholinyl,
N-methyl-piperazinyl, N-methyl-piperidinyl, N-methyl-pyrazolidinyl
and N-methyl-pyrrolidinyl are included within the definition of
"cycloheteroalkyl." A cycloheteralkyl group may be attached to the
remainder of the molecule via a ring carbon atom or a ring
heteroatom.
[0045] "Dialkylamino" or "Monoalkylamino," by themselves or as part
of other substituents, refer to radicals of the formula --NRR and
--NHR, respectively, where each R is independently selected from
the group consisting of alkyl and cycloalkyl, as defined herein.
Representative examples of dialkylamino groups include, but are not
limited to, dimethylamino, methylethylamino,
di-(1-methylethyl)amino, (cyclohexyl)(methyl)amino,
(cyclohexyl)(ethyl)amino, (cyclohexyl)(propyl)amino and the like.
Representative examples of monalkylamino groups include, but are
not limited to, methylamino, ethylamino, propylamino,
isopropylamino, cyclohexylamino, and the like.
[0046] "Electronegative" by itself or as part of another
substituent, refers to the tendency of a substituent to attract
valence electrons from neighboring atoms. Exemplary
electron-withdrawing groups include, acyl, formyl, sulfonyl,
alkoxy, carboxylate, haloalkyl, chloride, fluoride, cyano, nitro,
trifluoromethyl, difluoromethyl, fluoromethyl, trifluoromethoxy,
difluoromethoxy, fluoromethoxy, MeO, and the like
[0047] "Halogen" or "Halo," by themselves or as part of another
substituent, refer to a fluoro, chloro, bromo and/or iodo
radical.
[0048] "Haloalkyl," by itself or as part of another substituent,
refers to an alkyl group as defined herein in which one or more of
the hydrogen atoms is replaced with a halo group. The term
"haloalkyl" is specifically meant to include monohaloalkyls,
dihaloalkyls, trihaloalkyls, etc. up to perhaloalkyls. The halo
groups substituting a haloalkyl can be the same, or they can be
different. For example, the expression "(C.sub.1-C.sub.2)
haloalkyl" includes 1-fluoromethyl, 1-fluoro-2-chloroethyl,
difluoromethyl, trifluoromethyl, 1-fluoroethyl, 1,1-difluoroethyl,
1,2-difluoroethyl, 1,1,1-trifluoroethyl, perfluoroethyl, etc.
[0049] "Haloalkyloxy," by itself or as part of another substituent,
refers to a group of the formula --O-haloalkyl, where haloalkyl is
as defined herein.
[0050] "Heteroalkyl," "Heteroalkanyl," "Heteroalkenyl,"
"Heteroalkynyl," "Heteroalkyldiyl" and "Heteroalkyleno," by
themselves or as part of other substituents, refer to alkyl,
alkanyl, alkenyl, alkynyl, alkyldiyl and alkyleno groups,
respectively, in which one or more of the carbon atoms (and
optionally any associated hydrogen atoms), are each, independently
of one another, replaced with the same or different heteroatoms or
heteroatomic groups. Typical heteroatoms or heteroatomic groups
which can replace the carbon atoms include, but are not limited to,
O, S, N, Si, --NH--, --S(O)--, --S(O).sub.2--, --S(O)NH--,
--S(O).sub.2NH-- and the like and combinations thereof. The
heteroatoms or heteroatomic groups may be placed at any interior
position of the alkyl, alkenyl or alkynyl groups. Examples of such
heteroalkyl, heteroalkanyl, heteroalkenyl and/or heteroalkynyl
groups include --CH.sub.2--CH.sub.2--O--CH.sub.3,
--CH.sub.2--CH.sub.2--NH--CH.sub.3,
--CH.sub.2--CH.sub.2--N(CH.sub.3)--CH.sub.3,
--CH.sub.2--S--CH.sub.2, --CH.sub.3,
--CH.sub.2--CH.sub.2--S(O)--CH.sub.3,
--CH.sub.2--CH.sub.2--S(O).sub.2--CH.sub.3,
--CH.dbd.CH--O--CH.sub.3, --CH.sub.2--CH.dbd.N--O--CH.sub.3, and
--CH.sub.2--CH.sub.2--O--C.dbd.CH. For heteroalkyldiyl and
heteroalkyleno groups, the heteratom or heteratomic group can also
occupy either or both chain termini. For such groups, no
orientation of the group is implied.
[0051] "Heteroaryl," by itself or as part of another substituent,
refers to a monovalent heteroaromatic radical derived by the
removal of one hydrogen atom from a single atom of a parent
heteroaromatic ring systems, as defined herein. Typical heteroaryl
groups include, but are not limited to, groups derived from
acridine, .beta.-carboline, chromane, chromene, cinnoline, furan,
imidazole, indazole, indole, indoline, indolizine, isobenzofuran,
isochromene, isoindole, isoindoline, isoquinoline, isothiazole,
isoxazole, naphthyridine, oxadiazole, oxazole, perimidine,
phenanthridine, phenanthroline, phenazine, phthalazine, pteridine,
purine, pyran, pyrazine, pyrazole, pyridazine, pyridine,
pyrimidine, pyrrole, pyrrolizine, quinazoline, quinoline,
quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole,
thiophene, triazole, xanthene, and the like. Preferably, the
heteroaryl group comprises from 5 to 20 ring atoms (5-20 membered
heteroaryl), more preferably from 5 to 10 ring atoms (5-10 membered
heteroaryl). Preferred heteroaryl groups are those derived from
furan, thiophene, pyrrole, benzothiophene, benzofuran,
benzimidazole, indole, pyridine, pyrazole, quinoline, imidazole,
oxazole, isoxazole and pyrazine.
[0052] "Heteroarylalkyl" by itself or as part of another
substituent refers to an acyclic alkyl group in which one of the
hydrogen atoms bonded to a carbon atom, typically a terminal or
sp.sup.3 carbon atom, is replaced with a heteroaryl group. Where
specific alkyl moieties are intended, the nomenclature
heteroarylalkanyl, heteroarylakenyl and/or heteroarylalkynyl is
used. In preferred embodiments, the heteroarylalkyl group is a 6-21
membered heteroarylalkyl, e.g., the alkanyl, alkenyl or alkynyl
moiety of the heteroarylalkyl is (C1-C6) alkyl and the heteroaryl
moiety is a 5-15-membered heteroaryl. In particularly preferred
embodiments, the heteroarylalkyl is a 6-13 membered
heteroarylalkyl, e.g., the alkanyl, alkenyl or alkynyl moiety is
(C1-C3) alkyl and the heteroaryl moiety is a 5-10 membered
heteroaryl.
[0053] "Parent Aromatic Ring System" refers to an unsaturated
cyclic or polycyclic ring system having a conjugated .pi. electron
system. Specifically included within the definition of "parent
aromatic ring system" are fused ring systems in which one or more
of the rings are aromatic and one or more of the rings are
saturated or unsaturated, such as, for example, fluorene, indane,
indene, phenalene, etc. Typical parent aromatic ring systems
include, but are not limited to, aceanthrylene, acenaphthylene,
acephenanthrylene, anthracene, azulene, benzene, chrysene,
coronene, fluoranthene, fluorene, hexacene, hexaphene, hexalene,
as-indacene, s-indacene, indane, indene, naphthalene, octacene,
octaphene, octalene, ovalene, penta-2,4-diene, pentacene,
pentalene, pentaphene, perylene, phenalene, phenanthrene, picene,
pleiadene, pyrene, pyranthrene, rubicene, triphenylene,
trinaphthalene and the like.
[0054] "Parent Heteroaromatic Ring System" refers to a parent
aromatic ring system in which one or more carbon atoms (and
optionally any associated hydrogen atoms) are each independently
replaced with the same or different heteroatom. Typical heteroatoms
to replace the carbon atoms include, but are not limited to, N, P,
O, S, Si, etc. Specifically included within the definition of
"parent heteroaromatic ring system" are fused ring systems in which
one or more of the rings are aromatic and one or more of the rings
are saturated or unsaturated, such as, for example, benzodioxan,
benzofuran, chromane, chromene, indole, indoline, xanthene, etc.
Typical parent heteroaromatic ring systems include, but are not
limited to, arsindole, carbazole, .beta.-carboline, chromane,
chromene, cinnoline, furan, imidazole, indazole, indole, indoline,
indolizine, isobenzofuran, isochromene, isoindole, isoindoline,
isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole,
oxazole, perimidine, phenanthridine, phenanthroline, phenazine,
phthalazine, pteridine, purine, pyran, pyrazine, pyrazole,
pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine,
quinazoline, quinoline, quinolizine, quinoxaline, tetrazole,
thiadiazole, thiazole, thiophene, triazole, xanthene and the
like.
[0055] "Pharmaceutically acceptable salt" refers to a salt of a
compound of the invention which is made with counterions understood
in the art to be generally acceptable for pharmaceutical uses and
which possesses the desired pharmacological activity of the parent
compound. Such salts include: (1) acid addition salts, formed with
inorganic acids such as hydrochloric acid, hydrobromic acid,
sulfuric acid, nitric acid, phosphoric acid, and the like; or
formed with organic acids such as acetic acid, propionic acid,
hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic
acid, lactic acid, malonic acid, succinic acid, malic acid, maleic
acid, fumaric acid, tartaric acid, citric acid, benzoic acid,
3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,
methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic
acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid,
4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,
4-toluenesulfonic acid, camphorsulfonic acid,
4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic
acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary
butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic
acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic
acid and the like; or (2) salts formed when an acidic proton
present in the parent compound is replaced by a metal ion, e.g., an
alkali metal ion, an alkaline earth ion, or an aluminum ion; or
coordinates with an organic base such as ethanolamine,
diethanolamine, triethanolamine, N-methylglucamine, morpholine,
piperidine, dimethylamine, diethylamine and the like. Also included
are salts of amino acids such as arginates and the like, and salts
of organic acids like glucurmic or galactunoric acids and the like
(see, e.g., Berge et al., 1977, J. Pharm. Sci. 66:1-19).
[0056] "Pharmaceutically acceptable vehicle" refers to a diluent,
adjuvant, excipient or carrier with which a compound of the
invention is administered.
[0057] "Protecting group" refers to a group of atoms that, when
attached to a reactive functional group in a molecule, mask, reduce
or prevent the reactivity of the functional group. Typically, a
protecting group may be selectively removed as desired during the
course of a synthesis. Examples of protecting groups can be found
in Greene and Wuts, Protective Groups in Organic Chemistry,
3.sup.rd Ed., 1999, John Wiley & Sons, NY and Harrison et al.,
Compendium of Synthetic Organic Methods, Vols. 1-8, 1971-1996, John
Wiley & Sons, NY. Representative amino protecting groups
include, but are not limited to, formyl, acetyl, trifluoroacetyl,
benzyl, benzyloxycarbonyl ("CBZ"), tert-butoxycarbonyl ("Boc"),
trimethylsilyl ("TMS"), 2-trimethylsilyl-ethanesulfonyl ("SES"),
trityl and substituted trityl groups, allyloxycarbonyl,
9-fluorenylmethyloxycarbonyl ("FMOC"), nitro-veratryloxycarbonyl
("NVOC") and the like. Representative hydroxyl protecting groups
include, but are not limited to, those where the hydroxyl group is
either acylated (e.g., methyl and ethyl esters, acetate or
propionate groups or glycol esters) or alkylated such as benzyl and
trityl ethers, as well as alkyl ethers, tetrahydropyranyl ethers,
trialkylsilyl ethers (e.g., TMS or TIPPS groups) and allyl
ethers.
[0058] "Prodrug" refers to a derivative of an active compound
(drug) that undergoes a transformation under the conditions of use,
such as within the body, to release an active drug. Prodrugs are
frequently, but not necessarily, pharmacologically inactive until
converted into the active drug. Prodrugs are typically obtained by
masking a functional group in the drug believed to be in part
required for activity with a progroup (defined below) to form a
promoiety or "progroup" which undergoes a transformation, such as
cleavage, under the specified conditions of use to release the
functional group, and hence the active drug. The cleavage of the
promoiety may proceed spontaneously, such as by way of a hydrolysis
reaction, or it may be catalyzed or induced by another agent, such
as by an enzyme, by light, by acid, or by a change of or exposure
to a physical or environmental parameter, such as a change of
temperature, or combination thereof. The agent may be endogenous to
the conditions of use, such as an enzyme present in the cells to
which the prodrug is administered or the acidic conditions of the
stomach, or it may be supplied exogenously.
[0059] A wide variety of progroups suitable for masking functional
groups in active compounds to yield prodrugs are well-known in the
art. For example, a hydroxyl functional group may be masked as a
sulfonate, ester or carbonate promoiety, which may be hydrolyzed in
vitro to provide the hydroxyl group. An amino functional group may
be masked as an amide, imine, phosphinyl, phosphonyl, phosphoryl or
sulfenyl promoiety, which may be hydrolyzed in vivo to provide the
amino group. A carboxyl group may be masked as an ester (including
silyl esters and thioesters), amide or hydrazide promoiety, which
may be hydrolyzed in vivo to provide the carboxyl group. Other
specific examples of suitable progroups and their respective
promoieties will be apparent to those of skill in the art.
[0060] "Progroup" refers to a type of protecting group that, when
used to mask a functional group within an active drug, converts the
drug into a prodrug. Progroups are typically attached to the
functional group of the drug via bonds that are cleavable under
specified conditions of use.
[0061] "Substituted," when used to modify a specified group or
radical, means that one or more hydrogen atoms of the specified
group or radical are each, independently of one another, replaced
with the same or different substituent(s). Substituent groups
useful for substituting saturated carbon atoms in the specified
group or radical include, but are not limited to --R.sup.a, halo,
--O.sup.-, .dbd.O, --OR.sup.b, --SR.sup.b, --S.sup.-, .dbd.S,
--NR.sup.cR.sup.c, .dbd.NR.sup.b, .dbd.N--OR.sup.b, trihalomethyl,
--CF.sub.3, --CN, --OCN, --SCN, --NO, --NO.sub.2, .dbd.N.sub.2,
--N.sub.3, --S(O).sub.2R.sup.b, --S(O).sub.2O.sup.-,
--(CH.sub.2).sub.0-4S(O).sub.2OR.sup.b, --OS(O).sub.2R.sup.b,
--OS(O).sub.2O.sup.-, --OS(O).sub.2OR.sup.b, --P(O)(O.sup.-).sub.2,
--P(O)(OR.sup.b)(O.sup.-), --P(O)(OR.sup.b)(OR.sup.b),
--C(O)R.sup.b, --C(S)R.sup.b, --C(NR.sup.b)R.sup.b, --C(O)O.sup.-,
--C(O)OR.sup.b, --C(S)OR.sup.b, --C(O)NR.sup.cR.sup.c,
--C(NR.sup.b)NR.sup.cR.sup.c, --OC(O)R.sup.b, --OC(S)R.sup.b,
--OC(O)O.sup.-, --OC(O)OR.sup.b, --OC(S)OR.sup.b,
--NR.sup.bC(O)R.sup.b, --NR.sup.bC(S)R.sup.b,
--NR.sup.bC(O)O.sup.-, --NR.sup.bC(O)OR.sup.b,
--NR.sup.bC(S)OR.sup.b, --NR.sup.bC(O)NR.sup.cR.sup.c,
--NR.sup.bC(NR.sup.b)R.sup.b and
--NR.sup.bC(NR.sup.b)NR.sup.cR.sup.c, where R.sup.a is selected
from the group consisting of alkyl, cycloalkyl, heteroalkyl,
cycloheteroalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl;
each R.sup.b is independently hydrogen or R.sup.a; and each R.sup.c
is independently R.sup.b or alternatively, the two R.sup.cs are
taken together with the nitrogen atom to which they are bonded form
a 5-, 6- or 7-membered cycloheteroalkyl which may optionally
include from 1 to 4 of the same or different additional heteroatoms
selected from the group consisting of O, N and S. As specific
examples, --NR.sup.cR.sup.c is meant to include --NH.sub.2,
--NH-alkyl, N-pyrrolidinyl and N-morpholinyl.
[0062] Similarly, substituent groups useful for substituting
unsaturated carbon atoms in the specified group or radical include,
but are not limited to, --R.sup.a, halo, --O.sup.-, --OR.sup.b,
--SR.sup.b, --S.sup.-, --NR.sup.cR.sup.c, trihalomethyl,
--CF.sub.3, --CN, --OCN, --SCN, --NO, --NO.sub.2, --N.sub.3,
--S(O).sub.2R.sup.b, --S(O).sub.2O.sup.-, --S(O).sub.2OR.sup.b,
--OS(O).sub.2R.sup.b, --OS(O).sub.2O.sup.-, --OS(O).sub.2OR.sup.b,
--P(O)(O.sup.-).sub.2, --P(O)(OR.sup.b)(O.sup.-),
--P(O)(OR.sup.b)(OR.sup.b), --C(O)R.sup.b, --C(S)R.sup.b,
--C(NR.sup.b)R.sup.b, --C(O)O.sup.-, --C(O)OR.sup.b,
--C(S)OR.sup.b, --C(O)NR.sup.cR.sup.c,
--C(NR.sup.b)NR.sup.cR.sup.c, --OC(O)R.sup.b, --OC(S)R.sup.b,
--OC(O)O.sup.-, --OC(O)OR.sup.b, --OC(S)OR.sup.b,
--NR.sup.bC(O)R.sup.b, --NR.sup.bC(S)R.sup.b,
--NR.sup.bC(O)O.sup.-, --NR.sup.bC(O)OR.sup.b,
--NR.sup.bC(S)OR.sup.b, --NR.sup.bC(O)NR.sup.cR.sup.c,
--NR.sup.bC(NR.sup.b)R.sup.b and
--NR.sup.bC(NR.sup.b)NR.sup.cR.sup.c, where R.sup.a, R.sup.b and
R.sup.c are as previously defined.
[0063] Substituent groups useful for substituting nitrogen atoms in
heteroalkyl and cycloheteroalkyl groups include, but are not
limited to, --R.sup.a, --O.sup.-, --OR.sup.b, --SR.sup.b,
--S.sup.-, --NR.sup.cR.sup.c, trihalomethyl, --CF.sub.3, --CN,
--NO, --NO.sub.2, --S(O).sub.2R.sup.b, --S(O).sub.2O.sup.-,
--S(O).sub.2OR.sup.b, --OS(O).sub.2R.sup.b, --OS(O).sub.2O.sup.-,
--OS(O).sub.2OR.sup.b, --P(O)(O.sup.-).sub.2,
--P(O)(OR.sup.b)(O.sup.-), --P(O)(OR.sup.b)(OR.sup.b),
--C(O)R.sup.b, --C(S)R.sup.b, --C(NR.sup.b)R.sup.b, --C(O)OR.sup.b,
--C(S)OR.sup.b, --C(O)NR.sup.cR.sup.c,
--C(NR.sup.b)NR.sup.cR.sup.c, --OC(O)R.sup.b, --OC(S)R.sup.b,
--OC(O)OR.sup.b, --OC(S)OR.sup.b, --NR.sup.bC(O)R.sup.b,
--NR.sup.bC(S)R.sup.b, --NR.sup.bC(O)OR.sup.b,
--NR.sup.bC(S)OR.sup.b, --NR.sup.bC(O)NR.sup.cR.sup.c,
--NR.sup.bC(NR.sup.b)R.sup.b and
--NR.sup.bC(NR.sup.b)NR.sup.cR.sup.c, where R.sup.a, R.sup.b and
R.sup.c are as previously defined.
[0064] Substituent groups from the above lists useful for
substituting other specified groups or atoms will be apparent to
those of skill in the art.
[0065] The substituents used to substitute a specified group can be
further substituted, typically with one or more of the same or
different groups selected from the various groups specified
above.
[0066] "Syk Kinase" refers to the well-known 72 kDa non-receptor
(cytoplasmic) spleen protein tyrosine kinase expressed in B-cells
and other hematopoetic cells. Syk kinase includes two consensus
Src-homology 2 (SH2) domains in tandem that bind to phosphorylated
immunoreceptor tyrosine-based activation motifs ("ITAMs"), a
"linker" domain and a catalytic domain (for a review of the
structure and function of Syk kinase see Sada et al, 2001, J.
Biochem. (Tokyo) 130:177-186); see also Turner et al., 2000,
Immunology Today 21: 148-154). Syk kinase has been extensively
studied as an effector of B-cell receptor (BCR) signaling (Turner
et al., 2000, supra). Syk kinase is also critical for tyrosine
phosphorylation of multiple proteins which regulate important
pathways leading from immunoreceptors, such as Ca.sup.2+
mobilization and mitogen-activated protein kinase (MAPK) cascades
and degranulation. Syk kinase also plays a critical role in
integrin signaling in neutrophils (see, e.g., Mocsai et al. 2002,
Immunity 16:547-558).
[0067] As used herein, Syk kinase includes kinases from any species
of animal, including but not limited to, homosapiens, simian,
bovine, porcine, rodent, etc., recognized as belonging to the Syk
family. Specifically included are isoforms, splice variants,
allelic variants, mutants, both naturally occurring and man-made.
The amino acid sequences of such Syk kinases are well known and
available from GENBANK. Specific examples of mRNAs encoding
different isoforms of human Syk kinase can be found at GENBANK
accession no. gi|21361552|ref|NM.sub.--003177.2|,
gi|496899|emb|Z29630.1|HSSYKPTK[496899] and
gi|15030258|gb|BC011399.1|BC011399[15030258].
[0068] As used herein, the term "subject" encompasses mammals and
non-mammals. Examples of mammals include, but are not limited to,
any member of the Mammalian class: humans, non-human primates such
as chimpanzees, and other apes and monkey species; farm animals
such as cattle, horses, sheep, goats, swine; domestic animals such
as rabbits, dogs, and cats; laboratory animals including rodents,
such as rats, mice and guinea pigs, and the like. Examples of
non-mammals include, but are not limited to, birds, fish and the
like. The term does not denote a particular age or gender.
[0069] As used herein, the terms "treat" or "treatment" are used
interchangeably and are meant to indicate a postponement of
development of a disease and/or a reduction in the severity of such
symptoms that will or are expected to develop, where the disease is
associated with the functioning of a kinase. The terms further
include ameliorating existing symptoms, preventing additional
symptoms, and ameliorating or preventing the underlying metabolic
causes of symptoms.
[0070] The compounds of the present invention may be used to
inhibit or reduce the activity of kinases. In these contexts,
inhibition and reduction of activity of kinases refers to a lower
level of the measured activity relative to a control experiment in
which the cells or the subjects are not treated with the test
compound. In particular aspects, the inhibition or reduction in the
measured activity is at least a 10% reduction or inhibition. One of
skill in the art will appreciate that reduction or inhibition of
the measured activity of at least 20%, 50%, 75%, 90% or 100%, or
any number in between, may be preferred for particular
applications.
The Compounds
[0071] As described in the Summary, the instant disclosure provides
prodrugs of biologically active 2,4-pyrimidinediamine compounds,
such as the various 2,4-pyrimidinediamine compounds described in
U.S. application Ser. No. 10/355,543 filed Jan. 31, 2003
(US2004/0029902A1), international application Serial No.
PCT/US03/03022 filed Jan. 31, 2003 (WO 03/063794), U.S. application
Ser. No. 10/631,029 filed Jul. 29, 2003, international application
Serial No. PCT/US03/24087 (WO2004/014382), U.S. application Ser.
No. 10/903,263 filed Jul. 30, 2004 (US2005/0234049), and
international application Serial No. PCT/US2004/24716
(WO2005/016893). Prodrugs of the pyrimidine-2,4-diamine compounds
are of particular interest, as these compounds inhibit kinases,
such as inhibiting upstream Fc receptor signaling cascades as well
as Syk kinase and Syk kinase-dependent signaling cascades. The
prodrugs generally include such active 2,4-pyrimidinediamine
compounds in which one or more of the available primary or
secondary amine groups is masked with a progroup R.sup.P that
metabolizes in vivo by way of the corresponding hydroxy, thio- or
amino-methylamine intermediated to yield the active
2,4-pyrimidinediamine drug.
[0072] The invention provides novel compounds containing the
pyrimidine-2,4-diamine moiety, and compositions comprising the
compounds. In one aspect, the compounds of the invention have the
formula (I) ##STR4## wherein R.sub.3 is aryl or heteroaryl that is
optionally substituted; X.sub.3 and X.sub.4 are independently
selected from CH or N; X.sub.5 is selected from the group
consisting of CR.sub.12R.sub.13, O, S, SO, SO.sub.2, and NR.sub.14,
wherein R.sub.12 and R.sub.13 are independently selected from H,
OH, or lower alkyl; R.sub.4 is an electronegative group; R',
R.sub.5, R.sub.6 and R.sub.14 are independently selected from H,
lower alkyl, a progroup, cycloalkyl or aryl, and wherein at least
one of R', R.sub.5, R.sub.6 or R.sub.14 is the aforementioned
progroup linked via a constitutent carbamate, a thiocarbamate, a
dithiocarbamate, a urea, or a thiourea linkage; R.sub.7 and R.sub.8
are independently selected from the group consisting of H, halogen,
lower alkyl, cycloalkyl, aryl, and heteroaryl; and R.sub.9 and
R.sub.10 are independently selected from the group consisting of H,
halogen, --OH, -alkoxy, lower alkyl, cycloalkyl, aryl, and
heteroaryl; wherein R.sub.7 and R.sub.8, or R.sub.9 and R.sub.10
together form an oxo group, and provided R.sub.9 or R.sub.10 are
not --OH or alkoxy when X.sub.5 is NR.sub.14.
[0073] The nature of the progroup can vary, and will depend upon,
among other factors, the desired water solubility of the prodrug,
its intended mode of administration and/or its intended mechanism
or site of metabolism to the active 2,4-pyrimidinediamine compound.
The identity of the R.sup.3 group can also be selected so as to
impart the prodrug with desirable characteristics. For example,
lipophilic groups can be used to decrease water solubility and
hydrophilic groups can be used to increase water solubility. In
this way, prodrugs specifically tailored for selected modes of
administration can be obtained. The R.sup.3 group can also be
designed to impart the prodrug with other properties, such as, for
example, improved passive intestinal absorption, improved
transport-mediated intestinal absorption, protection against fast
metabolism (slow-release prodrugs), tissue-selective delivery,
passive enrichment in target tissues, targeting-specific
transporters, etc. Groups capable of imparting prodrugs with these
characteristics are well-known, and are described, for example, in
Ettmayer et al. (2004) J. Med. Chem. 47: 2393-2404. All of the
various groups described in these references can be utilized in the
prodrugs described herein.
[0074] The suitability of any particular progroup for a desired
mode of administration can be confirmed in biochemical assays. For
example, if a prodrug is to be administered by injection into a
particular tissue or organ, and the identities of the various
enzyme(s) expressed in the tissue or organ are known, the
particular prodrug can be tested for metabolism in biochemical
assays with the isolated enzyme(s). Alternatively, the particular
prodrug can be tested for metabolism to the active
2,4-pyrimidinediamine compound with tissue and/or organ extracts.
Using tissue and/or organ extracts can be of particular convenience
when the identity(ies) of the enzymes expressed in the target
tissues or organs are unknown, or in instances when the isolated
enzymes are not conveniently available. Skilled artisans will be
able to readily select progroups having metabolic properties (such
as kinetics) suitable for particular applications using such in
vitro tests. The specific prodrugs could also be tested for
suitable metabolism in in vitro animal models.
[0075] In another aspect, the present invention provides compounds
of formula (II) ##STR5## wherein X.sub.1 is O, S, or NR.sub.11;
X.sub.2 is selected from the group consisting of O and S; X.sub.3
and X.sub.4 are independently selected from CH or N; X.sub.5 is
selected from the group consisting of CR.sub.12R.sub.13, O, S, SO,
SO.sub.2, and NR.sub.14 wherein R.sub.12 and R.sub.13 are
independently selected from H, OH, lower alkyl, or together form an
oxo; and R.sub.14 is H or lower alkyl; R is selected from the group
consisting of straight or branched, saturated or unsaturated alkyl,
allyl, cycloalkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl,
prenylalkaryl and heteroarylalkyl, each of which is optionally
substituted; R.sub.1 and R.sub.2 are each independently selected
from the group consisting of H, OH, --OR.sub.11, NR.sub.15R.sub.15,
halo, lower alkyl, --C(O)O-alkyl, --C(O)OH,
--OP(.dbd.O)(OR.sub.11).sub.2, --OC(.dbd.O)OR.sub.11,
--OC(.dbd.O)R.sub.11, cycloalkyl, aryl, heteroaryl or together form
an oxo, wherein each R.sub.15 is independently selected from H,
lower alkyl, prenyl, allyl, --C(O)O-alkyl, cycloalkyl, aryl,
heteroaryl, alkaryl and alkheteroaryl, or two of R.sub.15 combine
to form an optionally substituted cycloheteroalkyl; R.sub.3 is aryl
or heteroaryl, each optionally substituted; each R.sub.11 is
independently H or lower alkyl; R.sub.4 is an electronegative group
such as NO.sub.2, fluorine, halogen, CN, haloalkyl, alkoxy,
carboxylate, CF.sub.3, CHF.sub.2, CH.sub.2F, CF.sub.3O--, and the
like; R.sub.5 and R.sub.6 are independently selected from H, lower
alkyl, cycloalkyl or aryl; R.sub.7, R.sub.8, R.sub.9, and R.sub.10
are independently selected from the group consisting of H, OH,
halogen, lower alkyl, cycloalkyl, aryl, and heteroaryl, or wherein
R.sub.7 and R.sub.8 or R.sub.9 and R.sub.10 together form an oxo
group; and n is an integer from 0 to 10.
[0076] In another aspect, the present invention provides compounds
of formula (III): ##STR6## wherein X.sub.1 is O or NR.sub.11; R is
selected from the group consisting of straight or branched,
saturated or unsaturated alkyl, allyl, cycloalkyl,
cycloheteroalkyl, aryl, and heteroaryl; and n is and integer
between 0 and 10. Thus, for example, R can be morpholine,
1-methylpiperidine, piperazine, 4-(3-propane-1-sulfonic acid)
piperazin-1-yl, dimethylamino, tryptamine,
N-tert-butylaceyltryptamine, phosphate, methyl phosphate, dimethyl
phosphate, phosphonate, and the like. Exemplary prodrugs are
described in Examples 1-10 and in FIG. 1.
[0077] In examples 1-10, (also as depicted in Schemes 1-3) a
carbamoyl chloride is produced which is then used to make prodrugs
of the invention. Also, as depicted in FIG. 1, haloalkyl carbamate
intermediates, for example but not limited to intermediate 10 where
the carbamate is on the N2-nitrogen, can be made at various
positions on the 2,4-pyrimidinediamine and further converted into
prodrugs of the invention. For example, chloromethyl ether
carbamates can be reacted with imines, like pyridine, to make
pyridinium salt prodrugs, 11. In another example, chloromethyl
ether carbamates can be reacted with alcohols or alkoxides to make
acetal-carbamate prodrugs, 12. In another example, chloromethyl
ether carbamates can be reacted with silver phosphonate salts to
make mixed phosphate-acetal-carbamate prodrugs, 13. In another
example, chloromethyl ether carbamates can be reacted with
carboxylate salts to make ester-acetal-carbamate prodrugs, 14.
[0078] Those of skill in the art will appreciate that the compounds
of the invention described herein may include functional groups
that can be masked with progroups to create prodrugs. Such prodrugs
are usually, but need not be, pharmacologically inactive until
converted into their active drug form. In the prodrugs of the
invention, any available functional moiety may be masked with a
progroup to yield a prodrug. Myriad progroups suitable for masking
such functional groups to yield promoieties that are cleavable
under the desired conditions of use are known in the art.
Methods of Synthesis
[0079] The compounds of the invention comprise isoxazoloanthrones,
as described above. The compounds can be obtained from commercial
sources, such as Aldrich Chemical Co. (Milwaukee, Wis.), Sigma
Chemical Co. (St. Louis, Mo.), or Maybridge (Cornwall, England), or
the compounds can be synthesized. The compounds of the present
invention, and other related compounds having different
substituents identified by any of the methods described above can
be synthesized using techniques and materials known to those of
skill in the art, such as described, for example, in March,
ADVANCED ORGANIC CHEMISTRY 4.sup.th Ed., (Wiley 1992); Carey and
Sundberg, ADVANCED ORGANIC CHEMISTRY 3.sup.rd Ed., Vols. A and B
(Plenum 1992), and Green and Wuts, PROTECTIVE GROUPS IN ORGANIC
SYNTHESIS 2.sup.nd Ed. (Wiley 1991). Starting materials useful for
preparing compounds of the invention and intermediates thereof are
commercially available or can be prepared by well-known synthetic
methods (see, e.g., Harrison et al., "Compendium of Synthetic
Organic Methods", Vols. 1-8 (John Wiley and Sons, 1971-1996);
"Beilstein Handbook of Organic Chemistry," Beilstein Institute of
Organic Chemistry, Frankfurt, Germany; Feiser et al., "Reagents for
Organic Synthesis," Volumes 1-21, Wiley Interscience; Trost et al.,
"Comprehensive Organic Synthesis," Pergamon Press, 1991;
"Theilheimer's Synthetic Methods of Organic Chemistry," Volumes
1-45, Karger, 1991; March, "Advanced Organic Chemistry," Wiley
Interscience, 1991; Larock "Comprehensive Organic Transformations,"
VCH Publishers, 1989; Paquette, "Encyclopedia of Reagents for
Organic Synthesis," 3d Edition, John Wiley & Sons, 1995). Other
methods for synthesis of the compounds described herein and/or
starting materials are either described in the art or will be
readily apparent to the skilled artisan. Alternatives to the
reagents and/or protecting groups may be found in the references
provided above and in other compendiums well known to the skilled
artisan. Guidance for selecting suitable protecting groups can be
found, for example, in Greene & Wuts, "Protective Groups in
Organic Synthesis," Wiley Interscience, 1999. Accordingly, the
synthetic methods and strategy presented herein are illustrative
rather than comprehensive.
[0080] The compounds and intermediates described herein can be
synthesized via a variety of different synthetic routes using
commercially available starting materials and/or starting materials
prepared by conventional synthetic methods. Suitable exemplary
methods that may be routinely used and/or adapted to synthesize
active 2,4-pyrimidinediamine compounds can be found in U.S. Pat.
No. 5,958,935, U.S. application Ser. No. 10/355,543 filed Jan. 31,
2003 (US2004/0029902A1), international application Serial No.
PCT/US03/03022 filed Jan. 31, 2003 (WO 03/063794), U.S. application
Ser. No. 10/631,029 filed Jul. 29, 2003, international application
Serial No. PCT/US/24087 (WO2004/014382), U.S. application Ser. No.
10/903,263 filed Jul. 30, 2004 (US2005/0234049), and international
application Serial No. PCT/US2004/24716 (WO2005/016893). These
active 2,4-pyrimidinediamine compounds can be used as starting
materials to synthesize the prodrugs.
[0081] Thus, for example, the compounds of the invention having
carbamate, thiocarbamate, or urea linkages can be synthesized using
the reactions shown in Scheme 1 below: ##STR7## ##STR8##
[0082] The compounds of the invention having thiocarbamate,
dithiocarbamate, or thiourea linkages can be synthesized using the
reactions shown in Scheme 2 below: ##STR9## ##STR10##
[0083] Alternative methods for the synthesis of compounds of the
invention having thiocarbamate, dithiocarbamate, or thiourea
linkages is shown in Scheme 3 below: ##STR11## ##STR12##
[0084] The procedures described herein for synthesizing the
compounds of the invention may include one or more steps of
protection and deprotection (e.g., the formation and removal of
acetal groups). In addition, the synthetic procedures disclosed
below can include various purifications, such as column
chromatography, flash chromatography, thin-layer chromatography
(TLC), recrystallization, distillation, high-pressure liquid
chromatography (HPLC) and the like. Also, various techniques well
known in the chemical arts for the identification and
quantification of chemical reaction products, such as proton and
carbon-13 nuclear magnetic resonance (.sup.1H and .sup.13C NMR),
infrared and ultraviolet spectroscopy (IR and UV), X-ray
crystallography, elemental analysis (EA), HPLC and mass
spectroscopy (MS) can be used as well. Methods of protection and
deprotection, purification and identification and quantification
are well known in the chemical arts.
Indications
[0085] Compounds of the present invention are useful for, but not
limited to, the prevention or treatment of cancer and related
diseases. The compounds of the invention have kinase inhibitory
activity, such as Syk kinase inhibitory activity, Src kinase
inhibitory activity, IGF-1R inhibitory activity, and the like. The
compounds of the invention are useful in therapy as antineoplasia
agents.
[0086] In vitro and cellular assays suitable for confirming the
activity of a particular 2,4-pyrimidinediamine compound are
described in detail in U.S. application Ser. No. 10/355,543 filed
Jan. 31, 2003 (US2004/0029902A1), international application Serial
No. PCT/US03/03022 filed Jan. 31, 2003 (WO 03/063794), U.S.
application Ser. No. 10/631,029 filed Jul. 29, 2003, international
application Serial No. PCT/US03/24087 (WO2004/014382), U.S.
application Ser. No. 10/903,263 filed Jul. 30, 2004
(US2005/0234049), and international application Serial No.
PCT/US2004/24716 (WO2005/016893).
[0087] The ability of a particular prodrug to metabolize to an
active 2,4-pyrimidinediamine compound under the desired conditions
of use can be confirmed in in vitro and/or in vivo assays, as
previously described.
[0088] Compounds of the invention can be useful for the treatment
of neoplasia including cancer and metastasis, including, but not
limited to: carcinoma such as cancer of the bladder, breast, colon,
kidney, liver, lung (including small cell lung cancer), esophagus,
gall-bladder, ovary, pancreas, stomach, cervix, thyroid, prostate,
and skin (including squamous cell carcinoma); hematopoietic tumors
of lymphoid lineage (including leukemia, acute lymphocitic
leukemia, acute lymphoblastic leukemia, B-cell lymphoma,
T-cell-lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy
cell lymphoma and Burkett's lymphoma); hematopoietic tumors of
myeloid lineage (including acute and chronic myelogenous leukemias,
myelodysplastic syndrome and promyelocytic leukemia); tumors of
mesenchymal origin (including fibrosarcoma and rhabdomyosarcoma,
and other sarcomas, e.g. soft tissue and bone); tumors of the
central and peripheral nervous system (including astrocytoma,
neuroblastoma, glioma and schwannomas); and other tumors (including
melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderoma
pigmentosum, keratoctanthoma, thyroid follicular cancer and
Kaposi's sarcoma). The compounds of the present invention are also
useful in the treatment of cancer related indications such as solid
tumors, sarcomas (especially Ewing's sarcoma and osteosarcoma),
retinoblastoma, rhabdomyosarcomas, neuroblastoma, hematopoietic
malignancies, including leukemia and lymphoma, tumor-induced
pleural or pericardial effusions, and malignant ascites.
[0089] The compounds of the present invention can also be useful
for promoting apoptosis.
[0090] The compounds of this invention can also act as inhibitors
of other protein kinases, e.g. ErbB, KDR, CDK-2, LCK, CDK-5, IKK,
JNK3, and thus be effective in the treatment of diseases associated
with other protein kinases.
[0091] Besides being useful for human treatment, these compounds
are also useful for veterinary treatment of companion animals,
exotic animals and farm animals, including mammals, rodents, and
the like. More preferred animals include horses, dogs, and cats. As
used herein, the compounds of the present invention include the
pharmaceutically acceptable derivatives thereof.
Uses and Administration
[0092] The compounds of the invention and/or compositions thereof
find particular use in the treatment and/or prevention diseases in
animals and humans caused by kinases. When used in this context,
the compounds may be administered per se, but are typically
formulated and administered in the form of a pharmaceutical
composition. The exact composition will depend upon, among other
things, the method of administration and will apparent to those of
skill in the art. A wide variety of suitable pharmaceutical
compositions are described, for example, in Remington's
Pharmaceutical Sciences, 20.sup.th ed., 2001).
[0093] Pharmaceutical compositions may take a form suitable for
virtually any mode of administration, including, for example,
topical, ocular, oral, buccal, systemic, nasal, injection,
transdermal, rectal, vaginal, etc., or a form suitable for
administration by inhalation or insufflation.
[0094] Formulations suitable for oral administration can consist of
(a) liquid solutions, such as an effective amount of the active
compound suspended in diluents, such as water, saline or PEG 400;
(b) capsules, sachets or tablets, each containing a predetermined
amount of the active ingredient, as liquids, solids, granules or
gelatin; (c) suspensions in an appropriate liquid; and (d) suitable
emulsions. Tablet forms can include one or more of lactose,
sucrose, mannitol, sorbitol, calcium phosphates, corn starch,
potato starch, microcrystalline cellulose, gelatin, colloidal
silicon dioxide, talc, magnesium stearate, stearic acid, and other
excipients, colorants, fillers, binders, diluents, buffering
agents, moistening agents, preservatives, flavoring agents, dyes,
disintegrating agents, and pharmaceutically compatible carriers.
Lozenge forms can comprise the active ingredient in a flavor, e.g.,
sucrose, as well as pastilles comprising the active ingredient in
an inert base, such as gelatin and glycerin or sucrose and acacia
emulsions, gels, and the like containing, in addition to the active
ingredient, carriers known in the art.
[0095] The compound of choice, alone or in combination with other
suitable components, can be made into aerosol formulations (i.e.,
they can be "nebulized") to be administered via inhalation. Aerosol
formulations can be placed into pressurized acceptable propellants,
such as dichlorodifluoromethane, propane, nitrogen, and the
like.
[0096] Suitable formulations for rectal administration include, for
example, suppositories, which consist of the packaged nucleic acid
with a suppository base. Suitable suppository bases include natural
or synthetic triglycerides or paraffin hydrocarbons. In addition,
it is also possible to use gelatin rectal capsules which consist of
a combination of the compound of choice with a base, including, for
example, liquid triglycerides, polyethylene glycols, and paraffin
hydrocarbons.
[0097] Formulations suitable for parenteral administration, such
as, for example, by intraarticular (in the joints), intravenous,
intramuscular, intradermal, intraperitoneal, and subcutaneous
routes, include aqueous and non-aqueous, isotonic sterile injection
solutions, which can contain antioxidants, buffers, bacteriostats,
and solutes that render the formulation isotonic with the blood of
the intended recipient, and aqueous and non-aqueous sterile
suspensions that can include suspending agents, solubilizers,
thickening agents, stabilizers, and preservatives. In the practice
of this invention, compositions can be administered, for example,
by intravenous infusion, orally, topically, intraperitoneally,
intravesically or intrathecally. Parenteral administration, oral
administration, subcutaneous administration and intravenous
administration are the preferred methods of administration. A
specific example of a suitable solution formulation may comprise
from about 0.5-100 mg/ml compound and about 1000 mg/ml propylene
glycol in water. Another specific example of a suitable solution
formulation may comprise from about 0.5-100 mg/ml compound and from
about 800-1000 mg/ml polyethylene glycol 400 (PEG 400) in
water.
[0098] A specific example of a suitable suspension formulation may
include from about 0.5-30 mg/ml compound and one or more excipients
selected from the group consisting of: about 200 mg/ml ethanol,
about 1000 mg/ml vegetable oil (e.g., corn oil), about 600-1000
mg/ml fruit juice (e.g., grapefruit juice), about 400-800 mg/ml
milk, about 0.1 mg/ml carboxymethylcellulose (or microcrystalline
cellulose), about 0.5 mg/ml benzyl alcohol (or a combination of
benzyl alcohol and benzalkonium chloride) and about 40-50 mM
buffer, pH 7 (e.g., phosphate buffer, acetate buffer or citrate
buffer or, alternatively 5% dextrose may be used in place of the
buffer) in water.
[0099] A specific example of a suitable liposome suspension
formulation may comprise from about 0.5-30 mg/ml compound, about
100-200 mg/ml lecithin (or other phospholipid or mixture of
phospholipids) and optionally about 5 mg/ml cholesterol in
water.
[0100] The formulations of compounds can be presented in unit-dose
or multi-dose sealed containers, such as ampules and vials.
Injection solutions and suspensions can be prepared from sterile
powders, granules, and tablets of the kind previously
described.
[0101] The pharmaceutical preparation is preferably in unit dosage
form. In such form the preparation is subdivided into unit doses
containing appropriate quantities of the active component. The unit
dosage form can be a packaged preparation, the package containing
discrete quantities of preparation, such as packeted tablets,
capsules, and powders in vials or ampoules. Also, the unit dosage
form can be a capsule, tablet, cachet, or lozenge itself, or it can
be the appropriate number of any of these in packaged form. The
composition can, if desired, also contain other compatible
therapeutic agents, discussed in more detail, below.
[0102] In therapeutic use, the compounds utilized in the
pharmaceutical method of the invention are administered to patients
at dosage levels suitable to achieve therapeutic benefit. By
therapeutic benefit is meant that the administration of compound
leads to a beneficial effect in the patient over time.
[0103] Initial dosages suitable for administration to humans may be
determined from in vitro assays or animal models. For example, an
initial dosage may be formulated to achieve a serum concentration
that includes the IC.sub.50 of the particular compound being
administered, as measured in an in vitro assay. Alternatively, an
initial dosage for humans may be based upon dosages found to be
effective in animal models of the disease. As one example, the
initial dosage may be in the range of about 0.01 mg/kg/day to about
200 mg/kg/day, or about 0.1 mg/kg/day to about 100 mg/kg/day, or
about 1 mg/kg/day to about 50 mg/kg/day, or about 10 mg/kg/day to
about 50 mg/kg/day, can also be used. The dosages, however, may be
varied depending upon the requirements of the patient, the severity
of the condition being treated, and the compound being employed.
The size of the dose also will be determined by the existence,
nature, and extent of any adverse side-effects that accompany the
administration of a particular compound in a particular patient.
Determination of the proper dosage for a particular situation is
within the skill of the practitioner. Generally, treatment is
initiated with smaller dosages which are less than the optimum dose
of the compound. Thereafter, the dosage is increased by small
increments until the optimum effect under circumstances is reached.
For convenience, the total daily dosage may be divided and
administered in portions during the day, if desired.
Combination Therapy
[0104] In certain embodiments of the present invention, the
compounds of the invention and/or compositions thereof can be used
in combination therapy with at least one other therapeutic agent. A
compound of the invention and/or composition thereof and the
therapeutic agent can act additively or, more preferably,
synergistically.
[0105] 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 compounds of the invention or 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.
[0106] Co-administration of a compound of the present invention and
another pharmaceutical agent is intended to embrace administration
of each agent in a sequential manner in a regimen that will provide
beneficial effects of the drug combination, and is intended as well
to embrace co-administration of these agents in a substantially
simultaneous manner, such as in a single capsule having a fixed
ratio of these active agents or in multiple, separate capsules for
each agent.
[0107] Specifically, 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
neoplasia, such as with radiation therapy or with cytostatic or
cytotoxic agents.
[0108] If formulated as a fixed dose, such combination products
employ the compounds of this invention within the accepted dosage
ranges. Compounds of Formulae I, II, or III can also be
administered sequentially with known anticancer or cytotoxic agents
when a combination formulation is inappropriate. The invention is
not limited in the sequence of administration; compounds of formula
I may be administered either prior to or after administration of
the known anticancer or cytotoxic agent.
[0109] 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 neoplasia by
combination drug chemotherapy. Such antineoplastic agents fall into
several major categories, namely, antibiotic-type agents,
alkylating agents, antimetabolite agents, hormonal agents,
immunological agents, interferon-type agents and a category of
miscellaneous agents.
[0110] In one aspect, the compounds of the invention can be
co-administered with antimetabolite-type/thymidilate synthase
inhibitor antineoplastic agents. Suitable antimetabolite
antineoplastic agents can be selected from but not limited to the
group consisting of 5-FU-fibrinogen, acanthifolic acid,
aminothiadiazole, brequinar sodium, carmofur, Ciba-Geigy CGP-30694,
cyclopentyl cytosine, cytarabine phosphate stearate, dezaguanine,
dideoxycytidine, dideoxyguanosine, didox, doxifluridine,
fazarabine, floxuridine, isopropyl pyrrolizine, methotrexate,
uricytin, and the like
[0111] In another aspect, the compounds of the invention can be
co-administered with alkylating-type antineoplastic agents.
Suitable alkylating-type antineoplastic agents can be selected from
but not limited to the group consisting of altretamine, anaxirone,
bestrabucil, budotitane, carboplatin, carmustine, chlorambucil,
cisplatin, cyclophosphamide, diplatinum cytostatic, elmustine,
fotemustine, tauromustine, temozolomide, teroxirone, tetraplatin,
trimelamol, and the like.
[0112] In one aspect, the compounds of the invention can be
co-administered with antibiotic-type antineoplastic agents.
Suitable antibiotic-type antineoplastic agents can be selected from
but not limited to the group consisting of aclarubicin, actinomycin
D, actinoplanone, anthracycline, bleomycin sulfate, bryostatin-1,
calichemycin, chromoximycin, dactinomycin, daunorubicin,
doxorubicin, doxorubicin-fibrinogen, erbstatin, esorubicin,
glidobactin, herbimycin, idarubicin, illudins, oxalysine,
oxaunomycin, sparsomycin, thrazine, zorubicin, and the like.
[0113] In one aspect, the compounds of the invention can be
co-administered with other 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 .alpha.-carotene,
.alpha.-difluoromethyl-arginine, acitretin, amonafide, ankinomycin,
antineoplaston A5, asparaginase, Avarol, bromofosfamide,
caracemide, claviridenone, cytochalasin B, cytarabine, cytocytin,
dacarbazine, paclitaxel, Efamol porphyrin, spirogermanium, taxol,
thaliblastine, vinblastine sulfate, and the like.
[0114] The invention relates to inhibitors of enzymes that catalyze
phosphoryl transfer and/or that bind ATP/GTP nucleotides,
compositions comprising the inhibitors, and methods of using the
inhibitors and inhibitor compositions. The inhibitors and
compositions comprising them are useful for treating or modulating
disease in which phosphoryl transferases, including kinases, may be
involved, symptoms of such disease, or the effect of other
physiological events mediated by phosphoryl transferases, including
kinases. The invention also provides for methods of making the
inhibitor compounds and methods for treating diseases in which one
or more phosphoryl transferase, including kinase, activities is
involved.
[0115] Alternatively, the present compounds can also be used in
co-therapies with other anti-neoplastic agents, such as other
kinase inhibitors including p38 inhibitors and CDK inhibitors, TNF
inhibitors, metallomatrix proteases inhibitors (MMP), EGFR
inhibitors such as Iressa, KDR inhibitors, COX-2 inhibitors
including celecoxib, rofecoxib, parecoxib, valdecoxib, and
etoricoxib, NSAID's, SOD mimics or .alpha.v.beta..sub.3
inhibitors.
[0116] As yet another specific example, the compounds of the
invention and/or compositions thereof may be administered in
combination with both ribovirin and an interferon.
EXAMPLES
[0117] The following examples are provided by way of illustration
only and not by way of limitation. Those of skill in the art will
readily recognize a variety of noncritical parameters that could be
changed or modified to yield essentially similar results.
[0118] The examples are offered for illustrative purposes only, and
are not intended to limit the scope of the present invention in any
way. Efforts have been made to ensure accuracy with respect to
numbers used (e.g., amounts, temperatures, etc.), but some
experimental error and deviation should, of course, be allowed
for.
Example 1
Synthesis of
N2-Chlorocarbonyl-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-f-
luoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine
[0119] ##STR13##
[0120] To pale yellow stirring mixture of
N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-tr-
imethoxyphenyl)-2,4-pyrimidinediamine (2.5 g, 5.31 mmol) and
triphosgene (1.67 g, 5.62 mmol) in dicholoroethane (20 mL) at 0
.sup.0C, NEt.sub.3 (1.08 g, 1.5 mL, 10.76 mmol) in dichloroethane
(10 mL) was added dropwise under nitrogen atmosphere for 10 min.
The orange reaction mixture was allowed to stir for 15 min at 0
.sup.0C followed by refluxing at 90 .sup.0C overnight. The
heterogeneous tan orange reaction mixture was cooled to room
temperature. The reaction mixture was diluted with EtOAc (75 mL).
Precipitated white solid formed was filtered. The white solid was
collected, treated with water, filtered and dried to provide
N2-chlorocarbonyl-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-f-
luoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (1.75 g,
61%). .sup.1H NMR (DMSO-d.sub.6): .delta. 11.08 (s, 1H), 9.97 (s,
1H), 8.44 (d, 1H, J=3.2 Hz), 7.35 (d, 1H, J=8.5 Hz), 7.24 (d, 1H,
J=8.5 Hz), 6.77 (s, 1H), 3.72 (s, 6H), 3.66 (s, 3H), 1.40 (s, 6H).
LCMS: ret. time: 12.53 min.; purity: 95%; MS (m/e): 534
(MH.sup.+).
General Procedure for the Preparation of Carbamates and
Thiocarbamates:
[0121]
N2-chlorocarbonyl-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6--
yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (1
eq) prepared in Example 1 was dissolved in dry CH.sub.2Cl.sub.2
(4.8 mL/mmol), alcohol (for carbamates) or thiol (for
thiocarbamates) (2 eq), NEt.sub.3 (7 eq) and DMAP (0.1 eq) were
added successively under nitrogen atmosphere at room temperature.
Contents were allowed to stir at room temperature and progress of
the reaction mixture was monitored by LC % MS. The reaction mixture
was concentrated upon consumption of carbamoylchloride. The crude
concentrate was treated with aq. NaHCO.sub.3 and the resulting
solid precipitated was filtered, washed with water, dried and
purified by either silica gel column chromatography or preparative
HPLC.
Example 2
Synthesis of
N4-(2,2-Dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-[[2-(morp-
holin-4-yl)ethoxy]carbonyl]-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediam-
ine
[0122] ##STR14##
[0123]
N4-(2,2-Dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-[[-
2-(morpholin-4-yl)ethoxy]carbonyl]-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimid-
inediamine was prepared from 4-(2-hydroxyethyl)morpholine and
N2-chlorocarbonyl-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-f-
luoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine. The crude
solid, obtained after concentration of the reaction mixture
followed by treatment with aq. NaHCO.sub.3, was purified by
NEt.sub.3 treated silica gel column chromatography. .sup.1H NMR
(CDCl.sub.3): .delta. 10.32 (s, 2H), 8.89 (s, 1H), 8.18 (d, 1H,
J=2.9 Hz), 7.49 (d, 1H, J=8.8 Hz), 7.06 (d, 1H, J=8.8 Hz), 6.52 (s,
2H), 4.29 (m, 2H), 3.81 (s, 3H), 3.74 (s, 6H), 3.57 (m, 4H), 2.56
(m, 2H), 2.33 (m, 4H), 1.48 (s, 6H). LCMS: ret. time: 8.30 min.;
purity: 92%; MS (m/e): 628 (MH.sup.+).
Example 3
Synthesis of
4-(2,2-Dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-[[1-methyl-
-piperidin-2-yl)methoxy]carbonyl]-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidi-
nediamine
[0124] ##STR15##
[0125]
4-(2,2-Dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-[[1-
-methyl-piperidin-2-yl)methoxy]carbonyl]-N2-(3,4,5-trimethoxyphenyl)-2,4-p-
yrimidinediamine was prepared from
N2-chlorocarbonyl-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-f-
luoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine and
1-methyl-2-piperidinemethanol in the similar manner as described in
the general procedure. The crude off white solid obtained after the
general workup was subjected to HPLC purification. .sup.1H NMR
(DMSO-d.sub.6): .delta. 11.03 (s, 1H), 9.67 (s, 1H), 8.33 (d, 1H,
J=3.0 Hz), 7.41 (d, 1H, J=8.5 Hz), 7.17 (d, 1H, J=8.5 Hz), 6.56 (s,
2H), 4.10 (d, 2H, J=4.7 Hz), 3.80 (s, 6H), 3.64 (s, 3H), 2.70-2.66
(m, 1H), 2.09 (s, 3H), 1.97-1.92 (m, 2H), 1.58-1.07 (m, 12H). LCMS:
ret. time: 8.54 min.; purity: 92%; MS (m/e): 627 (MH.sup.+).
Example 4
Synthesis of
2S--N2-[[2-(t-Butoxycarbonyl)amino-3-(1H-indol-3-yl)]propoxycarbonyl]-N4--
(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimet-
hoxyphenyl)-2,4-pyrimidinediamine
[0126] ##STR16##
[0127]
2S--N2-[[2-(t-Butoxycarbonyl)amino-3-(1H-indol-3-yl)]propoxycarbon-
yl]-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-
-trimethoxyphenyl)-2,4-pyrimidinediamine was prepared from
N.sub..alpha.-(t-butoxycarbonyl)-L-tryptophanol and
N2-chlorocarbonyl-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-f-
luoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine. The crude
white solid collected after the workup was purified by NEt.sub.3
treated silica gel column chromatography. .sup.1H NMR
(DMSO-d.sub.6): .delta. 11.00 (s, 1H), 10.76 (s, 1H), 9.66 (s, 1H),
8.32 (d, 1H, J=3.2 Hz), 7.36 (d, 1H, J=8.8 Hz), 7.31-7.27 (m, 2H),
7.08 (d, 1H, J=8.5 Hz), 7.03-6.99 (m, 2H), 6.91-6.86 (m, 1H), 6.78
(d, 1H, J=8.2 Hz), 6.65 (s, 2H), 4.12-4.08 (m, 1H), 3.99-3.94 (m,
1H), 3.86-3.82 (m, 1H), 3.69 (s, 6H), 3.63 (s, 3H), 2.74 (m, 2H),
1.38 (s, 6H), 1.29 (s, 9H). LCMS: ret. time: 13.63 min.; purity:
91%; MS (m/e): 787 (MH.sup.+).
Example 5
Synthesis of
2S--N2-[[2-Amino-3-(1H-indol-3-yl)]propoxycarbonyl]-N4-(2,2-dimethyl-3-ox-
o-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-py-
rimidinediamine
[0128] ##STR17##
[0129] Trifluoracetic acid (0.04 mL, 59 mg, 0.519 mmol) was added
to the stirring solution of
2S--N2-[[2-Amino-3-(1H-indol-3-yl)]propoxycarbonyl]-N4-(2,2-dimethyl-3-ox-
o-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-py-
rimidinediamine (93 mg, 0.118 mmol) in CH.sub.2Cl.sub.2 (5 mL) at 0
.sup.0C. Progress of the reaction was monitored by LC/MS. Reaction
mixture was concentrated after 1 hr of stirring the reaction
mixture at 0 .sup.0C. The crude was triturated with anhydrous
Et.sub.2O. Ethereal layer was decanted and dried to provide off
white solid. The solid obtained was purified by HPLC to give 26 mg
(32%) of
N2-[[[(2S)-2-amino-3-(1H-indol-3-yl)]propoxy]carbonyl]-N4-(2,2-dimethyl-3-
-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-
-pyrimidinediamine as a white solid. LCMS: ret. time: 9.34 min.;
purity: 92%; MS (m/e): 687 (MH.sup.+).
Example 6
Synthesis of
N4-(2,2-Dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-[2-[4-(3--
sulfopropyl)piperizin-1-yl]ethoxycarbonyl]-N2-(3,4,5-trimethoxyphenyl)-2,4-
-pyrimidinediamine
[0130] ##STR18##
[0131]
N4-(2,2-Dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-[2-
-[4-(3-sulfopropyl)piperizin-1-yl]ethoxycarbonyl]-N2-(3,4,5-trimethoxyphen-
yl)-2,4-pyrimidinediamine was prepared in the similar manner as
described in the general procedure from
N2-chlorocarbonyl-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-f-
luoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine and
4-(2-hydroxyethyl)-piperazinepropanesulfonic acid (EPPS) in
CH.sub.3CN. Reaction mixture was concentrated and diluted with
water. The solid precipitated was filtered, dried and purified by
preparative HPLC. .sup.1H NMR (DMSO-d.sub.6): .delta. 11.03 (s,
1H), 9.68 (s, 1H), 8.35 (d, 1H, J=3.2 Hz), 7.34 (d, 1H, J=8.8 Hz),
7.15 (d, 1H, J=8.8 Hz), 6.57 (s, 2H), 4.19 (m, 2H), 3.69 (s, 6H),
3.65 (s, 3H), 3.30-2.86 (m, 8H), 2.57-2.52 (m, 4H), 2.37-2.26 (m,
2H), 1.93-1.91 (m, 2H), 1.39 (s, 6H). LCMS: ret. time: 8.32 min.;
purity: 98%; MS (m/e): 749 (MH.sup.+).
Example 7
Synthesis of
N2-[2-(Dimethylamino)ethoxycarbonyl]-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1-
,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine
[0132] ##STR19##
[0133]
N2-[2-(Dimethylamino)ethoxycarbonyl]-N4-(2,2-dimethyl-3-oxo-4H-5-p-
yrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidine-
diamine was prepared from
N2-chlorocarbonyl-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-f-
luoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine and
N,N-dimethylethanolamine. The crude solid obtained was purified by
preparative HPLC. .sup.1H NMR (DMSO-d.sub.6): .delta. 11.04 (s,
1H), 9.68 (s, 1H), 8.33 (d, 1H, J=3.5 Hz), 7.39 (d, 1H, J=8.5 Hz),
7.16 (d, 1H, J=8.5 Hz), 6.54 (s, 2H), 4.17 (t, 2H, J=5.8 Hz), 3.68
(s, 6H), 3.64 (s, 3H), 2.45 (t, 2H, J=5.8 Hz), 2.08 (s, 6H), 1.39
(s, 6H). LCMS: ret. time: 8.87 min.; purity: 99%; MS (m/e): 586
(MH.sup.+).
Example 8
Synthesis of
1S--N2-[[-1-(t-Butoxycarbonyl)-2-methylpropyl]aminocarbonyl]-N4-(2,2-dime-
thyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxypheny-
l)-2,4-pyrimidinediamine
[0134] ##STR20##
[0135]
1S--N2-[[-1-(t-Butoxycarbonyl)-2-methylpropyl]aminocarbonyl]-N4-(2-
,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimetho-
xyphenyl)-2,4-pyrimidinediamine was prepared from
N2-chlorocarbonyl-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-f-
luoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine and
L-valine t-butyl ester hydrochloride in the similar manner as
described in the general procedure. .sup.1H NMR (DMSO-d.sub.6):
.delta. 10.97 (s, 1H), 10.45 (d, 1H, J=7.6 Hz), 8.35 (d, 1H, J=3.5
Hz), 6.75 (d, 1H, J=8.8 Hz), 6.71 (d, 1H, J=8.8 Hz), 6.52 (s, 2H),
4.15 (dd, 1H, J=4.7 and 6.7 Hz), 3.71 (s, 3H), 3.66 (s, 6H), 2.15
(m, 1H), 1.42 (s, 9H), 1.38 (s, 6H), 0.93 (dd, 6H, J=1.7 and 6.7
Hz). LCMS: ret. time: 14.87 min.; purity: 93%; MS (m/e): 670
(MH.sup.+).
Example 9
Synthesis of
N2-[2-(Carboxymethyl)aminocarbonyl]-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,-
4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine
[0136] ##STR21##
[0137]
N2-[2-(Carboxymethyl)aminocarbonyl]-N4-(2,2-dimethyl-3-oxo-4H-5-py-
rido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidined-
iamine was prepared in the similar as described in the general
procedure from glycine and
N2-chlorocarbonyl-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-f-
luoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine. The crude
concentrated reaction mixture was treated with 1N aq. HCl. The
solid precipitated was dried and purified by preparative HPLC.
.sup.1H NMR (DMSO-d.sub.6): .delta. 10.99 (s, 1H), 10.06 (t, 1H,
J=5.0 Hz), 8.26 (d, 1H, J=3.8 Hz), 6.78 (app s, 2H), 6.51 (s, 2H),
3.85 (d, 2H, J=5.0 Hz), 3.71 (s, 3H), 3.67 (s, 6H), 1.37 (s, 6H).
LCMS: ret. time: 9.74 min.; purity: 97%; MS (m/e): 572
(MH.sup.+).
Example 10
Synthesis of
(+/-)-N4-(2,2-Dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-[[1-
(1-pyridinium)ethoxy)carbonyl]]-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidine-
diamine iodide salt
[0138] ##STR22##
[0139] The intermediate
(+/-)-N2-(1-Chloroethoxycarbonyl)-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]-
oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine
was first synthesized. To a stirring mixture of
N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-tr-
imethoxyphenyl)-2,4-pyrimidinediamine (250 mg, 0.53 mmol) and
i-Pr.sub.2NEt (0.14 mL, 102 mg, 0.78 mmol) in dicholoroethane (10
mL) at -78 .sup.0C, 1-chloroethyl chloroformate (0.07 mL, 90 mg,
0.638 mmol) was added dropwise under nitrogen atmosphere over 5
min. After 1 h, the reaction mixture was diluted with EtOAc (10 mL)
at -78 .sup.0C. Reaction mixture was allowed to warm to room
temperature while stirring. Solid precipitated from pale brown
transparent reaction mixture after stirring the contents at room
temperature for 1 h. Reaction mixture was concentrated and diluted
with water (15 mL). The precipitated solid was filtered and dried
to provide
(+/-)-N2-(1-chloroethoxycarbonyl)-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]-
oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine
(250 mg, 81%). .sup.1H NMR (DMSO-d.sub.6): .delta. 11.04 (s, 1H),
9.78 (s, 1H), 8.37 (d, 1H, J=3.2 Hz), 7.39 (d, 1H, J=8.5 Hz), 7.17
(d, 1H, J=8.5 Hz), 6.64 (qt, 1H, J=5.7 Hz), 6.57 (s, 2H), 3.69 (s,
6H), 3.65 (s, 3H), 1.65 (d, 3H, J=5.7 Hz), 1.39 (s, 6H). LCMS: ret.
time: 10.35 min.; purity: 95%; MS (m/e): 578 (MH.sup.+).
Synthesis of
(+/-)-N4-(2,2-Dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-[[1-
(1-pyridinium)ethoxy)carbonyl]]-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidine-
diamine iodide salt
[0140]
(+/-)-N2-(1-Chloroethoxycarbonyl)-N4-(2,2-dimethyl-3-oxo-4H-5-pyri-
do[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinedia-
mine (50 mg, 0.086 mmol), pyridine (34 mg, 0.43 mmol) and NaI (129
mg, 0.86 mmol) in acetone were stirred at room temperature for 24
h. The reaction mixture was concentrated, diluted with water (5 mL)
and EtOAc (5 mL). The precipitate (pale brown) was filtered and
dried to provide the desired product,
(+/-)-N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-[[1-
(1-pyridinium)ethoxy)carbonyl]]-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidine-
diamine iodide salt. LCMS: ret. time: 8.82 min.; purity: 90%; MS
(m/e): 620 (M+). The remaining impurity was characterized as
N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-tr-
imethoxyphenyl)-2,4-pyrimidinediamine.
Example 11
Pharmacokinetics and Metabolism of the Compounds
[0141] Compounds were administered to rats orally at a dose of 4-5
mg/kg using PEG-400 as a vehicle. Selected compounds were also
administered as an IV bolus at a dose of 1 mg/kg. Plasma samples
were obtained from either the portal or jugular veins and analyzed
by LC/MS/MS for both the parent pyrimidine-2,4-diamine and the
prodrug compounds synthesized in the examples above.
Bioavailability was calculated using the AUC of
pyrimidine-2,4-diamine in jugular vein samples and the AUC of an IV
bolus dose of pyrimidine-2,4-diamine. For selected compounds,
portal vein samples were evaluated for both prodrug and
pyrimidine-2,4-diamine and the information was used to determine
the percent absorption of the orally administered dose. Results
from the in vivo evaluation of the compounds in rats are shown in
Table 1. TABLE-US-00001 TABLE 1 Summary of the pharmacokinetics of
the compounds in sprague-dawley rats. Cmax of Clearance rate
Compound Mode of pyrimidine- of prodrug, % of prodrug disclosed in:
Study # administration % F.sup.1 2,4-diamine.sup.2 ml/min/kg
absorbed.sup.3 Example 2 VO40197 IV and PO 3.1 41.6 72 20 Example 3
VO40219 PO 3 26.3 -- -- Example 5 VO40219 PO 27 237 -- -- Example 6
VO40219 PO 0 0 -- -- Example 9 VO40219 PO 3 26.3 -- -- .sup.1% F
calculated based on pyrimidine-2,4-diamine concentrations in
jugular vein samples. .sup.2Highest observed concentration of
pyrimidine-2,4-diamine in plasma following a 4 mg/kg oral dose or
prodrug .sup.3Calculated based on the following formula: %
absorption = (AUC of prodrug in portal vein following oral
administration/AUC or prodrug in jugular vein following IV
administration) * (IV dose/e) * 100
[0142] Many of the prodrugs evaluated orally in rats show the
presence of pyrimidine-2,4-diamine in systemic circulation as shown
in Table 1 (% F and Cmax). Thus, the in vivo studies demonstrate
that the prodrug moiety is enzymatically cleaved in vivo and
results in systemic circulation of pyrimidine-2,4-diamine parent
molecule.
[0143] Selected compounds were incubated in rat and human hepatic
microsomes (with and without NADPH) and analyzed by LC/MS/MS for
both the prodrug and pyrimidine-2,4-diamine. The results of the
compounds evaluated in vitro in hepatic microsome studies are
listed in Table 2. TABLE-US-00002 TABLE 2 Metabolic stability of
prodrugs in hepatic microsomes. Results Compound CYP450
Pyrimidine-2,4- disclosed in: System T.sub.1/2.sup.4
dependent.sup.5? diamine produced? Example 2 Rat and Human
<5/<5 Y Y microsomes Example 3 Rat and Human <5/<5 Y Y
microsomes Example 8 Rat and Human 35/10 Y Y microsomes
.sup.4Half-life of prodrug in the presence of NASDPH
.sup.5Determined by incubating compounds in microsomes in the
absence of NADPH. All of the compounds in the list were stable in
the absence of NADPH.
[0144] Studies were conducted in microsomes to determine whether
the prodrug moieties could be hydrolyzed by CYP450 enzymes. The
results from the use of compounds prepared in Examples 2, 3, and 8
provide clear evidence for P450 dependent cleavage of the prodrug
moiety. Since microsomes lack many of the cytosolic enzymes present
in rat and human liver, failure to detect pyrimidine-2,4-diamine in
microsomal incubations does not preclude conversion in vivo.
[0145] All publications and patent applications cited in this
specification are herein incorporated by reference as if each
individual publication or patent application were specifically and
individually indicated to be incorporated by reference.
[0146] Although the foregoing invention has been described in some
detail by way of illustration and example for purposes of clarity
of understanding, it will be readily apparent to one of ordinary
skill in the art in light of the teachings of this invention that
certain changes and modifications may be made thereto without
departing from the spirit or scope of the invention as defined in
the appended claims.
* * * * *