U.S. patent application number 17/492282 was filed with the patent office on 2022-09-08 for pyrazolo[1,5-a][1,3,5]triazine and pyrazolo[1,5-a]pyrimidine derivatives as cdk inhibitors.
This patent application is currently assigned to AURIGENE DISCOVERY TECHNOLOGIES LIMITED. The applicant listed for this patent is AURIGENE DISCOVERY TECHNOLOGIES LIMITED. Invention is credited to Rajeev Goswami, Subhendu Mukherjee, Ramulu Poddutoori, Susanta Samajdar.
Application Number | 20220281871 17/492282 |
Document ID | / |
Family ID | 1000006322366 |
Filed Date | 2022-09-08 |
United States Patent
Application |
20220281871 |
Kind Code |
A1 |
Samajdar; Susanta ; et
al. |
September 8, 2022 |
Pyrazolo[1,5-A][1,3,5]Triazine and Pyrazolo[1,5-A]Pyrimidine
Derivatives as CDK Inhibitors
Abstract
The present invention provides substituted pyrazolo[1,3,5]
triaziric and pyrazolo[1,5a] pyrimidine derivatives of formula (I),
which are therapeutically useful, particularly as selective
transcriptional CDK inhibitors including CDK7, CDK9, CDK12, CDK13
and CDK15, more particularly transcriptional CDK7 inhibitors
wherein X, ring A, ring B, L1, L2, R1,R2, R3, R4, R6, m, n and p
have the meanings given in the specification and pharmaceutically
acceptable salts thereof that are useful in the treatment and
prevention of disease or disorder associated with selective
transcriptional CDKs in a mammal. The present invention also
provides preparation of the compounds and pharmaccutical
formulations comprising at least one of the substituted pyrazolo
[1.5-a] [1-3,5] triazine and pyrazolo[1,5-a] pyrimidine derivatives
of formula (I) or a pharmaceutically acceptable salt thereof or a
stercoisomer liner thereof. ##STR00001##
Inventors: |
Samajdar; Susanta;
(Bangalore, IN) ; Poddutoori; Ramulu; (Karimnagar,
IN) ; Mukherjee; Subhendu; (Hooghly, IN) ;
Goswami; Rajeev; (Deharadun, IN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AURIGENE DISCOVERY TECHNOLOGIES LIMITED |
BANGALORE |
|
IN |
|
|
Assignee: |
AURIGENE DISCOVERY TECHNOLOGIES
LIMITED
BANGALORE
IN
|
Family ID: |
1000006322366 |
Appl. No.: |
17/492282 |
Filed: |
October 1, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15557028 |
Sep 8, 2017 |
11186576 |
|
|
PCT/IB2016/051302 |
Mar 8, 2016 |
|
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17492282 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 487/04 20130101;
A61K 31/53 20130101; A61K 45/06 20130101 |
International
Class: |
C07D 487/04 20060101
C07D487/04; A61K 45/06 20060101 A61K045/06; A61K 31/53 20060101
A61K031/53 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 9, 2015 |
IN |
1128/CHE/2015 |
Claims
1. A compound of formula (I): ##STR00313## or a pharmaceutically
acceptable salt or a stereoisomer thereof wherein, X is CH or N;
Ring A is monocyclic or bicyclic aryl, heteroaryl or
heterocycloalkyl; Ring B is cycloalkyl, heterocycloalkyl, aryl,
heteroaryl or absent; R.sub.1 is hydrogen, alkyl or cycloalkyl;
R.sub.2 is: ##STR00314## R.sub.3 at each occurrence independently
is halo, alkyl, hydroxy, alkoxy, amino, alkylamino, cyano, nitro or
haloalkyl; R.sub.4 at each occurrence independently is halo, alkyl,
hydroxy, alkoxy, --(NH).sub.q--S(O).sub.2--CH=CH.sub.2,
--(NH).sub.q--CH.sub.2--CH=CH--C(O)--NR.sub.aR.sub.b, ##STR00315##
wherein R.sub.5 and R.sub.5''at each occurrence independently are
hydrogen or alkyl; R.sub.5' is hydrogen, halo, alkyl, alkoxyalkyl
or --CH.sub.2--NR.sub.aR.sub.b; R.sub.6 is hydrogen or alkyl;
R.sub.a and R.sub.b are independently hydrogen or alkyl; or R.sub.a
and R.sub.b along with the nitrogen atom to which they are attached
form an optionally substituted heterocyclic ring having 0-2
additional heteroatoms selected from O, S and N; wherein the
optional substituent is one or more alkyl or halo; L.sub.1 is
--O--, --S--, --NH-- or absent; L.sub.2 is absent or optionally
substituted C.sub.1-C.sub.6 alkylene, wherein one or more methylene
units of the alkylene is optionally and independently replaced with
--C(O)--, --O----, --N(R.sub.7)-- or cycloalkylene; wherein R.sub.7
is hydrogen or alkyl; m is 0 to 1; n is 0, 1 or 2; p is 1, 2 or 3;
and q is 0 to 1.
2. The compound of claim 1, wherein the compound of formula (I) is
a compound of formula (IA): ##STR00316## or a pharmaceutically
acceptable salt thereof or a stereoisomer thereof
3. The compound of claim 1, wherein the compound of formula (I) is
a compound of formula (TB): ##STR00317## or a pharmaceutically
acceptable salt thereof or a stereoisomer thereof
4. The compound of claim 1, wherein the compound of formula (I) is
a compound of formula (IC): ##STR00318## or a pharmaceutically
acceptable salt thereof or a stereoisomer thereof
5. The compound of claim 1, wherein the compound of formula (I) is
a compound of formula (ID): ##STR00319## or a pharmaceutically
acceptable salt thereof or a stereoisomer thereof
6. The compound of claim 1, wherein the compound of formula (I) is
a compound of formula (IE): ##STR00320## or a pharmaceutically
acceptable salt thereof or a stereoisomer thereof
7. The compound of claim 1, wherein the compound of formula (I) is
a compound of formula (IF): ##STR00321## or a pharmaceutically
acceptable salt thereof or a stereoisomer thereof
8. The compound according to claim 1, wherein ring A is phenyl,
pyridyl or piperidinyl.
9. The compound according to claim 1, wherein ring B is cycloalkyl,
aryl, heterocycloalkyl or heteroaryl.
10. The compound according to claim 1, wherein L.sub.1 is NH or
O.
11. (canceled)
12. The compound according to claim 1, wherein R.sub.2 is
cyclohexyl, N-methyl-4-piperidinyl or tetrahydro-4-pyranyl.
13. The compound according to claim 1, wherein R.sub.1 is hydrogen
or alkyl.
14. The compound according to claim 1, wherein R.sub.4 is
##STR00322## wherein R.sub.5, R.sub.5' and R.sub.5'' are as defined
in claim 1.
15. The compound according to claim 14, wherein R.sub.5' is
--CH.sub.2--NR.sub.aR.sub.b; wherein R.sub.a and R.sub.b are
independently hydrogen or alkyl.
16. The compound according to claim 15, wherein R.sub.a and R.sub.b
along with the nitrogen atom to which they are attached form an
optionally substituted heterocyclic ring having 0-2 additional
heteroatoms selected from O, S and N.
17. A compound selected from the group consisting of:
TABLE-US-00030 Compound No: IUPAC name .sup. 40A.
3-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyraz-
olo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-methylbenzamide; .sup.
40B.
5-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyraz-
olo[1,5-
a][[1,3,5]triazin-4-yl)amino)methyl)phenyl)-2-methylbenzamide;
.sup. 40C.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazol-
o[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-6-methylpiperidine-2-carboxami-
de; 47.
3-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)py-
razolo[1,5- a][1,3,5]triazin-4-yl)amino)methyl)phenyl)benzamide;
48.
2-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)py-
razolo[1,5- a][1,3,5]triazin-4-yl)amino)methyl)phenyl)benzamide;
.sup. 48A.
3-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyraz-
olo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-2-methylbenzamide; 49.
(E)-4-((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1-
,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-N,N-dimethylbut-2-enamid-
e; 50.
(E)-4-(diethylamino)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 51.
(E)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-morpholinobut-2-enamide;
52.
(E)-4-(4-ethylpiperazin-1-yl)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-py-
ran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 53.
(E)-4-(dimethylamino)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 54.
(E)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-(pyrrolidin-1-yl)but-2-enami-
de; 55.
(E)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-(piperidin-1-yl)but-2-enamid-
e; 56.
(E)-4-(3-fluoropyrrolidin-1-yl)-N-(3-(((8-isopropyl-2-((tetrahydro-2H--
pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 57.
(E)-4-(3,3-difluoropyrrolidin-1-yl)-N-(3-(((8-isopropyl-2-((tetrahydro-
-2H-pyran-
4-yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-
enamide; 58.
(E)-4-(3-fluoropiperidin-l-yl)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-p-
yran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 59.
(E)-4-(dimethylamino)-N-(3-fluoro-5-(((8-isopropyl-2-((tetrahydro-2H-p-
yran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 60.
(E)-4-(dimethylamino)-N-(2-fluoro-3-(((8-isopropyl-2-((tetrahydro-2H-p-
yran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 61.
(E)-N-(3-fluoro-5-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)py-
razolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-morpholinobut-2-enamide;
62.
(E)-4-(dimethylamino)-N-(2-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl-
)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-enamide; 63.
(E)-N-(4-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-(pyrrolidin-1-yl)but-2-enami-
de; 79.
N-(2-((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-2-oxoethyl)acrylamide;
80.
N-(1-((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-1-oxopropan-2-yl)acrylam-
ide; 81.
N-(2-((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-2-oxoethyl)-N-methyl
acrylamide; 82.
N-(1-((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-2-methyl-1-oxopropan-2-
yl)acrylamide; 125. .sup.
4-acrylamido-N-(3-((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1-
,5- a][1,3,5]triazin-4-yl)amino)phenyl)benzamide; 130. .sup.
(E)-4-(4-(dimethylamino)but-2-enamido)-N-(3-((8-isopropyl-2-((1-methylpip-
eridin-
4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)phenyl)benzamide;
137. .sup.
(E)-4-(dimethylamino)-1-(4-(2-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperazin-
- 1-yl)but-2-en-1-one; 138. .sup.
(E)-1-(4-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)piperidin-1-yl)-4-(pyrrolidin-1-yl)but-
-2- en-1-one; and
or a pharmaceutically acceptable salt or a stereoisomer
thereof.
18. A compound [[is]] selected from the group consisting of:
TABLE-US-00031 Compound No: Structure 140 ##STR00323## 141
##STR00324## 142 ##STR00325## 143 ##STR00326## 144 ##STR00327## 145
##STR00328## 146 ##STR00329## 147 ##STR00330## 148 ##STR00331## 149
##STR00332## 150 ##STR00333## 151 ##STR00334## 152 ##STR00335## 153
##STR00336## 154 ##STR00337## 155 ##STR00338## 156 ##STR00339## 157
##STR00340## 158 ##STR00341## 159 ##STR00342## 160 ##STR00343## 161
##STR00344##
or a pharmaceutically acceptable salt or a stereoisomer
thereof.
19. A pharmaceutical composition comprising a compound of claim 1,
or a pharmaceutically acceptable salt or a stereoisomer thereof,
and at least one pharmaceutically acceptable carrier or
excipient.
20-23. (canceled)
24. A method of treating selective transcriptional CDKs mediated
disorders or diseases or condition in a subject comprising
administering a therapeutically effective amount of a compound
according to claim 1.
25. A method of inhibiting selective transcriptional CDKs in a
subject, comprising administering to the subject a compound of
claim 1.
26. (canceled)
27. The method of claim 24, wherein the selective transcriptional
CDKs mediated disorder or disease or condition is selected from the
group consisting of a cancer, an inflammatory disorder, an
auto-inflammatory disorder or an infectious disease.
28. The method of claim 27, wherein the cancer is selected from the
group consisting of a carcinoma, including that of the breast,
liver, lung, colon, kidney, bladder, including small cell lung
cancer, non-small cell lung cancer, head and neck, thyroid,
esophagus, stomach, pancreas, ovary, gall bladder, cervix, prostate
and skin, including squamous cell carcinoma; hematopoietic tumors
of lymphoid lineage, including leukemia, acute lymphoblastic
leukemia, acute lymphocytic leukemia, Hodgkins lymphoma,
non-Hodgkins lymphoma, B-cell lymphoma, T- cell lymphoma, hairy
cell lymphoma, myeloma, mantle cell lymphoma and Burkett's
lymphoma; hematopoietic tumors of myeloid lineage, including acute
and chronic myelogenous leukemias, myelodysplastic syndrome and
promyelocytic leukemia; tumors of masenchymal origin, including
fibrosarcoma and rhabdomyosarcoma; tumors of the central and
peripheral nervous system, including astrocytoma, neuroblastoma,
glioma and schwannomas; and other tumors, including seminoma,
melanoma, osteosarcoma, teratocarcinoma, keratoctanthoma,
xenoderoma pigmentosum, thyroid follicular cancer and Kaposi's
sarcoma.
29. The method of claim 24 any one of the claims 24, comprising an
additional step of administering to the subject in need thereof one
or more additional chemotherapeutic agents independently selected
from anti-proliferative agents, anti-cancer agents,
immunosuppressant agents and pain-relieving agents.
30. The method of claim 24, wherein selective transcriptional CDKs
are CDK7, CDK9, CDK12, CDK13 or CDK 18.
31-32. (canceled)
Description
This application claims the benefit of Indian provisional
application number 1128/CHE/2015 filed on 9Mar. 2015, which hereby
incorporated by reference.
FIELD OF THE INVENTION
[0001] This invention pertains to compounds which inhibit the
activity of selective transcriptional cyclin dependent kinases
(CDKs) including CDK7, CDK9, CDK 12, CDK 13 and CDKI8, more
particularly transcriptional cyclin dependent kinase-7 (CDK7). The
invention also provides pharmaceutically acceptable compositions
comprising compounds of the present invention and methods of using
said compositions in the treatment of diseases or disorder
associated with selective transcriptional CDKs.
BACKGROUND OF THE INVENTION
[0002] One of the most important and fundamental processes in
biology is the division of cells mediated by the cell cycle. This
process ensures the controlled production of subsequent generations
of cells with defined biological function. It is a highly regulated
phenomenon and responds to a complex set of cellular signals both
within the cell and from external sources. A complex network of
tumor promoting and suppressing gene products arc key components of
this cellular signalling process. Oyer-expression of
tumor-promoting components or the subsequent loss of the
tumor-suppressing products will lead to unregulated cellular
proliferation and the generation of tumors (Pardee, Science
246:603-608, 1989).
[0003] Kinases are important cellular enzymes that perform
essential cellular functions such as regulating cell division and
proliferation, and also appear to play a decisive role in many
disease states that are characterized by uncontrolled proliferation
and differentiation of cells. These disease states encompass a
variety of cell types and maladies such as cancer. atherosclerosis,
restenosis and other proliferative disorders (Kris MG et al.,:
"Efficacy of gefitinib, an inhibitor of the epidermal growth factor
receptor tyrosine kinase, in symptomatic patients with non-small
cell lung cancer: a randomized trial". JAMA 290 (16): 2149-58,
October 2003).
[0004] Cyclin-dependent kinases (CDKs) are relatively small
proteins, with molecular weights ranging from 34 to 40 kDa, and
contain little more than the kinase domain. CDK binds a regulatory
protein called a cyclin. Without cyclin, CDK has little kinase
activity only the cyclin-CDK complex is an active kinase. CDKs
phosphorylate their substrates on serines and threonines, so they
are serine-threonine kinases (Morgan, David O.,The Cell Cycle:
Principle.s of Control. London: New Science Press, 1st edition,
(2007)).
[0005] The members of the cyclin-dependent kinase (CDK) family play
critical regulatory roles in cell proliferation. There are
currently 20 known mammalian CDKs. While CDK7-13 and 18 have been
linked to transcription, only CDK 1, 2, 4 and 6 show demonstrable
association with the cell cycle. Unique among the mammalian CDKs,
CDK7 has consolidated kinase activities, regulating both the cell
cycle and transcription. In the cytosol, CDK7 exists as a
hetcrotrimeric complex and is believed to function as a
CDK1/2-activating kinase (CAK), whereby phosphorylation of
conserved residues in CDK1/2 by CDK7 is required for full catalytic
CDKs activity and cell cycle progression (Desai et al., Mol. Cell
Biol. 15, 345-350 (1995)).
[0006] CDK7, which complexes with cyclin II and MAT1 ,
phosphorylates the cell cycle CDKs in the activation of T-loop, to
promote their activities (see, e.g., Fisher et al., 1994). As such,
it has been proposed that inhibiting CDK7 would provide a potent
means of inhibiting cell cycle progression, which may be especially
relevant given that there is compelling evidence from gene knockout
studies in mice for lack of an absolute requirement for CDK2, CDK4
and CDK6 for the cell cycle, at least in most cell types (see,
e.g., Malumbres er al., 2009), whilst different tumors appear to
require some, but be independent of other interphase CDKs (CDK2)
CDK4, CDK6). Recent genetic and biochemical studies have confirmed
the importance of CDK7 for cell cycle progression (see, e.g.,
Larochelle er al., 2007: Gannza et al., 2012).
[0007] Cyclin-dependent kinase 7 (CDK7) activates cell cycle CDKs
and is a member of the general Transcription factor II Human
(TFIIH) CDK7 also plays a role in transcription and possibly in DNA
repair. The trimeric Cak complex CDK7/CyclintII/MAT I is also a
component of TFIIII, the general transcription/DNA repair factor
IIII (reviewed in Morgan, D. O., Annu Rev Cell Dec Biol 13, 261-91,
(1997)). As a TFIIII subunit, CDK7 phosphorylates the CTD
(Carboxy-Terminal-Domain) of the largest subunit of RNA polymerase
II (pol II). The CTD of mammalian pol II consists of 52 heptad
repeats with the consensus sequence .sup.1YSPTSPS.sup.7 and the
phosphorylation status of the Ser residues at positions 2 and 5 has
been shown to be important in the activation of RNAP-II indicating
that it is likely to have a crucial role in the function of the
CID. CDK7, which primarily phosphorylates Ser-5 (PS5) of RNAP-II at
the promoter as part of transcriptional initiation (Gomers et al.,
2006), incontrast with CDK9, which phosphorylates both Ser-2 and
Ser-5 of the CID heptad (Pinhero et al., 2004).
[0008] In addition to CDK7, other CDKs have been reported to
phosphorylate and regulate RNA pol (11) CTD. The other CDKs
include, Cdk9/ Cyclin T1 or T2 that constitute the active form of
the positive transcription elongation factor (P-TEFb) (Peterlin and
Price, 2006) and Cdk12/Cyclin K and Cdk13/Cyclin K as the latest
members of RNAPII CTD kinases (Bartkowiak et al., 2010; Blazek et
al., 2011).
[0009] Disruption of RNAP II CTD phosphorylation has been shown to
preferentially effect proteins with short half-lives, including
those of the anti-apoptotic BCL-2 family. (Konig et al., "The novel
cyclin-dependent kinase inhibitor flavopiridol downregulates Bcl-2
and induces growth arrest and apoptosis in chronic B-cell leukemia
lines." (Blood 1, 4307-4312 (1997) Gojo et al.) The transcriptional
non-selective cyclin-dependent kinase inhibitor flavopiridol
induces apoptosis in multiple myeloma cells through transcriptional
repression and down-regulation of Mel- 1; (Clin. Cancer Res. 8,
3527-3538 (2002)).
[0010] This suggests that the CDK7 enzyme complexes arc involved in
multiple functions in the cell; cell cycle control, transcription
regulation and DNA repair. It is surprising to find one kinase
involved in such diverse cellular processes, some of which are even
mutually exclusive. It also is puzzling that multiple attempts to
find cell cycle dependent changes in CDK7 kinase activity remained
unsuccessful. This is unexpected since activity and phosphorylation
state of its substrate, CDC2, fluctuate during the cell cycle
(Larochelle, S. et al. Genes Dev 12,370-81, (1998)). Indeed,
flavopiridol, a non-selective pan-CDK inhibitor that targets CTD
kinases, has demonstrated efficacy for the treatment of chronic
lymphocytic leukemia (CLL), but suffers from a poor toxicity
profile (Lin et al., "Phase II study of flavopiridol in relapsed
chronic lymphocytic leukemia demonstrating high response rates in
genetically high-risk disease." .J. Clin. Oncol. 27, 6012-6018
(2009); Christian et al., "Flavopiridol in chronic lymphocytic
leukemia: a concise review." Clin. Lymphoma Myelomu 9 Suppl. 3,
S179-S185 (2009)).
[0011] In-vitro studies revealed substrate preferences for the
different CDK7 complexes, indicating that CDK7 may form different
complexes with different substrate specificity and presumably
different in-vivo functions (Frit, P. et al., Biochimie 81, 27-38,
(1999): Schutz, P. et al. Cell 102, 599-607, (2000)).
[0012] Various CDK inhibitors have been reported in the literature
including WO2006052936 A2, WO2007038314A2, WO2008119792A1,
WO2013169401A1, US20020091263A1, WO2008151304A1, WO2010103486A1,
WO2010003133A2, WO2005026129A1, WO2012045195A 1, WO2007038314A2
etc.
[0013] There is a need for new compounds, formulations, treatments
and therapies to treat diseases and/or disorders associated with
selective transcriptional CDKs including CDK7, CDK 9, CDK 12, CDK
I3 and CDK 18; more particularly CKD7. It is, therefore, an object
of this invention to provide compounds useful in the treatment
and/or prevention or amelioration of such diseases and/or
disorders.
BRIEF DESCRIPTION OF ACCOMPANYING DRAWINGS:
[0014] FIG. 1: In vivo antitumor activity of CDK7 inhibitor in the
MV4-11 AML xenograft model in athymic nude mice.
SUMMARY OF THE INVENTION
[0015] Provided herein are pyrazolo [1,5-a][1,3,5 ] triazine and
pyrazolo[1,5-a] pyrimidine derivatives and pharmaceutical
compositions thereof, which arc useful as selective transcriptional
CDK inhibitors.
[0016] In one aspect of the present invention, it comprises
compounds of formula (I):
##STR00002##
or a pharmaceutically acceptable salt or a stereoi smiler thereof;
wherein.
[0017] X is CH or N;
[0018] Ring A is monocyclic or bicyclic aryl, hctcroaryl or
hcterocycloalkyl:
[0019] Ring B is cycloalkyl, heterocycloalkyl, aryl, heteroaryl or
absent;
[0020] R.sub.1 is hydrogen, alkyl or cycloalkyl:
[0021] R.sub.2 is an optionally substituted alkyl, cycloalkyl or
heterocycloalkyl; wherein the optional substituents are amino,
halo, hydroxy, alkoxy, alkoxyalkoxy, alkylamino, cyano, nitro or
haloalkyl:
[0022] R.sub.3 at each occurrence independently is halo, alkyl,
hydroxy, alkoxy, amino, alkylamino, cyano, nitro or haloalkyl;
[0023] R.sub.4 at each occurrence independently is halo, alkyl,
hydroxy, alkoxy, --(NH).sub.q--S(O).sub.2,--
##STR00003##
CH.dbd.CH.sub.2,
--(NH).sub.q--CH.sub.2--CH.dbd.CH--C(O)--NR.sub.aR.sub.b, wherein
R.sub.5 and R.sub.5''at each occurrence independently are hydrogen
or alkyl; R.sub.5' is hydrogen, halo, alkyl, alkoxyalkyl or
--CH.sub.2--NR.sub.aR.sub.b:
[0024] R.sub.6 is hydrogen or alkyl;
[0025] R.sub.a and R.sub.b are independently hydrogen or alkyl or
R.sub.a and R.sub.b, along with the nitrogen atom to which they arc
attached form an optionally substituted heterocyclic ring having
0-2 additional heteroatoms selected from O, S and N: wherein the
optional substituent is one or more alkyl or halo:
[0026] L.sub.1 is --O--, --S--, --NH-- or absent:
[0027] L.sub.2 is absent or optionally substituted alkylene,
wherein one or more methylene units of the alkylene is optionally
and independently replaced with --C(O)--, --O--, --N(R.sub.7)-- or
cycloalkylene: wherein R.sub.7 is hydrogen or alkyl;
[0028] m is 0 to 1:
[0029] n is 0, 1 or 2:
[0030] p is 1, 2 or 3: and
[0031] q is 0 to 1.
[0032] In another aspect, the present invention provides a
pharmaceutical composition comprising the compound of formula (I)
and atleast one pharmaceutically acceptable excipient (such as a
pharmaceutically acceptable carrier or diluent).
[0033] In yet another aspect, the present invention relates to the
preparation of compounds of formula (I).
[0034] In yet another aspect of the present invention, provided
herein are pyrazolo[1,5-a][1,3,5] ltriazine and pyrazolo[1,5-a]
plyrimidine derivatives of formula (I), which are useful for
medical use. In particular for the treatment or prevention of
diseases and/or disorders where selective transcriptional CDK
inhibition is desired.
DETAILED DESCRIPTION OF THE INVENTION
[0035] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as is commonly understood by one
of skill in art to which the subject matter herein belongs. As used
in the specification and the appended claims, unless specified to
the contrary, the following terms have the meaning indicated in
order to facilitate the understanding of the present invention.
[0036] As used herein, the term "optionally substituted" or
"suitable groups" refers to replacement of one or more hydrogen
radicals in a given structure with a radical of a specified
substituent including, but not limited to: halo, alkyl, alkenyl,
alkynyl, aryl, hcterocyclyl, thiol, alkylthio, arylthio,
alkylthioalkyl, arylthioalkyl, alkylsulfnnyl, alkylsulfonylalkyl,
arylsulfonylalkyl, alkoxy, aryloxy, aralkoxy, aminocarbonyl,
alkylaminocarhonyl, arylaminocarbonyl, alkoxycarhonyl,
aryloxycarbonyl, haloalkyl, amino, cyano, nitro, alkylamino,
arylamino, alkylaminoalkyl, arylaminoalkyl, aminoalkylamino,
hydroxy, alkoxyalkyl, carboxyalkyl, alkoxycarbonylalkyl,
aminocarbonylalkyl, acyl, aralkoxycarbonyl, carboxylic acid,
sulfonic acid, sulfonyl, phosphonic acid, aryl, heteroaryl and
heterocyclic. It is understood that the substituent may be further
substituted.
[0037] As used herein, unless otherwise defined the term "alkyl"
alone or in combination with other (erm(s) means saturated
aliphatic hydrocarbon chains, including C.sub.1-C.sub.10, straight
or C.sub.1,-C.sub.10, branched alkyl groups. Examples of "alkyl"
include hut are not limited to methyl, ethyl, propyl, isopropyl,
butyl, isobutyl, sec-butyl, tort-butyl, isopentyl or neoperayl and
the like.
[0038] As used herein, the term "halo" or "halogen" alone or in
combination with other lerm(s) means fluorine, chlorine, bromine or
iodine.
[0039] The term "hydroxy" or "hydroxyl" refers to OH group.
[0040] The term "amino" refers to --NH.sub.2 group.
[0041] As used herein, the (erm "alkoxy" or "alkoxyl" refers to the
group alkyl-O-- or --O-alkyl, where alkyl groups arc as defined
above. Exemplary C.sub.1-C.sub.10 alkyl group containing alkoxy
groups include but arc not limited to methoxy, ethoxy, n-propoxy,
n-butoxy, t-hutoxy and the like. An alkoxy group can be
unsubstituted or substituted with one or more suitable groups.
[0042] As used herein, the term "alkoxylalkoxy" refers to the group
alkyl-O-alkoxy- or -alkoxy-O-alkyl, where alkoxy groups are defined
above. Exemplary alkoxylalkoxy groups include but are not limited
to methoxyethoxy, ethoxyethoxy, methoxypropoxy, ethoxymethoxy,
ethoxypropoxy, propoxymethoxy, propoxyethoxy and the like.
[0043] As used herein, the term "cycloalkyl" alone or in
combination with other term(s) means C.sub.3-C.sub.10 saturated
cyclic hydrocarbon ring. A cycloalkyl may be a single ring, which
typically contains from 3 to 7 carbon ring atoms. Examples of
single-ring cycloalkyls include but are not limited to cyclopropyl,
cyclohutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. A
cycloalkyl may alternatively be polycyclic or contain more than one
ring. Examples of polycyclic cycloalkyls include bridged, fused and
spirocyclic carbocyclyls and the like.
[0044] As used herein, the term "aryl" is optionally substituted
monocyclic, bicyclic or polycyclic aromatic hydrocarbon ring system
of about 6 to 14 carbon atoms. Examples of a C.sub.6-C.sub.11 aryl
group include, hut arc not limited to phenyl, naphthyl, biphenyl,
anthryl, tctrahydronaphthyl, fluorcnyl, indanyl, biphenylenyl and
acenaphthyl. Aryl group can be unsubstituted or substituted with
one or more suitable groups.
[0045] The term "heterocycloalkyl" refers to a non-aromatic,
saturated or partially saturated, monocyclic or polycyclic ring
system of 3 to 15 members having at least one heteroatom or
heterogroup selected from O, N, S, S(O), S(O).sub.2, NH or C(O)
with the remaining ring atoms being independently selected from the
group consisting of carbon, oxygen, nitrogen and sulfur. Examples
of "Heterocycloalkyl" include, but are not limited to azetidinyl,
oxetanyl, irnidazolidinyl, pyrrolidinyl, oxazolidinyl,
thiazolidinyl, pyrazolidinyl , tetrahydrofuranyl, piperidinyl,
piperazinyl, tetrahydropyranyl, morpholinyl, oxapiperazinyl,
oxapiperidinyl, tetrahydrofuryl, tetrahydropyranyl,
tetrahydrothiophenyl, di h ydropyranyl, indolinyl, indolinylmethyl,
azepanyl and their N-oxides thereof. Attachment of a
heterocycloalkyl substituent can occur via either a carbon atom or
a heteroatom. A heterocycloalkyl group can be optionally
substituted with one or more suitable groups by one or more
aforesaid groups.
[0046] As used herein, the term "hetcroaryl" alone or in
combination with other term(s) means a completely unsaturated ring
system containing a total of 5 to 14 ring atoms. At least one of
the ring atoms is a heteroatom (i.e., oxygen, nitrogen or sulfur),
with the remaining ring atoms/groups being independently selected
from the group consisting of carbon, oxygen, nitrogen or sulfur. A
heteroaryl may be a single-ring (monocyclic), bicyclic or
polycyclic ring system. Examples of "heteroaryl" include but are
not limited to pyridyl, pyridine- 1-oxide, indolyl, benzimidazolyl,
benzothiazolyl and the like. The term heteroaryl includes their
N-oxides thereof.
[0047] The term "heteroatom" as used herein designates a sulfur,
nitrogen or oxygen atom.
[0048] As used herein, the term "compound(s)" comprises ale
compounds disclosed in the present invention.
[0049] As used herein, the term "comprise" or "comprising" is
generally used in the sense of include, that is to say permitting
the presence of one or more features or components.
[0050] As used herein, the term "or" means "and/or" unless stated
otherwise.
[0051] As used herein, the term "including" as well as other forms,
such as "include", "includes" and "included" is not limiting.
[0052] As used herein, the term "composition" is imended lo
encompass a product comprising the specified ingredients in the
specified amounts, as well as any product which results, directly
or indirectly, from combination of the specified ingredients in the
specified amounts. By "pharmaceutically acceptable" it is meant the
carrier, diluent or excipient must he compatible with the other
ingredients of the formulation and not deleterious to the recipient
thereof.
[0053] As used herein, the term "treat", "treating" and "treatment"
refer to a method of alleviating or abrogating a disease and/or its
attendant symptoms.
[0054] As used herein, the term "prevent", "preventing" and
"prevention" refer to a method of preventing the onset of a disease
and/or its attendant symptoms or barring a subject from acquiring a
disease. As used herein, "prevent", "preventing" and "prevention"
also include delaying the onset of a disease and/or its attendant
symptoms and reducing a subject's risk of acquiring a disease.
[0055] As used herein, the "subject" to which administration is
contemplated includes, but are not limited to humans and/or
non-human animals; for example, mammals and birds. In certain
embodiments the animal is mammal. A non-human animal may be
transgenic animal.
[0056] As used herein, the term "administer", "administering" adm i
n i st i on refers to implanting, absorbing, ingesting, injecting,
inhaling or otherwise introducing a compound of formula (I) or a
pharmaceutical composition thereof.
[0057] As used herein, the term "therapeutically effective amount"
refers to that amount of the compound being administered sufficient
to prevent development of or alleviate to some extent one or more
of the symptoms of the condition or disorder being treated.
[0058] "Pharmaceutically acceptable" means that, which is useful in
preparing a pharmaceutical composition that is generally safe,
non-toxic and neither biologically nor otherwise undesirable and
includes that which is acceptable for veterinary as well as human
pharmaceutical use.
[0059] The term "stereoisomers" or "isomers" refers to any
enantiomers, diastereoisomers or geometrical isomers of the
compounds of formula (I), (IA), (IB), (IC), (ID), (IE) and (IF)
wherever they are chiral or when they bear one or more double
bonds. When the compounds of the formula (I), (IA), (IB), (IC),
(ID), (IE) and (IF): and related formulae are chiral, they can
exist in racemic or in optically active form. It should be
understood that the invention encompasses all stereochemical
isomeric forms, including diastereomeric, enantiomeric and epimeric
forms, as well as d-isomers and l-isomers and mixtures thereof.
Individual stereoisomers of compounds can be prepared synthetically
from commercially available starting materials which contain chiral
centers or by preparation of mixtures of enantiomeric products
followed by separation such as conversion lo a mixture of
diaslereomers followed by separation or recrystallizabon,
chromatographic techniques, direct separation of enantiomers on
chiral chromatographic columns or any other appropriate method
known in the art. Starting compounds of particular stereochemistry
are either commercially available or can be made and resolved by
techniques known in the art. Additionally, the compounds of the
present invention may exist as geometric isomers. The present
invention includes all cis, trans, syn, anti, enigegen (E) and
zusammen (Z) isomers as well as the appropriate mixtures
thereof.
[0060] As used hercn the term "CDK" refers to a cyclin-dependent
kinase. A CDK binds a cyclin (e.g. , Cyclin II), which is a
regulatory protein. CDKs phosphorylate their substrates at serines
and threonines. CDKs include CDK1, CDK2, CDK2, CDK4, CDK5, CDK7,
CDK8, CDK9, CDK10, CDK11, CDK12, CDK13, CDK14, CDK16, CDK18 and
CDK20. CDK7 is a CDK wherein the substrate is Cyclin II, MAT1
(e.g., MNAT1) or Cyclin II and MAT1 complex. The term CDK inhibitor
refers to selective transcriptional CDK inhibitor.
[0061] The present invention provides substituted
pyrazolo[1,5-a][1,3,5]triazine and pyrazolo[1,5-a]pyrimidine
derivatives of formula (I), which are useful for the inhibition of
selective transcriptional CDKs, particularly selective
transcriptional CDK7, CDK9, CDK12. CDK13 or CDK 18, more
particularly selective transcriptional CDK7.
[0062] The present invention further provides pharmaceutical
compositions comprising the said substituted pyrazolo[1 ,5-a ][1 ,3
,5]utriazine and pyrazolo[1,5-a]pyrimidine compounds and their
derivatives as therapeutic agents.
[0063] According to first embodiment, the present invention
provides compounds of formula (I)
##STR00004##
or a pharmaceutically acceptable salt or a stereoisomer thereof;
wherein.
[0064] X is CH or N;
[0065] Ring A is monocyclic or bicyclic aryl, heteroaryl or
heterocycloalkyl;
[0066] Ring B is cycloalkyl, heterocycloalkyl, aryl, heteroaryl or
absent;
[0067] R.sub.1 is hydrogen, alkyl or cycloalkyl;
[0068] R.sub.2 is an optionally substituted alkyl, cycloalkyl or
heterocycloalkyl; wherein the optional substituents are amino,
halo, hydroxy, alkyl, alkoxy, alkoxyalkoxy, alkylamino, cyano,
nitro or hak)alkyl;
[0069] R.sub.3 al each occurrence independently is halo, alkyl,
hydroxy, alkoxy, amino, alkylamino, cyano, nitro or haloalkyl;
[0070] R.sub.4 at each occurrence independently is halo, alkyl,
hydroxy, alkoxy, --(NH).sub.q--S(O).sub.2--CH.dbd.CH.sub.2,
--(NH).sub.q--CH.sub.2--CH.dbd.CH--C(O)--NR.sub.aR.sub.b,
##STR00005##
wherein R.sub.5 and R.sub.5''at each occurrence independently are
hydrogen or alkyl: R.sub.5' is hydrogen, halo, alkyl, alkoxyalkyl
or 13 CH.sub.2--NR.sub.aR.sub.b;
[0071] R.sub.6 is hydrogen or alkyl;
[0072] R.sub.a and R.sub.b are independently hydrogen or alkyl; or
R.sub.a and R.sub.b along with the nitrogen atom to which they are
attached form an optionally substituted heterocyclic ring having
0-2 additional heteroatoms selected from O, S and N; wherein the
optional substituent is one or more alkyl or halo:
[0073] L.sub.1 is --O--, --S--, --NH-- or absent;
[0074] L.sub.2 is absent or optionally substituted C.sub.1-C.sub.6,
alkylcne, wherein one or more methylene units of the alkylene is
optionally and independently replaced with --C(O)--, --O--,
--N(R.sub.7)-- or cycloalkylene: wherein R.sub.7 is hydrogen or
alkyl;
[0075] m is to 1:
[0076] n is 0, 1 or 2;
[0077] p is 1, 2 or 3; and
[0078] q is to 1.
[0079] In another embodiment of the present invention, the compound
of formula (I) is a compound of formula (IA):
##STR00006##
[0080] or a pharmaceutically acceptable salt thereof or a
stereoisomer thereof;
[0081] wherein,
[0082] ring A, ring B, L.sub.1, L.sub.2, R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.6, m, n and p are same as defined in formula
(I).
[0083] In yet another embodiment of the present invention, the
compound of formula (I) is a compound of formula (IB):
##STR00007##
or a pharmaceutically acceptable salt thereof or a stereoisomer
thereof; wherein,
[0084] X, ring B, L.sub.1, L.sub.2, R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.6, m, n and p are same as defined in formula
(I).
[0085] In yet another embodiment of the present invention, the
compound of formula (I) is a compound of formula (IC):
##STR00008##
or a pharmaceutically acceptable salt thereof or a stereoisomer
thereof; wherein.
[0086] R.sub.2 is optionally substituted cycloalkyl or optionally
substituted heterocycloalkyl;
[0087] X, ring A, ring B, L.sub.2, R.sub.1, R.sub.3, R.sub.4,
R.sub.6, m and p are same as defined in formula (I).
[0088] In another embodiment of the present invention, the compound
of formula (I) is a compound of formula (ID):
##STR00009##
or a pharmaceuticallY acceptable salt thereof or a stereoisomer
thereof; wherein.
[0089] R.sub.2 is optionally substituted cycloalkyl or optionally
substituted heterocycloalkyl;
[0090] X, ring B L.sub.2, R.sub.1, R.sub.3, R.sub.4, R.sub.6, m, n
and p are same as defined in formula (I).
[0091] In another embodiment of the present invention, the compound
of formula (I) is a compound of formula (IE):
##STR00010##
or a pharmaceutically' acceptable salt thereof or a stereoisomer
thereof; wherein,
[0092] X, ring B L.sub.2, R.sub.1, R.sub.3, R.sub.4, R.sub.6, m, n
and p are same as defined in formula (I).
[0093] In another embodiment of the present invention, the compound
of formula (I) is a compound of formula (IF):
##STR00011##
or a pharmaceutically acceptable salt thereof or a stereoisomer
thereof; wherein,
[0094] X, L.sub.1, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.6 , m,
n and p are same as defined in formula (I).
[0095] According to one embodiment, the present invention provides
compounds of formula (I) and (IA): wherein, ring A is aryl; and
ring B, X, L.sub.1, L.sub.2, R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.6, m, n and p are same as defined in formula (I).
[0096] According to another embodiment the present invention
provides compounds of formula (I) and (IA) wherein ring A is
monocyclic or bicyclic heteroaryl or heterocycloalkyl and their
N-oxides thereof; and ring B, X, L.sub.1, L.sub.2, R.sub.1,
R.sub.2, R.sub.3, R.sub.4, R.sub.6, m, n and p are same as defined
in formula (I).
[0097] According to yet another embodiment the present invention
provides compounds of formula (I). (IB), (IC), (ID), (IE) and (IF);
wherein X is N.
[0098] According to yet another embodiment the present invention
provides compounds of formula (I), (IA). (IB), (IC), (ID) and (IE);
wherein. L.sub.2, is absent or selected from --NHC(O)--,
--C(O)NH--, --OC(O)--,
##STR00012##
[0099] According to yet another embodiment the present invention
provides compounds of formula (I), (IA), (IB), (IC), (ID) and (IE);
wherein ring B is absent or selected from phenyl, cyclohexyl,
piperidinyl, pyrrolidinyl, azitidinyl, 1-methyl-1H-pyrazole,
piperazinyl and morpholinyl.
[0100] The embodiments below are illustrative of the present
invention and are not intended to limit the claims lo the specific
embodiments exemplified.
[0101] According to certain embodiments of the present invention,
specifically provided are compounds of formula (I), (IA), (IB).
(IC). (ID), (II) and (IF); wherein. R.sub.1 is hydrogen, alkyl or
cycloalkyl; preferably the said alkyl is ethyl or isopropyl and the
said cycloalkyl is cyclopropyl.
[0102] According to certain embodiments of the present invention,
specifically provided are compounds of formula (I), (IA), (IB)
(IC), (ID), (IE) and (IF); wherein, R.sub.7 is optionally
substituted cycloalkyl or heterocycloalkyl; preferably optionally
substituted cycloalkyl is
##STR00013##
and in particular optionally substituted heierocycloalkyl is
##STR00014##
[0103] According to certain embodiments of the present invention,
specifically provided are compounds of formula (I), (IA), (IB),
(IC). (ID), (IE) and (IF); wherein. R.sub.2 is alkyl optionally
substituted with amino, alkoxy or alkoxylalkoxy; preferably R.sub.2
is methyl, aminohutyl, tnethoxyethyl, isohutanyl and
methoxyethoxyethyl.
[0104] According to certain embodiments of the present invention,
specifically provided are compounds of formula (I), (IA), (IB),
(IC), (ID), (IE) and (IF); wherein, R.sub.6 is hydrogen or alkyl;
preferably the said alkyl is methyl.
[0105] According to certain embodiments of the present invention,
specifically provided are compounds of formula (I), (IA) and (IC);
wherein, ring A is phenyl, piperidinyl, pyridyl and
pyridine-N-oxide.
[0106] According to certain embodiments of the present invention,
specifically provided are compounds of formula (I), (IA), (IB),
(IC). (ID), (IE) and (IF); wherein. R.sub.3 is halo or alkyl;
preferably the said halo is fluoro and tile said alkyl is methyl or
ethyl.
[0107] According to certain embodiments of the present invention,
specifically provided are compounds of formula (I), (IA), (IB),
(IC). (ID), (IE) and (IF); wherein. R.sub.4 is
--(NH).sub.q--S(O).sub.2--CH.dbd.CH.sub.2,
--(NH).sub.4--CH.sub.2--CH.dbd.CH--C(O)--NR.sub.aR.sub.b,
##STR00015##
and R.sub.5, R.sub.5', R.sub.5'', R.sub.a, R.sub.b and q are same
as defined in formula (I).
[0108] According to the preceding embodiment, specifically provided
are compounds of formula (I), (IA), (IB), (IC), (ID), (IE) and
(IF); wherein, R.sub.1 is
##STR00016## ##STR00017##
[0109] According lo certain embodiments, the present invention
provides compounds of the formula (I) in which `m` and `n` are
independently 0 or 1: and `p` is 1.
[0110] According to certain embodiments, the present invention
provides a compound selected from the group consisting of;
TABLE-US-00001 Compound No: IUPAC name 1.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-3-carboxamide;
2.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)azetidine-2-carboxamide;
3.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro)-2H-pyran-4-yl)amino)pyr-
azolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-2-carboxamide:
4.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide;
5. Isomer-1:
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)
pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-
carboxamide; 6. Isomer-2:
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)
pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-
carboxamide; 7.
(1,4-cis)-4-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexa-
ne-1- carboxamide; 8.
1-acryloyl-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)amino)pyrazo-
lo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-3-carboxamide;
9.
(1,4-cis)-4-acrylamido-N-(3-(((2-(((1r,4r)-4-hydroxycyclohexyl)amino)--
8-
isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexa-
ne-1- carboxamide; 10. Isomer-1:
4-acrylamido-N-(3-(((2-((3-hydroxycyclohexyl)amino)-8-
isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexa-
ne-1- carboxamide; 11. Isomer-2:
4-acrylamido-N-(3-(((2-((3-hydroxycyclohexyl)amino)-8-
isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexa-
ne-1- carboxamide; 12. Isomer-1:
1-acryloyl-N-(3-(((2-((3-hydroxycyclohexyl)amino)-8-
isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidi-
ne-3- carboxamide; 13. Isomer-2:
1-acryloyl-N-(3-(((2-((3-hydroxycyclohexyl)amino)-8-
isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidi-
ne-3- carboxamide; 14.
1-acryloyl-N-(3-(((2-(((3S,4S)-3-fluoropiperidin-4-yl)amino)-8-
isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidi-
ne-3- carboxamide; 15.
1-acryloyl-N-(3-(((2-((3-aminocyclohexyl)amino)-8-isopropylpyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide;
16.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-3-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxamide;
17.
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)
phenyl)-1-(3-methylbut-2-enoyl)azetidine-2- carboxamide; 18.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)azetidine-3-carboxamide;
19. (R)-1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)pheny1)azetidine-
-2- carboxamide; 20. (S)-l-acry1oy1-N-(3-(((8-isopropyl-2-
telrahydro-2H-pyran-4- yl)amino)pyrazolo| 1,5-a][ 1,3,5
|triazin-4-yl)amino)methyl)phenyl)aze(idine-2- carboxamide; 21.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxamide;
22. 3-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexa-
ne-1- carboxamide; 23.
4-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)morpholine-2-carboxamide;
24.
4-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)morpholine-3-carboxamide;
25.
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-propioloylmorpholine-3-
carboxamide; 26.
4-acryloyl-N-(4-ethyl-3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)am-
ino)pyrazolo]1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)morpholine-3-carboxamide;
27.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)-4-methylphenyl)azetidine-2-carboxamid-
e; 28.
1-acryloyl-N-(5-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)-2-methylphenyl)azetidine-2-carboxamid-
e; 29.
1-acryloyl-N-(4-fluoro-3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)azetidine-2-carboxamide;
30.
1-acryloyl-N-(2-fluoro-3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1,5-
a][1,3.5]triazin-4-yl)amino)methyl)phenyl)azetidine-2-carboxamide;
31.
1-acryloyl-N-(3-fluoro-5-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)azetidine-2-carboxamide;
32.
1-acryloyl-N-(4-fluoro-3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide;
33.
1-acryloyl-N-(3-fluoro-5-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide;
34.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)-4-methylphenyl)piperidine-2-carboxami-
de; 35.
1-acryloyl-N-(2-fluoro-3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxamide;
36.
1-acryloyl-N-(3-fluoro-5-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxamide;
37.
1-acryloyl-N-(4-fluoro-3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxamide;
38.
(1,4-cis)-4-acrylamido-N-(2-fluoro-3-(((8-isopropyl-2-((tetrahydro-2H--
pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexa-
ne-1- carboxamide; 39.
(1,4-cis)-4-acrylamido-N-(4-fluoro-3-(((8-isopropyl-2-((tetrahydro-2H--
pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexa-
ne-1- carboxamide; 40.
4-acryloyl-N-(5-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)ammo)methyl)pyridin-3-yl)morpholine-3-carboxamide;
.sup. 40A.
3-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyraz-
olo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-methylbenzamide; .sup.
40B.
5-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyraz-
olo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-2-methylbenzamide; .sup.
40C.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazol-
o[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-6-methylpiperidine-2-carboxami-
de; .sup. 40D.
1-acryloyl-N-(3-(((8-isopropyl-2-(((S)-tetrahydro-2H-pyran-3-yl)amino)pyr-
azolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxamide;
.sup. 40E.
1-acryloyl-N-(3-(((8-isopropyl-2-(((R)-tetrahydro-2H-pyran-3-yl)amino)pyr-
azolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxamide;
41.
(S)-N-(1-acryloylpiperidin-3-yl)-3-(((8-isopropyl-2-((tetrahydro-2H-py-
ran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)benzamide;
42.
1-acryloyl-N-(4-fluoro-3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)azetidine-
-2- carboxamide; 43.
1-acryloyl-N-(2-fluoro-5-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide;
44.
1-acryloyl-N-(2-fluoro-5-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxamide;
45. 1-acryloyl-N-(3-(1-((8-isopropyl-2-((tetrahydro-2H-pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)ethyl)phenyl)piperidine-
-3- carboxamide; 46.
3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl
4-acryloylpiperazine-1-carboxylate; 47.
3-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)py-
razolo[1,5- a][1,3,5]triazin-4-yl)amino)methyl)phenyl)benzamide;
48.
2-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)py-
razolo[1,5- a][1,3,5]triazin-4-yl)amino)methyl)phenyl)benzamide;
.sup. 48A.
3-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyraz-
olo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-2-methylbenzamide; 49.
(E)-4-((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1-
,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-N,N-dimethylbut-2-enamid-
e; 50.
(E)-4-(diethylamino)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 51.
(E)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-morpholinobut-2-enamide;
52.
(E)-4-(4-ethylpiperazin-1-yl)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-py-
ran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 53.
(E)-4-(dimethylamino)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide: 54.
(E)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-(pyrrolidin-1-yl)but-2-enami-
de; 55.
(E)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-(piperidin-1-yl)but-2-enamid-
e; 56.
(E)-4-(3-fluoropyrrolidin-1-yl)-N-(3-(((8-isopropyl-2-((tetrahydro-2H--
pyran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 57.
(E)-4-(3,3-difluoropyrrolidin-1-yl)-N-(3-(((8-isopropyl-2-((tetrahydro-
-2H-pyran-
4-yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-
enamide; 58.
(E)-4-(3-fluoropiperidin-1-yl)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-p-
yran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 59.
(E)-4-(dimethylamino)-N-(3-fluoro-5-(((8-isopropyl-2-((tetrahydro-2H-p-
yran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 60.
(E)-4-(dimethylamino)-N-(2-fluoro-3-(((8-isopropyl-2-((tetrahydro-2H-p-
yran-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-ena-
mide; 61.
(E)-N-(3-fluoro-5-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)py-
razolo[1,5-
a][1,3,5](triazin-4-yl)amino)methyl)phenyl)-4-morpholinobut-2-enamide;
62.
(E)-4-(dimethylamino)-N-(2-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl-
)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-enamide; 63.
(E)-N-(4-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-4-(pyrrolidin-1-yl)but-2-enami-
de; 64.
(E)-1-(4-(dimethylamino)but-2-enoyl)-N-(2-fluoro-5-(((8-isopropyl-2-
((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide; 65.
(E)-1-(but-2-enoyl)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)azetidine-2-carboxamide;
66.
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-1-propioloylazetidine-2-carbox-
amide; 67.
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-1-methacryloylazetidine-2-
carboxamide; 68.
(E)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-1-(4-methoxybut-2-enoyl)pyrrol-
idine-3- carboxamide; 69.
(E)-1-(4-(dimethylamino)but-2-enoyl)-N-(3-(((8-isopropyl-2-((tetrahydr-
o-2H- pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide; 70.
(E)-1-(4-(dimethylamino)but-2-enoyl)-N-(4-fluoro-3-(((8-isopropyl-2-
((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide; 71.
(E)-1-(4-(dimethylamino)but-2-enoyl)-N-(3-fluoro-5-(((8-isopropyl-2-
((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide; 72.
(E)-1-(4-(dimethylamino)but-2-enoyl)-N-(3-(1-((8-isopropyl-2-((tetrahy-
dro-2H- pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)ethyl)phenyl)piperidine-3-carboxamide; 73.
(E)-4-(4-(dimethylamino)but-2-enoyl)-N-(3-(((8-isopropyl-2-((tetrahydr-
o-2H- pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)methyl)phenyl)morpholine-3-carboxamide; 74.
(E)-1-(4-(dimethylamino)but-2-enoyl)-N-(3-(((8-isopropyl-2-((tetrahydr-
o)-2H- pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)methyl)phenyl)piperidine-2-carboxamide; 75.
3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl(E)-4-(4-(dimethylamino)but-
2-enoyl)piperazine-1-carboxylate; 76.
(E)-1-(4-(dimethylamino)-4-oxobut-2-en-1-yl)-N-(3-(((8-isopropyl-2-((t-
etrahydro- 2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)methyl)phenyl)azetidine-2-carboxamide; 77.
(E)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-1-(4-(pyrrolidin-1-yl)but-2-
enoyl)piperidine-2-carboxamide; 78.
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-1-(vinylsulfonyl)piperidine-2-
carboxamide; 79.
N-(2-((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-2-oxoethyl)acrylamide;
80.
N-(1-((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-1-oxopropan-2-yl)acrylam-
ide; 81.
N-(2-((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-2-oxoethyl)-N-methyl
acrylamide; 82.
N-(1-((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-2-methyl-1-oxopropun-2-
yl)acrylamide; 83.
1-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)py-
razolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclopropane-1-carboxamide;
84.
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-1-(N-methylacrylamido)cyclopro-
pane- 1-carboxamide; 85.
4-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)py-
razolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-1-methyl-1H-pyrazole-3-
carboxamide; 86.
1-acryloyl-N-(3-((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyraz-
olo[1,5-
a][1,3,5]triazin-4-yl)amino)phenyl)azetidine-2-carboxamide; 87.
(1,4-cis)-4-acrylamido-N-(3-((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl-
)amino)pyrazolo[1,5
-a][1,3,5]triazin-4-yl)amino)phenyl)cyclohexane-1-carboxamide; 88.
1-acryloyl-N-(3-(((8-ethyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-
-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-4-carboxamide;
89.
1-acryloyl-N-(3-(((2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,-
5]triazin-4- yl)amino)methyl)phenyl)piperidine-3-carboxamide; 90.
(1,4-cis)-4-acrylamido-N-(3-(((8-ethyl-2-((1-methylpiperidin-4-yl)oxy)-
pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexane-1-carboxamide;
91.
(1,4-trans)-4-acrylamido-N-(3-(((8-isoproprayl-2-((1-methylpiperidin-4-
-
yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexane-
- 1-carboxamide; 92. Isomer-1:
4-acrylamido-N-(3-(((2-((3-fluoro-1-methylpiperidin-4-yl)amino)-8-
isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexa-
ne-1- carboxamide; 93. Isomer-2:
4-acrylamido-N-(3-(((2-((3-fluoro-1-methylpiperidin-4-yl)amino)-8-
isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexa-
ne-1- carboxamide; 94.
1-acryloyl-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo-
[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)azetidine-2-carboxamide;
95.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)oxy)pyrazo-
lo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide;
96.
1-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)oxy)pyrazo-
lo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-3-carboxamide;
97.
(1,4-cis)-4-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-y-
l)oxy)pyrazolo]1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexane-1-carboxamide;
98. Isomer-1:
4-acrylamido-N-(3-(((2-((3-fluorotetrahydro-2H-pyran-4-yl)oxy)-8-
isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexa-
ne-1- carboxamide; 99. Isomer-2:
4-acrylamido-N-(3-(((2-((3-fluorotetrahydro-2H-pyran-4-yl)oxy)-8-
isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexa-
ne-1- carboxamide; 100.
(1,4-cis)-4-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydrofuran-3-yl)-
oxy)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexane-1-carboxamide;
101.
(1,4-cis)-4-acrylamido-N-(3-(((8-isopropyl-2-((1-methylpyrrolidin-3--
yl)oxy)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexane-1-carboxamide;
102.
1-acryloyl-N-(3-(((2-isobutoxy-8-isopropylpyrazolo[1,5-a][1,3,5]tria-
zin-4- yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide; 103.
1-acryloyl-N-(3-(((8-isopropyl-2-(2-methoxyethoxy)pyrazolo[1,5-a][1,-
3,5]triazin- 4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide;
104.
1-acryloyl-N-(3-(((8-isopropyl-2-(2-(2-methoxyethoxy)ethoxy)pyrazolo-
[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide;
105.
(1,4-cis)-4-acrylamido-N-(3-(((8-cyclopropyl-2-((1-methylpiperidin-4-
-yl)oxy)pyrazolo[1,5-
a][1,3,5|triazin-4-yl)amino)methyl)phenyl)cyclohexane-1-carboxamide;
106.
(1,4-cis)-4-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-
-yl)amino)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexane-1-carboxamide;
107.
1-acryloyl-N-(3-(((2-(4-aminobutoxy)-8-isopropylpyrazolo[1,5-a][1,3,-
5]triazin-4- yl)amino)methyl)phenyl)piperidine-4-carboxamide; 108.
1-acryloyl-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazo-
lo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide;
109. Isomer-1:
1-acryloyl-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide;
110. Isomer-2:
1-acryloyl-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,-
5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide;
111.
1-acryloyl-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazo-
lo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-4-carboxamide;
112.
4-acryloyl-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazo-
lo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)morpholine-2-carboxamide;
113.
4-acryloyl-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazo-
lo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)morpholine-3-carboxamide;
114.
3-acrylamido-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexane-1-carboxamide;
115.
(1,4-cis)-4-acrylamido-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-y-
l)oxy)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexane-1-carboxamide;
116.
1-acryloyl-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazo-
lo[1,5-
a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-3-carboxamide;
117.
1-acryloyl-N-(2-fluoro-5-(((8-isopropyl-2-((1-methylpiperidin-4-
yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine--
3- carboxamide; 118.
(1,4-cis)-4-acrylamido-N-(2-fluoro-5-(((8-isopropyl-2-((1-methylpipe-
ridin-4-
yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexane-
-1- carboxamide; 119.
1-acryloyl-N-(3-(1-((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyra-
zolo[1,5-
a][1,3,5]triazin-4-yl)amino)ethyl)phenyl)piperidine-3-carboxamide;
120.
2-(1-acryloylpiperidine-3-carboxamido)-5-(((8-isopropyl-2-((1-methyl-
piperidin-4-
yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)pyridine
1-oxide; 121.
3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,-
5]triazin-4- yl)amino)methyl)phenyl
4-acryloylpiperazine-1-carboxylate; 122.
(E)-1-(4-(dimethylamino)but-2-enoyl)-N-(3-(((8-isopropyl-2-((1-methy-
lpiperidin-
4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidin-
e-3- carboxamide; 123.
(E)-1-(4-(dimethylamino)but-2-enoyl)-N-(2-fluoro-5-(((8-isopropyl-2--
((1- methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)methyl)phenyl)piperidine-3-carboxamide; 124.
(1,4-cis)-4-((E)-4-(dimethylamino)but-2-enamido)-N-(2-fluoro-5-(((8--
isopropyl-
2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)methyl)phenyl)cyclohexane-1-carboxamide; 125.
4-acrylamido-N-(3-((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyraz-
olo[1,5- a][1,3,5]triazin-4-yl)amino)phenyl)benzamide; 126.
(1,4-cis)-4-acrylamido-N-(3-((8-isopropyl-2-((1-methylpiperidin-4-yl-
)oxy)pyrazolo[1,5-
a][1,3,5]triazin-4-yl)amino)phenyl)cyclohexane-1-carboxamide; 127.
1-acryloyl-N-(3-((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazol-
o[1,5- a][1,3,5]triazin-4-yl)amino)phenyl)piperidine-4-carboxamide;
128.
N-(1-acryloylpiperidin-4-yl)-3-((8-isopropyl-2-((1-methylpiperidin-4-
- yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)benzamide; 129.
N-(1-acryloylpiperidin-3-yl)-3-((8-isopropyl-2-((1-methylpiperidin-4-
- yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)benzamide; 130.
(E)-4-(4-(dimethylamino)but-2-enamido)-N-(3-((8-isopropyl-2-((1-meth-
ylpiperidin-
4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)phenyl)benzamide;
131.
(1,4-cis)-4-((E)-4-(dimethylamino)but-2-enamido)-N-(3-((8-isopropyl--
2-((1- methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)phenyl)cyclohexane-1-carboxamide; 132.
(1,4-Trans)-4-((E)-4-(dimethylamino)but-2-enamido)-N-(3-((8-isopropy-
l-2-((1- methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)phenyl)cyclohexane-1-carboxamide; 133.
(E)-N-(1-(4-(dimethylamino)but-2-enoyl)piperidin-3-yl)-3-((8-isoprop-
yl-2-((1-
methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)benzami-
de; 134.
(E)-N-(1-(4-(dimethylamino)but-2-enoyl)piperidin-4-yl)-3-((8-isoprop-
yl-2-((1-
methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)benzami-
de;
135.
(1,4-cis)-4-(((E)-4-(dimethylamino)-4-oxobut-2-en-1-yl)amino)-N-(3-(-
((8-
isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-
yl)amino)methyl)phenyl)cyclohexane-1-carboxamide; 136.
1-acryloyl-N-(3-(((8-isopropyl-2-methoxypyrazolo[1,5-a][1,3,5]triazi-
n-4- yl)amino)methyl)phenyl)piperidine-4-carboxamide; 137.
(E)-4-(dimethylamino)-1-(4-(2-(((8-isopropyl-2-((tetrahydro-2H-pyran-
-4-
yl)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperazin-
- 1-yl)but-2-en-1-one; 138.
(E)-1-(4-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[-
1,5-
a][1,3,5]triazin-4-yl)amino)methyl)piperidin-1-yl)-4-(pyrrolidin-1-yl)but-
- 2-en-1-one; and 139.
1-acryloyl-N-(3-(((3-isopropyl-5-((tetrahydro-2H-pyran-4-yl)amino)py-
razolo[1,5-
a]pyrimidin-7-yl)amino)methyl)phenyl)azetidine-2-carboxamide,
or a pharmaceutically acceptable salt thereof or a stereoisomer
thereof.
[0111] In certain embodiments, the present invention provides a
pharmaceutical composition comprising a compound of formula (I),
(IA), (IB), (IC), (ID), (IE) and (IF) or a pharmaceutically
acceptable salt thereof or a stereoisomer thereof as described
herein and at least one pharmaceutically acceptable excipient (such
as a pharmaceutically acceptable carrier or diluent). Preferably,
the pharmaceutical composition comprises a therapeutically
effective amount of at least one compound described herein. The
compounds described in the present patent application may be
associated with a pharmaceutically acceptable excipient (such as a
carrier or a diluent) or he diluted by a carrier or enclosed within
a carrier which can be in the form of a capsule, sachet, paper or
other container.
[0112] In yet another embodiment, the compounds of the present
invention are thought to he kinase inhibitors. In certain
embodiments, the compounds of the present invention are CDK
inhibitors. In certain embodiments, the compounds of the present
invention are CDK7 inhibitors. In certain embodiments, the
compounds of the present invention arc selective CDK inhibitors
(e.g. , being more active in inhibiting a CDK than a non-CDK
kinase). In certain embodiments, the compounds of the present
invention are selective CDK7 inhibitors (e.g., being more active in
inhibiting CDK7 than a non-CDK7 kinase). In certain embodiments,
the compounds of the present invention are selective CDK9
inhibitors. In certain embodiments, the compounds of the present
invention are selective CDK 12 inhibitors. In certain embodiments,
the compounds of the present invention are selective CDK 13
inhibitors. In certain embodiments, the compounds of the present
invention are selective CDK18 inhibitros.
[0113] In another embodiment, the present invention provides
pharmaceutical composition for use in treating and/or preventing a
disease and/or disorder associated with selective transcritptional
CDKs. In another embodiment, the present invention provides
pharmaceutical composition for use in treating a subject suffering
from a disease or condition associated with aberrant activity of
selective transcriptional CDKs.
[0114] In another embodiment, the present invention provides
pharmaceutical composition comprising the compound of formula (I),
(IA), (IB), (IC), (ID), (IE) and (IF) for use in treating and/or
preventing a disease and/or disorder associated with s selective
transcriptional CDK inhibitors, in particularly selective
transcriptional inhibitor is CDK7, CDK9, CDK12, CDK13 or CDK18:
more particularly CDK7.
[0115] In another embodiment, the present invention provides
pharmaceutical composition comprising the compound of formula (I),
(IA), (IB), (IC), (ID), (IE) and (IF) for use in treating a subject
suffering from a disease or condition associated with aberrant
activity of selective transcriptional CDK inhibitors, in
particularly selective transcriptional inhibitor is CDK7, CDK9,
CDK12, CDK13 or C18; more particularly CDK7.
[0116] In yet another embodiment, the present invention provides a
method of treating disorders or diseases or condition mediated by
CDK in a subject comprising administering a therapeutically
effective amount of a compound of the present invention, in
particularly CDK is CDK7, CDK.sub.9, CDK 12, CDK13 or CDK18 , more
particularly selective trasnscriptional CDK7.
[0117] In yet another embodiment, the present invention provides a
method of inhibiting selective transcriptional CDK inhibitors, in
particularly selective transcriptional inhibitor is CDK7, CDK9, CDK
12, CDK 13 or CDK18: more particularly CDK7, in a subject in need
thereof by administering to the subject one or more compounds
described herein in the amount effective to cause inhibition of
such receptor.
[0118] In another aspect of the present invention relates to
methods of inhibiting the activity of a kinase in a biological
sample or subject. In certain embodiments, the kinase is selective
transcriptional CDK. certain embodiments, the kinase is selective
transcriptional CDK7. In other embodiments, the kinase is selective
transcriptional CDK9, CDK12, CDK 13 or CDK 18.
[0119] In other embodiments, the activity of the kinase is aberrant
activity of the kinase. In other embodiments, the inhibition of the
activity of the kinase is irreversible. In other embodiments, the
inhibition of the activity of the kinase is reversible. In other
embodiments, the methods of inhibiting the activity of the kinase
include attaching a compound of formula (I) to the kinase. In yet
another embodiment, the compounds of present invention are
selective transcrilpional CDK inhibitors, which are key regulators
of the cell cycle.
[0120] In yet another embodiment, the compounds of present
invention are selective inhibitors of transcriptional CDKs, which
are key regulators of the cell cycle, inhibits both cell cycle
progression and transcription.
[0121] In yet another embodiment, the compounds of present
invention are selective transcriptional CDK inhibitors, which are
key regulators of the cell cycle, inhibits both cell cycle
progression and transcription: wherein the selective
transcriptional CDK inhibitor is CDK7, CDK9, CDK 12, CDK13 or CDK
18.
[0122] In yet another embodiment, the compounds of present
invention are selective transcriptional CDK inhibitors, which are
key regulators of ale cell cycle, inhibits both cell cycle
progression and transcription; wherein ale selective
transcriptional CDK inhibitor is CDK7.
[0123] In yet another embodiment, the compounds of present
invention also inhibits the phosphorylation of ser5 of RNA
Polymerase II CID, consistent with mechanism-based inhibition of
CDK7.
[0124] In yet another embodiment, the compounds of present
invention also inhibits the phosphorylation of ser2 and/or ser7 of
RNA Polymerase IICTD, consistent with mechanism-based inhibition of
transcriptional CDKs.
[0125] In yet another embodiment, the compounds of present
invention (selective translational CDK inhibitors) when
administered in vivo apoptotic response indicated by PARP
cleavage.
[0126] In yet another embodiment, the compounds of present
invention (selective translational CDK inhibitors) after invivo
administration may show pro-apoptotic effects the down-regulation
of short half-life survival proteins like Mcl-I.
[0127] The compounds of the invention arc typically administered in
the form of a pharmaceutical composition. Such compositions can be
prepared using procedures well known in the pharmaceutical art and
comprise at least one compound of the present invention. The
pharmaceutical composition of the present invention comprises one
or more compounds described herein and one or more pharmaceutically
acceptable excipients. Typically, the pharmaceutically acceptable
excipients are approved by regulatory authorities or are generally
regarded as safe for human or animal use. The pharmaceutically
acceptable excipients include, hut are not limited to, carriers,
diluents, glidants and lubricants, preservatives, buffering agents,
chelating agents, polymers, gelling agents, viscosifying agents,
solvents and the like.
[0128] The pharmaceutical composition can be administered by oral,
parenteral or inhalation routes. Examples of the parenteral
administration include administration by injection, percutaneous,
transmucosal, transnasal and transpulmonary administrations.
[0129] Examples of suitable carriers include, hut are not limited
to, water, salt solutions, alcohols, polyethylene glycols, peanut
oil, olive oil, gelatin, lactose, terra alba, sucrose, dextrin,
magnesium carbonate, sugar, amylose, magnesium stearate, talc,
gelatin, agar, pectin, acacia, slearic acid. lower alkyl ethers of
cellulose, silicic acid, fatly acids, fatty acid amines, fatly acid
monoglycerides and diglycerides, fatty acid esters and
polyoxyethylene.
[0130] The pharmaceutical composition may also include one or more
pharmaceutically acceptable auxiliary agents, wetting agents,
suspending agents, preserving agents, buffers, sweetening agents,
flavouring agents, colorants or any combination of the
foregoing.
[0131] The pharmaceutical compositions may be in conventional
forms, for example, tablets, capsules, solutions, suspensions,
injectables or products for topical application. Further, the
pharmaceutical composition of the present invention may be
formulated so as to provide desired release profile.
[0132] Administration of the compounds of the invention, in pure
form or in an appropriate pharmaceutical composition, can be
carried out using any of the accepted routes of administration of
pharmaceutical compositions. The route of administration may be any
route which effectively transports the active compound of the
patent application to the appropriate or desired site of action.
Suitable routes of administration include, but are not limited to,
oral, nasal, buccal, dermal, intradermal, transdermal, parenteral,
rectal, subcutaneous, intravenous, intraurethral, intramuscular or
topical.
[0133] Solid oral formulations include, hut are not limited to,
tablets, capsules (soft or hard gelatin), dragees (containing the
active ingredient in powder or pellet form), troches and
lozenges.
[0134] Liquid formulations include, but are not limited to, syrups,
emulsions and sterile injectable liquids, such as suspensions or
solutions.
[0135] Topical dosage forms of the compounds include ointments,
pastes, creams, lotions, powders, solutions, eye or ear drops,
impregnated dressings and may contain appropriate conventional
additives such as preservatives, solvents to assist drug
penetration.
[0136] The pharmaceutical compositions of the present patent
application may be prepared by conventional techniques known in
literature.
[0137] Suitable doses of the compounds for use in treating the
diseases or disorders described herein can be determined by those
skilled in the relevant art. Therapeutic doses are generally
identified through a dose ranging study in humans based on
preliminary evidence derived from the animal studies. Doses must be
sufficient to result in a desired therapeutic benefit without
causing unwanted side effects. Mode of administration, dosage forms
and suitable pharmaceutical excipients can also he well used and
adjusted by those skilled in the art. All changes and modifications
are envisioned within the scope of the present patent
application.
[0138] In one embodiment the compounds as disclosed in the present
invention are formulated for pharmaceutical administration.
[0139] Yet another embodiment of the present invention provides use
of the compounds as disclosed in the present invention in the
treatment and prevention of diseases or disorder associated with
selective transcriptional CDK inhibitors, in particularly selective
transcriptional CDK inhibitor is CDK7, CDKY, CDK 12, CDK13 or
CDK18: more particularly CDK7. Yet another embodiment of the
present invention provides use of the compound or a
pharmaceutically acceptable salt thereof, in treating and/or
preventing a disease for which the symptoms thereof arc treated,
improved, diminished and/or prevented by inhibition of selective
transcriptional CDK inhibitors, in particularly selective
transcriptional inhibitor is CDK7, CDKY, CDK12, CDK13 or CDK18:
more particularly CDK7.
[0140] According to yet another embodiment, the selective
transcriptional CDK's mediated disorder or disease or condition is
proliferative diseases or disorder or condition.
[0141] According to aforesaid embodiment, the proliferative
diseases or disorders or conditions are selected from hut are not
limited to the group consisting of a cancer, an inflammatory
disorder, an auto-inflammatory disorder or an infectious
disease.
[0142] In other embodiments, the proliferative disease to be
treated or prevented using the compounds of formula (I) will
typically be associated with aberrant activity of CDKs, more
particularly with CDK7, CDK9, CDK 12, CDK13 or 18. Aberrant
activity of CDK7, CDK9, CDK12, CDK13 or CDK18 may be an elevated
and/or an inappropriate (e.g., abnormal) activity of CDK7, CDK9,
CDK12, CDK13 or CDK18. In certain embodiments, CDK7, CDK9, CDK12,
CDK13 or CDK18 are not overexpressed, and the activity of CDK7,
CDK9, CDK12, CDK13 or CDK18 arc elevated and/or inappropriate. In
certain other embodiments, CDK7, CDK9, CDK12, CDK13 or CDK18 arc
overexpressed, and the activity of CDK7, CDK9, CDK12, CDK13 or
CDK18 are elevated and/or inappropriate. The compounds of formula
(I), and pharmaceutically acceptable salts or stereoisomers, and
compositions thereof, may inhibit the activity of CDK7, CDK9,
CDK12, CDK13 or CDK18 are been useful in treating and/or preventing
proliferative diseases.
[0143] According to yet another embodiment, the compounds of the
present invention are expected to be useful in the therapy of
proliferative diseases such as viral diseases, fungal diseases,
neurological/neurodegenerative disorders, autoimmune, inflammation,
arthritis, anti-proliferative (e.g., ocular retinopathy), neuronal,
alopecia and cardiovascular diseases.
[0144] According to yet another embodiment, the compounds of the
present invention are useful in the treatment of a variety of
cancers, including hut not limited to carcinoma, including that of
the breast, liver, lung, colon, kidney, bladder, including small
cell lung cancer, non-small cell lung cancer, head and neck,
thyroid, esophagus, stomach, pancreas, ovary, gall bladder, cervix,
prostate and skin, including squamous cell carcinoma; hemampoietic
tumors of lymphoid lineage, including leukemia, acute lymphoblastic
leukemia, acute lymphocytic leukemia,
[0145] I lodgkins lymphoma, non-Hodgkins lymphoma, B-cell lymphoma,
T- cell lymphoma, hairy cell lymphoma, myeloma, mantle cell
lymphoma and Burkell'S lymphoma; hematopoietic tumors of myeloid
lineage, including acute and chronic myelogenous leukemias,
myelodysplastic syndrome and promyelocytic leukemia; tumors of
masenchymal origin, including fibrosarcoma and rhabdomyosarcoma;
tumors of the central and peripheral nervous system, including
astrocytoma, neuroblastoma, glioma and schwannomas; and other
tumors, including seminoma, melanoma, osteosarcoma,
teratocarcinoma, keratoctanthoma, xenoderoma pigmentosum, thyroid
follicular cancer and Kaposi's sarcoma.
[0146] According to vet another embodiment, the subject is a mammal
including human.
[0147] According to vet another embodiment the present invention
provides compounds fir use as a medicament.
[0148] According to yet another embodiment the invention provides
the use of the compounds of the present invention in the
manufacture of a medicament.
[0149] According to yet another embodiment the invention provides
the use of the compounds of the present invention in the
manufacture of a medicament for the treatment of diseases and/or
disorder associated with selective transcriptional CDK
inhibition.
[0150] According to yet another embodiment the present invention
provides compounds for use as a medicament for the treatment of
diseases and/or disorder associated with selective transcriptional
CDK inhibition.
[0151] According to yet another embodiment the present invention
comprises an additional step of administering to the subject in
need thereof one or more additional chemotherapeutic agents
independently selected from anti-proliferative agents, anti-cancer
agents, immunosuppressant agents and pain-relieving agents.
[0152] The method(s) of treatment of the present invention
comprises administering a safe and effective amount of a compound
according to formula (I) or a pharmaceutically acceptable salt
thereof to a patient (particularly a human) in need thereof.
[0153] Compounds of the invention arc indicated both in the
therapeutic and/or prophylactic treatment of the above-mentioned
conditions. For the above-mentioned therapeutic uses the dosage
administered will, of course, vary with the compound employed, the
mode of administration, the treatment desired and the disorder or
disease indicated.
[0154] The compounds of the present invention may be used as single
drug or as a pharmaceutical composition in which the compound is
mixed with various pharmacologically acceptable materials.
[0155] According to one embodiment, the compounds of the present
invention can also contain unnatural proportions of atomic isotopes
at one or more of the atoms that constitute such compounds. For
example, the present invention also embraces isotopically-labeled
variants of the present invention which are identical to those
recited herein, but for the fact that one or more atoms of the
compound are replaced by an atom having the atomic mass or mass
number different from the predominant atomic mass or mass number
usually found in nature for the ;nom. All isotopes of any
particular atom or element as specified are contemplated within the
scope of the compounds of the invention and their uses. Exemplary
isotopes that can be incorporated in to compounds of the invention
include isotopes of hydrogen, carbon, nitrogen, oxygen,
phosphorous, sulfur, fluorine, chlorine and iodine, such as
.sup.2H("D"), .sup.3H, .sup.11C, .sup.13C, .sup.14C, .sup.13N,
.sup.15N, .sup.15O, .sup.17O, .sup.18O, .sup.32P, .sup.33P,
.sup.35S, .sup.18F, .sup.36Cl, .sup.123I and .sup.125I.
Iisotopically labeled compounds of the present invention can
generally be prepared by following procedures analogous to those
disclosed in the schemes and/or in the examples herein below, by
substituting an isotopically labeled reagent for a non-isotopically
labeled reagent.
General Procedures:
[0156] Compounds of the present invention may be prepared by
synthetic chemical processes, examples of which arc shown herein.
It is meant to be understood that the order of the steps in the
processes may be varied, that reagents, solvents and reaction
conditions may be substituted for those specifically mentioned and
that vulnerable moieties may be protected and deprotected, as
necessary.
[0157] A general approach for the synthesis of compounds of general
formula (I) is depicted in below schemes. As used herein the below
schemes the terms `R.sub.1`, `R.sub.2`. `R.sub.3`,
`R.sub.4`,`R.sub.6`, `A`, `B`, `L.sub.1`, `L.sub.2``m` `n` and `p`
are same as described in compound of formula (I).
[0158] General synthesis for the preparation of key intermediates
(formula 1.5) and (formula 3.4) were described in the given
scheme-a and scheme-b respectively:
##STR00018##
[0159] The general procedure for the preparation of key
intermediate (1.5) was synthesized in two routes by using compound
of formula -1.0 as starting material.
Route-a:
[0160] The compound of formula-1.0 can be treated with formula-1.1
in presence of suitable base such as DIPEA, TEA and the like, in
presence of suitable polar solvents such as ACN, 1,4-dioxane, DMSO,
DCE and the like at a temperature of about 20.degree. C. to
35.degree. C. for about 2 to 24 h to provide the compounds of
formula-1.2.sub.-- Compounds of formula-1.2 can be further treated
with mCPBA in presence of suitable solvents such as DCM,
CHCl.sub.3, DCE and the like at a temperature of about 0.degree.C
to 35.degree.C for about 2 to 24 h to provide the compounds of
formula-1.3. The compounds of formula 1.4 (wherein L.sub.11=NH) can
be synthesized by treating compounds of formula 1.3 with
appropriate amine with or without solvent (solvents such as NMP and
the like) at a temperature of about 100.degree.C to 1.50.degree.C
for about 1 to 24 h. Alternatively, the compounds of formula 1.4
(wherein L.sub.1=O) can be synthesized by treating compounds of
formula 1.3 with appropriate alcohol in the presence of suitable
base such as NaH, LiH, KH, K.sub.2CO.sub.3 or Cs.sub.2CO.sub.3 and
the like in suitable solvents such as THF, DMSO, DMF, 1,4-dioxane
or diethyl ether and the like, at a temperature of about
-30.degree. C. to 100.degree. C. for about 1-24 h. Compounds of
formula-1.4 undergoes reduction of nitro group in presence of
suitable reagents like zinc dust/NH.sub.4Cl or Fe/NH.sub.4Cl or
Zn/aq. NH.sub.4Cl with appropriate combinational ratios of solvents
such as THF:MeOH:Water, THF;EOh: water, methanol:water,
ethanol:water, methanol or ethanol and the like at a temperature of
about 20.degree. C. to 120.degree. C. for about 1-24 h to provide
the compounds of formula 1.5.
Route-b:
[0161] The compound of formula 1.5 can be prepared by using
compound of formula-1.0, on reacting with the compound of formula
2.1 and proceeded till the formation of compound of formula 2.4 by
using the procedure similar to the preparation of compound of
formula 1.4 as depicted in route-a. The resultant compound of
formula 2.4 is further undergoes deprotection of Boc in presence of
suitable reagents such as TFA and the like, in presence of suitable
solvent such as DCM, chloroform, THF or 1,4-dioxane and the like,
at a temperature of about 20.degree. C. to 35.degree. C. for about
2 to 24 h to provide the compounds of formula-1.5.
[0162] The general procedure for the preparation of key
intermediate (3.4) was synthesized according to scheme-b:
##STR00019##
[0163] The compounds of general formula-3.4 can be prepared by
using compound of formula 1.0 and reacting with compound of formula
3.1 and further proceeded to the formation of compound of formula
3.4 according to the procedure depicted in route-b of scheme-a.
General Scheme for the Preparation of Compound of Formula (I):
[0164] The compound according to general formula (I) was prepared
according to the procedure depicted in routc-a and routc-b of
schemc-I and schcmc-II by using compound of formula -1.5 as
starting material.
##STR00020##
Route-a:
[0165] The compound of formula (I) can be synthesized by treating
compound of formula-1.5 with appropriate acid in the presence of
suitable reagents such as HATU, EDC.HCl-HOBt and the like, in the
presence of suitable base such as DIPEA or TEA and the like, in a
suitable solvent such as DMF, THY, DMSO or DCM and the like at a
temperature of about 20.degree. C. to 35.degree. C. for about 1-24
h.
Route-b:
[0166] The compound of formula 3.4 was prepared according to the
procedure depicted in route-a a of scheme-I. The resulting compound
of formula-3.4 undergoes dcprotection of Boc in presence of
suitable reagent TFA and the like, in presence of suitable solvent
such as DCM, chloroform, THF or 1,4-dioxane and the like, at a
temperature of about 20.degree. C. to 35.degree. C. for about 2 to
24 h to provide the compounds of formula-3.5. The compound of
formula-3.5 can be treated with appropriate acid chloride in
presence of suitable base TEA or DIPEA and the like, in presence of
suitable solvent such as DCM, chloroform, THF or 1,4-dioxane and
the like, at a temperature of about 20.degree. C. to 35.degree. C.
for about 2 to 24 h to provide the compounds of formula-I.
[0167] Alternatively, the compound of formula-I can be synthesized
by treating formula-3.5 with appropriate acid in the presence of
suitable reagents such as HATU, EDC.HCl-HOBt and the like, in the
presence of suitable base such as DIPEA or TEA and the like, in a
suitable solvent such as DMF, THF, DMSO or DCM and the like at a
temperature of about 20.degree. C. to 35.degree. C. for about 1-24
h.
[0168] Alternatively, the compound of formula (II) of compound of
formula (I), wherein ring B is absent was prepared according to the
procedure depicted in scheme-Il by using compound of formula -1.5
as starting material.
##STR00021##
[0169] The compounds of formula-If of formula (I) can be prepared
by treating compound of formula-1.5 with respective acid chloride
(prepared from respective haloalkenoic acid in presence of suitable
reagents such as oxalyl chloride in a suitable solvent such as DMF
or THF and the like) in presence of base such as DIPEA or TEA in
presence of suitable solvent such as DCM, chloroform, THF or
1,4-dioxane and the like, at a temperature of about 20.degree. C.
to 35.degree. C. for about 2 to 24 h. The obtained compound can be
treated with various types of amines in presence of base such as
K.sub.2CO.sub.3, Na.sub.2CO.sub.3 and the like in a suitable
solvent such as ACN, THF, DMF, DMSO and the like, at a temperature
of about 20.degree. C. to 100.degree. C. for about 2 to 24 h.
Abbreviations :
[0170] The following abbreviations refer respectively to the
definitions herein: LDA (Lithium diisopropylamide); K.sub.2CO.sub.3
(Potassium carbonate) PdCl.sub.2(dppf).sub.2-DCM
(1,1'-Bis(diphenylphosphino)ferrocencdichloropalladium(II);
dichloromethane complex), DHP (3,4-Dihydro-2H-Pyran) PTSA
(p-Tolucnesulfonic acid); EDCI (1
-Ethyl-3-(3-dimethylaminopropyl)carbodiimide; Dikis
(Bis(triphenylphosphine)palladium(II) dichloride); NH.sub.3
solution (Ammonia solution); Prep Column (Preparative column); Prep
(Preparative Thin layer Chromatography); rt (Retention time): RT
(Room temperature); DMF (Dimethylformamide); h (hour); LC-MS
(Liquid chromatography mass spectroscopy): NaOH (Sodium hydroxide);
Na.sub.2SO.sub.4 (Sodium sulphate); ACN/CH.sub.3CN (Acetonitrile);
HCl (Hydrochloric acid); THF (tetrahydrofuran); DCM
(Dichloromethane); TFA (Trifluoroacetic acid); TLC (Thin layer
chromatography); DIPEA (Diisopropyl Ethyl amine): DMSO-d.sub.6
(Dimethyl sulfoxide-d); HATU
(1-[Bis(dimethylarnino)methylene]-1II-1,
2,3-triazolo[4,5-b]pyridinium 3-oxid-hexafluorophosphate):
Boc.sub.2O (Ditert-butyl dicarhonate): HPLC (High pressure liquid
chromatography); NaIICO.sub.3 (Sodium bicarbonate); Nal I (Sodium
hydride); SEM Chloride (2-(Trimcthylsilyl)ethoxy methylchloride):
Cs.sub.2CO.sub.3 (Cesium carbonate): BINAP
(2,2'-Bis(diphenylphosphino)-1, 1 '-binaphthyl):
Pd.sub.2(dha).sub.3 (Tris(dibenzyli deneacetonc)dipalladium(0));
TEA (triethyl amine), TPP (Triphenyl phosphine), DIAD
(Diisopropylazodicarboxylate), LiBII4 (Lithium borohydride), TMSCl
(Chlorotrimethylsilane).
EXAMPLES
[0171] Although the invention has been illustrated by certain of
the preceding examples, it is not to be construed as being limited
thereby; but rather, the invention encompasses the generic area as
hereinbefore disclosed. Various modifications and embodiments can
be made without departing from the spirit and scope thereof.
The MS data provided in the examples described below were obtained
as follows: Mass spectrum: LC/MS Agilent 6120 Quadrupole LC/MS. The
NMR data provided in the examples described below were obtained as
follows: .sup.1H-NMR: Varian 400 MHz.
[0172] The microwave chemistry was performed on a CliM
lixplorer.
Synthesis of Intermediates:
[0173] The procedure for the compounds of formula (I) are detailed
herein below stepwise including the general synthesis of various
intermediates involved in process of manufacture of the compounds
according to the present invention.
Intermediate-1: Synthesis of
4-chloro-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine
##STR00022##
[0175] This intermediate was prepared from the procedure depicted
in US2008/045536 by using
2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4(31I)-one (prepared
according to US2006/106019) as starting material: LCMS: m/z=200.9
(M.+-.H).
Intermediate-2: Synthesis of
4-chloro-8-ethyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine
Step-1: Synthesis of 2-formylbutanenitrile
##STR00023##
[0177] LDA 2.0M solution in THF (55 mL ) was added to a stirred
solution of butyronitrile (7.5 g, 108.6 mmol) in dry THF (50 mL )
at -78.degree. C. The resulting reaction mixture was stirred at
-78.degree. C. for 15min. Eth,T1formate (8.03 g, 108.6 mmol) was
added at -78.degree. C. and then allowed to stir the reaction
mixture for overnight. After completion of the reaction, the
reaction mixture was quenched with ice water, adjusted to pII 4 by
using 2NIICl and extracted with ethyl acetate (2.times.100 mL). The
combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated to afford the title
compound (5.6 g crude). The obtained product was taken for next
step without purification.
Step-2: Synthesis of 4-ethyl-1H-pyrazol-5-amine
##STR00024##
[0179] Hydrazine hydrate (5.6 mL) was added to a solution of
2-formylbulanenitrile (5.6 g, 57.73 mmol) in ethanol (112 mL ) at
room temperature followed by acetic acid (0.5 mL ). Then the
reaction mixture was heated at 90.degree. C. for 6h. After
completion of the reaction, cooled to room temperature and quenched
with ice-water, adjusted the pH 9 by using K.sub.2CO.sub.3 and the
reaction mixture was extracted with ethyl acetate (2.times.25mI.).
The combined organic phase was washed with brine, dried over
Na.sub.7SO.sub.4, filtered and concentrated to afford the title
compound (5.6 g crude) taken for next step without purification
LCMS: m/z=120.3 (M+H).sup.-.
Step-3: Synthesis of
8-ethyl-2-thioxo-2,3-dihydropyrazolo[1,5-a][1,3,5]triazin-4(1H)-one
##STR00025##
[0181] To a solution of 4-ethyl-1H-pyrazol-5-amine (5.0 g, 44.6
mmol) in DCM (20 mL) was added O-ethyl carbonisothiocyanatidate
(5.9 g, 45.03 mmol) drop wise at 0.degree. C. The reaction mixture
was allowed to stirred at room temperature for 1 h. The solid
obtained was filtered and dried to afford the intermediate of
N-((4-ethyl-1II-pyrazol-5-yl)carbamothioyl)butyramide (3.5 g,
32.40%) LCMS: mlz=242.8 (M+H).sup.-. The intermediate formed was
further dissolved in acetonitrile (100 mL), added K.sub.2CO.sub.3
(6.2 g, 44.9 mmol) and the reaction mixture was heated at
60.degree. C. for 3 h. After completion of the reaction, the
reaction mixture quenched with water and acidified with 2NCl. The
solid obtained was filtered and dried to afford the title compound
(2.6 g, 91.22%): LCMS: nt/z =197.0 (M+H).sup.-.
Step-4: Synthesis of
8-ethyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4(3H)-one
##STR00026##
[0183] 2M NaOH (10 mL, 20 mmol) was added to a stirred solution of
8-ethyl-2-thioxo-2,3-dillydropyrazolo[1,5-a ][1,3,5 ]triazin-4(1
H)-one (2.0 g, 10.19 mmol) in ethanol (40 mL) at 0.degree. C. The
resulting reaction mixture was stirred at 0.degree. C. for 10 min.
Added methyl iodide (1.5 g, 10.50 mmol) at 0.degree. C.; after
completion of the addition, the reaction mixture was maintained at
room temperature for 4 h. After completion of the reaction,
volatiles were removed under reduced pressure and obtained residue
was diluted with ice-cold 2N HCl, the solid precipitated was
filtered and dried under vacuum to afford the title compound (1.7
g, 79.43%). I1NMR (DMSO-d.sub.6): .delta. 12.73 (s, 1II), 7.90 (s,
1II), 2.58-2.51 (q, 2H), 2.50 (s, 3H), 1.23-1.19 (t, 3H). LCMS: mlz
=210.9 (M+H).sup.-.
Step-5: Synthesis of
4-chloro-8-ethyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazinc
##STR00027##
[0185] N,N-Diethyl aniline (3.6 g, 24.1 mmol) was added to a
stirred solution of
8-ethyl-2-(rnethylthio)pyrazolo[1,5-a][1,3,5]triazin-4(3H)-one (1.7
g, 8.08 mmol) in POCl.sub.3 (35 mL) at 0.degree.C. The reaction
mixture was heated at 90.degree.C. for 4 h. After completion of the
reaction, the reaction mixture was concentrated under vacuum and
diluted with ice-cold water. The aqueous layer was extracted with
ethyl acetate (2.times.50 mL). The combined organic phase was
washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified using
combiflash (0-20%EtOAc/Hexane) to afford desired title compound
(1.5 g, 81.52%); LCMS: m/z=228.9 (M+H).sup.+.
[0186] The below intermediates-3 and 4 were prepared according to
the above protocol (Intermediate-2) by using appropriate reactants,
reagents at suitable conditions. The characterization data of the
intermediates are summarized herein.
TABLE-US-00002 Int Analytical No. Structure Data 3 ##STR00028##
LCMS: m/z = 192.0 (M - 51).sup.-. 4 ##STR00029## LCMS: m/z = 243.1
(M + 1).sup.+
Intermediate-5: Synthesis of tert-butyl
(3-(aminomettyl)-4-ethylphenyl)carbamate
Step-1: Synthesis of tert-butyl
(4-bromo-3-cyanophenyl)carbamate
##STR00030##
[0188] Di-tert-butyl dicarbonate (1.14 g, 4.8 mmol) was added to a
solution of 5-amino-2-bromobenzonitrile (0.8 g, 4 mmol) and DMA
(0.58 g, 4.8 mmol) in DCM (20 mL ) and the resulting reaction
mixture was stirred for 8 h at room temperature. After completion
of reaction, the reaction mixture was diluted with ice cold water
and extracted with DCM (3.times.50 mL). The combined organic phase
was washed with brine, dried over sodium sulphate, filtered and
concentrated under reduced pressure. The residue was purified by
100-200 mesh silica column by eluting with 15% ethyl acetate-hexane
to afford the title compound (1 g, 83%). LCMS: mfz=297.15
(M+H).sup..
Step-2: Synthesis of tert-butyl(3-cyano-4-ethylphenyl)carbamate
##STR00031##
[0190] Tert-butyl (4-bromo-3-cvanophenvl)carbarnate (4.0 g, 18.6
mmol), Ethyl boronic acid (1 g, 3.3 mmol), Cs.sub.2CO.sub.3 (3.21
g, 9.9 mmol) were taken in a pressure vessel. A mixture of toluene
(10 mL) and ethanol (1mL) was added. The suspension was degassed
and flushed with nitrogen gas for 15 min. Then Pd(PPh.sup.3).sub.4
(0.38 g, 0.33 mmol) was added, the pressure vessel was sealed and
heated overnight at 110.degree. C. After completion of the
reaction, cooled to room temperature and was quenched with water
and diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.25 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated. The residue was purified by 100-200 silica gel
column chromatography to afford desired title compound (0.28 g,
66%); LCMS: m/z=247.2 (M+H).sup..
Step-3: Synthesis of tert-butyl
(3-(aminomethyl)-4-ethylphenyl)carbarnate
##STR00032##
[0192] Nickel chloride hexahydrate (0.17 g. 1.2 mmol) was added to
a cooled solution of tert-butyl (3-cyano-4-ethylphenyl)carbarnate
(0.5 g, 2 mmol) in methanol at 0.degree. C. and stirred the
reaction mixture for 10 min and added sodium borohydridc (0.53 g,
14 mmol) lot wise at 0.degree. C. The resulting reaction mixtures
was stirred for 1 h at room temperature and cooled to 0.degree. C.
and added diethylenc triamine (0.24 g, 2.4 mmol) and stirred for 1
h at room temperature and concentrated under vacuum. The residue
was diluted with water, precipitated solid was filtered ut and
dried. Purified the crude using cornbiflash t afford the title
compound (0.35 g, 65%) LCMS: m/z=251.1 (M+HI).sup.+.
[0193] The below intermediates-6 and 7 were prepared according to
the above protocol (step-1 & 3 of intermediate-5) by using
appropriate reactants, reagents at suitable conditions. The
characterization data of the intermediates are summarized
herein.
TABLE-US-00003 Int Analytical No. Structure Data 6 ##STR00033##
LCMS: m/z = 241 (M + H).sup.+. 7 ##STR00034## LCMS: m/z = 224 (M +
H) . indicates data missing or illegible when filed
Intermediate-8: Synthesis of 1-acryloylpiperidine-4-carboxylic
acid
##STR00035##
[0195] 2M NaOH (7.8 mL, 15.5 mmol) was added to a solution f
piperidine-4-carboxylic acid (1.0 g, 7.81 mmol) in (20 mL) of
THF:Water (6:4) at 0.degree. C. and then stirred for 10 min. Added
acryloyl chloride (0.7 g, 7.69=101) at 0.degree. C., then the
reaction mixture was allowed to room temperature and stirred for 1
h. After completion of the reaction, the reaction mixture was
quenched with ice water, adjusted pH to 4 by citric acid and
extracted with ethyl acetate (2.times.100 mL ). The combined
organic phase was washed with brine, dried over Na.sub.2SO.sub.4,
filtered and concentrated. The obtained product was triturated with
diethyl ether, solid was filtered and dried to afford the title
compound (0.45 g, 32.14%). .sup.1HNMR (DMSO-d.sub.6): .delta. 12.35
(s, 1H), 6.82-6.73 (m, 1H), 6.08-6.02 (dd, 1 H), 5.615-5.61 (dd, 1
H), 4.24-4.19 (d, 1 H), 3.96-3.91 (d, 1 H), 3.15-3.05 (t, 1H),
2.82-2.74 (t, 1H), 2.53-2.44 (m, 1H), 1.84-1.79 (m, 2H), 1.43-1.35
(in. 2H): LCMS: m/z=184 (M+H).sup.1.
[0196] The below intermediates-9 to 14 were prepared according to
the above protocol (intermediate-8) by using appropriate reactants,
reagents at suitable conditions. The characterization data of the
intermediates arc summarized herein.
TABLE-US-00004 Int Analytical No. Structure Data 9 ##STR00036##
LCMS: m/z = 156.05 (M + H) . 10 ##STR00037## LCMS: m/z = 170.00 (M
+ H) . 11 ##STR00038## LCMS: m/z = 170.00 (M + H).sup.+ 12
##STR00039## LCMS: m/z = 185.21 (M + H).sup.-. 13 ##STR00040##
LCMS: m/z = 198.21 (M + H).sup.+. 14 ##STR00041## LCMS: m/z =
198.21 (M + H).sup.-. indicates data missing or illegible when
filed
Intermediate 15: Synthesis of
N-(3-aminophenyl)-4-nitrohenzamide
##STR00042##
[0198] To a cooled solution of 4-nitro benzoic acid (0.3 g, 1.85
mmol) in DMF (2 mL ) at 0.degree. C. was added HATU (0.84 g, 2.22
mmol) followed by DIPEA (0.47m13.7 mmol) and finally added
1,3-diaminobenzene (0.2. g, 1.85 mmol). The reaction mixture was
stirred for 1 h at room temperature. The reaction mixture was
quenched with ice-water and diluted with ethyl acetate. The aqueous
layer was separated and extracted with ethyl acetate (2.times.25
mL). The combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated the crude residue (0.2
g crude). LCMS; rn/z=258.10 (M+H).sup.1.
Intermediate-16: Synthesis of
tert-butyl(S)-3-(3-(aminomethyl)benzamido)piperidine-1-carboxylate
Step- 1 : Synthesis of
tert-butyl(S)-3-(3-cyanobenzamido)piperidine-1-carboxylate
##STR00043##
[0200] I IATU (1.94 g, 5.1 mmol) and DIPEA (1.25 mL, 6.8 mmol) were
added to a cooled solution of 3-cyanobenzoic acid (0.5 g, 5.98
mmol) in dry DMF (5 mL) at 0.degree. C. Tert-butyl
(S)-3-aminopiperidine-1-carboxylate (0.66 g, 3.4 mmol) was to the
reaction mixture and stirred for 2 h at room temperature. After
completion of reaction, the reaction mixture was diluted with
ice-water and diluted with ethyl acetate. The aqueous layer was
separated and extracted with ethyl acetate (2.times.25(nI,). The
combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated the crude residue ((1.8
g crude). LCMS: m/z=330 (M.times.H).sup.+.
Step-2: Synthesis of tert-butyl
(S)-3-(3-(arninornethyl)benzamido)piperidine-1-carboxylate
##STR00044##
[0202] Nickel(II) chloride hexahydrate (0.29 g, 1.23trunol) was
added to a solution of tert-butyl
(S)-3-(3-cyanobenzamido)piperidine-1-carboxylate (0.8 g, 2.47 mmol)
in MeOH (10 mL) at 0.degree. C, followed by NaBII.sub.4 (0.73 g,
19.32 mmol) was added portion wise. The resulting reaction mixture
was stirred at ambient temperature for lh, then cooled to 0.degree.
C. and added diethylenetriamine (0.25 g, 2.46mtnol) and allowed to
stir at ambient temperature for 1 h. After completion of the
reaction, the reaction mixture was concentrated under vacuum,
diluted with ethyl acetate (30 mL), washed with water and brine
solution. The organic layer was dried over Na.sub.2SO.sub.4,
filtered and concentrated the crude residue (0.5 g, crude). LCMS:
m/z=334 (M+H).sup.+.
[0203] The below Intermediate-17 was prepared according to the
above protocol (Intermediate-16) by using appropriate reactants,
reagents at suitable conditions. The characterization data of the
intermediates are summarized herein.
TABLE-US-00005 Int No. Structure Analytical Data 17 ##STR00045##
LCMS: m/z = 334.2 (M + H) . indicates data missing or illegible
when filed
Intermediate-18: Synthesis of (tert-butyl
3((5-(aminomethyl)-2-fluorophenyl)carbamoyl)
piperidine-1-carboxylate
Step-1: Synthesis of tert-butyl
3((5-cyano-2-fluorophenyl)carbamoyl)piperidine-1-carboxylate
##STR00046##
[0205] To a cooled solution of
1-(tert-butoxycarbonyl)piperidine-3-carboxylic acid (1 g, 4.3 mmol)
in pyridine (5m1,) at 0.degree. C. was added
3-amino-4-fluorolienzonitrile (0.59 g, 4.3 mmol) followed by
POCl.sup.3 (0.68 g, 4.3 mmol), The reaction mi xture was stirred
for 2 h at room temperature. The reaction mixture was quenched with
ice-water and diluted with ethyl acetate. The aqueous layer was
separated and extracted with ethyl acetate (2.times.25 mL ). The
combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated to crude residue (1.1
g) LMCS: rn/z=346. 5(M-H).sup.-.
Step-2: Synthesis of
tert-butyl3-((5-(aminomethyl)-2-fluorophenyl)carbamoyl)piperidine-1-carbo-
xylate
##STR00047##
[0207] Nickel chloride hexahydrate (0.29 g, 1.21 mmol) was added to
a cooled solution of tert-butyl
3-((5-cyano-2-fluorophenyl)carhamoyl)piperidine-1-carboxylate (1.1
g, 3.1 mmol) in methanol at 0.degree. C. and stirred the reaction
mixture for 10 min and added sodium borohvdride (0.84 g, 22.1 mmol)
lot wise at 0.degree. C., stirred the reaction mixture for 1 h at
room temperature, further cooled to 0.degree. C. and added
diethylenetriamine (0.32 g, 3.1 mmol) and stirred for 1 h at room
temperature and concentrated under vacuum. Diluted the reaction
mixture with water, solid separated was filtered and dried. The
obtained crude was purified by using combiflash to afford the title
compound (0.8 g, 72%). LCMS: m/z=352 (M+H).sup..
[0208] The below Intermediates-19 to 20 were prepared according to
the above protocol (Intermediate-18) by using appropriate
reactants, reagents at suitable conditions. The characterization
data of the intermediates are summarized herein.
TABLE-US-00006 Int No. Structure Analytical Data 19 ##STR00048##
LCMS: m/z = 336.15 (M - 1).sup.-. 20 ##STR00049## LCMS: m/z = 335.1
(M + H).sup.-.
Intermediate-21: Synthesis of
tert-butyl3-((3-(1-aminoethyl)phenyl)carbamoyl)piperidine-1-carboxylate
Step-1: Synthesis of tert-butyl
3-((3-acetylphenyl)carbamoyl)piperidine-1-carboxylate
##STR00050##
[0210] The process of this step was adopted from intermediate-15
(1.2 g). LCMS: m/z=347.1 (M+H).sup.-.
Step-2: Synthesis of tert-butyl
(E)-3((3-(1-(hydroxyimino)ethyl)phenyl)carbamoyl)
piperidine-1-carboxylate
##STR00051##
[0212] To a solution of
tert-butyl3((3-acetylphenyl)carbamoy)piperidinc-1-carboxylatc (1.2
g, 3.4rrno1) in EtOH(20 mL) was added 50% Aq. NH.sub.2OH solution
(0.57mg,17.3 mmol) at RT. The reaction mixture was stirred at
80.degree. C. for 2 h. The reaction mixture was concentrated to
afford desired title compound (1.1 g, 88%). LCMS: m/z=362.1
(M+H).sup.-.
Step-3: Synthesis of tert-butyl 3-((3-(1
-aminoethyl)phenyl)carbamoyl)piperidine-1-carboxylate
##STR00052##
[0214] To a solution of tert-butyl
(E)-3-((3-(1-(hydroxyimino)ethyl)phenyl)carbamoyl)
piperidine-1-carboxylatc (1 g,2.7 mmol) in acetic acid (5 mL) was
added zinc (0.905 g, 13.8 mmol). The reaction mixture was stirred
at room temperature for 4 h. After completion of the reaction the
reaction mixture was filtered through celite and diluted with ethyl
acetate. The aqueous layer was separated and extracted with ethyl
acetate (2.times.25 mL). The combined rganic phase was washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to
afford desired title compound (0.9 g, crude). LCMS: m/z=347.7
(M+H).sup.1.
Intermediate-22: Synthesis of tert-butyl
4-(3-aminobenzamido)piperidine-1-carboxylate
Step-1: Synthesis of
tert-butyl4-(3-nitrobenzamido)piperidine-1-carboxylate
##STR00053##
[0216] The process of this step was adopted from intermediate-15
(1.2 g crude).
Step-2: Synthesis of tert-butyl
4-(3-aminobenzamido)piperidine-1-carboxylate
##STR00054##
[0218] NH.sub.4Cl (3.67 g, 68.7 mmol) and Zinc (2.24 g,34.3 mmol)
was sequentially added to a solution of
tert-butyl4-(3-nitrobenzarnido)piperidine-1-carboxylate (1.2 g, 3.4
mmol) in mixture of THF:McOH:water (2:1:1, 40 mL) at 0.degree. C.
and the resulting reaction mixture was stirred at ambient
temperature for 3 h. After completion of the reaction, the reaction
mixture was filtered through cclite pad and washed with ethyl
acetate. Filtrate was concentrated and quenched with ice-water and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.25 mL ). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated the crude residue (0.9 g). LCMS: m/z=264.20
(M-tert-Bu+1).sup.1.
Intermediate-23: Synthesis of 3-fluoro-1 -methylpiperidin-4-ol
Step-1 : Synthesis of tert-butyl
3-fluoro-4-hydroxypiperidine-1-carboxylate
##STR00055##
[0220] To a solution of tert-butyl 3- fluoro-4-oxopiperidine-
1-carboxylate (10 g, 45.87 mmol, WO2015/022662) in MeOH (200 mL)
was added sodiumborohydride (2.5 g, 69.44 mmol) lot wise at
0.degree. C.. Then the reaction mixture was allowed to stir at room
temperature for 1 h. After completion of the reaction, the reaction
mixture was concentrated under vacuum and diluted with ice-cold
saturated ammonium chloride solution. The aqueous layer was
extracted with ethyl acetate (2.times.50 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to afford the title
compound (9.1 g, crude). LCMS; m/z=220.0 (M+H).sup.+.
Step-2: Synthesis of tert-butyl
4-(benzyloxy)-3-fluoropiperidine-1-carboxylate
##STR00056##
[0222] Nall (60%) (2.5 g, 104.16 mmol) was added to a solution of
tert-butyl3-fluoro-4-hydroxypiperidinc-1 -carboxylate (9.0 g, 0.041
mmol) in DMF 180 mL) under inert atmosphere at and stirred for 25
min. Benzylbromide (7.0 g, 0.040 mmol) was added, the resulting
reaction mixture was allowed to RT and stirred for 5 h. After
completion of the reaction, cooled to room temperature and was
quenched with ice-water and diluted with ethyl acetate (25 mL). The
aqueous layer was separated and extracted with ethyl acetate
(2.times.100 mL). The combined organic phase was washed with brine,
dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue
was purified by column chromatography (0-10% EtOAc/hexane) to
afford title compound (10.5 g, 83.33%). LCMS: m/z=310.3
(M+H).sup.1.
Step-3: Synthesis of 4-(bentyloxy)-3-fluoropiperidine
##STR00057##
[0224] To a solution of
tert-butyl4-(benzyloxy)-3-fluoropiperidine-1-carboxylate (10.0 g,
32.32 mmol) in DCM (30 mL) was added TFA (15.0 mL) drop wise at
0.degree. C. Then the reaction mixture was allowed to stirred at
room temperature for 5 h. After completion of the reaction, the
reaction mixture was concentrated under vacuum and treated with
ice-cold NaHCO.sub.3 solution. The aqueous layer was extracted with
ethyl acetate (2.times.50 mL). The combined organic phase was
washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to afford the title compound
(7.5 g, crude). LCMS; m/z=209.7(M.+-.H).sup.1.
Step-4: Synthesis of 4-(benzyloxy)-3-fluoro-1-rnethylpiperidine
##STR00058##
[0226] To a solution of 4-(benzyloxy)-3-fluoropiperidine (8.0 g,
38.27 mmol) in THF (160 mL ) was added TEA (3.9 g, 38.61 mmol),
followed by dimethylsulfate (4.9 g, 38.84rnmol) at 0.degree. C..
Then the reaction mixture was allowed to stirred at room
temperature for 2 h. After completion of the reaction, the reaction
mixture was concentrated under vacuum and the residue was diluted
with water and ethyl acetate, separated the organic phase. The
organic phase was washed with brine, dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to afford the
title compound (2.5 g, crude). LCMS: m/z=225.4(M+H).sup..
Step-5: Synthesis of 3-fluoro-l-methylpiperidin-4-ol
##STR00059##
[0228] Pd/C (10%) (0.9 g) was added to a solution of
4-(benzyloxy)-3-fluoro-1-methylpiperidine (2.5 g, 11.19 mmol) in
ethanol was taken in par shaker (250 mL), applied 50 psi hydrogen
pressure, maintained for 24 h at RT. After completion of the
reaction, the reaction mixture was filtered through celite pad and
the filtrate was concentrated under reduced pressure to afford the
title compound (2.0 g, crude). LCMS: m/z=134.1 (M+H).sup.-.
Intermediate-24: Synthesis of
8-ethyl-2-(methylthio)-N-(3-nitrobenzyl)pyrazolo[1,5-a][3,5]triazin-4-ami-
ne.
##STR00060##
[0230] DIPEA (2.0 g, 15.5 mmol) was added to a stirred solution of
(3-nitrophenyl) methanamine.HCI (1.45 g, 7.7 1 mmol) in
acetonitrile (50 mL) at 0.degree. C.. Then added
4-chloro-8-ethyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine (1.7
g, 7.43 ininol intermediate-2). The resulting reaction mixture was
stirred at ambient temperature for 2 h. After completion of the
reaction, concentrated the reaction mixture to remove volatiles and
the residue was diluted with ice-cold water, the solid obtained was
filtered and dried to afford the title compound (2.4 g, 94.11%).
LCMS: m/z=345.2 (M+H).sup.+.
[0231] The below Intermediates-25 to 28 were prepared according to
the above protocol (intermediate-24) by using appropriate
reactants, reagents at suitable conditions. The characterization
data of the intermediates arc summarized herein.
TABLE-US-00007 Int No. Structure Analytical Data 25 ##STR00061##
LCMS: m/z = 316.9 (M + H) . 26 ##STR00062## LCMS: m/z = 359.05 (M +
H) . 27 ##STR00063## LCMS: m/z = 457.1 (M + H).sup.+. 28
##STR00064## LCMS: m/z = 429 (M + H) . indicates data missing or
illegible when filed
Intermediate-29: Synthesis of
N4-(5-amino-2-ethylbenzyl)-8-isopropyl-N2-(letrallydro-2H-pyran-4-yl)pyra-
zolo[1 ,5-a][1,3,5]triazine-2,4-diamine
Step-1: Synthesis of tert-butyl
(4-ethyl-3-(((8-isopropyl-2-(methylsulfonyl)pyrazolo[1,5-a[]1.3,5]triazin-
-4-yl)amino)methyl)phenyl)carbamate
##STR00065##
[0233] m-CPBA (0.45 g, 0.26 (mmol) was added portion wise to a
solution of tert-butyl
(4-ethyl-3-(((8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4-y-
l)amino)methyl1) phenyl)carbamate (0.4 g, 0.87 mmol,
intermediate-27) in DCM (10 mL). The resulting reaction mixture was
stirred at ambient temperature for 4 h. After completion of the
reaction, the reaction inixture was concentrated under vacuum and
purified the residue in 100-200 mesh silica column by eluting with
40-50% ethyl acetate-hexane to afford the title compound (0.38 g,
83%). LCMS: m/z.=489.1 (M+H).sup..
Step-2: Synthesis of
tert-butyl(4-ethyl-3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)py-
razolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)carbamate
##STR00066##
[0235] A mixture of 4-aminotetrahvdropyran (0.44 g, 4.36 mmol) and
tert-butyl
(4-ethyl-3-(((8-isopropyl-2-(methylsulfonyl)pvrazolo[1,5-a][1,3,5]triazin-
-4-yl)amino)methyl)phenyl) carbamate (0.38 g, 0.72 mmol) were
heated at 100.degree. C. for 2 h. After completion of the reaction,
cooled to room temperature and was quenched with ice-water and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.25 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated. The residue was purified by 100-200 silica gel
column chromatography to afford desired title compound (0.28 g,
66%). LCMS: m/z=510.3 (M+H).sup.1.
Step-3: Synthesis of
N4-(5-amino-2-cthylbenzyl)-8-isopropyl-N2-(tetrahydro-2H-pyran-4-yl)pyraz-
olo[1,5-a][1.3.5]triaz inc-2.4-diamine.
##STR00067##
[0237] TFA (0.5mI,) was added to a solution of tert-butyl
(4-ethyl-3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-
-a][1,3,5]triazin-4-yl)amino)methyl)phenyl) carbamate (0.28 g, 0,48
mmol) in DCM (2.5 mL ) at 0.degree. C. The reaction mixture allowed
to stir at room temperature for 2 h. After completion of the
reaction, the reaction mixture was concentrated under vacuum to
afford desired title compound (0.22 g of TFA salt ) LCMS: m/z=410.2
(M+H).sup..
[0238] The below Intermediates-30 to 32 were prepared according to
the above protocol (Intermediate-29) by using appropriate
reactants, reagents at suitable conditions. The characterization
data of the intermediates arc summarized herein.
TABLE-US-00008 Int No. Structure Analytical Data 30 ##STR00068##
LCMS: m/z = 400 (M + H) . 31 ##STR00069## LCMS: m/z = 383.1 (M + H)
. 32 ##STR00070## LCMS: m/z = 382 (M + H).sup.+. indicates data
missing or illegible when filed
Intermediate-33: Synthesis of
N4-(3-aminobenzyl)-8-isopropyl-N2-(tetrahydro-2H-pyran-4-yl)pyrazolo[1.5--
a][1,3.5]triazine-2,4-diamine
Step-1: Synthesis of
8-isopropyl-2-(methylsulfonyl)-N-(3-nitrobenzyl)pvrazolo[1,5-a][1,3,5]tri-
azin-4-amine
##STR00071##
[0240] mCPBA (1.73 g, 10.05 mmol) was added portion wise to a
solution of
8-isopropyl-2-(methylthio)-N-(3-nitrohenzyl)pyrazolo[1,5-a][1,3,5]triazin-
-4-amine (1.2 g, 3.35 mmol) in DCM (15OrnI). After completion of
the reaction, the reaction mixture was extracted with 2M aq. NaOH
and DCM. The organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated to afford the tide
compound (1 g, 76%). LCMS: m/z=391.1 (M+H).sup.1.
Step-2: Synthesis of
8-isopropyl-N4-(3-nitrobenzyl)-N2-(telrahydro-2H-pyran-4-yl)pyrazolo
[1.5-a][1 .3.5]triazine-2,4-diamine
##STR00072##
[0242] A mixture of 4-aminotetrahydropyran (0.26 g, 2.56 mmol) and
8-isopropyl-2-(methylsulfonyl)-N-(3-nitrobenzyl)pyrazolo[1,5-a][1,3,5]tri-
azin-4-amine (0.2 g, 0.51 mmol) were allowed to heat at 100.degree.
C. for 2-12 h. After completion of the reaction, the reaction
mixture was cooled to room temperature, quenched with ice-water and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.25 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated. The residue was purified by 100-200 silica gel
column chromatography to afford desired title compound (0.2 g,
95%). LCMS: m/z=412.49 (M+H).sup..
Step-3: Synthesis of
N4-(3-aminobenzyl)-8-isopropyl-N2-(tetrahydro-2H-pyran-4-yl)
pyrazolo[1,5-a][1,3,5]triazine-2,4-diamine
##STR00073##
[0244] To a solution of
8-isopropyl-2((1-methylpiperidin-4-yl)oxy)-N-(3-nitrobenzyl)pyrazolo[1,5--
a][1,3,5]triazin-4-amine (0.65 g, 1.52 mmol) in THF: MeOH:Water
(3:2:1) were added zinc (0.5 g. 7.64 mmol) and ammonium chloride
(0.4 g, 7.64 mmol). The reaction mixture was stirred at room
temperature for 4 h. After completion of reaction the reaction
mixture filtered through celitc and diluted with ethyl acetate. The
aqueous layer was separated and extracted with ethyl acetate
(2.times.25 mL ). File combined organic phase was washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated. The
residue was purified by 100-200 silica gel column chromatography to
afford desired title compound (0.5 g, 83%). LCMC m/z=382.4
(M+H).sup.-.
[0245] The below intermediates-34 to 38 were prepared according to
the above protocol (intermediate-33) by using appropriate
reactants, reagents at suitable conditions. The characterization
data of the intermediates are summarized herein.
TABLE-US-00009 Int No. Structure Analytical Data 34 ##STR00074##
LCMS: m/z = 381 (M + H) . 35 ##STR00075## LCMS: m/z = 395.21 (M +
H) . 36 ##STR00076## LCMS: m/z = 396.65 (M + H) . 37 ##STR00077##
LCMS: m/z = 396.35 (M + H) . 38 ##STR00078## LCMS: m/z = 499.2 (M +
H) . indicates data missing or illegible when filed
Intermediate-39: Synthesis of
N4-(3-aminophenyl)-8-isopropyl-N2-(tetrahydro-2H-pyran-4-yl)pyrazolo[1,5--
a][1,3,5]triazine-2,4-diamine
Step-1: Syn thesis of 8-isopropyl-2-(methylthio)-N-(3
-nitrophenyl)pyrazolo[1.5-a][1,3.5]triazin-4-amine
##STR00079##
[0247] The process of this step was adopted from intermediate-24 (2
g, 47%). LCMS: m/z=345.0 (M+H).sup..
Step-2: Synthesis of
8-isopropyl-2-(methylsulfonyl)-N-(3-nitrophenyl)pyrazolo[1,5-a][1,3,5]tri-
azin-4-amine
##STR00080##
[0249] The process of this step was adopted from step-1 of
intermediate-29 (2.0 g, 93%). LCMS: m/z=376.95 (M+H).sup.+.
Step-3: Synthesis of
8-isopropyl-N4-(3-nitrophenyl)-N2-(tetrahydro-2H-pyran-4-yl)pyrazolol[5-a-
][1.3.5]triazine-2,4-diamine
##STR00081##
[0251] The process of this step was adopted from step-2 of
intermediate-29 (I .2 g, 94%). LCMS: m/z=398 (M+H).sup..
Step-4: Synthesis of
N-4-(3-aminophenyl)-8-isopropyl-N2-(tetrallydro-2H-pyran-4-yl)pyrazolo[5--
a][1,3.5]triazine-2,4-diamine
##STR00082##
[0253] The process of this step was adopted from step-3 of
intermediate-33 (0.9 g, 75%). LCMS: m/z=368.5 (M+H).sup.+.
Intermediate-40: Synthesis of
N-(3-aminobenzyl)-8-ethyl-2-((1-methylpiperidin-4-yl)oxy)
pyrazolo[5-a][1,3,5]triazin-4-amine.
Step-1: Synthesis of
8-ethyl-2-((1-rnethylpiperidin-4yl)oxy)-N-(3-nitrobenzyl)pyrazolo[1,5-a][-
1,3,5]triazin-4-amine
##STR00083##
[0255] The process of this step was adopted from step-1 of
intermediate-29 (1.98 g, 72.52%). LCMS: m/z=377.2 (M+H).sup.+.
Step-2: Synthesis of 8-ethyl-2(1
-methylpiperidin-4-yl)oxy)-N-(3-nitrobenzyl)pyrazolo[1,5-a][1,3,5]triazin-
-4-amine
##STR00084##
[0257] NaH (0.25 g, 10.6 mmol) was added to a solution of
1-methylpiperidin-4-ol (1.2 g, 10.4 mmol) in THF (40 mL ) under
inert atmosphere at 0.degree. C. and allowed to stir for 25 min.
Then
8-ethyl-2-(methylsulfonv1)-N-(3-nitrobenzvl)pyrazolo[1,5-a][1,3,5]triazin-
-4-amine (1.0 g, 2.65 mmol) was added at same temperature and the
resulting reaction mixture was allowed to stir at RT for 2 h. After
completion of the reaction, reaction mixture was cooled to room
temperature, quenched with ice-water and diluted with ethyl acetate
(25 mL). The aqueous layer was separated and extracted with ethyl
acetate (2.times.100 mL). The combined organic phase was washed
with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated.
The residue was purified by combiflash (0-10% MeOH/DCM) to afford
title compound (0.6 g, 55.04%). LCMS: m/z=412.9 (M+H).sup.1.
Step-3: Synthesis of
N-(3-aminobenzyl)-8-ethyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo
[1,5-a][1,3,5]triazin-4-amine
##STR00085##
[0259] The process of this step was adopted from step-3 of
intermediate-33 (0.49 g, 81_66%). LCMS: m/z=381.48 (M+H).sup.+.
[0260] The below intermediates-41 to 51 were prepared according to
the above protocol (intermediate-40) by using appropriate
reactants, reagents at suitable conditions.sub.-- The
characterizati(m data of the intermediates are summarized
herein.
TABLE-US-00010 Int No. Structure Analytical Data 41 ##STR00086##
LCMS: m/z = 396.21 (M + H) . 42 ##STR00087## LCMS: m/z = 482.40 (M
+ H).sup.+. 43 ##STR00088## LCMS: m/z = 401.8 (M + H) . 44
##STR00089## LCMS: m/z = 414.1 (M + H) . 45 ##STR00090## LCMS: m/z
= 355.1 (M + H) . 46 ##STR00091## LCMS: m/z = 357.4(M + H) . 47
##STR00092## LCMS: m/z = 399.15(M - H) . 48 ##STR00093## LCMS: m/z
= 369.1 (M + H) . 49 ##STR00094## LCMS: m/z = 383.21 (M + H) . 50
##STR00095## LCMS: m/z = 382.15 (M + H) . 51 ##STR00096## LCMS: m/z
= 470.35 (M + H).sup.+. indicates data missing or illegible when
filed
[0261] The present invention is further exemplified, but not
limited, by the following examples that illustrate the preparation
of compounds according to the invention
Example-1: Synthesis of 1
-acryloyl-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo-
[5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-3-carboxamide
(Compound-I)
##STR00097##
[0263] To a cooled solution of 1-acryloylpiperidine-3-carboxylic
acid (0.086 g, 0.47 mmol, intermediate-12) in DMF (4 mL) at
0.degree. C. was added HATU (0.22 g, 0.59 mmol) and DIPEA (0.2mL
1.18mrnol) and finally added
N4-(3-aminobenzyl)-8-isopropyl-N2-(tetrahydro-2H-pyran-4-yl)pyrazol-
o[1,5-a][1,3,5]triazine-2,4-diamine (0.15 g. 0.39 mmol,
intermediate-33). The reaction mixture was stirred for 2 h at room
temperature. After completion of the reaction, the reaction mixture
was quenched with ice-water and diluted with ethyl acetate. The
aqueous layer was separated and extracted with ethyl acetate
(2.times.25 mL). The combined organic phase was washed with brine,
dried over Na.sub.2SO.sub.4, filtered and concentrated to crude
residue. The residue was purified by combiflash to afford the title
compound (0.05 g, 25%). .sup.1HNMR (DMSO-d.sub.6, 400 MHz): .delta.
9.99-9.96 (d, 1H), 8.95-8.70 (m, 1H), 7.72 (s, 1H), 7.51 (s, 2H),
7.25-7.21 (t, 1H), 7.03-7.01 (d, 1 H), 6.90-6.79 (m, 2H), 6.H-6.07
(d, 1H), 5.68-5.63 (t, 1H) , 4.59-4.46 (m, 2H), 4.32-4,28 (d, 1 H),
4.10-4.00 (m, 2H), 3.82 (s, 3H), 3.18-3.16 (d, 1H), 3.07-3.00 (t,
1H), 2.91 (s, 1 H), 2.78-2.67 (m, 1 H), 2.46 (m, 1H), 1.95-1.92
(rn, 1 H), 1.84 (s, 1 H), 1.73-1.63 (m, 3H), 1.46-1.35 (rn, 3H),
L23-1.22 (d, 6H); LCMS; m/z=547.9 (M+H).sup.-.
[0264] The below compounds were prepared by procedure similar to
the one described in Example-1 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvents
at suitable reaction conditions. The physicochemical
characteristics of the compounds are summarized herein below
table.
TABLE-US-00011 Compd No Compound structure Analytical data 2
##STR00098## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
10.18-10.17 (d, 1H), 7.87-7.34 (m, 3H), 7.27-7.21 (t, 1H),
7.08-7.06 (d, 1H), 6.36-6.29 (m, 1H), 6.13-6.05 (m, 1H) 5.73-5.70
(m, 1H), 5.59-5.56 (t, 1H), 5.06-4.79 (m, 1H), 4.59 (s, 2H),
4.21-4.15 (m, 1H), 3.90-3.80 (m, 3H), 3.37-3.31 (m, 2H), 2.95-2.88
(m, 1H), 2.66-2.58 (m, 1H), 2.21-2.12 (m, 1H), 1.90-1.65 (m, 4H),
1.43 (s, 2H), 1.20 (s, 6H): LCMS: m/z = 519.30 (M + H).sup.-. 3
##STR00099## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.14 (s,
1H), 10.02 (s, 1H), 8.92 (s, 1H), 8.65 (s, 1 H),7.70 (s, 1H), 7.67-
7.48 (m, 3H), 7.24-7.20 (m, 1H), 7.05-7.02. (s, 1H), 6.81 (s, 1H),
6.66-6.54 (m, 1H), 6.14-6.08 (m, 1H), 5.70-5.67 (m, 1H), 4.62-4.58
(m, 2H), 4.47-4.46 (m, 1H), 3.82 (s, 2H), 3.71-3.56 (m, 2H), 2.90
(s, 2H), 2.15-2.08 (m, 2H), 2.02-1.84 (m, 2H), 1.59 (s, 1H),
1.44-1.37 (m, 2H), 1.30-1.23 (in, 6H): LCMS: m/z = 533.3 (M +
H).sup.- . 4 ##STR00100## .sup.1H NMR (DMSO-d.sub.6 400 MHz):
.delta. 10.04-10.03 (d, 1H), 8.9 (s, 1H), 7.67 (s, 1H), 7.46 (m,
3H), 7.21-7.17 (t, 1H). 6.99-6.97 (d. 1H), 6.78 (s, 2H), 6.57-6.49
(m, 1H), 6.11-6.06 (m, 1H), 6.06 (s, 1H), 5.63-5.60 (m, 1H), 4.55
(s, 1H), 3.78-3.74 (m, 3H), 3.67-3.63 (m, 2H), 3.58-3.52 (m, 2H),
3.25-3.17 (m, 1H), 3.15-3.13 (m, 1H), 2.86 (s, 2H), 2.16-2.06 (m,
3H), 1.93 (s, 1H), 1.53-1.33 (m, 3H), 1.18 (s, 6H); LCMS: m/z =
533.3 (M + H).sup.+. 5 ##STR00101## Isomer-1: .sup.1H NMR
(DMSO-d.sub.6, 400 MHz); .delta. 10.07-10.06 (d, 1H), 8.90 (s, 1H),
7.71 (s, 1H), 7.51 (m, 2H), 7.23-7.21 (t, 1H), 7.03-7.01 (d, 1H),
6.89-6.80 (m, 1H), 6.61-6.53 (m, 1H), 6.15-6.11 (m, 1H), 5.67-5.63
(m, 1H), 4.59 (s, 2H), 3.82-3.62 (m, 3H), 3.58-3.50 (m, 2H),
3.39-3.37 (m, 1H), 3.23-3.06 (m, 2H), 2.89 (s, 2H), 2.19-1.83 (m,
2H), 1.59-1.38 (m, 3H), 1.27-1.23 (d, 2H), 1.11-1.02 (d, 6H); LCMS:
m/z = 533.2 (M + H).sup.+. 6 ##STR00102## Isomer-2: .sup.1H NMR
(DMSO-d.sub.6, 400 MHz); .delta. 10.08- 10.06 (d, 1H), 8.90 (s,
1H), 7.71 (s, 1H), 7.50 (m, 2H), 7.25-7.21 (t, 1H), 7.03-7.01 (d,
1H), 6.89-6.80 (m, 1H), 6.61-6.53 (m, 1H), 6.15-6.11 (m, 1H),
5.67-5.63 (m, 1H), 4.58 (s, 2H), 3.82-3.62 (m, 5H), 3.58-3.50 (m,
2H). 3.39-3.37 (m, 1H), 3.23-3.07 (m, 1H), 2.89 (s, 1H), 2.19-1.99
(m, 4H) 1.59-1.38 (m, 2H), 1.27-1.23 (d, 2H) 1.23 (s, 6H); LCMS:
m/z = 533.25 (M + H).sup.- . 7 ##STR00103## .sup.1H NMR
(DMSO-d.sub.6 400 MHz): .delta. 9.80 (s, 1H), 8.90 (s, 1H), 8.65
(s. 1H), 8.02-8.00 (d, 1H), 7.71 (s, 1H) 7.50 (s, 2H) 7.23-7.19 (t,
1H) 6.99-6.97 (d, 1H) 6.82 (s, 1H) 6.39-6.32 (m, 1H) 6.08-6.03 (d,
1H), 5.56-5.52 (m, 1H), 4.59 (s, 2H) 4.05-4.00 (m, 1H) 3.87-3.83
(m, 4H), 2.90 (s, 1H) 2.38-2.36 (m, 1H), 1.82-1.69 (m, 6H),
1.59-1.53 (m, 6H), 1.19 (s, 6H); LCMS: m/z = 561.4 (M + H).sup.-. 8
##STR00104## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
10.00-9.97 (d, 1H), 8.91-8.64 (rn, 1H), 7.71 (s, 1H), 7.51 (s, 2H),
7.25-7.23 (t, 1H), 7.02 (s, 1H), 6.90-6.75 (m, 2H), 6.11-6.07 (d,
1H), 5.69 5.63 (t, 1H), 4.60 (s, 2H), 4.49-4.28 (m, 1H), 4.13-4.00
(m, 2H), 3.65 (s, 3H), 3.23-3.16 (m, 2H), 3.07-3.01 (t, 1H), 2.90
(s, 1H), 2.78-2.67 (m, 3H), 2.42 (s, 1H), 2.20 (s, 3H), 2.04-1.85
(m, 4H), 1.73-1.67 (m, 3H), 1.49-1.34 (m, 4H), 1.23 (s, 6H); LCMS:
m/z = 560.5 (M + H).sup.- . 9 ##STR00105## .sup.1H NMR
(DMvSO-d.sub.6, 400 MHz): .delta. 9.80 (s, 1H) 8.02- 8.00 (d, 1H)
7.87 (s, 1H) 7.72 (s, 1H) 7.61 (t, 1H) 7.50 (s, 2H), 7.23-7.19 (t,
2H), 7.08 (s, 1H) 6.96 (s, 2H), 6.38-6.31 (m, 1H), 6.08-6.03 (m,
1H), 5.55-5.52 (m, 1H) 4.59-4.57 (m, 2H), 3.87 (s, 1H) 3.38 (s,
2H), 2.91 (s, 2H), 2.67 (s, 2H), 2.32 (s, 2H), 1.90-1.68 (m, 6H),
1.53-1.40 (m, 3H), 1.22-1.20 (d, 7H); LCMS: m/z = 575.0 (M +
H).sup.-. 10 & 11 ##STR00106## Isomer-1: .sup.1H NMR
(DMSO-d.sub.6, 400 MHz): .delta. 9.80 (s, 1H), 8.60 (s, 1H),
8.02-8.00 (d, 1H), 7.70 (s, 1H), 7.50 (m, 2H). 7.23-7.19 (t, 1H),
6.98 (s, 1H), 6.55 (m, 1H), 6.39-6.32 (rn, 1H), 6.08-6.03 (m, 1H),
5.56-5.53 (d, 1H), 4.60 (s, 4H), 4.11-4.09 (m, 1H), 3.89 (s, 1H),
3.69 (s, 1H), 3.43-3.42 (m, 1H), 3.17-3.16 (d, 2H), 2.91 (s, 2H),
2.67 (s, 2H), 2.38-2.37 (m, 1H), 1.83-1.69 (m, 5H), 1.60-1.50 (m,
4H), 1.23 (s, 6H): LCMS: m/z = 575.40 (M + H) . Isomer-2: .sup.1H
NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.79 (s, 1H), 8.60 (s, 1H),
8.02-8.01 (d, 1H), 7.69 (s, 1H), 7.55- 7.53 (m, 2H), 7.23-7.19 (t,
1H), 6.98 (s, 1H), 6.55 (m, 1H), 6.39-6.32 (m, 1H), 6.08-6.04 (in,
1H), 5.56-5.53 (d, 1H), 4.60 (s, 2H), 4.38 (s, 1H), 4.13-4.09 (m,
2H), 3.90 (m, 2H), 3.17-3.16 (d, 2H), 2,91 (s, 1H), 2.67 (s, 2h),
2,38 (s, 1H), 1.80-1.68(m, 4H), 1.60-1.36 (m, 7H), 1.23(s, 6H);
LCMS: m/z = 575.30 (M + H) . 12 & 13 ##STR00107## Isomer-1:
.sup.1H NMR-d.sub.6, 400 MHz): .delta. 10.03 (s, 1H), 8.57 (s, 1H),
7.66 (s, 1H), 7.53-7.40 (m, 2H), 7.21-7.18 (t, 1H), 6.99 (s, 1H),
6.69 (s, 1H), 6.58-6.50 (m, 1H), 6.11-6.07 (m, 1H), 5.64-5.60 (m,
1H), 4.56 (s, 2H), 3.79-3.59 (m, 3H), 3.55-3.39 (m, 2H), 3.20-3.04
(m, 3H), 2.85 (s, 2H), 2.17-1.91 (m, 4H), 1.73-1.61 (m, 5H), 1.18
(s, 6H): LCMS: m/z =546.90 (M + H) . Isomer-2: .sup.1H NMR
(DMSO-d.sub.6, 400 MHz): .delta. 10.04- 10.02 (d, 1H), 8.54 (s,
1H), 7.65 (s, 1H), 7.50-7.48 (m, 2H), 7.21-7.18 (t, 1H), 6.98 (s,
1H), 6.57-6.51 (m, 2H), 6.11-6.06 (m, 1H), 5.64-5.60 (m, 1H),
4.56-4.35 (m, 3H), 4.09-4.05 (m, 1H), 3.89 (s, 1H), 3.79-3.59 (m,
2H), 3.55-3.42 (m, 2H), 3.18-3.04 (m, 2H), 2.16-1.91 (m, 3H),
1.77-1.53 (m, 4H), 1.41-1.32 (m, 3H), 1.19 (s, 6H); LCMS: m/z
=547.20 (M + H) . indicates data missing or illegible when
filed
Example-2: Synthesis of
1-acryloyl-N-(3-(((2-(((3R,4R)-3-fluoropiperidin-4-yl)amino)-8-isopropylp-
yrazolol[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carbo-
xamide (Compound-14).
Step-1: Synthesis of tert-butyl
(3R,4R)-4-((4-((3-(1-acryloylpyrrolidine-3-carboxamido)
benzyl)amino)-8-isopropylpyrazolo[1,3,5]triazin-2-yl)amino)-3-fluoropiper-
idine-1-carboxylate
##STR00108##
[0266] To a cooled solution of 1-acryloylpyrrolidine-3-carhoxylic
acid (0.05 g, 0.30 mmol, intermediate-10) in DMF (2 mL) at
0.degree. C. was added HATU (0.11 g, 0.30 mmol) and DIPEA (0.07mL,
%0.40 mmol) and finally added tert-butyl
(3R,4R)-44(44(3-arninobentyl)arnino)-8-isopropylpyrazolo[1,5-a][1,3,5]tri-
azin-2-yl)amino)-3 -fluoropiperidine-1 -carboxylate (0.1 g, 0.20
mmol, intermediate-38). The reaction mixture was stirred for 2 h at
room temperature. The reaction mixture was quenched with ice-water
and diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.20 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated to crude residue. The residue was purified by
combiflash to afford desired title compound (0.1 g, 76%).
Step-2: Synthesis cif
1-acrylovl-N-(3-(((2-(((3R,4R)-3-fluoropiperidin-4--v1)amino)-8-isopropyl-
pyrazolo[1.5-a][1,3.5]triazin-4-yl)arnino)rneth
yl)phenyl)pyrrolidine-3-carboxamide
##STR00109##
[0268] TFA (1 mL) was added to a solution of tert-butyl
(3R,4R)-4((4-((3-(1-acryloylpyrrolidine-3-carboxamido)henzyparnino)-8-iso-
propylpyrazolo[1,5-a ][1,3,5]triazin
-2-yl)amino)-3-fluoropiperidine-1-carboxylate (0.1g, 0.15 mmol)in
DCM (5 mL) at 0.degree. C. Then the reaction mixture allowed
stirring at room temperature for 12 h. After completion of the
reaction, the reaction mixture was evaporated to afford amine. The
residue was purified by prep. HPLC (Method: A: 0.1% TFA, B:
Acetonitrile:MeOH, Column: XBRIDGE C-18 (19 mm*150 mm, 5 .mu.m)) to
afford desired title compound (0.04 g, 50%). .sup.1HNMR
(DMSO-d.sub.6, 400 MHz): .delta. 10.09-10.07 (d, 1H), 8.40 (s, 3H),
7.79 (s, 1H), 7.62-7.51 (m, 2H), 7.26-7.22 (t, 1H), 7.09-6.96 (m,
1H), 6.61-6.54 (m, 1H), 6.15-6.10 (tn, 1H), 5.75-5.65 (m, 2H), 4.63
(s, 2H), 4.25 (m, 2H), 3.82-3.63(m, 3H), 3.66-3.45 (in, 2H),
3.37-3.10 (m, 4H), 2.93-2.90 (t, 2H), 2.22-1.92 (m, 2H), 1.71 (s,
2H), 1.39 (s, 6H): LCMS: m/z=550.65 (M+H).sup.+.
[0269] The below compounds were prepared by procedure similar to
the one described in Example-2 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvents
at suitable reaction conditions. The physicochemical
characteristics of the compounds are summarized herein below
table.
TABLE-US-00012 Compd No Compound structure Analytical data 15
##STR00110## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
10.07-10.05 (d, 1H), 8.54 (s, 1H), 7.78-7.70 (m, 4H), 7.48 (m, 2H),
7.23- 7.19 (m, 1H), 7.07 (s, 1H), 6.99 (s, 1H), 6.57-6.51 (m, 1H),
6.12-6.07 (d, 1H), 5.65-5.62 (m, 1H), 4.57 (s, 3H), 3.36-3.33 (m,
3H), 3.19-3.05 (m, 3H), 2.92-2.87 (m, 2H), 2.14-2.04 (m, 2H),
1.96-1.28 (m, 4H), 1.19 (s, 8H): LCMS: m/z = 546.25 (M + H)- .
indicates data missing or illegible when filed
Example-3: Synthesis of
1-acryloyl-N-(3-(((8-isopropyl-2-(((S)-tetrahydro-2H-pyran-3-yl)amino)pyr-
azolo[1,5-a][1,3,5]triazin-4-yl)atnino)methyl)phenyl)piperidine-2-carboxam-
ide (Compound-16)
Step-1: Synthesis of
tert-butyl2(3-(((8-isopropyl-2-(((S)-tetrahydro-2H-pyran-3-yl)amino)pvraz-
olo[1.5-a][1,3.5]triazin-4-yl)amino)methyl)phenyl)carbamoyl)piperidine-1-c-
arboxvlate
##STR00111##
[0271] To a cooled solution of
1-(tert-butoxycarbonyl)piperidine-2-carboxylic acid (0.13 g, 0.577
mmol) in DMF (10 mL) at 0.degree. C. was added HATU (0.26 g, 0.68
mmol) followed by DIPEA (0.2 mL, 1.04 mmol) and finally added
(S)-N4-(3-aminobenzyl)-8-isopropyl-N2-(tctrahydro-2H-pyran-3-yl)pyrazolo[-
1,5-a][1,3,5 ]triazine-2,4-diamine (0.2 g, 0.524 mmol,
intermediate-34). The reaction mixture was stirred for lh at room
temperature. The reaction mixture was quenched with ice-water and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.25 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated the crude residue was purified by 100-200 silica
gel column chromatograph to afford desired title compound (0.14 g,
42%). LCMS: m/z=593 (M+H).sup.+.
Step-2: Synthesis of
N-(3-(((8-isopropyl-2-(((S)-tetrahydro-2H-pyran-3-yl)amino)pYrazolo[1,5-a-
][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxamide.
##STR00112##
[0273] TFA (0.5rnL) was added to a solution of
tert-butyl2-((3-(((8-isopropyl-2-(((S)-tetrahydro-2H-pyran-3-yl)amino)pyr-
azolo[1,5-a][1,3,5 ]triazin-4-yparnino)methypphenyl)
carbamoyl)piperidine-1-carboxylate (0.14 g, 0.23 mmol) in DCM (5 mL
) at 0.degree. C. Then the reaction mixture allowed to stir at room
temperature for 2 h. After completion of the reaction, the reaction
mixture was concentrated under vacuum, to afford desired title
compound (0.12 g, crude). 1,LCMS: mit 493 (M+H).sup..
Step-3: Synthesis of
1-acryloyl-N-(3-(((8-isopropyl-2-(((S)-tetrahydro-2H-pyran-3-yl)amino)pyr-
azolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxami-
de.
##STR00113##
[0275] To a cooled solution of
N-(3-(((8-isopropyl-2-(((S)-tetrahydro-2H-pvran-3-yl)amino)pyrazolo[1.5-a-
][1,3,5]triazin-4-yl)arnino)methyl)phenyl)piperidine-2-carboxarnide
(0.12 g, 0.24 mmol) in DCM (5 mL ) added TEA (0.1rnI 0.72 mmol) and
stirred for 5min and added acryloyl chloride (0.022 g, 0.24 mmol)
diluted in DCM (1mI,) drop wise at 0.degree. C. The reaction
mixture was allowed to stir at room temperature for 1 h. After
completion of the reaction, the reaction mixture quenched with
water and then diluted with DCM (10 mL). Organic layer washed with
water followed by brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated under vacuum and purified by preparative I IPLC
(Method: A: water, B: Acetonitrile, Column: Kinctex EVC) C-18 (150
mm*21.2, 5 .mu.m) to afford desired title compound (0.03 g, 30%) as
free base. .sup.1HNMR (DMSO-d.sub.6, 400 MHz): d 9.92 (s, 1H),
8.90-8.68 (d, 1H), 7.71 (s, 1H), 7.50 (s, 2H), 7.25-7.21 (t, 1H),
7.02 (s, 1H), 6.88-6.82 (m, 1H), 6.68 (s, 1H), 6.10-6.06 (d, 1H),
5.68-5.66 (d, 1H), 5.12 (s, 1H), 4.58 (s, 2H), 3.98-3.71 (m, 4H),
3.51-3.46 (t, 2H), 3.21 (t, 1H), 3.02 (t, 1H), 3.02-2.91 (m, 3H),
2.12-2.09 (d, 1H), 1.66-1.52 (m, 4H), 1.38 (s. 2H), 1.23 (s, 6H);
LCMS: m/z =547.3 04(M+H).sup.-.
[0276] The below compounds were prepared by procedure similar to
the one described in lxample-3 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvents
at suitable reaction conditions. The physicochemical
characteristics of the compounds are summarized herein below
(able.
TABLE-US-00013 Compd No Compound structure Analytical data 17
##STR00114## .sup.1H NMR (DMSO-d.sub.6, 400 MHz = .delta. 7.69 (s,
1H), 7.57 (m, 1H), 7.44 (m, 1H), 7.24 (m, 1H), 7.04 (m, 1H), 5.65
(s, 1H), 5.41 (s, 1H), 4.88 (m, 1H), 4.56 (m, 2H), 4.08 (m, 1H),
3.78 (m, 3H), 3.42 (m, 3H), 2.88 (m, 2H), 2.13 (m, 1H), 2.09 (m,
3H), 1.96 (m, 3H), 1.63 (m, 2H), 1.42 (m, 2H), 1.20 (d, 6H), 1.08
(m, 2H); LCMS: m/z = 347.7 (M + H).sup.- . 18 ##STR00115## .sup.1H
NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.08 (s, 1H), 8.70 (s, 1H),
7.71 (s, 1H), 7.5-7.48 (m, 2H), 7.26 (t, 1H), 7.04 (d. 1H). 6.90
(m, 1H), 6.34 (m, 1H), 6.12 (d, 1H), 5.68 (d, 1H), 4.59-4.40 (m,
2H), 4.37 (t, 1H), 4.35 (t, 1H). 4.08 (m, 1H), 4.03 (m, 1H), 3.94
(m, 3H), 3.52 (m, 2H), 2.90 (m, 2H), 1.98-1.92 (m, 2H), 1.6-1.4 (m,
2H), 1.23-1.07 (d, 6H): LCMS: m/z = 518.7(M + H)- . 19 ##STR00116##
.sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.15 (s, 1H), 8.70
(s, 1H), 7.70 (s, 1H), 7.54 (s, 2H), 7.26-7.22 (t, 1H), 7.05-7.03
(d, 1H), 6.81 (s, 1H), 6.36-6.29 (m, 1H), 6.13-6.05 (m, 1H),
5.72-5.55 (m, 1H), 5.05-4.78 (m, 1H), 4.58 (s, 2H), 4.20-4.15 (m,
1H), 3.90-3.81 (m, 4H), 2.49-2.44 (m, 2H), 2.21-2.07 (m, 1H),
1.81-1.72 (m, 2H), 1.44-1.37 (m, 4H), 1.21 (s, 6H); LCMS: m/z =
519.20 (M + H).sup.+. 20 ##STR00117## .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 10.15 (s, 1H), 8.70 (d, 1H), 7.70 (s, 1H),
7.52-7.48 (s, 2H), 7.23 (t, 1H), 7.05 (d, 1H), 6.90-6.75 (m, 1H),
6.36-6.30 (m, 1H), 6.13-6.05 (m, 1H) 5.72-5.55 (m, 1H), 5.05-4.78
(m, 1H), 4.58 (brs, 2H), 4.20-4.15 (m, 1H), 3.90-3.81 (m, 3H), 2.89
(brs, 1H), 2.49-2.44 (m, 2H), 2.21-2.12 (m, 1H), 1.81-1.74 (m, 2H),
1.65-1.30 (m, 4H), 1.22 (s, 6H): LCMS: m/z = 519.15 (M + H) . 21
##STR00118## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 8.66-8.62
(m, 2H), 7.71 (s, 1H), 7.64-7.53 (m, 2H), 7.25-7.21 (t, 1H), 7.03
(m, 1H), 6.89-6.82 (m, 2H), 6.10 (d, 1H), 5.69-5.66 (d, 1H),
5.12-5.11 (d, 2H), 4.58 (m, 2H), 3.98-3.95 (d, 1H), 3.82 (m, 3H),
3.51-3.46 (m, 1H), 2.99-2.90 (m, 2H), 2.12-2.09 (d, 1H), 1.82 (m,
1H), 1.66-1.62 (m, 4H), 1.38 (m, 4H), 1.23 (d, 6H); LCMS: m/z =
547.0 (M + H) 22 ##STR00119## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 9.87 (s, 1H), 8.9- 8.98 (m, 2H), 7.71 (s, 1H), 7.50 (m,
2H), 7.22 (t, 1H), 7.0 (d, 1H), 6.90 (m, 1H), 6.81 (m, 1H), 6.21
(m, 1H), 6.08 (d, 1H), 5.57 (d, 1H), 4.57 (m, 2H), 3.82-3.65 (m,
5H), 2.9 (m, 1H), 1.90-1.87 (m, 6H), 1.62-1.45 (m, 2H), 1.36-1.29
(m, 3H), 1.27-1.23 (d, 6H), 1.1-1.08 (m, 2H); LCMS: m/z = 561.7 (M
+ H).sup.-. 23 ##STR00120## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 9.85 (d, 1H), 8.80 (d, 1H), 7.72 (s, 1H), 7.62-7.57 (m,
2H), 7.25 (t, 1H), 7.07 (d, 1H), 6.85-6.79 (m, 2H), 6.19-6.11 (m,
1H), 5.74-5.72 (d, 1H), 4.60-4.51 (m, 3H), 4.12-3.83 (m, 4H),
3.90-3.70 (m, 3H), 3.55 (l, 1H), 3.10-2.80 (m, 3H), 1.82 (brs, 1H),
1.65-1.35 (m, 4H), 1.24 (s, 6H): LCMS: m/z = 549.70 (M + H) . 24
##STR00121## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.96 (s;
1H), 8.70 (d, 1H), 7.71 (s, 1H), 7.65-7.45 (m, 2H), 7.24 (r, 1H),
7.04 (brs, 1H), 6.89-6.82 (m, 2H) 6.17-6.13 (m, 1H), 5.75-5.66 (m,
1H), 4.87-4.85 (m, 1H), 4.59 (brs, 2H), 4.29-4.26 (d, 1H),
3.89-3.65 (m, 6H), 3.60-3.45 (m, 1H), 2.90 (brs, 1H), 2.45-2.55 (m,
2H), 1.82 (brs, 1H), 1.70-1.30 (m, 4H), 1.22 (d, 6H); LCMS: m/z =
549.40 (M + H).sup.-. 25 ##STR00122## .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 10.14-10.09 (d, 1H), 8.95 (s, 1H), 8.70 (s, 1H),
7.71 (s, 1H), 7.49 (m, 2H), 7.27-7.23 (t, 1H), 7.06 (s, 1H), 6.81
(s, 1H), 4.80- 4.79 (d, 1H), 4.70 (s, 1H), 4.60 (s, 2H), 4.35-4.26
(m, 1H), 4.03-3.99 (t, 1H), 3.90-3.73 (m, 4H), 3.69-3.45 (m, 1H),
3.49-3.43 (m, 1H), 2.67 (s, 1H), 1.82 (s, 2H), 1.63 (s, 1H), 1.45
(m, 3H), 1.22 (s, 6H); LCMS: m/z = 547.70 (M + H) . 26 ##STR00123##
.sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.63 (s, 1H), 8.13 (s,
1H), 7.67 (s, 1H), 7.53-7.51 (m, 1H), 7.42-7.41 (d, 1H), 7.15-7.12
(d, 1H), 6.41-6.39 (m, 1H), 6.12-6.07 (m, 1H), 5.69-5.66 (d, 1H),
4.80 (s, 1H), 4.68-4.66 (d, 2H), 4.31-4.28 (m, 2H), 3.86-3.64 (m,
6H), 3.68-3.64 (m, 1H), 3.46-3.31 (m, 3H), 3.0-2.95 (m, 2H), 2.75-
2.67 (m, 2H), 1.93-1.81 (m, 2H), 1.55-1.46 (m, 2H), 1.18 (s, 6H),
1.18-1.16 (m, 3H); LCMS: m/z = 577.5 (M + H).sup.-. 27 ##STR00124##
.sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.06 (s, 1H), 8.60
(s, 1H), 7.73-7.72 (m, 1H), 7.58 (s, 1H), 7.33 (s, 1H), 7.11-7.09
(d, 1H), 6.79 (s, 1H), 6.55 (s, 1H), 6.34-6.27 (m, 1H), 6.11-6.04
(m, 1H), 5.71-5.68 (m, 1H), 5.56- 5.53 (t, 1H), 5.01-4.97(m, 1H),
4.78-4.75 (m, 1H), 4.56-4.54 (d, 2H), 4.19-4.10 (m, 1H), 3.89-3.81
(m, 2H), 3.17-3.15 (d, 1H), 2.91 (s, 2H), 2.59-2.41 (m, 2H), 2.38
(s, 3H), 2.05-2.17 (m, 1H), 2.08 (m, 1H), 1.81 (s, 1H), 1.44 (s,
2H),. 1.24 (s, 6H): LCMS: m/z = 532.9 (M + H).sup.+. 28
##STR00125## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.64 (s,
1H), 8.90- 8.64 (m, 1H), 7.69 (s, 1H), 7.57 (m, 1H), 7.33 (m, 1H),
7.14-7.13 (m, 2H), 6.83 (m, 1H), 6.37 (m, 1H), 6.18- 6.04 (m, 1H),
5.75 (d, 1H), 5.61 (d, 1H), 5.10 (m, 1H), 4.95 (t, 1H), 4.53 (m,
2H), 4.21-4.17 (t, 2H), 3.93-3.85 (m, 4H), 2.89 (m, 1H), 2,14 (s,
3H), 1.81-1.67 (m, 2H). 1.44 (m, 2H). 1.21 (d, 6H): LCMS: m/z =
533.2 (M + H) . 29 ##STR00126## .sup.1H NMR (DMSO-d.sub.6, 400
MHz): .delta. 10.19 (s, 1H), 8.66 (d, 1H), 7.74 (s, 1H), 7.58 (m,
2H), 7.18-7.13 (t, 1H), 6.83 (m, 1H), 6.54 (s, 1H), 6.35-6.29 (m,
1H), 6.13- 6.05 (m, 1H), 5.72-5.69 (m, 1H), 5.58-5.56 (m, 1H),
5.02-4.98 (m, 1H), 4.77-4.74 (m, 1H), 4.65 (s, 2H), 4.21-4.14 (m,
1H), 3.89-3.83 (m, 3H), 2.41 (m, 1H), 2.20-2.18 (m, 1H), 1.83 (m,
2H), 1.59-1.479 (m, 3H), 1.24 (s, 6H); LCMS: m/z = 537 (M +
H).sup.-. 30 ##STR00127## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 10.05 (d, 1H), 9.00- 8.52 (m, 1H), 7.9 (m, 1H), 7.72 (s,
1H), 7.10 (m, 2H), 6.89 (m, 1H), 6.39 (m, 1H), 6.21 (d, 1H), 6.10
(d, 1H), 5.78 (d, 1H), 5.06 (m, 1H), 4.67 (m, 2H), 4.22 (t, 2H),
3.91-3.81 (m, 4H), 2.9 (m, 2H), 2.33 (m, 1H), 1.81 (m, 1H), 1.44
(m, 3H), 1.22 (d, 6H); LCMS; m/z = 537.65 (M + H) . 31 ##STR00128##
.sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.36 (s, 1H), 8.81
(d, 1H), 7.72 (s, 1H), 7.58-7.20 (m, 2H), 6.91-6.80 (m, 2H),
6.36-6.29 (m, 1H), 6.13-6.06 (m, 1H), 5.73-5.56 (m, 1H), 5.05-4.759
(m, 1H), 4.58 (brs, 2H), 4.21-4.10 (m, 1H), 3.90-3.83 (m, 3H),
3.17-3.15 (m, 2H), 2.90 (brs, 1H), 2.32-2.08 (m, 3H), 1.92-1.82 (m,
1H), 1.56- 1.37 (m, 3H), 1.22 (s, 6H): LCMS: m/z = 537.3 (M +
H).sup.-. 32 ##STR00129## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 10.13 (s, 1H), 8.7 (d, 1H), 7.70-7.40 (m, 3H), 7.12 (t,
1H), 6.83 (m, 1H), 6.39-6.52 (m, 1H), 6.13-6.09 (m, 1H), 5.63-5.62
(m, 1H), 4.64 (s, 2H), 3.79-3.59 (m, 5H), 3.49-3.43 (m, 2H),
3.33-3.04(m, 2H), 3.04(s, 2H) 2,16-1.80 (m, 4H), 1.55-1.34 (m, 3H),
1.23 (s, 6H); LCMS; m/z = 551.45 (M + H) . 33 ##STR00130## .sup.1H
NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.25 (d, 1H), 8.8 (d, 1H),
7.69 (s, 1H), 7.49-7.16 (m, 2H), 7.00-6.70 (m, 2H), 6.56-6.49 (m,
1H), 6.11-6.06 (m, 1H), 5.64-5.60 (m, 1H), 4.54 (s, 2H), 3.77-3.73
(m, 3H), 3.67-3.43(m, 4H), 3.30-3.12(m, 2H), 3.05(s, 1H) 2.15-1.90
(m, 4H), 1.52-1.33 (m, 3H), 1.19 (s, 6H); LCMS: m/z = 551.3(M +
H).sup.+. 34 ##STR00131## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 9.82 (s, 1H), 8.56 (d, 1H), 7.72 (s, 1H), 7.56 (m, 2H),
7.27 (m, 2H), 7.08- 7.06 (d, 2H), 6.85-6.79 (m, 3H), 6.00 (d, 1H),
5.66- 5.63 (d, 1H), 5.07 (s, 1H), 4.55-4.53 (d, 3H), 4.30- (m, 1H),
3.94-3.91 (d, 1H), 3.82 (m, 4H), 3.46 (m, 1H), 3.16-2.91 (m, 2H),
2.31-2.25 (m, 3H), 2.13-1.97 (m, 2H), 1.82 (s, 2H), 1.63-1.54 (m,
2H), 1.23-1.22 (d, 6H); LCMS: m/z = 561.1 (M + H) . 35 ##STR00132##
.sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.70 (d, 1H), 8.60-
8.90 (d, 1H), 7.70 (s, 1H), 7.50 (s, 1H), 7.06 (m, 2H), 6.85 (m,
2H), 6.09-6.05 (d, 1H), 5.67-5.64 (d, 1H), 5.24 (s, 1H), 4.63 (s,
2H), 3.90 (d, 1H), 3.78 (m, 4H), 2.90 (m, 1H), 2.29-2.28 (d, 2H),
1.8 (m, 1H), 1.62 (m, 4H), 1.37 (m, 4H), 1.19 (s, 6H); LCMS: m/z =
565.45 (M + H).sup.-. 36 ##STR00133## .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 10.14 (s, 1H), 8.70 (s, 1H), 7.71 (s, 1H),
7.50-7.28 (m, 2H), 6.87-6.80 (m, 3H), 6.53 (s, 2H), 6.09-6.05 (d,
1H) 5.68-5.65 (m, 1H), 5.08-5.07 (d, 1H), 4.56 (s, 2H), 3.97-3.81
(m, 4H), 3.45 (m, 1H), 3.32 (s, 1H), 2.89 (s, 2H), 2,09-2.06 (d,
1H), 1.81 (s, 1H), 1.65-1.58 (m, 3H), 1.37 1.35 (d, 3H), 1.22 (s,
6H); LCMS: m/z = 565.75 (M + H) . 37 ##STR00134## .sup.1H NMR
(DMSO-d.sub.6, 400 MHz): .delta. 9.97 (s, 1H), 8.96- 8.65 (d. 1H),
7.75-7.43 (m, 3H), 7.16-7.12 (t, 1H), 6.94-6.82 (m, 2H), 6.10-6.03
(m, 1H), 5.69-5.67 (d, 1H), 5.09 (s, 1H), 4.80-4.68 (m, 2H),
3.98-3.83 (m, 4H), 2.93 (m, 2H), 2.19-1.99 (m, 2H), 1.84 (s, 1H),
1.66-1.62 (m, 3H), 1.38-1.25 (m, 8H): LCMS: m/z = 565.2 (M +
H).sup.+. 38 ##STR00135## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 9.55 (s, 1H), 9.00- 8.52 (m, 1H), 8.01-7.99 (d, 1H), 7.68
(s, 1H), 7.60 (m, 1H), 7.02 (m, 2H), 6.90 (m, 1H), 6.35-6.28 (m,
1H), 6.05-6.00 (dd, 1H), 5.52-5.49 (dd, 1H), 4.63 (s, 2H),
3.84-3.77 (m, 3H), 3.36-3.33 (m, 2H), 3.25 (m, 1H), 2.80 (m, 1H),
2.51-2.48 (m, 1H), 1.79-1.74 (m, 3H), 1.67-1.65 (m, 2H), 1.60-1.49
(m, 3H), 1.40 (m, 2H), 1.18 (d, 6H), 1.05-1.03 (t, 2H); LCMS: m/z =
579.3 (M + H) . 39 ##STR00136## .sup.1H NMR (DMSO-d.sub.6, 400
MHz): .delta. 9.84 (s, 1H), 8.7 (d, 1H), 8.00 (d, 1H), 7.74 (s,
1H), 7.07-7.04 (m, 2H), 7.10 (t, 1H), 6.90 (m, 1H), 6.37-6.31 (m,
1H), 6.07- 6.02 (m, 1H), 5.55-5.52 (m, 1H), 4.63 (s, 2H), 3.86-
3.60 (m, 4H), 3.20 (s, 1H), 2.90 (s, 2H), 2.67-2.52 (s, 3H),
1.90-1.60 (m, 5H), 1.59-1.40 (m, 5H), 1.23 (s, 6H): LCMS: m/z =
579.35 (M + H).sup.-. 40 ##STR00137## .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 10.06 (s, 1H), 8.76(s, 1H), 8.61 (s, 1H), 8.47
(s, 1H), 8.18 (s, 1H), 7.80 (s, 1H), 6.9-0-6.80 (m, 1H), 6.60-6.58
(d, 1H), 6.27-6.23 (d, 1H), 5.85-5.82 (d, 1H), 5.00 (d, 1H),
4.79-4.77 (d, 2H), 4.51-4.47 (d, 1H), 4.04-3.97 (m, 5H), 3.86-3.82
(m, 1H), 3.62-3.48 (m, 3H), 3.14-3.06 (m, 2H), 1.97-1.94 (d, 2H),
1.68-1.63 (m, 2H), 1.40- 1.38 (d, 6H): LCMS: m/z = 550.3 (M + H) .
40A ##STR00138## LCMS: m/z = 569.5 (M + H) . 40B ##STR00139## LCMS:
m/z = 569.5 (M + H).sup.-. 40C ##STR00140## LCMS: m/z = 561.25 (M +
H) . 40D ##STR00141## LCMS: m/z = 547.30 (M + H).sup.-. 40E
##STR00142## LCMS: m/z = 547.70 (M + H).sup.-. indicates data
missing or illegible when filed
Example-4: Synthesis of
(S)--N-(1-acryloylpiperidin-3-yl)-3-(((8-isopropyl-2-((tetrallyclro-2H-py-
ran-4-yl)arnino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)benzamide
(Compound-41).
Step-1: Synthesis of tert-butyl
(S)-3-(3-(((8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4-yl)-
amino)methyl)benzamido)piperidine-1-carboxylate
##STR00143##
[0278] DIPEA (0.5 mL .,2.47 mmol) was added to a solution of
4-chloro-8-isopropyl-2-(rnethylthio)pyrazolo[1,5-a][1,3,5]triazine
(0.3 g, 1.23 mmol, intermediate-4) and
tert-butyl(S)-3-(3-(aminomethyl)benzamido)piperidine-1-carboxylate
(0.495 g, 1.48 rn mol, Intermediate-16) in acetonitrile (10 mL) at
0.degree. C.. The resulting reaction mixture was stirred at ambient
temperature for 4 h. After completion of the reaction, the reaction
mixture was concentrated under vacuum and purified by column
chromatography using 100-200 silica gel to afford the title
compound (0.4 g, 66%). LCMS: m/z=540(M+H).sup.+.
Step-2: Synthesis of ten-butyl
(S)-3-(3-(((S-isopropyl-2-(methylsulfonyl)pyrazolo[5-a][1.3,5]triazin-4-y-
l)amino)methyl)benzamido)piperidine -1 -carboxylate
##STR00144##
[0280] mCPBA (1.53 g, 5.79 mmol) was added portion wise to a
solution of tent-butyl
(S)-3-(3(((8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4-ypar-
nino)methyl)henzamido) piperidine-1-carboxylate (0.8 g, 1.44 mmol)
in DCM (10 mL) and stirred for overnight at RT. After completion of
the reaction, the reaction mixture was extracted with 2M aq. NaOH
and DCM (10 mL). The organic phase was washed with brine, dried
over Na.sub.2SO.sub.4, filtered and concentrated to afford the
title compound (0.7 g, crude). LCMS: m/z=572(M+H).sup..
Step-3: Synthesis of tert-butyl (S)-3-(3(((8-i
sopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pvrazolo[1 ,5 -a][1
,3,5]triazin4-yl)amino)methyl)benzamido)piperidine-1-carboxylate
##STR00145##
[0282] A mixture of 4-aminotetrahydropyran (0.25 g, 2.45 mmol) and
tert-butyl
(S)-3-(3-(((8-isopropyl-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazin-4-
-yl)arnino)methyl)benzamido) piperidine-1-carboxylate (0.35 g,
0.614 mmol) were allowed to heat at 100.degree. C. for 2 h. After
completion of the reaction, the reaction mixture was cooled to room
temperature and quenched with ice-water and diluted with ethyl
acetate. The aqueous layer was separated and extracted with ethyl
acetate (2.times.25mL). The combined organic phase was washed with
brine, dried over Na.sub.7S01, filtered and concentrated. The
residue was purified by 100-200 silica gel column chromatography to
afford desired title compound (0.16 g, 44%). LCMS: m/z=592.9
(M.times.H).sup..
Step-4: Synthesis of (S)-3-(((S-
isopropyl-2(tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1.3,5]triazin--
4-yl)amino)methyl)-N-(piperidin-3-yl)benzamide
##STR00146##
[0284] The process of this step was adopted from step-2 of
example-3 (0,10 g crude). LCMS: m/z=493.4 (M+H).sup.1.
Step-5: Synthesis of (S)-N-(1-acryloylpiperidin-3-yl
)-3-(((8-isopropyl-24(tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,3,-
5]triazin-4-yl)amino)rnethyl)benzamide
##STR00147##
[0286] The process of this step was adopted from step-3 of
example-3. The obtained crude compound was purified by preparative
HPLC (Method; A:water, 13: Acetonitrile, Column: X Bridge C-18 (150
mm*21,2, 5.mu.m) to afford the desired title compound (0.014 g,
20%). .sup.1HNMR (DMSO-d.sub.6, 400 MHz): .delta. 9.00 (s, 1 H),
8.75 (s, 1 H), 8.32-8.30 (m, 1 H), 7.84 (s, 1 H), 7.71-7.68 (m,
2H), 7.51-7.49 (m, 1H), 7.41-7.37 (rn, 1H), 6.92-6.67 (m, 2H),
6.07-6.03 (t, 1H), 5.65-5.61 (m, 1 H), 4.69 (s, 2H), 3.96-3.79 (m,
6H), 3.17-3.15 (d, 1H), 2.89 (s, 2H), 1.92-1.76 (m, 3H), 1.64-1.57
(m, 2H), 1.42 (s, 2H), 1.28-1.21 (m, 7H): LCMS: m/z 546.8
(M+H).sup..
[0287] The below compounds were prepared by procedure similar t the
one described in Example-4 with appropriate variations in
reactants, quantities of reagents in presence of suitable solvents
at suitable reaction conditions. The physicochemical
characteristics of the compounds arc summarized herein below
table.
TABLE-US-00014 Compd No Compound structure Analytical data 42
##STR00148## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.09 (d,
1H), 9.02- 8.4 (m, 1H), 8.22-7.90 (m, 1H), 7.69 (s, 1H), 7.22-7.14
(m, 2H), 7.10-6.4 (m, 1H), 6.36 (m, 1H), 6.18 (m, 1H), 5.75 (d,
1H), 5.03 (m, 1H), 4.84 (m, 2H), 4.20 (t, 1H), 3.88-3.82 (m, 3H),
3.33 (m, 1H), 2.88 (m, 1H), 2.66 (m, 1H), 2.29 (m, 1H), 1.82-1.62
(m, 2H), 1.63-1.43 (m, 2H), 1.20 (d, 6H), 1.08 (m, 2H); LCMS: m/z =
537.7(M + H) . 43 ##STR00149## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 9.88 (s, 1H), 8.9 (m, 1H), 7.93-7.85 (m, 1H), 7.70 (s, 1H),
7.21-7.14 (m, 2H), 6.92-6.89 (m, 1H), 6.61-6.53 (m, 1H), 6.15-6.10
(dd, 1H), 5.68-5.64 (dd. 1H), 4.53 (s, 2H), 3.83-3.79 (m, 3H),
3.71-3.66 (m, 2H), 3.55-3.51 (m, 2H), 3.46- 3.27 (m, 3H), 2.88 (m,
1H), 2.22-2.15 (m, 2H), 1.98- 1.97 (m, 2H), 1.83-1.64 (m, 2H),
1.43-1.20 (dd, 6H): LCMS: m/z = 551.3 (M + H).sup.+. 44
##STR00150## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.72-9.67
(d, 1H), 8.70 (s, 1H), 7.69 (s, 2H), 7.20-7.15 (m, 3H), 6.87-6.80
(m, 2H), 6.54 (s, 3H), 6.11-6.06 (d, 1H), 5.69-5.65 (m, 1H),
5.24-5.23 (d, 1H), 4.53 (s, 2H), 4.04 (s, 2H), 3.98- 3.95 (d, 1H),
3.82 (s, 2H), 2.88 (s, 1H), 2.16-2.12 (d, 1H), 1.81 (s, 1H), 1.64
(m, 3H), 1.41-1.38 (m, 3H), 1.20-1.19 (d, 6H): LCMS: m/z = 565.75
(M + H).sup.- ; 45 ##STR00151## .sup.1H NMR (DMSO-d.sub.6, 400
MHz): .delta. 9.95-9.93 (d, 1H), 8.50 (m, 1H), 7.66 (s, 1H), 7.41
(m, 1H), 7.18 (m, 1H), 7.07-7.06 (m, 1H), 6.85-6.74 (m, 2H),
6.06-6.02 (d, 1H), 5.64-5.59 (t, 1H), 5.1 (m, 1H), 4.45-4.42 (d,
1H), 4.28-4.24 (d, 1H), 4.06-3.96 (m, 1H), 3.81-3.67 (m, 3H), 3.36
(m, 1H), 2.99-2.84 (m, 3H), 2.67-2.50 (m, 1H), 1.91-1.69 (m, 4H),
1.51-1.45 (m, 3H), 1.39-1.36 (m, 3H), 1.30 (d, 6H), 1.06-1.02 (m,
2H); LCMS: m/z = 561.35 (M + H).sup.+. indicates data missing or
illegible when filed
Example-5: Synthesis of
3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)arnino)pyrazolo[1,5-a][1,3,-
5]triazin-4-yl)annino)methyl)phenyl
4-acryloylpiperazine-1-carboxylate (Compound-46)
Step-1: Synthesis of
3-(((8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1
,3,5]triazin-4-yl)amino)methyl)phenol
##STR00152##
[0289] The process of this step was adopted from step-1 of
example-4 (0.5 g, 73%). LCMS: m/z=330.2 (M+H).sup..
Step-2: Synthesis of
3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4yl)amino)pyrazolo[1.5-a][1
.3.5]triazin-4-yl)amino)methyl)phenol
##STR00153##
[0291] The process of this step was adopted from step-2 of
example-4 (0.35 g, 63%). LCMS: m/z=362.3 (M+H).sup.-.
Step-3: Synthesis of
3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5-
]triazin-4-yl)annino)methyl)phenol
##STR00154##
[0293] The process of this step was adopted from step-3 of
example-4 (0.22 g, 70%). LCMS: m/z =383.2 (M+H).sup..
Step-4: Synthesis of 1-(tert-butyl)
4-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)
pvrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)piperazine-1,4-dicarboxyl-
ate
##STR00155##
[0295] To a solution of
3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5-
]triazin-4-yl)amino)methyl)phenol (0.227 g, 0.59 mmol) in 10 DCM,
was added DMAP (0.086 g, 0.70 mmol), followed by
tert-butyl4-(chlorocarbonyl)piperazine 1 carboxylate (0.147 g, 0.59
mmol, synthesized according t procedure depicted in
US2007/270433A1). Reaction mixture was stirred for 4 h at RT. After
completion of reaction, reaction mixture was quenched to ice cold
water and partitioned between water and DCM. The product was
extracted with DCM (3.times.25m1), washed with brine, dried over
sodium sulphate, filtered and concentrated to dryness. The product
was purified hyl00-200 mesh silica column by eluting with 30%-50%
ethyl acetate-hexane to afford the title compound (0. 15 g, 42%).
LCMS: m/z=595.8 (M+H).sup.-.
Step-5: Synthesis of
3-4(8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5]-
triazin-4-yl)amino)methyl)phenyl piperazine-1-carboxylate
##STR00156##
[0297] The process of this step was adopted from step-2 of
example-3 (0.1 g of TFA salt). LCMS: m/z=495.4 (M+H).sup.1.
Step-6: Synthesis of
3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)arnino)pyrazolo[1
,5-a][1 ,3,5
]triazin-4-yl)arnino)methyl)phenyl4-acryloylpiperazine-1-carboxylate
##STR00157##
[0299] The process of this step was adopted from step-3 of
example-3. The obtained crude compound was purified by preparative
HPLC (0.01% NH.sub.4OH in water, B: Acetonitrile, Column: Gemini NX
C-18 (21.2 mm*150 mm, 5 .mu.m)) to afford desired title compound
(0.035 g, 26%). .sup.1HNMR (DMSO-d.sub.6, 400 MHz): .delta.
5.95-5.66 (d, 1H), 7.68 (s, 1H), 7.31-7.14 (m, 1H), 7.17 (s, 1H),
7.08 (s, 1H), 7.98-7.96 (d, 1 H), 6.90-6.87 (m, 1 H). 6.81-6.74 (m,
1 H), 6.52 (s, 1H), 6.13-6.08 (m, 1H). 5.70-5.67 (m, 1H). 4.62-4.56
(rn, 2H), 3.80-3.78 (m, 4H). 3.60-1.56 (rn, 6H), 3.40-3.31 (m, 2H),
2.86 (s, 1H), 1.80-1.60 (rn, 1H), 1.50-1.38 (m 4H) 1.18 (s, 6H);
LCMS: m/z=549.6 (M+H).sup.-.
Example-6: Synthesis of
3-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyraz-
olo[1,5 -a][1,3,5]triazin-4-yl)amino)methyl)phenyl)benzamide
(Compound-47)
Step-1: Synthesis of
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,-
3,5]triazin-4-yl)amino)methyl)phenyl)-3-nitrobenzamide
##STR00158##
[0301] The process of this step was adopted from step-1of example-1
(0.38 g). LCMS: m/z=511.60 (M+H).sup.-.
Step-2: Synthesis of
3-amino-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)
pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)benzamide
##STR00159##
[0303] To a solution of
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo
[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-3-ni trobentamide
(0.38 g, 0.71 mmol) in THE;MeOH Water (3;2;1) were added zinc
(0.465 g, 7.1 mmol) and ammonium chloride (0.76 g, 14.3 mmol). The
reaction mixture was stirred at roorn temperature for 4 h. After
completion of reaction, the reaction mixture filtered through
celite and diluted with ethyl acetate. The aqueous layer was
separated and extracted with ethyl acetate (2.times.25 mL). The
combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by 100-200 silica gel column chromatography to afford
desired title compound (0.3 g, 70%). LCMS: m/z=501.20
(M+H).sup.1.
[0304] Step-3: Synthesis of
3-acrylamido-N-(34(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)
pyrazolo[1,5-a][1,3.5]triazin-4-yl)amino)methyl)phenyl)benzamide
##STR00160##
[0305] The process of this step was adopted from step-3 of
example-3. The obtained crude compound was purified by preparative
HPLC (Method: A: water, B: Acetonitrile-Methanol, Column:ZORBAX XDB
C-18 (21.1 mm*150 mm, 5 .mu.m) to afford desired title compound
(0.022 g, 25%)..sup.1HNMR (DMSO-d.sub.6, 400 MHz): .delta. 10.33
(s, 1H), 10.26 (s, 1H), 8.11 (s, 1H), 7.91 (d, 1H), 7.70) (rn, 2H),
7.61-7.59 (d, 2H), 7.47 (t, 1H), 7.28 (t, 1H), 7.08 (m, 1H), 6.70
(m, 1H), 6.53 (m, 1 H), 6.30 (d, 1H), 5.79-5.75 (m, 2H), 4.62 (m,
2H), 3.81 (m, 3H), 2.90 (rn, 1 H), 1.81-1.50 (m, 3H), 1.49-1.2 (rn,
3H), 1.21 (d, 6H); LCMS: m/z=555.2 (M+H).sup.-.
[0306] The below compound was prepared by procedure similar to the
one described in Ixample-6 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvents
at suitable reaction conditions. The physicochemical
characteristics of the compounds arc summarized herein below
table.
TABLE-US-00015 Compd No Compound structure Analytical data 48
##STR00161## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.63 (s,
1H), 10.44 (s, 1H), 8.18 (d, 1H). 7.74 (m, 2H); 7.54 (t, 2H), 7.31
(m, 1H), 7.25 (m, 1H), 7.12 (m, 2H), 6.40 (m, 1H), 6.21 (d, 1H),
5.76 (d, 1H), 4.62 (m, 2H), 3.88-3.79 (m, 4H), 2.96 (m, 2H), 1.81
(m, 3H), 1.45 (m, 3H), 1.20 (d, 6H); LCMS: m/z = 555.20 (M +
H).sup.-. 48A ##STR00162## LCMS: m/z = 569.30 (M + H).sup.+.
Example-7: Synthesis of (E)-4((3
-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)
pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-N,N-di
methylbut-2-enamide (Compound-49)
##STR00163##
[0308] To a solution of
N4-(3-aminobenzyl)-8-isopropyl-N2-(tetraltydro-2H-pyran-4-y1)pyrazolo[1,5-
-a][1,3,5]triazine-2,4-diamine (0.1 g. 0.26 mmol, intermediate-33)
and (E)-4-bromo-N,N-dimethylbut-2-enamide (0.061 g, 31.4 mmol) in
CAN (3 mL) was added K.sub.2CO.sub.3 (0.157 g, 1.136 mmol) at RT,
then stirred at 80.degree. C. for 4 h. After reaction completion,
the reaction mixture was cooled to room temperature, quenched with
water and diluted with ethyl acetate.
[0309] The aqueous layer was separated and extracted with ethyl
acetate (2.times.15 mL). The combined organic phase was washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated. The
residue was purified by preparative HPLC (Method: A: 0.02% NH.sub.3
in water, B:ACN, Column: Gemini NX C18 : 150 mm*21.2mm) to afford
desired title compound (0.028 g, 20%). .sup.1HNMR (DMSO-d.sub.6,
400MHz): .delta. 8.40-8.80 (d, 1H), 7.70 (s, 1H), 7.02-6.99 (t,
1H), 6.84 (s, 1H), 6.66-6.63 (m, 2H), 6.60-6.54 (d, 2H), 6.44-6.42
(d, 1H), 5.99 (s, 1H), 4.44 (s, 2H), 3.89 (s, 5H), 3.30 (m, 2.89
(s. 4H). 2.82 (s. 3H), 1.89 (s, 1 H), 1.23 (s, 1H), 1.10 (s, 6H);
LCMS; m/z=493.6 (M+H).sup.+.
Example-8: Synthesis of
(E)-4-(diethylamino)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)ami-
no)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-enamide
(Compound-50)
Step-1: Synthesis of
(E)-4-bromo-N-(3-(((8-isopropyl-2-((tetruhydro-2H-pyran-4-yl)amino)
pyrazolol[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)but-2-enamide
##STR00164##
[0311] To a cooled solution of (E)-4-bromobut-2-cnoic acid (0.86 g,
5.24mrno1) in DCM (10 mL) was added 2 drops of DMF and followed by
oxalyl chloride (0.9 mL10.49 mmol) at 0.degree. C., then stirred at
RT for 1h, evaporated the solvent completely under vacuum and
dissolved again in DCM (5mL). In another flask, a solution of
N4-(3-aminolienzyl)-8-isopropyl-N2-(tetrahydro-2H-pyran-4-yl)pyrazolo[1,5-
-a][1,3,5]triazine-2,4-diamine (1 g, 2.62 mmol, Intermediate-33) in
DCM (10 mL) and DIPEA (1.45 mL 7.87 mmol) was cooled to 0.degree.
C. To this the above acid chloride in DCM was added slowly drop
wise, reaction mixture was allowed to stirred at RT for 1h. After
completion of the reaction, the reaction mixture quenched with
water and then diluted with DCM (20 mL). The organic layer washed
with water followed by brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated, the residue was purified by 100-200 mesh silica
gel column chromatography to afford desired title compound (0.7 g,
60%). LCMS: m/z=530.05 (M+H).sup.1.
Step-2: Synthesis of
(E)-4-(diethylamino)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)ami-
no)pvrazolo[1,5-a][1,3,5]triazin -4-yl
)amino)methyl)phenyl)but-2-enamide
##STR00165##
[0313] To a stirred solution of
(E)-4-hromo-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazo-
lo[1,5-a][1,3,5]triazin4yl)amino)methyl)phenyl)butenamide (0.15 g,
0.28 mmol) and diethylamine (0.042. g, 0.568 mmol) in ACN (3 mL )
was added K.sub.2CO.sub.3 (0.1 g, 0.71 mmol) at RT, then stirred at
80.degree. C. for 2 h. After completion of reaction, the reaction
mixture was cooled to room temperature, quenched with water and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.15 mL ). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated. The residue was purified by preparative
H1.sup.31,C (Method; A: 0.1% Il'A in water, 13; Acetonitrile,
Column; X Bridge ; 150 mm*19.0 mm) to afford desired title compound
as TFA salt, after that passed this TFA salt through Vari pure
basic resin column to remove TFAto afford final compound (0.027 g,
54%). .sup.1HNMR (DMSO-d.sub.6, 400 MHz ): (DMSO-d.sub.6, 400 MHz):
.delta. 10.35 (s, 1H), 9.70 (s. 1H), 9.60 (s, 1 H). 8.90 (s, 1 H),
7.95 (s, 1 H), 7.75 (s, 1 H), 7.55 (m. 1H), 7.34-7.30 (t, 1 H),
7.15 (s, 1H), 6.81-6.73 (m, 1H), 6.53-6.49 (d, 1H), 4.66 (s, 2H),
4.02-4.00 (t, 2H), 3.92-3.91 (m, 2 H), 3.82 (m, 3H), 3.20 (s, 2H),
3.18-3.14 (m, 4H), 3.02-2.97 (m, 1H), 1.69 (s, 1H), 1.48 (s, 2.H),
1.27-1.23 (m, 10H): LCMS; =521.65 (M+H).sup.31 .
[0314] The below compounds were prepared by procedure similar to
the one described in Example-8 with appropriate variations in
reactants, quantities of reagents in presence of suitable solvents
at suitable reaction conditions. The physicochemical
characteristics of the compounds are summarized herein below
table.
TABLE-US-00016 Compd No Compound structure Analytical data 51
##STR00166## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
10.30-10.10 (d, 1H), 9.50 (s, 1H), 8.74 (s, 2H), 7.86-7.54 (m, 3H),
7.30 (t, 1H), 7.12 (s, 1H), 6.90-6.73 (m, 2H), 6.47-6.43 (d, 1H),
4.63 (s, 2H), 4.01 (s, 3H), 3.90-3.70 (m, 5H), 3.12 (s, 3H), 2.94
(s, 2H), 2.00 (s, 1H), 1.84 (s, 1H), 1.66 (s, 1H), 1.46 (s, 2H),
1.24 (s, 6H): LCMS: m/z = 535.7 (M + H) . 52 ##STR00167## .sup.1H
NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.03 (s, 1H), 7.71 (s, 1H),
7.77-7.75 (m, 2H), 7.26-7.22 (t, 1H), 7.03-7.02 (d, 1H), 6.9-6.8
(m, 1H), 6.72-6.65 (m, 1H), 6.24-6.21 (d, 1H), 4.60 (d, 2H),
3.90-3.80 (m, 3H), 3.35-3.30 (m, 3H), 3.08-3.07 (d, 2H), 2.907 (s,
1H), 2.67 (s, 1H), 2.45-2.33 (m, 5H), 2.32-2.28 (m, 2H), 1.83 (s,
1H), 1.60-1.45 (m, 2H), 1.23 (s, 7H), 0.99-0.95 (t, 3H); LCMS: m/z.
= 562.35 (M + H).sup.-. 53 ##STR00168## .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 9.96 (s, 1H), 8.90 (s, 1H), 8.6 (s, 1H), 7.67 (s,
1H), 7.60-7.50 (m, 1H), 7.22- 7.18 (m, 1H), 6.99-6.97 (m, 1H),
6.88-6.77 (m, 1H), 6.69-6.64 (m, 1H), 6.20-6.16 (m, 1H), 4.58-4.52
(m, 2H), 3.80-3.72 (m, 3H), 3.44-3.33 (m, 2H), 2.95-2.92 (m, 2H),
2.88-2.82 (m, 1H), 2.22-2.20 (m, 6H), 1.81- 1.75 (m, 1H), 1.60-1.50
(m, 2H), 1.35-1.49 (m, 2H), 1.18 (s, 6H); LCMS: m/z = 493.6 (M + H)
. 54 ##STR00169## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
10.03 (s, 1H), 8.65 (s, 1H), 7.71 (s, 1H), 7.56 (m, 2H), 7.25-7.21
(t, 1H), 7.02-7.01 (d, 1H), 6.83 (s, 1H), 6.77-6.71 (m, 1H),
6.26-6.22 (d, 1H), 4.59 (s, 2H), 3.82 (s, 3H), 3.20-3.16 (m, 2H),
2.90 (s, 1H), 2.54-2.32 (m, 4H), 1.82 (s, 1H), 1.72-1.68 (m, 4H),
1.59 (s, 1H), 1.45 (s, 2H), 1.37 (s, 2H), 1.22 (s, 6H); LCMS: m/z =
519.30 (M +H).sup.+.. 55 ##STR00170## .sup.1HNMR (DMSO-d.sub.6, 400
MHz): .delta. 10.23 (s, 1H), 8.90 (s, 1H), 8.65 (s, 1H), 7.71 (s,
1H), 7.55 (m, 2H), 7.25- 7.21 (t, 1H), 7.03 (s, 1H), 6.83 (s, 1H).
6.73-6.66 (m, 1H), 6.23-6.19 (d, 1H), 4.59 (s, 2H), 3.82 (s, 4H),
3.05- 3.03 (m, 2H), 2.90 (s, 1H), 2.32 (s, 4H), 1.82 (s, 1H),
1.52-1.47 (m, 6H), 1.39-1.37 (d, 3H), 1.22 (s, 6H); LCMS: m/z =
533.20 (M + H).sup.- . 56 ##STR00171## .sup.1HNMR (DMSO-d.sub.6,
400 MHz): .delta. 10.05 (s, 1H), 8.65 (s, 1H), 7.71 (s, 1H), 7.56
(m, 2H), 7.25-7.21 (t, 1H), 7.03-7.01 (d, 1H), 6.83 (s, 1H),
6.76-6.70 (m, 1H), 6.27-6.23 (d, 1H), 5.26-5.11 (m, 1H), 4.59 (s,
2H), 3.82 (s, 3H), 3.24-3.18 (m, 4H), 2.87-2.78 (m, 3H), 2.66- 2.58
(m, 2H), 2.34-2.28 (m, 1H), 2.17-2.08 (m, 1H), 1.93-1.81 (m, 2H),
1.59-1.37 (m, 2H), 1.22 (s, 6H); LCMS: m/z = 537.60 (M + H).sup.-.
57 ##STR00172## .sup.1HNMR (DMSO-d.sub.6, 400 MHz): .delta. 10.05
(s, 1H), 8.92 (s, 1H), 8.65 (s, 1H), 7.71 (s, 1H), 7.58-7.54 (m,
2H), 7.26-7.22 (t, 1H), 7.03-7.01 (d, 1H), 6.82 (s, 1H), 6.73- 6.67
(m, 1H), 6.27-6.23 (d, 1H), 4.59 (s, 2H), 3.82 (s, 3H), 3.26-3.25
(m, 2H), 2.93-2.87 (t, 3H), 2.74-2.70 (m, 2H), 2.33-2.19 (m, 2H),
1.83 (s, 1H), 1.58 (s, 1H), 1.45 (s, 1H), 1.36 (s, 2H), 1.22 (s,
6H); LCMS: m/z = 555.60 (M + H).sup.1. 58 ##STR00173## .sup.1HNMR
(DMSO-d.sub.6, 400 MHz): .delta. 10.05 (s, 1H), 8.66 (s, 1H), 7.71
(s, 1H), 7.60-7.50 (m, 2H), 7.27-7.23 (t, 1H), 7.05 (s, 1H),
6.89-6.82 (m, 1H), 6.73-6.68 (m, 1H), 6.32 (s, 1H), 4.60 (s, 2H),
3.81 (s, 4H), 3.20-3.16 (m, 2H), 2.67-2.55 (m, 2H), 2.33-2.32 (m,
2H), 2.90 (s, 2H), 1.81 (s, 4H), 1.58 (s, 2H), 1.44 (s, 2H), 1.36
(s, 1H), 1.22 (s, 6H); LCMS: m/z = 551.20 (M + H).sup.+. 59
##STR00174## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.13 (s,
1H), 8.95 (s, 1H), 8.70 (s, 1H), 7.72 (s, 1H), 7.61 (s, 1H), 7.30-
7.20 (m, 1H), 6.87-6.83 (d, 2H), 6.75-6.68 (m, 1H), 6.22-6.1 8 (d,
1H), 4.59 (s, 2H), 3.82-3.75 (d, 3H), 3.03- 3.01 (d, 2H). 2.90 (s,
1H), 2.66 (s, 1H), 2.32 (s, 1H), 2.14 (s, 5H), 1.82 (s, 2H), 1.60
(s, 2H), 1.45 (s, 2H), 1.22 (s, 6H): LCMS: m/z = 511.20 (M +
H).sup.- . 60 ##STR00175## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 9.84 (s, 2H), 8.90 (s, 1H), 8.65 (s, 2H), 7.80 (s, 2H),
7.72 (s, 1H), 7.08 (s, 3H), 6.90 (s, 2H), 6.75-6.70 (m, 2H),
6.45-6.41 (d, 2H), 4.67 (s, 2H), 3.90-3.70 (m, 5H), 3.03-3.00 (d,
2H), 2.67 (s, 2H), 2.20 (s, 6H), 2.12-1.97 (m, 2H), 1.81 (s, 2H),
1.55-1.45 (m, 4H), 1.23 (s, 8H); LCMS: m/z = 511.2(M + H).sup.-. 61
##STR00176## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.13 (s,
1H), 8.95 (s, 1H), 8.70 (s, 1H), 7.72 (s, 1H), 7.61 (s, 1H), 7.30-
7.20 (m, 1H), 6.87-6.85 (d, 2H), 6.75-6.68 (m, 1H), 6.23-6.20 (d,
1H), 4.59 (s, 2H), 3.82-3.75 (d, 3H), 3.58- 3.56 (m, 4H), 3.34-3.29
(m, 1H), 3.10-3.08 (m, 2H), 2.90 (s, 1H), 2.36 (s, 4H), 1.86 (s,
1H), 1.53-1.35 (m, 3H), 1.22 (s, 6H); LCMS: m/z = 533.30 (M + H) .
62 ##STR00177## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.70
(s, 1H), 8.85 (s, 1H), 8.58 (s, 1H), 7.71 (s, 1H), 7.51-7.49 (d,
1H), 7.33- 7.31 (d, 1H), 7.26-7.22 (t, 1H), 7.13 (s, 1H), 6.87 (s,
1H), 6.77-6.71 (m, 1H), 6.49-6.45 (d, 1H), 4.60-4.59 (d, 2H), 3.82
(s, 3H), 2.91-2.90 (m, 1H), 2.66 (s, 1H), 2.40 (s, 6H), 2.33 (s,
2H), 1.83 (s, 1H), 1.46 (s, 2H), 1.23-1.21 (d, 7H): LCMS: m/z =
493.20 (M + H).sup.-. 63 ##STR00178## .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 10.03 (s, 1H), 8.80- 8.60 (d, 1H), 7.69 (s, 1H),
7.58-7.56 (d, 2H), 7.30-7.27 (d, 2H), 6.90-6.71 (m, 2H), 6.27-6.22
(d, 1H), 4.53 (s, 2H), 3.85-3-82 (d, 2H), 3.43 (s, 2H), 3.21-3.19
(d, 3H), 2.90 (s, 1H), 1.83 (s, 1H), 1.69 (s, 6H), 1.44 (m, 3H),
1.22 (s, 8H); LCMS: m/z = 519.25 (M + H).sup.- . indicates data
missing or illegible when filed
Example-9: Synthesis of (E)-
1-(4-(dimethylamino)but-2-enoyl)-N-(2-fluoro-5-(((8-isopropyl-2-((tetrahy-
dro-2H-pyran-4-yl)amino)pyraiolo[1,5-a][1,3,5]triazin-4-yl)amino)
methyl)phenyl) pyrrolidine-3-carboxamide (Compound-64)
##STR00179##
[0316] To a cooled solution of (E)-4-(dimethylamino)but-2-enoic
acid hydrochloride (0.050 g, 0.3 mmol) in DMF (2 mL) at 0.degree.
C. was added HATU (0.137 g 0.30 mmol) followed by DIPEA (0.1 mL 0.6
mmol) and finally added
N-(2-fluoro-5-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyr-
azolo[1,5-a][1,3,5]triaz
in-4-yl)arnino)methyl)phenyl)pyrrolidine-3-carboxamide (0.150 g,
0.3 mmol; this was prepared by using the procedure of example-3).
The reaction mixture was stirred for 2h at room temperature. The
reaction mixture was quenched with ice-water and diluted with ethyl
acetate. The aqueous layer was separated and extracted with ethyl
acetate (2.times.25mI,). The combined organic phase was washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated and
purified by preparative HPLC (Method: A: 0.01% in ammonia in water,
B: Acetonitrile-MeOH, Column: EVO C-18 (150 mm*21.2, 5 .mu.m) to
afford desired title compound (0.04 g, 24%). .sup.1HNMR
(1DMSO-d.sub.6, 400 MHz): .delta. 9.98-9.88 (s, 1 H), 9.00 (s, 1
H), 8.60 (s, 1 H), 7.85 (m, 1H), 7.70 (s, 1H), 7.22-7.15 (m, 2H),
6.64 (m, 1H), 6.63-6.58 (m, 1H), 6.38-6.33 (dd, 1H), 4.54 (s, 2H),
3.82-3.78 (m, 3H), 3.67-3.60 (m, 2H), 3.54 (m, 2H), 3.53-3.51 (m
1H), 3.34-3.27 (m, 1H), 3.02-3.01 (d, 2 H), 2.89 (m, 1H), 2.20-2.14
(in, 7H), 2.00-1.98 (m, 1H), 1.83 (m, 1H), 1.64 (m, 1H), 1.45 (in,
2H ), 1.21 (d, 6H): LCMS: m/z=608.75 (M+H).sup.+.
[0317] The below compounds were prepared by procedure similar to
the one described in Example-9 with appropriate variations in
reactants, quantities of reagents in presence of suitable solvants
at suitable reaction conditions. The physicochemical
characteristics of the compounds arc summarized herein below
table.
TABLE-US-00017 Compd No Compound structure Analytical data 65
##STR00180## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
10.23-10.20 (m, 1H), 8.88-8.72 (s, 1H), 7.76 (s, 1H), 7.60 (s, 2H),
7.32-7.30 (m, 1H), 7.10 (s, 1H), 6.90 (s, 1H), 6.72 (m, 1H),
6.09-6.05 (d, 1H), 5.80 (t, 1H), 5.10-4.89 (m, 1H), 4.64 (s, 2H),
4.20-4.19 (m, 1H), 3.90-3.86 (m, 3H), 2.95 (s, 2H), 2.57-2.50 (m,
2H), 2.22 (m, 2H), 1.90-1.88 (d, 3H), 1.78-1.76 (d, 1H), 1.50 (s,
3H), 1.28 (s, 6H); LCMS; m/z = 533.2 (M + H).sup.- . 66
##STR00181## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
10.19-10.14 (d, 1H), 8.9-8.6 (d, 1H), 7.72 (s, 1H), 7.55 (m, 2H),
7.28-7.24 (t, 1H), 7.10-7.06 (m, 1H), 6.90-6.60 (m, 1H), 6.66 (s,
3H), 4.98-4.77 (m, 1H), 4.60 (s, 2H), 4.31 (s, 1H), 4.16-4.12 (m,
1H), 3.93-3.82 (m, 3H), 2.91 (m, 1H), 2.23-2.14 (m, 1H), 1.84-1.80
(m, 2H), 1.46-1.37 (m, 2H), 1.24 (s, 6H); LCMS: m/z = 517.1 (M + H)
. 67 ##STR00182## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
10.06 (s, 1H), 9.0-8.65 (m, 2H), 7.70 (s, 1H), 7.50 (m, 2H), 7.26
(t, 1H), 7.04 (d, 1H), 6.81 (m, 1H), 5.46 (d, 1H), 5.18 (d, 1H),
4.95 (tn, 1H), 4.77 (m, 1H), 4.58 (m, 2H), 4.18 (m, 1H), 3.93-3.80
(m, 4H), 2.89 (m, 1H), 2.15 (m, 2H), 1.81 (m, 3H), 1.68 (m, 2H),
1.45 (m, 2H), 1.22 (d, 6H); LCMS: m/z = 533.25 (M + H).sup.-. 68
##STR00183## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.05 (s,
1H), 8.90 (s, 1H), 8.60 (s, 1H), 7.71 (s, 1H), 7.50 (s, 1H), 7.25
(t, 1H), 7.03 (s, 1H), 6.80 (s, 1H), 6.68- 6.64 (d, 1H), 6.39-6.38
(d, 1H), 4.58 (s, 2H), 4.05 (s, 2H), 3.81-3.76 (m, 3H), 3.65-3.62
(t, 2H), 3.55- 3.53 (m, 2H). 3.35-3.36 (m, 3H), 3.10 (m, 2H), 2.90
(s, 1H), 2.10 (m, 2H), 2.80 (m, 2H), 1.62-1.45 (m, 3H), 1.23 (s,
6H); LCMS: m/z = 577.2 (M + H) . 69 ##STR00184## .sup.1H NMR
(DMSO-d.sub.6, 400 MHz): .delta. 10.06 (s, 1H), 8.9 (s, 1H), 7.71
(s, 1H), 7.51 (m, 2H), 7.25-7.21 (t, 1H), 7.03-7.01 (d, 1H), 6.85
(s, 1H), 6.62-6.56 (m, 1H), 6.68-6.33 (m, 1H), 4.59 (s, 2H),
3.81-3.76 (m, 4H), 3.67-3.61 (m, 2H), 3.54-3.43 (m, 2H), 3.21- 3.08
(m, 2H), 30.3-3.01 (d, 2H), 2.90 (s, 1H), 2.18- 2.13 (m, 7H),
1.98-1.90 (m, 2H), 1.60-1.49 (m, 3H), 1.22 (s, 6H); LCMS: m/z =
590.5 (M + H).sup.- . 70 ##STR00185## .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 10.04 (s, 1H), 8.90-8.60 (m, 2H), 7.70 (s, 2H),
7.51 (s, 1H), 7.35 (s, 1H), 7.11-7.06 (t, 1H), 6.79 (s, 2H),
6.59-6.53 (m, 1H), 6.32-6.27 (d, 1H), 4.61 (s, 2H), 3.77-3.70 (m,
4H), 3.63-3.60 (m, 2H), 3.58-3.53 (m, 1H), 3.48-3.38 (m, 2H),
3.29-3.08 (m, 2H), 3.01-2.95 (m, 2H), 2.86 (s, 2H), 2.11-1.90 (m,
6H), 1.87-1.77 (m, 1H), 1.52-1.19 (m, 3H), 1.19 (s, 6H); LCMS: m/z
= 608.3 (M + H).sup.- . 71 ##STR00186## .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 10.23 (s, 1H), 8.90-8.65 (d, 1H), 7.68 (s, 1H),
7.50 (s, 1H), 7.16 (s, 1H), 6.84-6.81 (d, 2H), 6.60-6.53 (m, 1H),
6.32- 6.28 (d, 1H), 4.55 (s, 2H), 3.76-3.71 (m, 3H). 3.60- 3.56 (m,
1H), 3.51-3.45 (m, 1H), 3.43-3.46 (m, 1H), 3.29-3.13 (m, 1H),
3.05-3.02 (m, 2H), 2.87 (s, 1H), 2.63 (s, 2H), 2.28 (s, 2H),
2.14-1.89 (m, 7H), 1.79 (s, 1H); 1.52-1.32 (s, 3H), 1.19 (s, 6H);
LCMS: m/z = 608.3 (M + H).sup.-. 72 ##STR00187## .sup.1H NMR
(DMSO-d.sub.6, 400 MHz): .delta. 9.97 (s, 1H), 8.6 (s, 1H), 8.5 (s,
1H), 7.71 (s, 1H), 7.52-7.42 (m, 3H), 7.22 (m, 1H), 7.12-7.10 (d,
1H), 6.80 (m, 1H), 6.62-6.59 (m, 2H), 5.40 (s, 1H), 5.10 (s, 1H),
4.5 (m, 1H), 4.32 (m, 1H), 4.04-4.01 (m, 1H), 3.85- 3.72 (m, 3H),
3.33-2.97 (m, 3H), 2.88 (m, 2H), 2.13-2,09 (m, 6H), 1.92 (m, 1H),
1.74-1.68 (m, 3H), 1.56-1.54 (d, 2H), 1.49-1.41 (m, 2H), 1.21 (d,
6H), 1.10-1.06 (t, 2H); LCMS: m/z = 618.5 (M + H).sup.-. 73
##STR00188## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.04 (s,
1H), 8.95-8.65 (m, 1H), 7.70 (s, 1H), 7.48 (m, 2H), 7.26- 7.22 (t,
1H), 7.04 (s, 1H), 6.64 (s, 1H), 4.85-4.84 (d, 1H), 4.59 (s, 1H),
4.26-4.23 (d, 1H), 3.85-3.82 (d, 4H), 3.69-3.63 (d, 1H), 3.34 (s,
1H), 3.03 (s, 2H), 2.95-2.90 (m, 1H), 2.55-2.54 (m, 2H), 2.45 (s,
2H), 2.33-2.32 (m, 2H), 2.13 (s, 4H), 2.04 (s, 2H), 1.81 (s, 2H),
1.60 (s, 1H), 1.45 (s, 2H), 1.21 (s, 6H); LCMS: m/z = 606.30 (M +
H).sup.-. 74 ##STR00189## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 9.92 (s, 1H), 8.94 (s, 1H), 8.60 (s, 1H), 7.71 (s, 1H),
7.49 (s, 1H), 7.28 (t, 2H), 7.02 (s, 1H), 6.88-6.81 (m, 1H),
6.66-6.44 (m, 1H), 5.11 (s, 1H), 4.80-4.58 (m, 3H), 3.96-3.83 (m,
4H), 3.02-3.01 (d, 1H), 2.94-2.90 (d, 2H), 2.67 (s, 3H), 2.32 (s,
3H), 2.13 (s, 4H), 2.04- 1.98 (s, 1H), 1.82 (s, 1H), 1.66-1.61 (d,
3H), 1.44- 1.37 (m, 3H), 1.22 (s, 6H); LCMS: m/z = 604.25 (M +
H).sup.- . 75 ##STR00190## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 8.37 (s, 1H), 8.21 (s, 1H), 7.66 (s, 1H), 7.35-7.31 (m,
1H), 7.24- 7.22 (m, 1H), 7.18 (s, 1H), 7.04-7.01 (d, 1H), 6.67-
6.61 (m, 1H), 6.51-6.47 (m, 1H), 4.65 (s, 2H), 3.86- 3.84 (m, 3H),
3.64-3.63 (m, 4H), 3.56-3.52 (m, 4H), 3.42-3.35 (m, 4H), 3.10-3.06
(m, 1H)), 2.18 (s, 6H), 1.84-1.81 (m, 2H), 1.55-1.46 (m, 2H),
1.26-1.24 (d, 6H); LCMS: m/z = 606.2 (M + H).sup.+. indicates data
missing or illegible when filed
Example-10: Synthesis of
(E)-1-(4-(dirnethylamino)-4-oxobut-2-en-1-yl)-N-(3-(((8-isopropyl-2-((tei-
rahydro-2H-pyran-4-yl)amino)pyrazolo[1
,5-a][1,3,5]triazin-4-yl)amino)
methyl)phenyl)azetidine-2-carboxamide (Compound-76)
##STR00191##
[0319] To a stirred solution of
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)arnino)pyrazolo[1,5-a][1
,3,5]triazin-4-yl)amino)rnethyl)phenyl)azetidine-2-carboxamide (0.1
g, 0.21 mmol) in ACN (3 mL) was added DIPEA (0.07 mL, 0.43 mmol)
and (E)-4-brorno-N,N-dimethylbut-2-enamide (0.05 g, 0.25 mmol) at
0.degree. C. The reaction mixture was stirred for 12H at RT. Water
was added and extracted with dichloromethane. Me aqueous layer was
separated and extracted with dichloromethane (2.times.25m1,). The
combined organic phase was washed with hrine, dried oVer
Na.sub.2SO.sub.4, filtered and concentrated to afford crude
residue. The residue was purified by prep. HPLC (Method: A: 0.02%
Ammonia in water. B: Acetonitrile, Column: WATER X BRIDGE (19
mm*150 mm, 5.mu.m)) to afford desired title compound (0.05 g, 41%).
.sup.1HNMR (DMSO-d.sub.6, 400 MHz): .delta. 9.63 (s, 1H), 8.90-8.60
(m. 1H), 7.70-7.53 (m, 3H), 7.25-7.21 (t, 1H), 7.04 (s. 1H), 6.84
(s. 1H), 6.55 (s. 2H), 4.58 (s. 2H), 3.80 (s. 3H), 3.67-3.63 (t.
1H), 3.35-3.19 (m 5H), 2.96-2.90 (m, 5H), 2.79 (s, 3H), 2.66 (s,
1H), 2.32-2.65 (m, 1H), 2.13-2.09 (M, 1H), 1.82 (s, 2H), 1.45-1.32
(m, 2H), 1.22 (s, 6H): LCMS: m/z=576.45 (M+H).sup..
Example-11: Synthesis of
(E)-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)
pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-1-(4-(pyrrolid-1y-
l)but-2-enoyl)piperidine-2-carboxamide (Compound-77)
##STR00192##
[0321] To a cooled solution of (E)-4-bromobut-2-cnoic acid (0.28 g,
1.71 mmol) in DMF (10 mL) at 0.degree. C. was added HATU (0.65 g,
1.71 mmol) followed by DIPEA (0.31 mL, 1.71 mmol) and finally added
N-(3
-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5]-
triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxamide (0.42 g,
0.855 mmol). The reaction mixture was stirred for 1 h at room
temperature. The reaction mixture was quenched with ice-water and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.25 mL ). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated the crude residue was purified by 100-200 silica
gel column chromatography to afford desired intermediate (0.3 g,
46%). LCMS: m/z=641 (M+2H).sup.. The intermediate was dissolved in
ACN (5mL) and pyrrolidine (0.067 g, 0.93 mmol) and
K.sub.2CO.sub.3(0.13 g, 0.931 mmol) was added at RT, stirred at
80.degree. C. for 2 h. After reaction completion, the reaction
mixture was cooled to room temperature, quenched with water and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.15 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated. The residue was purified by preparative HPLC
(Method: A: 0.01% TFA in water, B MeOH;CAN (1;1), Column; Zorbax
XDB C18: 150 mm*21.2 mm) to afford desired title compound as TFA
salt, after that passed this TFA salt through Vari pure basic resin
column to removal the TFA from compound to afford final compound
(0.04 g, 26%). .sup.1HNMR (DMSO-d.sub.6, 400 MHz): 5 9.91 (s. 1H),
8.95-8.66 (d, 1H), 7.70 (s, 1H), 7.64 (s, 7.54-7.48 (m. 1H),
7.24-7.20 (t. 1H), 7.03 (s, 1H), 6.89-6.81 (d. 2H), 6.63 (s, 1H),
5.11 (s. 1H), 4.57 (m, 2H), 3.95-3.92 (m, 4H), 3.40-3.39 (m, 2H),
3.19-3.18 (d, 1H), 3.00 (s, 2.89 (s, 1 H), 2.67 (s, 1H), 2.42 (s,
3H), 2.33-2.32 (d, 1H), 2.H-2.08 (d, 1H) 1.98 (s, 1H), 1.83-1.61
(m, 6 H), 1.53 (s, 3H), 1.37 (s,3H), 1.23 (s, 6H): LCMS: m/z=630.3
(M+H).sup.-.
Example-12: Synthesis of
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo
[1,5-a][1
,3,5]triazin-4-yl)amino)rnethyl)plienyl)-1-(vinylsulfonyl)piperidine-2-ca-
rboxamide (Compound-78)
##STR00193##
[0323] To the solution of
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,-
3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-2-carboxamide (0.15
g, 0.3 mmol: prepared according to procedure of example-3) in DCM
(10 mL.) was added Et.sub.3N (0.123 mL 0.9 mmol) and
2-chloroethane-1-sulfonyl chloride (0.5 g, 0.3 mmol) at 0.degree.
C. After stirring 2 h at room temperature, the reaction mixture was
quenched with ice-water and diluted with DCM. The aqueous layer was
separated and extracted with DCM (2.times.25 mL ). The combined
organic phase was washed with brine, dried over Na.sub.2SO.sub.4,
filtered and concentrated the crude residue was purified by
preparative HPLC (Method; A: water, B; ACN, Column; Zorbax XDB C18
(21.2 mm.times.150 mm, 5.mu.) to afford desired title compound
(0.025 g, 10.5%). .sup.1HNMR DMSO-d.sub.6, 400 MHz): .delta. 9.99
(s, 1 H), 8.80 (d, 1 H), 7.71 (s, 1 H), 7.60-7.40 (m, 2H), 7.23 (t,
1 H), 7.04-7.02 (d, 1 H), 6.95-6.75 (m, 1 H), 6.66-6.60 (m, 1 H),
6.05-5.96 (m, 2H), 4.58 (hrs, 2H), 4.49-4.48 (d 1 H), 3.90-3.70 (m,
4H), 3.55-3.45 (m, 2H), 2.89 (brs. 1 H), 2.03-2.00 (m, 1 H),
1.85-1.58 (m, 6H), 1.39-133 (m, 4H), 1.22 (s. 6H); LCMS: m/z=583.0
(M+H).sup.1.
Example-13: Synthesis of
N-(24(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a]-
[1,3,5]triazin-4-yl)amino)methyl)phenyl)arnino)-2-oxoethyl)acrylamide
(Compound-79)
Step- 1 : Synthesis of
tert-butyl(2(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo-
[1,5-a][1,3.5]triazin-4-yl
)amino)methyl)phenyl)amino)-2-oxoethyl)carbarmate
##STR00194##
[0325] To a cooled solution of (tert-butoxycarhonyl)glycine (0.033
g, 0.3 mmol) in DMF (5 mL) at 0.degree. C. was added HATU (0.152 g,
0.4mnol) followed by DIPEA (0.1 mL, 0.62 mmol) and finally added
N4-(3-aminobenzyl)-8-i
sopropyl-N2-(tetrahydro-2H-pyran-4-yl)pyrazolo [1,5-a][1,3,5
]triazine-2,4-diamine (0.1 g, 0.26 mmol; Intermediate-33). The
reaction mixture was stirred for 1 h at room temperature. The
reaction mixture was quenched with ice-water and diluted with ethyl
acetate. The aqueous layer was separated and extracted with ethyl
acetate (2.times.25mI,), The combined organic phase was washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated the
crude residue was purified by 100-200 silica gel column
chromatography to afford desired title compound (0.12 g, crude).
LCMS: m/z=539 (M+H).sup.+.
Step-2: Synthesis of
2-amino-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4yl)amino)pyrazolo[1,-
5a][1,3,5]triazin-4-yl )amino)methyl)phenyl)acetamide
##STR00195##
[0327] TFA (1mL) was added to a solution of
tert-butyl(2((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazol-
o[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-2-oxoethyl)carbamat-
e (0.15 g, 0.34 mmol) in DCM (3mL) at 0.degree. C. Then the
reaction mixture allowed to stir at room temperature for 2 h. After
completion of the reaction, the reaction mixture was concentrated
under vacuum to afford desired title compound (0.15 g, crude).
LCMS;=439 (M+H).sup.1.
Step-3: Synthesis of
N-(2-((3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo
[1.5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)amino)-2-oxoethvl)acrylami-
de
##STR00196##
[0329] To a cooled solution of
2-amino-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1-
.5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)acetamide (0.127 g,
0.28 mmol) in DCM (5mL) added TEA (0.08mL 0.56 mmol) and stirred
for 5 min and added acryloyl chloride (0.026 g, 0.28 mmol) drop
wise at 0.degree. C.. The reaction mixture was allowed to stir at
room temperature for 1 h. After completion of the reaction, the
reaction mixture quenched with water and then diluted with DCVI (10
mL). Organic layer washed with water followed by brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated under vacuum and
purified by preparative HPLC (Method: A: 0.02% in ammonia in water,
B: MeOH : Acetonitrile(1 :1), Column: Gemini NX C-18 (150 mm*21.2,
5 .mu.m) to afford the title compound (0.043 g, 43%) as free base.
.sup.1HNMR DMSO-d.sub.6, 400 MHz); .delta. 10.08 (s. 1 H). 9.92 (s,
1 H). 8.50-8.47 (t, 7.76 (s, 1 H), 7.52 (s. 2 H), 7.31-7.27 (t, 1
H), 7.08 (s. 1 H). 6.85 (s, 1 H), 6.41-6.34 (m, 1 H), 6.17-6.12 (m,
5.68-5.65 (m, 1H), 4.64 (s, 2H), 3.99-3.97 (d, 2H), 3.87 (s. 3H).
2.95 (s, 3H), 1.87 (s, 2H), 1.50 (s. 2H). 1.28 (s. 6H); LCMS;
m/z=493.3 (M+H).sup.1.
[0330] The below compounds were prepared by procedure similar to
the one described in Example-13 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvents
at suitable reaction conditions. The physicochemical
characteristics of the compounds are summarized herein below
table.
TABLE-US-00018 Compd No Compound structure Analytical data 80
##STR00197## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.05 (s,
1H), 8.70 (s, 1H), 8.40-8.38 (d, 1H), 7.70 (s, 1H), 7.51 (s, 1H),
7.24-7.21 (t, 1H), 7.03-7.00 (d, 1H), 6.82 (s, 1H), 6.36-6.29 (m,
1H), 6.10-6.05 (m, 1H), 5.60-5.57 (m, 1H), 4.58 (s, 1H), 4.50-4.43
(m, 1H), 3.81 (s, 3H), 3.32 (s, 2H), 2.89 (s, 2H), 2.66 (s, 1H),
2.32 (s, 1H), 1.82 (s, 3H), 1.44 (m, 2H), 1.22 (s, 6H); LCMS: m/z =
507.20 (M + H).sup.- . 81 ##STR00198## .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 10.09 (d, 1H), 9.0- 8.50 (m, 1H), 7.70 (s, 1H),
7.48 (m, 2H), 7.23 (m, 1H), 7.02 (m, 1H), 6.84 (m, 2H), 6.62 (m,
1H), 6.14 (m, 1H), 5.72 (d, 1H), 4.58 (m, 2H), 4.23 (s, 2H), 3.81
(m, 3H), 3.32 (m, 1H), 3.10 (m, 2H), 2.87 (m, 2H), 1.82 (m, 2H),
1.44 (m, 2H), 1.22 (d, 6H); LCMS: m/z = 507.20 (M + H).sup.+. 82
##STR00199## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.39 (s,
1H), 9.0- 8.5 (m, 2H), 8.15 (s, 1H), 7.69 (s, 1H), 7.57-7.46 (m,
2H), 7.20 (t, 1H), 6.98 (m, 1H), 6.82 (m, 2H), 6.33 (m, 1H), 6.05
(d, 1H), 5.57 (d, 1H), 4.56 (m, 2H), 3.81 (m, 3H), 2.89 (m, 2H),
1.82 (m, 3H), 1.41 (d, 6H), 1.22 (m, 6H); LCMS: m/z = 521.2 (M +
H).sup.+. 83 ##STR00200## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 9.49 (s, 1H), 8.59 (s, 2H), 7.69 (s, 1H), 7.56 (m, 1H),
7.52 (m, 1H), 7.23 (t, 1H), 7.03 (m, 1H), 6.81 (m, 1H), 6.25 (m,
1H), 6.05 (d, 1H), 5.61 (d, 1H), 4.57 (m, 2H), 3.81 (m, 3H). 2.89
(m, 2H), 1.82 (m, 3H), 1.37 (m, 4H), 1.22 (d, 6H), 0.94 (m, 2H):
LCMS: m/z = 519.4 (M + H).sup.-. 84 ##STR00201## .sup.1H NMR
(DMSO-d.sub.6, 400 MHz): .delta. 7.69 (s, 1H), 7.57 (m, 2H), 7.42
(m, 1H), 7.23 (t, 1H), 7.06 (d, 1H), 6.7 (m, 1H), 6.64 (m, 1H),
6.15 (m, 1H), 5.67 (d, 1H), 4.55 (m, 2H), 3.79 (m, 3H), 3.35-3.29
(m, 3H), 2.91 (m, 4H), 1.77 (m, 2H), 1.57 (m, 2H), 1.44 (m, 4H),
1.19 (d, 6H): LCMS: m/z = 533.7 (M + H).sup.- . indicates data
missing or illegible when filed
Example-14: Synthesis of
4-acrylamido-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyraz-
olo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-1-methyl-1H-pvrazole-3--
carboxamide (Compound-85)
Step- 1 : Synthesis of
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1.-
3,5]triazin-4-yl)amino)methyl)phenyl)-1-methyl-4-nitro-1H-pyrazole-3-carbo-
xamide
##STR00202##
[0332] To a cooled solution of 1-methyl-4-nitro
1H-pyrazole-3-carboxylic acid (0.108 g, 0.62mmol: US 2009/156582A1)
in DMF (10 mL) at 0.degree. C. was added HATU (0.3 g, 0.78mmol)
followed by DIPEA (0.2 mL, 1.04 mmol) and finally added
N4-(3-arninohenzyl)-8isopr2-(tetrahydro-2H-pyran-4-yl)pyrazolo
[1,5-a][1,3,5]triazine-2,4-diarni ne (0.2 g. 0. 52 mmol:
intermediate-33). The reaction mixture was stirred for 1 h at room
temperature The reaction mixture was quenched with ice-water and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.25 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated the crude residue was purified by 100-200 silica
gel column chromatography to afford the title compound (0.23 g,
82%). LCMS: m/z=535.1 (M+H).sup.-.
Step-2: Synthesis of:
4-amino-N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl )amino)
pyrazolol
[1.5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-1-methyl-1H-pvr-
azole-3-carboxamide
##STR00203##
[0334] To a solution of
N-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,-
3,5]triazin-4-yl)amino)methyl)phenyl)-1-methyl-4-nitro-1H-pyrazole-3-carbo-
xamide(0.35 g, 0.655 mmol) in THF:MeOH:Water (3:2:1 ratio, 15 mL)
were added zinc (0.43 g, 6.554mrnol) and ammonium chloride (0.7 g,
13.1 mmol). The reaction mixture was stirred at room temperature
for 4 h. After completion of reaction the reaction mixture filtered
through celitc bed and diluted with ethyl acetate. The aqueous
layer was separated and extracted with ethyl acetate (2.times.25
mL). The combined organic phase was washed with brine, dried over
Na,S0..sub.1, filtered and concentrated. The residue was purified
by 100-200 silica gel column chromatography to afford desired title
compound (0.3 g, 90%). LCMS: m/z=504.9 (M+H).sup.+.
Step-3: Synthesis of 4- acrylamido-N
-(3-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo
[1.5-a][1,3.5]triazin
-4-yl)amino)methyl)phenyl)-1-methyl-1H-pyrazole-3-carboxamide
##STR00204##
[0336] To a cooled solution of
4-arnino-N-(3-(((8-isopropyl-2(tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,-
5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)-1-methyl-1H-pyrazole-3-carbox-
amide (0.2 g, 0.39 mmol) in DCM (5mL) was added TEA (0.01 mL, 0.79
mmol) and stirred for 5min and added acryloyl chloride (0.035 g,
0.39 mmol) in DCM(1mL) drop wise at 0.degree. C.. The reaction
mixture was allowed to stir at room temperature for 1 h. After
completion of the reaction, the reaction mixture quenched with
water and then diluted with DCM (10 mL). Organic layer washed with
water followed by brine, dried over Na.sub.2SO4, filtered and
concentrated under vacuum and purified by combiflash column by
using 100-200 silica gel and ethyl acetate and hexane as eluent to
afford the title compound (0.062 g, 28%) NMR (DMSO-d.sub.6, 400
MHz): .delta. 10.29 (s, 1H), 9.85 (s, 1H), 8.95-8.70 (m, 1H), 8.42
(s, 1H), 7.90 (s. 1H), 7.74 (s. 2H), 7.32-7.30 (t, 1H), 7.13 (s,
1H), 6.89 (s, 1 H), 6.68-6.64 (m, 1H), 6.31-6.26 (d, 1 H),
5.82-5.79 (d, 1 H), 4.67 (s, 2H), 4.00 (s, 4H), 3.87 (s, 4H), 2.93
(s, 2H), 1.90 (s. 2H), 1.52 (s. 2H), 1.28 (s, 6H): LCMS: m/z=559.1
(M+H).sup..
Example- 15: Synthesis of 1-acryloyl -N-(3((8-i
sopropyl-2-((tetrahyd ro-2H-pyran-4-yl)amino)pyrazolo[1 ,5-a][1
,3,5]triazin-4-yl)amino)phenyl)azetidine-2-carhoxamide
(Compound-86).
Step-1: Synthesis of tert-butyl 2-((3
(8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)
pvrazolo[1,5-a][1,3.5]triazin-4-yl)amino)phenyl)carbamoyl)azctidine-1-car-
boxylate.
##STR00205##
[0338] The process of this step was adopted from step-1 of
example-3 (0.18 g, 48%). LCMS: m/z =551.6 (M+H).sup..
Step-2: Synthesis of N-(3((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl
)arnino)pyrazolo[1,5-a][1.3,5]triazin-4-yl)amino)phenyl)azetidine-2-carbo-
xamide
##STR00206##
[0340] The process of this step was adopted from step-2 of
example-3 (0.11 g crude). LCMS: m/z.+-.451.3 (M+H).sup..
[0341] Step-3: Synthesis of
1-acryloyl-N-(3-((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)arnino)pyrazol-
o[1,5
-a][1,3,5]triazin-4-yl)atnino)phenyl)azetidine-2-carboxamide
##STR00207##
[0342] The process of this step was adopted from step-3 of
example-3. The obtained crude compound was purified by preparative
HPLC (Method: A: water, B: Acetonitrile-Methanol, Column: KINETEX
C18 : 21.2mm*150mm) to afford desired title compound (0.035 g,
35%). .sup.1H NMR (DMSO-d.sub.6, 400MHz): .delta. 10.3-10.1 (m,
1H), 7.82 (s, 1H), 7.50-7.70 (m, 1H), 7.34-7.30 (t, 1 H), 7.20-6.90
(d. 1 H). 6.40-6.33 (m, 1 H), 6_18-6.10 (m, 1 H). 5.77-5.62 (m, 1
H), 5.61-4.90 (m, 1H), 4.88-4.22 (m, 1H), 4.20-3.91 (m, 4H),
3.42-3.30 (m, 2H), 3.30-2.96 (m, 1H), 2.92-2.68 (m, 2H), 2.34-2.33
(m, 2H), 2.26 (s, 1H), 1.87 (s. 2H), 1.48 (s, 2H), 1.27-1.25 (d,
6H); LCMS:=505.3 (M+H).sup.-.
[0343] The below compound was prepared by procedure similar to the
one described in Example- 15 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvents
at suitable reaction conditions. The physicochemical
characteristics of the compounds are summarized herein below
table.
TABLE-US-00019 Compd No Compound structure Analytical data 87
##STR00208## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.1-9.87
(m, 2H), 8.25 (s, 1H), 8.05-8.04 (d, 1H), 7.81 (s, 2H), 7.52 (s,
1H), 7.29-7.25 (t, 1H), 7.20 (s, 1H), 6.90 (s, 1H), 5.57-5.54 (m,
1H), 3.91-3.85 (m, 4H), 2.96-2.93 (m, 2H), 2.67 (s, 1H), 2.33 (s,
2H), 1.84-1.74 (m, 6H), 1.63-1.48 (m, 6H), 1.26-1.24 (d, 6H); LCMS:
m/z = 547.75 (M + H).sup.+. ##STR00209##
Example-16: Synthesis of
1-acryloyl-N-(3-(((8-ethyl-2((1-methylpiperidin-4-yl)oxy)pyrazolo
[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-4-carboxamide
(Compound-88).
##STR00210##
[0345] HATU (0.15 g, 0.39 mmol) followed by DIPEA (0.15 g, 1.16
mmol) was added to a cooled solution of
1-acryloylpiperidine-4-carboxylic acid (0.08 g, 0.43 mmol) in dry
DMF (5 mL ) at 0.degree. C.
N-(3-aminobenzyl)-8-ethyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][-
1,3,5]triazin-4-amine (0.15 g, 0.39 mmol, intermediate-40) was
added to above reaction mixture and the resulting reaction mixture
was stirred for 1 h at room temperature. After completion of
reaction, the reaction mixture was quenched with ice-water and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.50 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated. The residue was purified by prep. HPLC
Column:XBRIDGE (19mm.times.150mm), 0.01% HCOOH (A), Acetonitrile
(B) to afford desired title compound (0.04 g, 18.69%). .sup.1HNMR
(DMSO-d.sub.6400 MHz): .delta. 9.93 (s, 1H), 9.30-9.27 (t, 1H),
7.94 (s, 1H), 7.55 (s, 1H), 7.51-7.49 (d, 1H), 7.25-7.21 (t, 1H),
7.01-6.99 (d, 1H), 6.84-6.77 (m, 1H), 6.H-6.06 (m, 1H), 5.68-5.64
(m, 1H). 4.90 (s. 1H). 4.61-4.60 (d, 2.H), 4.44-4_41 (d. 1H).
4.10-4M7 (d, 1 H). 3_15-3.05 (in. 2H), 2.74-2.52 (in. 4H),
2.35-2_25 (m, 4H), 1.95-1.85 (m, 2.H), 1.81-1.78 (d, 2H). 1.75-1.65
(m, 3H). 1.55-1.35 (m, 3H), 1.23-1.17 (m, 3H); LCMS; mit 548.1 (M+H
).sup.-.
[0346] The below compounds were prepared by procedure similar to
the one described in Example-16 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvents
at suitable reaction conditions. The physicochemical
characteristics of the compounds are summarized herein below
table.
TABLE-US-00020 Compd. No. Compound structure Analytical data 89
##STR00211## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
10.05-9.97 (d, 1H), 9.39-9.36 (t, 1H), 8.05 (s, 1H), 7.53-7.51 (m,
2H), 7.27-7.23 (t, 1H), 7.04-7.02 (d, 1H), 6.95-6.75 (m, 1H), 6.17
(s, 1H), 6.11-6.07 (d, 1H), 5.68-5.55 (t, 1H), 4.95-4.81 (m, 1H),
4.64-4.62 (d, 2H), 4.51-4.22 (m, 1H), 4.15-3.95 (m, 1H), 3.29-3.15
(m, 1H), 3.15- 2.95 (m, 1H), 2.85-2.55 (m, 2H), 2.49-2.35 (m, 1H),
2.25-2.05 (m, 4H), 1.95-1.85 (m, 2H), 1.80-1.55 (m, 3H), 1.45-1.15
(m, 3H), 0.88-0.84 (t, 1H); LCMS m/z = 519 (M + H).sup.+. 90
##STR00212## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.80 (s,
1H), 9.28- 9.25 (t, 1H), 8.03-8.01 (d, 1H), 7.94 (s, 1H), 7.56-
7.54 (d, 1H), 7.50 (s, 1H), 7.24-7.20 (t, 1H), 6.99- 6.97 (d, 1H),
6.38-6.31 (m, 1H), 6.08-6.01 (m, 1H), 5.55-5.52 (m, 1H), 4.95-4.75
(m, 1H), 4.61-4.60 (d, 2H), 3.88 (s, 1H), 2.67-2.52 (m, 3H),
2.49-2.18 (m, 1H), 2.11-1.92 (m, 4H), 1.90-1.49 (m, 11H), 1.29-
1.19 (m, 3H), 0.87-0.82 (m, 3H); LCMS: m/z = 561.2 (M + H).sup.+.
91 ##STR00213## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.77
(s, 1H), 7.95- 7.93 (d, 1H), 7.88 (s, 1H), 7.48-7.46 (m, 2H), 7.19-
7.15 (t, 1H), 6.95-6.93 (d, 1H), 6.16-6.10 (m, 1H), 6.03-5.98 (m,
1H), 5.52-5.49 (m, 1H), 4.78 (m, 2H), 4.55 (s, 4H), 2.94-2.90 (m,
2H), 2.55 (m, 2H), 2.28- 2.20 (m, 1H), 2.11 (s, 3H), 2.07-2.04 (m,
3H), 1.88- 1.79 (m, 3H), 1.58-1.56 (m, 2H), 1.44-1.41 (d, 2H),
1.22-1.13 (m, 8H); LCMS: m/z = 575.4 (M + H).sup.+. 92 & 93
##STR00214## Isomer-1: .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
9.81 (s, 1H), 9.38-9.37 (t, 1H), 8.03-8.01 (d, 1H), 7.96 (s, 1H),
7.57-7.55 (d, 1H), 7.50 (s, 1H), 7.25-7.21 (t, 1H), 7.00-6.99 (d,
1H), 6.39-6.32 (m, 1H), 5.56-5.53 (d, 1H), 4.95-4.90 (m, 1H),
4.75-4.61 (m, 3H), 3.89 (s, 1H), 3.00-2.95 (m, 2H), 2.67-2.60 (m,
1H), 2.38- 2.33 (m, 2H), 2.22 (s, 3H), 2.18-2.07 (m, 3H), 1.80-
1.54 (m, 9H), 1.27-1.25 (d, 6H); LCMS: m/z = 594.5 (M + H).sup.+.
Isomer-2: .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.81 (s,
1H), 9.37-9.34 (t, 1H), 8.03-8.01 (d, 1H), 7.96 (s, 1H), 7.57-7.55
(d, 1H), 7.52 (s, 1H), 7.25-7.21 (t, 1H), 7.00-6.99 (d, 1H),
6.39-6.32 (m, 1H), 5.56-5.53 (d, 1H), 4.98-4.91 (m, 1H), 4.79 (s,
1H), 4.63-4.61 (d, 2H), 3.89 (s, 1H), 3.01-2.88 (m, 2H), 2.67-2.62
(m, 1H), 2.28-2.23 (m, 2H), 2.18 (s, 3H), 2.17-2.07 (m, 2H),
1.95-1.56 (m, 10H), 1.27-1.25 (d, 6H); LCMS: m/z = 594.5 (M +
H).sup.+. 94 ##STR00215## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 10.16 (s, 1H), 9.31- 9.28 (t, 1H), 7.92 (s, 1H), 7.55-7.54
(m, 2H), 7.29- 7.25 (t, 1H), 7.05-7.03 (d, 1H), 6.37-6.30 (m, 1H),
6.14-6.06 (m, 2H), 5.73-5.74 (m, 1H), 4.83-4.79 (m, 2H), 4.62-4.61
(d, 2H), 4.19-4.18 (m, 2H), 3.90 (m, 1H), 2.98-2.94 (m, 1H),
2.67-2.59 (m, 2H), 2.15-2.06 (m, 5H), 1.93-1.91 (m, 2H), 1.63-1.60
(d, 2H), 1.26- 1.24 (d, 6H); LCMS: m/z = 533.2 (M + H).sup.+. 95
##STR00216## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
10.05-10.03 (d, 1H), 9.30-9.27 (t, 1H), 7.90 (s, 1H), 7.51-7.47 (m,
2H), 7.23-7.19 (t, 1H), 7.00-6.98 (d, 1H), 6.57-6.50 (m, 1H),
6.11-6.06 (m, 1H), 5.63-5.60 (d, 1H), 5.00- 4.95 (m, 1H), 4.56 (s,
2H), 3.79-3.74 (m, 2H), 3.68- 3.63 (m, 2H), 3.58-3.48 (m, 4H),
3.17-3.05 (m, 1H), 2.96-2.90 (m, 1H), 2.13-2.02 (m, 2H), 2.02-1.93
(m, 2H), 1.58-1.51 (m, 2H), 1.22-1.20 (d, 6H); LCMS: m/z = 534.3 (M
+ H).sup.+. 96 ##STR00217## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 10.00-9.98 (d, 1H), 9.34-9.31 (t, 1H), 7.95 (s, 1H), 7.51
(s, 2H), 7.27-7.22 (m, 2H), 7.10 (s, 1H), 7.03-7.01 (d, 1H), 6.97
(s, 1H), 6.90-6.79 (m, 1H), 6.11-6.07 (d, 1H), 5.68-5.63 (t, 1H),
5.02 (s, 1H), 4.62-4.61 (d, 2H), 4.49-4.28 (m, 2H), 4.10-4.00 (m,
2H), 3.84-3.80 (m, 1H), 3.23-3.17 (m, 1H), 3.07-2.94 (m, 2H),
2.78-2.67 (m, 1H), 2.33 (m 1H), 1.97-1.94 (m, 2H), 1.73-1.56 (m,
4H), 1.27- 1.25 (d, 6H); LCMS : m/z = 548.4 (M + H).sup.+. 97
##STR00218## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.81 (s,
1H), 9.31- 9.28 (t, 1H), 8.32 (s, 1H), 8.02-8.00 (d, 1H), 7.94 (s,
1H), 7.55-7.51 (m, 2H), 7.25-7.21 (t, 1H), 7.00-6.98 (d, 1H),
6.39-6.32 (m, 1H), 6.08-6.03 (m, 1H), 5.56- 5.53 (m, 1H), 5.04-4.99
(m, 1H), 4.61-4.60 (d, 2H), 3.88-3.79 (m, 3H), 3.46-3.40 (t, 2H),
3.05-2.94 (m, 1H), 2.38 (m, 1H), 1.98-1.94 (m, 2H), 1.80-1.70 (m,
4H), 1.63-1.50 (m, 5H), 1.26-1.25 (d, 6H); LCMS: m/z = 562.4 (M +
H).sup.+. 98 & 99 ##STR00219## Isomer-1: .sup.1H NMR
(CDCl.sub.3-d.sub.6, 400 MHz): .delta. 7.72 (s, 1H), 7.63 (s, 1H),
7.41-7.35 (m, 2H), 7.29-7.26 (m, 1H), 7.08-7.06 (d, 1H), 6.95-6.85
(m, 1H), 6.29-6.24 (m, 1H), 6.11-6.04 (m, 1H), 5.95-5.85 (d, 1H),
5.63- 5.60 (m, 1H), 5.40-5.25 (m, 1H), 4.85-4.74 (m, 2H), 4.15-3.85
(m, 3H), 3.65-3.55 (m, 2H), 3.09-3.06 (m, 1H), 2.38-2.36 (m, 2H),
1.88-1.80 (m, 8H), 1.29- 1.27 (d, 6H); LCMS: m/z = 580.1 (M +
H).sup.+. Isomer-2: .sup.1H NMR (CDCl.sub.3-d.sub.6, 400 MHz):
.delta. 7.71 (s, 1H), 7.56 (s, 1H), 7.47 (s, 1H), 7.43-7.41 (d,
1H), 7.27-7.23 (m, 3H), 7.04-7.02 (m, 2H), 6.28-6.23 (m, 1H),
6.10-6.03 (m, 1H), 5.94-5.84 (d, 1H), 5.62-5.59 (d, 1H), 5.35-4.90
(m, 2H), 4.76-4.73 (m, 2H), 4.25- 4.05 (m, 3H), 3.75-3.51 (m, 2H),
3.06-3.02 (m, 1H), 2.45-2.35 (m, 2H), 2.05-1.70 (m, 7H), 1.28-1.27
(d 6H); LCMS: m/z = 580.1 (M + H).sup.+. 100 ##STR00220## .sup.1H
NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.76 (s, 1H), 9.26 (t, 1H),
7.98-7.96 (d, 1H), 7.90 (s, 1H), 7.53-7.51 (d, 1H), 7.46 (s, 1H),
7.20-7.17 (m, 1H), 6.96-6.94 (d, 1H), 6.34-6.27 (m, 1H), 6.04-5.99
(dd, 1H), 5.51-5.48 (dd, 1H), 5.37-5.35 (m, 1H), 4.57-4.56 (d, 2H),
3.84- 3.80 (m, 2H), 3.77-3.66 (m, 3H), 2.15-2.12 (m, 2H), 1.98-1.96
(m, 2H), 1.75-1.64 (m, 4H), 1.51-1.45 (m, 4H), 1.23-1.21 (d, 6H);
LCMS: m/z = 548.1 (M + H).sup.+. 101 ##STR00221## .sup.1H NMR
(DMSO-d.sub.6, 400 MHz): .delta. 9.81 (s, 1H), 9.27 (t, 1H),
8.03-8.01 (d, 1H), 7.94 (s, 1H), 7.58-7.56 (d, 1H), 7.49 (s, 1H),
7.25-7.21 (t, 1H), 7.00-6.98 (d, 1H), 6.39-6.32 (m, 1H), 6.08-6.03
(dd, 1H), 5.55-5.52 (dd, 1H), 5.37 (m, 1H), 4.61-4.60 (d, 2H), 3.9
(m, 1H), 3.40-3.38 (m, 2H), 2.99-2.96 (m, 1H), 2.67 (m, 1H),
2.66-2.64 (m, 2H), 2.37-2.35 (m, 2H), 2.25 (m, 3H), 1.80-1.77 (m,
3H), 1.58-1.53 (m, 3H), 1.27-1.26 (d, 6H), 1.10-1.07 (t, 2H); LCMS:
m/z = 560.8 (M + H).sup.+. 102 ##STR00222## .sup.1H NMR
(DMSO-d.sub.6, 400 MHz): .delta. 10.08-10.07 (d, 1H), 9.27-9.24 (t,
1H), 7.94 (s, 1H), 7.54 (s, 2H), 7.27-7.23 (t, 1H), 7.05-7.03 (d,
1H), 6.61-6.54 (m, 1H), 6.15-6.10 (m, 1H), 5.76-5.64 (m, 1H),
4.63-4.62 (d, 2H), 4.03-4.01 (d, 2H), 3.80-3.63 (m, 2H), 3.57- 3.47
(m, 2H), 3.21-3.08 (m, 1H), 3.02-2.96 (m, 1H), 2.18-1.94 (m, 3H),
1.27-1.25 (m, 6H), 0.94-0.92 (d, 6H); LCMS: m/z = 506.4 (M +
H).sup.+. 103 ##STR00223## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 10.09 (d, 1H), 9.32 (t, 1H), 7.95 (s, 1H), 7.57 (d, 1H),
7.51 (d, 1H), 7.27 (t, 1H), 7.05 (d, 1H), 6.62 (m, 1H), 6.15 (d,
1H), 5.67 (d, 1H), 4.63-4.61 (d, 2H), 4.36-4.34 (d, 2H), 3.72- 3.67
(m, 3H), 3.59-3.53 (m, 1H), 3.46-3.26 (m, 6H), 3.20 (m, 1H),
2.18-2.08 (m, 2H), 1.27-1.25 (d, 6H); LCMS: m/z = 506.15 (M - H).
104 ##STR00224## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.08
(d, 1H), 9.32 (t, 1H), 7.95 (s, 1H), 7.57 (s, 1H), 7.50 (s, 1H),
7.27 (t, 1H), 7.04 (d, 1H), 6.61 (m, 1H), 6.15 (d, 1H), 5.6 (d,
1H), 4.63-4.61 (d, 2H), 4.35-4.33 (d, 2H), 3.82-3.5 (m, 3H),
3.53-3.42 (m, 3H), 3.41-3.30 (m, 2H), 3.21- 2.97 (m, 6H), 3.00 (m,
1H), 2.19-2.09 (m, 2H), 1.33- 1.25 (d, 6H); LCMS: m/z = 550.2 (M -
H); 105 ##STR00225## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta.
9.80 (s, 1H), 9.30- 9.15 (t, 1H), 8.02-8.01 (d, 1H), 7.85 (s, 1H),
7.55- 7.53 (d, 1H), 7.49 (s, 1H), 7.24-7.20 (t, 1H), 6.98- 6.96 (d,
1H), 6.38-6.32 (m, 1H), 6.08-6.03 (m, 1H), 5.56-5.52 (m, 1H),
4.81-4.80 (m ,1H), 4.60-4.59 (d, 2H), 3.88 (s, 1H), 2.67-2.58 (m,
2H), 2.45-2.25 (m, 1H), 2.16 (s, 3H), 2.10-2.05 (t, 2H), 1.92-1.85
(m, 2H), 1.80-1.50 (m, 10H), 1.10-1.07 (t, 1H), 0.84-0.74 (m, 4H);
LCMS: m/z = 572.9 (M + H).sup.+. ##STR00226##
Example-17: Synthesis of 1-acryloyl-N-(3-(((8-
isopropyl-2-(pipcridin-4-ylo y)pyrazolo[1,5-a][1,
3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-4-carboxamide
(Compound-106)
Step-1: Synthesis of
tert-butyl4((4((3-(1-acryloylpiperidine-4-carboxamido)benzyl)amino)-8-iso-
propvlpvrazolo[1.5-a][1,3.5]triazin 2
yl)oxy)piperidine-1-carboxylate
##STR00227##
[0348] The process of this step was adopted from example-16. The
obtained crude compound was purified by combi flash to afford the
title compound (0.1 g, 50%), LCMS: m/z=647.4 (M+H).sup.-.
Step-2: Synthesis of
1-acrvlovl-N-(3-(((8-isopropyl-2-(piperidin-4-yloxy)pyrazolo[1,5-a][1,3,5-
]triazin-4-yl )arnino)methyl)phenyl)piperidine-4-carboxamide
##STR00228##
[0350] TFA(1 mL.) was added to a solution of tert-butyl
4((4((3-(1-acryloylpiperidine-4-carboxamido)benzynamino)-8-isopropylpyraz-
olo[1,5-a][1,3,5]triazin-2-yl )oxy)piperidine-1-carboxylate(0.1 g,
0.15 mmol)in DCM (10 mL) at 0.degree. C. Then the reaction mixture
allowed to stir at room temperature for 2h. After completion of the
reaction, the reaction mixture was concentrated under vacuum and
purified by prep. HPLC (Method; A; 0.01% Ammonia in water, B:
Acetonitrile-Methanol, Column: EVA-C-18 (21.2 mm*150 mm, 5 .mu.m))
to afford desired title compound (0.06 g. 22%). .sup.1HNMR
(DMSO-d.sub.6, 400 MHz): .delta. 9.93 (s, 1H), 9.39-9.36 (t. 1H).
8.48 (s, 1H), 8.38 (s, 1H), 7.97 (s, 1H), 7.59 (s, 1 H), 7.48-7.46
(d, 1H), 7.25-7.21 (t, 1H), 7.02-7.00 (d, 1H), 6.84-6.77 (m, 1H),
6.H-6.07 (m, 1H), 5.68-5.65 (in, 1H), 5.11 (s, 1H), 4.63-4.61 (d,
2H), 4.45-4.42 (d, 1H), 4.H-4.08 (m, 2H), 3.23 (m, 2H), 3.H-3.08
(m, 3H), 3.01-2.94 (m, 1H), 2.70-2.64 (m, 1H), 2.33 (m, 1H), 2.08
(m, 2H), 1.86-1.78 (m, 3H), 1.49-1.40 (m, 2H), 1.27-1.25 (d, 5H);
LCMS: m/z=547.4 (M+H).sup.-.
[0351] The below compound was prepared by procedure similar to the
one described in Example-17 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvents
at suitable reaction conditions. The physicochemical
characteristics of the compounds are summarized herein below
table.
TABLE-US-00021 Compd. No. Compound structure Analytical data 107
##STR00229## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.92 (s,
1H), 8.36 (s, 1H), 7.95 (s, 1H), 7.53-7.52 (m, 2H), 7.25-7.21 (t,
1H), 7.01-6.99 (d, 1H), 6.84-6.77 (m, 1H), 6.11-6.06 (m, 1H),
5.67-5.64 (m, 1H), 4.60 (s, 2H), 4.44-4.41 (d, 1H), 4.26-4.23 (t,
2H), 4.10-4.07 (m, 2H), 3.6 (s, 2H), 3.16- 2.95 (m, 3H), 2.78-2.75
(t, 1H), 2.70-2.57 (m, 2H), 1.81-1.68 (m, 3H), 1.65-1.59 (m, 2H),
1.51-1.43 (m, 2H), 1.27-1.25 (d, 6H); LCMS: m/z = 535.4 (M +
H).sup.-.
Example-18: Synthesis of 1 -acryloyl -N-(3-(((8-isopropyl-2-((1
-methylpiperidin-4-yl)oxy)
pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carb-
oxamide (Compound-108)
Step-1: Synthesis of
tert-butyl3((3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-
-a][1.3,5]triazin-4-yl
)amino)methyl)phenyl)carbamoyl)pyrrolidine-1-carboxylate
##STR00230##
[0353] To a cooled solution of
1-(tert-hutoxycarlionyl)pvrrolidine-3-carboxylic acid (0.6 g, 0.75
mmol) in DMF (4 mL ) at 0.degree. C. was added HATU (0.36 g,
0.94mmol) and DIPEA (0.21 mL 1.26 mmol) and finally added
N-(3-aminobenzyl)-8-isopropyl-2((1-methylpiperidin
-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-amine (0.25 g, 0.63 mmol,
Intermediate-41). The reaction mixture was stirred for 2h at room
temperature. The reaction mixture was quenched with ice-water and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.25 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated to crude residue. The residue was purified by
combiflash to afford the title compound (0.1 g, 50%). LCMS:
m/z=593.4 (M+H).sup.+.
Step-2: Synthesis of N-(3-(((8-isopropyl-2-((1-
methylpiperidin-4-yl)oxy)pyrazolo[1.5-a][1,3,5]triazin-4-yl)amino)methyl)-
phenyl)pyrrolidine-3-carboxamide
##STR00231##
[0355] TFA (3 mL) was added to a solution of tert-butyl
3-((3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5-
]triazin-4-yl)amino)methyl)phenyl)
carbamoylpyrrolidine-1-carboxylate (0.3 g, 0.50 mmol) in DCM (10
mL) at 0.degree. C. Then the reaction mixture allowed to stir at
room temperature for 2h. After completion of the reaction, the
reaction mixture was concentrated under vacuum (0.25 g, Crude).
LCMS: m/z=493.4 (M+H).sup.+.
Step-3: Synthesis of
1-acryloyl-N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl
)oxy)pyrazolo
[1.5][1,3,5]triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide
##STR00232##
[0357] To a solution of
N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]-
triazin-4-yl)amino)methyl)phenyl)pyrrolidine-3-carboxamide (0.25 g,
0.50 mmol) in DCM (10 mL) was added DIPEA (0.17 mL, 1.01 mmol) and
acryloyl chloride (0.05 g, 0.55 mmol) at 0.degree. C. The reaction
mixture was stirred for 2h at rt. Water was added and extracted
with ethyl acetate. The aqueous layer was separated and extracted
with dichloromethane (2.times.25 mL ). combined organic phase was
washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated to afford crude and purified by prep. HPL C (Method:
A; 0.01% Ammonia in water, 13: Acetonitrile-Methanol, Column;
EVA-C-18 (21.2mm*150 mm, 5 .mu.m) to afford desired title compound
(0.06 g, 22%). LCMS: m/z=547.2.1 (M+H).sup.+. .sup.1HNMR
(DMSO-d.sub.6, 400 MHz); .delta. 10.50 (s, 1H), 9.95 (s, 1H), 8.18
(s, 2H), 8.06-8.02 (m, 2H), 7.94 (s, 1H), 7.50-7.48 (d, 1H),
7.34-7.30 (m, 2H), 6.36-6.33 (m, 1H), 6.09-6.04 (m, 1H), 5.56-5.53
(m, 1 H), 4.90 (s, 1 H), 3.91 (m, 2H), 3.03-2.99 (m, 2H), 2.71-2.67
(m, 3H), 2.01-1.99 (m, 3H), 1.84-1.57 (m, 8H), 1.29-1.27 (d, 6H):
LCMS: m/z=547.4 (M+H).sup..
[0358] The below compounds were prepared by procedure similar to
the one described in
[0359] Example-18 with appropriate variations in reactants,
quantities of reagents, in presence of suitable solvents at
suitable reaction conditions. The physicochemical characteristics
of the compounds arc summarized herein below table.
TABLE-US-00022 Compd. No. Compound structure Analytical data 109
& 110 ##STR00233## Isomer-1: .sup.1H NMR (DMSO-d.sub.6, 400
MHz): .delta. 10.09-10.07 (d, 1H), 9.31-9.28 (t, 1H), 7.93 (s, 1H),
7.53-7.50 (m, 2H), 7.26-7.22 (t, 1H), 7.02-7.01 (d, 1H), 6.60-6.53
(m, 1H), 6.14-6.10 (m, 1H), 5.67-5.64 (m, 1H), 4.85 (s, 1H), 4.59
(d, 2H), 3.79-3.77 (m, 1H), 3.69-3.62 (m, 2H), 3.58- 3.49 (m, 2H),
3.20-3.07 (m, 2H), 3.03-2.92 (m, 2H), 2.69 (s, 1H), 2.20-2.05 (m,
6H), 1.97-1.91 (m, 2H), 1.63 (s, 2H), 1.29-1.24 (m, 6H); LCMS: m/z
= 547.0 (M + H).sup.+. Isomer-2: .sup.1H NMR (DMSO-d.sub.6, 400
MHz): .delta. 10.09-10.07 (d, 1H), 9.30-9.27 (t, 1H), 7.93 (s, 1H),
7.53-7.50 (m, 2H), 7.26-7.22 (t, 1H), 7.02-7.00 (d, 1H), 6.60-6.53
(m, 1H), 6.14-6.10 (m, 1H), 5.66-5.64 (m, 1H), 4.84 (s, 1H),
4.61-4.59 (d, 2H), 3.79-3.77 (m, 1H), 3.69-3.62 (m, 2H), 3.58-3.48
(m, 3H), 3.20-3.07 (m, 1H), 2.97-2.92 (m, 2H), 2.69 (s, 2H),
2.19-2.07 (m, 5H), 1.99-1.91 (m, 2H), 1.64- 1.62 (m, 2H), 1.25-1.24
(m, 6H); LCMS: m/z = 547.0 (M + H).sup.+. 111 ##STR00234## .sup.1H
NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.90 (s, 1H), 9.27 (s, 1H),
7.86 (s, 1H), 7.51 (s, 2H), 7.23-7.19 (t, 1H), 6.99- 6.98 (d, 1H),
6.83-6.76 (t, 1H), 6.10-6.05 (m, 1H), 5.66- 5.63 (m, 1H), 4.83-4.79
(m, 1H), 4.58 (s, 2H), 4.43-4.40 (d, 1H), 4.10-4.06 (d, 1H), 3.15
(s, 1H), 3.09-3.03 (s, 1H), 2.98-2.91 (m, 1H), 2.66-2.53 (m, 4H),
2.14 (s, 2H), 2.09-2.04 (t, 2H), 1.91-1.81 (m, 2H), 1.89-1.80 (m,
2H), 1.63-1.56 (m, 2H), 1.50-1.40 (m, 2H), 1.24-1.23 (d, 6H); LCMS:
m/z = 561.4 (M + H).sup.+. 112 ##STR00235## .sup.1H NMR
(DMSO-d.sub.6,400 MHz): .delta. 9.89-9.81 (d, 1H), 9.27 (s, 1H),
7.93 (s, 1H), 7.63-7.58 (m, 2H), 7.28-7.24 (m, 1H), 7.06-6.04 (m,
1H), 6.85-6.78 (m, 1H), 6.18-6.11 (m, 1H), 5.75-5.73 (m, 1H),
4.85-4.80 (m, 1H), 4.61-4.50 (m, 2H), 4.11-3.94 (m, 1H), 3.57-3.39
(m, 1H), 3.42-3.35 (m, 1H), 3.0-2.95 (m, 1H), 2.60-2.58 (m, 2H),
2.25 (s, 3H), 2.22-2.1 (m, 2H), 1.95-1.75 (m, 2H), 1.63-1.60 (m,
2H), 1.18 (s, 6H); LCMS: m/z = 563.5 (M + H).sup.-. 113
##STR00236## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.05 (s,
1H), 9.32- 9.29 (t, 1H), 7.93 (s, 1H), 7.52-7.48 (m, 2H), 7.28-7.24
(t, 1H), 7.05-7.03 (d, 1H), 6.89-6.82 (m, 1H), 6.17-6.13 (m, 1H),
5.76-5.73 (d, 1H), 4.87-4.86 (m, 1H), 4.62-4.61 (d, 2H), 4.29-4.26
(d, 1H), 4.12-4.11 (d, 1H), 3.89-3.84 (t, 2H), 3.70-3.64 (m, 2H),
3.17-3.16 (d, 2H), 3.00-2.93 (m, 3H), 2.67-2.62 (m, 2H), 2.18-2.13
(m, 3H), 1.92 (s, 3H), 1.26-1.24 (d, 6H); LCMS: m/z = 563.70 (M +
H).sup.+. 114 ##STR00237## .sup.1H NMR (CDCl.sub.3-d.sub.6, 400
MHz): .delta. 7.83 (s, 1H), 7.71-7.67 (m, 2H), 7.36-7.34 (d, 1H),
7.05-7.03 (d, 1H), 6.96-6.94 (m, 1H), 6.27-6.23 (m, 1H), 6.13-6.07
(m, 2H), 5.62-5.59 (m, 1H), 5.15 (s, 1H), 4.72-4.71 (d, 2H),
3.96-3.90 (m, 2H), 3.05-2.94 (m, 2H), 2.49 (s, 3H), 2.45-2.42 (m,
1H), 2.30-2.20 (m, 4H), 2.10-1.65 (m, 9H), 1.51-1.44 (m, 2H),
1.28-1.20 (m, 6H); LCMS: m/z = 574.9 (M + H).sup.-. 115
##STR00238## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.13 (s,
1H), 9.31- 9.28 (t, 1H), 7.93 (s, 1H), 7.62-7.63 (m, 1H), 7.56 (s,
1H), 7.29-7.25 (t, 1H), 7.05-7.03 (d, 1H), 6.43-6.36 (m, 1H),
6.25-6.20 (m, 1H), 5.74-5.71 (m, 1H), 4.83-4.81 (m, 1H), 4.63-4.61
(d, 2H), 3.87 (s, 1H), 3.16 (d, 1H), 2.98- 2.93 (m, 1H), 2.67-2.60
(m, 2H), 2.35 (m, 2H), 2.16-2.10 (m, 4H), 1.92-1.90 (m, 2H),
1.79-1.52 (m, 10H), 1.26- 1.25 (d, 6H); LCMS: m/z = 575.1 (M +
H).sup.+. ##STR00239##
Example-19: Synthesis of
-acryloyl-N-(3-(((8-isopropyl-2-((1-methylpiperidin
-4-yl)oxy)pyrazolo[1 ,5-a][1
,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-3-carboxamide
(Compound-116)
Step-1: Synthesis of tert-butyl
3((3(((8-isopropyl-2-(methylthio)pyrazolo[1
,5-a][1,3,5]triazin-4-yl)arnino)
methyl)phenyl)carbamoyl)piperidine-1-carboxylate
##STR00240##
[0361] DIPEA (1.2 mL 7.23 mmol) was added to a stirred solution of
4-chloro-8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine
(0.35 g, I .44 mmol; intermediate-4) and
tert-butyl3-((3-(aminomethyl)phenyl)carbamoyl)piperidine-1-carboxyl
ate (0.48 g, 1.44 mmol, intermediate-17) in acetonitrile (10 rnL)
at 0.degree. C. The resulting reaction mixture was stirred at
ambient temperature for 4h. After completion of the reaction, the
reaction mixture was concentrated under vacuum and purified the
residue in 100-200 mesh silica column by eluting with 15% ethyl
acetate-hexane to afford the title compound (0.5 g, 64%). LC.MS:
m/z=540.1 (M+H).sup..
Step-2: Synthesis of tert-butyl 3 -((3-(((8-isopropyl-2methyl
sulfonyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)
methyl)phenyl)carbamoyl)piperidine-1-carboxylate
##STR00241##
[0363] mCPBA (0.48 g, 2.78 mmol) was added portion wise to a
stirred solution of tert-butyl
3-((3-(((8-isopropyl-2-(methylthio)pyrazolo[1,5-a
][1,3,5]triazin-4-yl)amino)methyl)phenyl) carbamoyl)
piperidine-1-carboxylate (0.5 g, 0.92 mmol) in DCM (100 mL ). After
reaction completion the reaction mixture was extracted with 2M aq
NaOH and DCM. The organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated to afford the title
compound (0.5 g, 94%). LCMS: m/z=572.2 (M+H).sup.-.
Step-3: Synthesis of tert-butyl
3-((3-(((8-isopropyl-2-((1-methylpiperidin-4-yl
)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl
)amino)methyl)phenyl)carbamoyl)piperidine-1-carboxylate
##STR00242##
[0365] Nall (0.042 g, 1.05 mmol) was added to DMSO (4mL) under
inert atmosphere and stirred for 15 min. 1-Methylpiperidin-4-ol
(0.12 g, 1.05 mmol) was added to the reaction mixture and stirring
continued for 10 min. Added tert-butyl
3-((3-(((8-isopropyl-2-(methylsulfonyppyrazolo[1,5-a][1,3,5]triazin-4-yl)-
amino)methyl)phenyl)carbamoyl) piperidine-1-carboxylate (0.2 g,
0.35 mmol) and stirring continued for 10 min. The reaction mixture
was heated to 60.degree. C. for 1h. After completion of the
reaction, the reaction mixture was cooled to room temperature,
quenched with ice-water and diluted with ethyl acetate. aqueous
layer was separated and extracted with ethyl acetate (2.times.25 mL
). combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by 100-200 mesh silica gel column chromatography to afford
desired title compound (0.2 g, 94%). LCMS: m/z=607.2
(M+H).sup.+.
Step-4: Synthesis of
N-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]-
triazin-4-yl)amino)methyl)phenyl)piperidine-3-carboxamide
##STR00243##
[0366] The process of this step was adopted from step-2 of
example-18 (0.15 g crude). LCMS: m/z=507.2 (M+H).sup.+.
Step-5: Synthesis of
1-acryloyl-N-(3-(((8-isopropyl-2((I-methylpiperidin-4-yl)oxy)pyrazolo
[1.5-a][1.3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-3-carboxamide
##STR00244##
[0368] The process of this step was adopted from step-3 of
example-18. The obtained crude compound was purified by preparative
HPLC (Method: A: 5mm ammonium acetate in water, B:
Acetonitrile-Methanol, Column: Zorbax XDB-C-18 (150 mm*21_2mm, 5
.mu.m)) to afford desired title compound as acetate salt (0.02. g,
18%). .sup.I HNMR (DMSO-d.sub.6, 400 MHz): .delta. 10.0 (d, 1 H),
7.93 (s, 1 H), 7.49-7.52 (m, 2H), 7.24 (t, 1 H), 7.0 (d, 1 H),
6.83-6.85 (m, 2H), 608 (m, 1 H), 5.63 (t, 1H), 4.82-4.83 (m, 1 H),
4.61 (s, 2H), 4.43-4.5 (rn, 1 H), 4.29-4.32 (m, 1 H), 4.0-4.1 (in,
1 H), 2.93-3.06 (in, 1H), 2.59-2.62 (m, 2H), 2.16 (s, 2H),
2.06-2.11 (m 2H), 1.91-1.93 (m, 2H), 1.84 (s, 6H), 1.60-1.84 (in,
3H), 1.26 (dd, 6H). LCMS: m/z=561.25 (M+H).sup..
[0369] The below compounds were prepared by procedure similar to
the one described in example-19 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvenst
at suitable reaction conditions. The physicochemical
characteristics of the compounds arc summarized herein below
table.
TABLE-US-00023 Compd. No. Compound structure Analytcial data 117
##STR00245## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.78-9.77
(d, 1H), 9.33-9.30 (t, 1H), 7.92 (s, 1H), 7.90-7.80 (m, 1H),
7.21-7.12 (m, 2H), 6.95-6.75 (m, 1H), 6.15-6.00 (m, 1H), 5.75-5.55
(t, 1H), 4.85-4.75 (m, 1H), 4.58-4.57 (d, 2H), 4.55-3.95 (m, 2H),
3.40-3.36 (m, 1H), 3.35- 3.15 (m, 1H), 3.05-2.95 (m, 1H), 2.74-2.60
(m, 4H), 2.16-2.08 (m, 4H), 1.95-1.91 (m, 3H), 1.71-1.60 (m, 4H),
1.45-1.35 (m, 1H), 1.25-1.24 (d, 6H); LCMS: m/z = 578.8 (M +
H).sup.+. 118 ##STR00246## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 9.56 (s, 1H), 9.32- 9.28 (t, 1H), 8.01-8.00 (d, 1H), 7.92
(s, 1H), 7.79-7.77 (d, 1H), 7.20-7.15 (t, 1H), 7.12-7.10 (m, 1H),
6.37-6.31 (m, 1H), 6.08-6.03 (dd, 1H), 5.55-5.52 (dd, 1H), 4.84 (m,
1H), 4.58-4.57 (d, 2H), 3.87 (m, 1H), 2.97-2.93 (m, 1H), 2.66-2.60
(m, 2H), 2.16-2.10 (m, 4H), 1.94-1.91 (m, 3H), 1.80-1.77 (m, 3H),
1.64-1.54 (m, 8H), 1.25- 1.23 (d, 6H); LCMS: m/z = 593.25 (M +
H).sup.+. 119 ##STR00247## .sup.1H NMR (DMSO-d.sub.6, 400 MHz):
.delta. 10.01-9.98 (d, 1H), 9.12 (s, 1H), 7.93 (s, 1H), 7.63-7.60
(d, 1H), 7.48-7.47 (d, 1H), 7.27-7.23 (t, 1H), 7.13-7.11 (d, 1H),
6.90-6.79 (m, 1H), 6.11-6.07 (d, 1H), 5.69-5.63 (t, 1H), 5.28-5.26
(m, 1H), 4.80 (m, 1H), 4.50-4.47 (m, 1H), 4.33-4.29 (m, 1H),
4.11-4.00 (m, 1H), 3.23-3.17 (m, 1H), 2.99-2.94 (m, 1H), 2.78-2.67
(m, 4H), 2.17 (s, 3H), 2.11-2.08 (m, 2H), 1.95 (m, 2H), 1.83 (m,
1H), 1.70-1.66 (m, 3H), 1.63-1.58 (d, 3H), 1.56 (m, 1H), 1.26-1.24
(d, 6H); LCMS: m/z = 575.35 (M + H).sup.+.
Example-20: Synthesis of
2-(1-acryloylpiperidine-3-carboxamido)-5-(((8-isopropyl-2-((1-methylpiper-
idin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)pyridine
1-oxide (Compound-120)
Step-1: Synthesis of tert-butyl
34(5-(((8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amin-
o)methyl)pyridin-2-yl)carbamoyl)piperidine-1-carboxylate
##STR00248##
[0371] The process of this step was adopted from step-1 of
example-19 (0.58 g, 52%). LCMS: rn/z =540.9 (M+H).sup.1.
[0372] Step-2: Synthesis of 2-(1-(tert-hatoxvcarbonyl)piperidine 3
carhoxamido)-5-M8-isopropyl-2-(methylsulfonyl)pvrazolo[1,5-a][1.3,5]triaz-
in-4-yl )arnino)rnethyl)pvridine 1-oxide
##STR00249##
[0373] The process of this step was adopted from step-2 of
example-19 (0.6 g). LCMS: m/z=589.35 (M+H).sup..
Step-3: Synthesis of
2-(1-(tert-hutoxycarbonyl)piperidine-3-carboxamido)-5-(((8-isopropyl-2-((-
1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl-
)pyridine 1-oxide
##STR00250##
[0375] The process of this step was adopted from step-3 of
example-19 (0.5 g, 5(l%). LCMS: m/z =624.45 (M+H).sup.-.
Step-4: Synthesis of 5(((8isopropyl2((1-methylpiperidin-4-yl
)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl
)amino)methyl)-2-(piperidine-3-carboxamido)pyridine 1-oxide
##STR00251##
[0377] The process of this step was adopted from step-2 of
example-18 (0.4 g crude). LCMS: m/z =524.1(M+H).sup.
Step-5: Synthesis of
1-acryloyl-N-(5-(((8-isopropyl-2-((1-melhylpiperidin-4-yl)oxy)pyrazolo[1.-
5-a][1,3,5]triazin-4-yl)amino)methyl)-1-(11-oxidanyl)-114-pyridin-2-yl)pip-
eridine-3-carboxamide
##STR00252##
[0379] The process of this step was adopted from step-3 of
example-18. The obtained crude compound was purified by preparative
HPLC (Method: A: 0.01%Ammonia in water, B: Acetonitrile, Column:
KINETEX EVO C-18 (21.1mm*150 mm, 5 .mu.m) to afford desired title
compound (0.007 g,10.2%). .sup.1HNMR DMSO-d.sub.6, 400 MHz);
.delta. 10.62-10.56 (rn, 1H), 9.26 (s, 1H), 8.36 (s, 1H), 8.22-8.20
(d, 1H), 7.93 (s, 1H), 7.40-7.38 (d, 1H), 6.88-6.78 (m, 1H),
6.1-6.03 (t, 1H), 5.68-5.61 (m, 1H), 4.84 (m, 1H), 4.58 (s, 1H),
4.46-4.43 (m, 1 H), 4.19-3.95 (m, 1H), 2.99-2.92 (m, 1 H),
2.85-2.79 (m, 1H), 2.66-2.59 (m, 2H), 2.16-2.13 (m, 3H), 1.95-1.92
(m, 2H), 1.67-1.62 (m, 2H), 1.36 (m, 1 H), 1.25-1.24 (d, 6H), 1.53
(m, 2H), 1.29-1.27 (d, 6H); LCMS: m/z =578.4 (M+H).sup.-.
Example-21: Synthesis of
3-(((8-isopropyl-2((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]tria-
zin-4-yl)amino)methyl)phen yl 4-aceyloylpiperazine-1 1-carboxylate
(Compound-121)
Step-1: Synthesis of
3-3-(((8-isopropyl-2((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]tr-
iazin-4-yl)amino)methyl)phenol
##STR00253##
[0381] The process of this step was adopted from step-3 of
example-19 (0.25 g, 60%). LCMS: m/z=397.4 (M+H).sup.1.
Step-2: Synthesis of 1-(tert-butyl)
4-(3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)
pvrazolo[1,5a][1,3,5]triazin-4-yl )amino)methyl)phenyl)piperazine-1
,4-dicarboxylate
##STR00254##
[0383] To a solution of
3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]tri-
atin-4-yl)arnino)methyl)phenol (0,25 g, 0,63mmol) in 10 mL MDC, was
added DMAP (0.092 g, 0.75 mmol), followed by (tert-butyl
4-(chlorocarbonyl)piperazine-1-carboxylate (Synthesized according
to literature procedure explained in US2007/270433A1) (0.157 g,
0.63 mmol), Reaction mixture was stirred for 4h at RT and then
mixture was quenched to ice cold water and partitioned between
water and DMC. The product was extracted three times with MDC
(25ml.times.3), washed with brine, dried over sodium sulphate,
filtered and concentrated to dryness. The product was purified by
combiflash column by eluting with 30%-50% ethyl acetate-hexane to
afford the title compound (0.22 g, 57%). LCMS: m/z=609.5
(M+H).sup.1.
Step-3: Synthesis of
3-(((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1.5-a][1,3,5]tri-
azin-4-yl)amino)methyl)phenyl piperazine-1-carboxylate
##STR00255##
[0385] The process of this step was adopted from step-2 of
example-18 (0.1 g TFA salt). LCMS: m/z=509.5 (M+H).sup.1.
Step-4: Synthesis of
3-(((8-isopropyl-24(1-methylpiperidin-4-yl)oxy)pyrazolo[1.5-a][1,3,5]tria-
zin-4-yl)amino)methyl)phenyl 4-acrvlovlpiperaiine-1carrboxylate
##STR00256##
[0387] The process of this step was adopted from step-3 of
example-18. The obtained crude compound was purified by preparative
HPI,C (0.01% NH40H in water, 13; Acetonitrile, Column; Gemini NX
C-18 (21.2mm*I50rnm, 5prn) to afford desired title compound ((1.25
g, 12%). .sup.IHNMR (DEMSO-d.sub.6, 400 MHz); .delta. 9.33-9.30
(rn, 1 H), 7.93 (s, 1H), 7.36-7.33 (m, 1 H), 7.22-7.20 (m, 1H),
7.12 (s, 1 H), 6.85-6.78 (rn, 1H), 6.17-6.12 (m, 1 H), 5.74-5.72
(m, 1H), 4.88-4.75 (rn, 1H), 4.65-4.64 (rn, 2H), 3.65-3.60 (rn,
5H), 3.44-3.38 (m, 4H), 3.30-2.93 (m, 1H), 2.71-2.60 (m, 2H), 2.20
(s, 3H), 2.0-1.90 (m, 2H), 1.72-1.61 (m, 2H), 1.18 (s, 6H): LCMS:
m/z 563.5 (M+H).sup..
Example-22: Synthesis of (E)- 1-(4-(dimethylamino)but-2-enoyl)-N-(3
-(((8-isopropyl-2(1-methyl-piperidine-4-yl)oxy)pyrazolo[1,5-a][1,3,5]tria-
zin-4-yl)amino)methyl)phenyl)-piperidine-3-carboxamide
(Compound-122)
##STR00257##
[0389] To a stirred solution of (E)-4-(dimethylamino)but-2-enoic
acid (0.02 g, 0.118 mmol) in (4mI,) was added HATU (0.067 g, 0_177
mmol) followed by DIPEA (0.04mI,, 0.23 mmol) and finally added
N-(3-(48-isopropyl-2-((1-methylpiperidin-4-.sub.3.sup.,1)oxy)pyrazolo[1,5-
-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)piperidine-3-carboxamide
(0.06 g, 0.118 mmol, step-4 product of example-19). The reaction
mixture was stirred for 1 h at room temperature. The reaction
mixture was quenched with ice-water and diluted with ethyl acetate.
The aqueous layer was separated and extracted with ethyl acetate
(2.times.25 mL ). The combined organic phase was washed with brine,
dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue
was purified by prep. HPLC (Method; A; 0.1% TFA in water, B
Acetonitrile-Methanol, Column; X bridge C-18 (150mm*19mm, 5 .mu.m))
to afford desired title compound(0.15 g, 23%),.sup.1HNMR
(DMSO-d(.sub.6, 400 MHz); .delta. 10.01 (s, 1H), 7.98 (s, 1 H),
7.47-7.57 (m, 2H), 7.26 (t, 1H), 7.03 (d, 1H), 6.94 (t, 1H),
6.56-6.57 (m, 1H), 5.21 (s, 1H), 5.02-5.03 (m, 1H), 4.65-4.66 (m,
2H), 4.47 (d, 1H), 4.25 (d, 1H), 4.06-4.07 (m, 2H), 3.87-3.89 (m,
3H), 3.40-3.41 (m, 4H), 2.97-2.99 (m, 2H). 2.76-2.77 (m, 6H), 2.2 g
(d, 2H), 2.28 (d, 2H), 1.99 (d, 2H), 1.80 (m, 3H), 1.66 (m, 1H),
1.26 (d, 6H). LCMS: tniz =618.25 (M+H).sup.1.
[0390] The below compounds were prepared by procedure similar to
the one described in Example-22 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvents
at suitable reaction conditions The plivsicochemical
characteristics of the compounds arc summarized herein below
table.
TABLE-US-00024 Compd. No. Compound structure Analytcial data 123
##STR00258## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 9.77 (s,
1H), 9.31 (s, 1H), 7.92 (s, 1H), 7.85 (m, 1H), 7.21-7.16 (t, 1H),
7.13-7.12 (m, 1H), 6.59 (m, 2H), 4.85-4.83 (m, 1H), 4.57 (s, 2H),
4.44 (d, 1H), 4.2 (d, 1H), 4.01 (t, 1H), 2.98-2.92 (m, 4H),
2.72-2.66 (m, 4H), 2.16 (s, 3H), 2.12-2.09 (m, 7H), 1.92 (m, 3H),
1.72-1.61 (m, 4H), 1.32 (m, 1H), 1.25-1.23 (d, 6H); LCMS: m/z =
636.40 (M + H).sup.+. 124 ##STR00259## .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 9.80 (s, 1H), 9.56 (s, 2H), 9.372-9.32 (m, 1H),
8.18-8.16 (d, 1H), 7.93 (s, 1H), 7.73-7.71 (d, 1H), 7.17-7.08 (m,
2H), 6.53-6.47 (m, 1H), 6.34-6.30 (d, 1H), 5.18 (m, 1H), 4.99 (m,
1H), 4.58-4.55 (t, 3H), 3.91-3.85 (m, 3H), 3.46-3.43 (m, 1H),
3.30-3.27 (m, 1H), 3.12- 3.10 (m, 3H), 2.95-2.93 (m, 1H), 2.78-2.73
(m, 5H), 2.45 (m, 1H), 2.24-2.20 (m, 1H), 2.14-2.11 (m, 1H), 1.96
(m, 1H), 1.72-1.64 (m, 4H), 1.57- 1.54 (m, 4H), 1.22-1.21 (d, 6H);
LCMS: m/z = 650.35 (M + H).sup.+.
Example-23: Synthesis of
4-acrylamido-N-(34(8-isopropyl-24(1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-
-a][1,3,5 ]triazin-4-yl)amino)phenyl)benzamide (Compound-125).
##STR00260##
[0391] Step- 1 : Synthesis of
N-(3((8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1
,3,5]triazin-4-yl)arnino)phenyl)-4-nitrohenzannide
##STR00261##
[0393] DIPEA1PLA (0.7mL 4.13 mmol) was added to a stirred solution
of
4-chloro-8-isopropyl-2-(methlthio)pyrazolo[1,5-a][1,3,5]triazine,
(0.2 g, 0.82 mmol; Intermediate-4) (Synthesized according to
procedure described in WO2013/128028) and
N-(3-aminophenyl)-4-nitrobenzamide (0.21 g, 0.82 mmol,
Intermediate-15) in acetonitrile (5 mL ) at 0.degree. C. The
resulting reaction mixture was stirred at ambient temperature for
4h. After completion of the reaction, the reaction mixture was
concentrated under vacuum and purified the residue in 100-200 mesh
silica column by eluting with 15% ethyl acetate-hexane to afford
the title compound (0.15 g, 40%). LCMS: (biz =464.05
(M+H).sup..
Step-2: Synthesis of N-(3(8-isopropyl-2
methylsulfonyl)pyrazolo[1.5-a][1,3,5]triazin-4-yl)amino)phenyl)-4-nitrobe-
nzamide.
##STR00262##
[0395] The process of this step was adopted from step-2 of
example-19 (0.15 g, 93%). LCMS: rn/z=496.1 (M+H).sup.1.
Step-3: Synthesis of
N-(3(8-isopropyl-2(1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triaz-
in-4-yl)arnino)phenyl)-4-nitrobenzamide
##STR00263##
[0397] The process of this step was adopted from step-3 of
example-19 (0.Ig, 66%). LCMS: m/z =531.15 (M+H) .sup.-.
Step-4: Synthesis of
4-Amino-N-(3((8-isopropyl-2((1-methylpiperidin-4-yl )oxy)pyrazolo
[1,5-a][1,3.5]triazin-4ly)amino)phenyl)benzamide
##STR00264##
[0399] 10% Pd/C (0.02 g) was added to a solution of
N-(34(8-isopropyl-24(1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]tri-
atin-4-yl)amino)phenyl)-4-nitrobenzamide (0.1 g, 0.18 mmol) in
methanol (5mL). The reaction mixture was stirred under hydrogen
balloon pressure for 4h at room temperature. The reaction mixture
was filtered over celite bed, celite bed washed with methanol. The
filtrate was concentrated to obtain title compound (0.08 g crude).
LCMS: m/z 501.15 (M+H).sup..
Step-5: Synthesis of
4-acrylamido-N-(4(8-isopropyl-2(1-methylpiperidin-4-yl)oxy)
pvrazolo[1,5-a][1.3,5]triazin-4-yl)amino)plenyl)benzamide
##STR00265##
[0401] The process of this step was adopted from step-3 of
example-18. The btained crude compound was purified by prep. HPLC
(Method: A: 10 mm ammonium acetate in water, B:
Acetonitrile-Methanol, Column: XDB-C-18 (250 mm*21.2 mm, 5 .mu.m))
to afford desired title compound as acetate salt(0.008 g, 10%).
.sup.1HNMR (DVSO-d.sub.6400 MHz): .delta. 10.5 (s, 1 H), 10.2 (s, 1
H), 8.25 (s, 1H), 8.03 (s, 1H), 7.98 (d, 2H), 7.81 (d, 2H),
7.53-7.54 (m, 1 H), 7.41 (d, 1H), 7.37 (d. HI), 6.44-6.52 (m, 2H),
6.30 (dd. HI). 5.81 (dd, 1 H), 4.9-4.92 (m, 1 H), 3.16 (s. 3H),
2.94-3.05 (m, 4H), 2.59-2.60 (m, 3H), 1.62-1.65 (m, 2H), 1.28 (d,
6H). LCMS: m/z=555.45 (M+H).sup..
[0402] The below compounds were prepared by procedure similar to
the one described in Example-23 with appropriate variations in
reactants, quantities of reagents at suitable reaction conditions.
The physicochemical characteristics of the compounds are summarized
herein below table.
TABLE-US-00025 Compd. No. Compound structure Analytical data 126
##STR00266## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.52 (s,
1H), 9.95 (s, 1H), 8.18 (s, 2H), 8.06-8.04 (m, 1H), 8.02 (m, 1H),
7.94 (s, 2H), 7.50-7.48 (d, 1H), 7.34-7.30 (m, 2H), 6.36-6.33 (m,
1H), 6.09-6.04 (d, 1H), 5.56-5.53 (d, 1H), 4.91 (m, 1H), 3.91 (m,
1H), 3.03-2.99 (m, 3H), 2.67-2.50 (m, 3H), 2.18-2.01 (m, 3H),
1.99-1.84 (m, 2H), 1.81-1.57 (m, 7H), 1.29-1.27 (d, 6H); LCMS: m/z
= 561.40 (M + H).sup.+. 127 ##STR00267## .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 10.52 (s, 1H), 10.05 (s, 1H), 8.01-7.99 (d, 1H),
7.45-7.43 (d, 1H), 7.37-7.35 (d, 1H), 7.32-7.28 (t, 1H), 6.86-6.79
(m, 1H), 6.12-6.07 (dd, 1H), 5.68-5.65 (dd, 1H), 4.89 (m, 1H), 4.5
(d, 1H), 4.1 (m, 1H), 3.38-3.36 (m, 2H), 3.10-3.02 (m, 1H),
3.0-2.98 (m, 1H), 2.7-2.62 (m, 4H), 2.15-2.11 (m, 2H), 2.11-2.06
(m, 2H), 1.99-1.97 (m, 2H), 1.86-1.83 (m, 2H), 1.66-1.64 (m, 2H),
1.53 (m, 2H), 1.29-1.27 (d, 6H); LCMS: m/z = 546.8 (M + H).sup.+.
##STR00268##
Example-24: Synthesis of
N-(1-acryloylpiperidin-4-yl)-4(8-isopropyl-2-((1-methylpiperidin-4-yl)oxy-
)pyrazolo[1 ,5-a ][1,3,5]triazin-4-yl)amino)benzamide
(Compound-128).
Step-1: Synthesis of
tert-bulyl-4-(3((8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin--
4-yl)amino)benzamido)piperine-1-carboxylate
##STR00269##
[0404] The process of this step was adopted from step-1 of
example-23 (0.5 g crude): LCMS: m/z526.40 (M+H).sup.1.
Step-2: Synthesis of tert-butyl
4-(3((8-isopropyl-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl)am-
ino)benzarnido)piperidine-1-carboxylate
##STR00270##
[0406] The process of this step was adopted from step-2 of
example-19 (0.4 g, 75%). LCMS: m/z =556.26 (M+H).sup.1 .
Step-3: Synthesis of tert-butyl
4-(3(8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1.5-a][1,3,5]tri-
azin-4-yl)amino)benzarnido)piperidine-1-caboxylate
##STR00271##
[0408] The process of this step was adopted from step-3 of
example-19 (0.2 g, 47%). LCMS: m/z =593.90 (M+H).sup.1.
Step-4: Synthesis of
3-((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1.3,5]tria-
zin-4-yl)amino)-N-(piperidin-4-yl)benzamide
##STR00272##
[0410] The process of this step was adopted from step-2 of
example-18 (0.15 g crude). LCMS: rn/z =493.40 (M+H).sup.1.
Step-5: Synthesis of N
-(1-acryloylpiperidin-4-yl)-3-isopropyl-2-((1-methylpiperidin-4-yl)oxy)py-
razo[1.5-a][1,3,5]triarin-4-yl)amino)benzarnide
##STR00273##
[0412] The process of this step was adopted from step-3 of
example-19. The obtained crude compound was purified by preparative
HPLC (Method: A: 0.01%. NH.sub.4OH in water, B: Acetonitrile,
Column: Kinetex EVO C- (21.2mtn*150mm, 5 .mu.m)) to afford desired
title compound (0.010 g,9%),.sup.1HNMR (DMSO-d.sub.6, 400 MHz): d
8.35-8.33 (d, 1H), 8.21 (s, 1H), 8.02 (s, 1H), 7.86-7.84 (d, 1H),
7.65-7.63 (d, 1H), 7.50-7.46 (t, 1H), 6.85-6.78 (in. 1H), 6.H-6.07
(m,
[0413] 1H), 5.69-5.66 (in, 1H), 4.87-4.84 (rn, 1H), 4.39-4.36 (m,
1H), 4.07-3.98 (m, 2H), 3.22-3.14 (m, 2H), 3.04-2.97 (in, 1H),
2.83-2.80 (m, 2I 1), 2.67-2.61 (m, 2I 1), 2.15-2.10 (m, 4H),
2.08-1.98 (m, 2H), 1.88 (m, 2H), 1.87-1.68 (m, 2H), 1.66-1.45 (m,
2H), 1.28-1.27 (d, 6H): LCIVIS: /mz=547.40 (M+H).sup.-.
[0414] The below compound was prepared by procedure similar to the
one described in Example-24 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvents
at suitable reaction conditions. physicochemical characteristics of
the compounds are summarized herein below table.
TABLE-US-00026 Compd. No. Compound structure Analytical data 129
##STR00274## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.65 (s,
1H), 8.31-8.30 (d, 1H), 8.13 (s, 1H), 7.94 (s, 1H), 7.78 (s, 1H),
7.56 (s, 1H), 7.42 (m, 1H), 6.81-6.50 (m, 1H), 6.06-6.02 (m, 1H),
5.64-5.61 (d, 1H), 4.81 (m, 1H), 3.99-3.90 (m, 3H), 3.75 (m, 1H),
2.99-2.92 (m, 3H), 2.69-2.59 (m, 2H), 2.07-2.04 (m, 2H), 2.03-1.95
(m, 2H), 1.95-1.92 (m, 2H), 1.73 (m, 2H), 1.63-1.61 (m, 2H), 1.39
(m, 2H), 1.25-1.23 (d, 6H); LCMS: m/z = 547.2 (M + H).sup.+.
##STR00275##
Example-25: Synthesis of
(E)-4-(4-(dimethylamino)but-2-enamido)-N-(3-((8-isopropyl-2-((1-methylpip-
eridin-4-yl)oxy)pyrazolo[1,5-a][1,3,5
]triazin-4-yl)amino)phenyl)benzamide (Compound-130)
##STR00276##
[0416] Oxalyl chloride (0.025 g, 0.2 mmol) was added to a solution
of bromocrotonoie acid (0.033 g, 0.2 mmol) in dry DCM (2mI,) was
added followed by one drop of DMF(5 mL) was added. The reaction
mixture was stirred for 2 hat room temperature and evaporated under
reduced pressure to obtain (E)-4-hromobut-2-enoyl chloride. This
was dissolved in dry DCM (5 mL) and added to a solution of
4-amino-N-(3-48-isopropyl-2-((1-methylpiperidin-4-yl)oxy)pyrazolo[1,5-a][-
1,3,5]triazin-4-yl)amino)phenyl)benzamide (0.05 g, 0.1rnmol; step-5
product of example-23) in acetonitrile (2 mL) at 0.degree. C. The
resulting mixture was stirred for 1 h at same temperature and
dimethyl amine (1 mL, 20 mmol) was added and continued stirring for
4h at room temperature. The reaction mixture was quenched with
ice-water and diluted with ethyl acetate. The aqueous layer was
separated and extracted with ethyl acetate (2.times.25 mL). The
combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by prep. HPLC (Method: A: Ammonium acetate. B:
Acetonitrile-Water (1:1), Column: Gemini-NX to afford desired title
compound (0.05 g, 20%). .sup.1HMNR (DMSO-d.sub.6, 400 MHz): .delta.
10.68-10.65 (d, 2H), 10.32-10.29 (d, 1 H), 9.91 (s, 1 H), 8.25-8.19
(d, 1 H), 8.08 (s, 1H), 8.00-7.98 (d, 2H), 7.84-7.82 (d, 2H),
7.55-7.52 (t, 1 H), 7.47-7.37 (m, 2H), 6.82-6.75 (m, 1 H),
6.51-6.48 (d, 1 H), 5.33 (s, 1 H), 5_11 (s. 1H), 3.96 (s, 3H),
3.51-3.48 (d, 2H), 3.35-3.32 (m, 2H), 3.17-102 (m, 4H), 2.82 (s,
7H), (m, 2H). 2.33 (s, 1 H), 2.24-2.21 (d, 1 H), 2.05-1.98 (t, 1H),
1.81-1.78 (m, 1 H), 1.31-1.29 (d, 6H); LCMS: m/z=612.4
(M+H).sup.1.
Example-26: Synthesis of
(1s,4)-4(E)-4-(dimethylarnino)but-2-enamido)-N-(3-((8-isopropyl-2-((1-met-
hylpiperidin-4-yl)oxy)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)phenyncyclo-
hexane-1-carboxamide (Compound-131)
##STR00277##
[0418] To a cooled solution of (E)-4-(dimethylarnino)but-2-enoic
acid hydrochloride (0.025 g, 0.15 mmol) in DMF (2 mL) at 0.degree.
C. was added HATU (0.068 g, 0.18 mmol) followed by DIPEA (0.52mL,
0.30 mmol) and added
(1,4-cis)-4-amino-N-(3-((8-isopropyl-2-((1-methylpiperidin-4-yl)oxy-
)pyrazolo[1,5-a][1,3,5]triazin-4-yparnino)plienyl)c yclohexane-1
-carboxamide (0.076 g, 0.15 mmol). The reaction mixture was stirred
for 2 hat room temperature. The reaction mixture was quenched with
ice-water and diluted with ethyl acetate. The aqueous layer was
separated and extracted with ethyl acetate (2.times.25 mL ). The
combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated (0.025 g, 26%).
.sup.1HNMR (DMSO-d.sub.6, 400 MHz): .delta. 10.52 (s, 1H), 9.94 (s,
1H), 8.02 (s, 1 H) 7.98-7.94 (m, 2H), 7.50-7.48 (d, 1H), 7.36-7.28
(m, 2H), 6.56-6.49 (m, 1 H), 6.21-6.17 (d, 1 H), 4.91-4.86 (m, 1H),
3.89 (m, 1 H), 3.41-3.40 (m, 2H), 3.17 (d, 1 H), 2.99-2.66 (m, 2H),
2.66-2.51 (m, 2H), 2.50-2.49 (m, 6 H), 2.44-2.32 (m, 2H), 2.09-2.00
(m, 7H), 1.98-1.97 (m, 2H) 1.86-1.81 (m, 2 H), 1.73-1.55 (m, 6H),
1.25-1.23 (d, 6 H), 1.10-1.07 (m, 2 H); LCMS: m/z=618.50
(M+H).sup.-.
[0419] The below compounds were prepared by procedure similar to
the one described in Example-26 with appropriate variations in
reactants, quantities of reagents, in presence of suitable solvents
at suitable reaction conditions. The physicochemical
characteristics of the compounds are summarized herein below
table.
TABLE-US-00027 Compd. No. Compound structure Analytical data 132
##STR00278## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.5 (s,
1H), 9.95 (s, 1H), 8.02-7.99 (d, 2H), 7.91 (d, 1H), 7.46 (d, 1H),
7.36-7.28 (m, 2H), 6.57-6.50 (m, 1H), 6.01 (d, 1H), 4.93 (m, 1H),
3.58 (m, 1H), 3.04-2.91 (m, 4H), 2.67-2.63 (m, 2H), 2.32-2.29 (m,
2H), 2.13-2.10 (m, 3H), 2.07-2.00 (m, 7H), 1.97-1.89 (m, 2H),
1.89-1.87 (m, 4H), 1.69-1.64 (m, 2H), 1.55-1.46 (m, 3H), 1.33-1.27
(d, 6H). LCMS: m/z = 618.45 (M + H).sup.+. 133 ##STR00279## .sup.1H
NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.76- 10.74 (d, 1H),
10.02-9.55 (m, 2H), 8.40 (m, 1H), 8.18 (s, 1H), 8.05 (s, 1H),
7.88-7.83 (t, 1H), 7.66-7.64 (d, 1H), 7.50- 7.46 (t, 1H), 7.22 (s,
2H), 7.09 (s, 3H), 6.96 (s, 1H), 6.88-6.83 (dd, 1H), 6.58-6.52 (m,
1H), 5.18 (m, 1H), 4.43-4.40 (m, 1H), 3.94-3.90 (d, 1H), 3.85-3.77
(m, 2H), 3.47 (m, 1H), 3.16 (m, 2H), 3.01-2.94 (m, 1H), 2.91 (m,
2H), 2.77-2.74 (m, 2H), 2.68 (s, 1H), 2.59 (m, 1H), 2.19 (m, 1H),
2.04- 2.00 (m, 1H), 1.91 (m, 1H), 1.79-1.77 (m, 2H), 1.61 (m, 1H),
1.41 (m, 1H), 1.27-1.25 (d, 6H); LCMS: m/z = 604.2 (M + H).sup.+.
134 ##STR00280## .sup.1H NMR (DMSO-d.sub.6, 400 MHz): .delta. 10.69
(s, 1H), 8.32-8.30 (d, 1H), 8.25 (s, 1H), 8.04 (s, 1H), 7.87-7.85
(d, 1H), 7.66-7.64 (d, 1H), 7.52-7.46 (t, 1H), 6.66-6.56 (m, 1H),
4.91- 4.90 (m, 1H), 4.39-4.36 (m, 1H), 4.06- 4.03 (m, 3H),
3.20-3.15 (m, 2H), 3.06- 3.00 (m, 3H), 2.79-2.66 (m, 3H), 2.32-
2.08 (m, 9H), 2.00-1.99 (m, 2H), 1.86 (m, 2H), 1.72-1.70 (m, 2H),
1.43-1.41 (m, 2H), 1.29-1.27 (d, 6H), 1.19-1.06 (m, 1H); LCMS: m/z
= 604.40 (M + H).sup.+. ##STR00281##
Example-27: Synthesis of (1s,
4s)-4-(((E)-4-(dimethylamino)-4-oxobut-2-cn-1
-yl)arnino)-N-(3-(((8-isopropyl-2-((1
-methylpiperidin-4-yl)oxy)pyrazolo
[1,5-a][1,3,5]triazin-4-yl)amino)rnethyl)phenyl)cyclohezane-1-carboxamide
(Compound-135)
Step-1: Synthesis of tert-butyl
((1s,4s)-4((3-(((8-isopropyl-2(1-methylpiperidin-4-yl)oxy)pyrazolo[1.5-a]-
[1,3,5]triazin-4-yl)amino)methyl)phenyl)carbamoyl)cyclohexyl)
carbamate
##STR00282##
[0421] The process of this step was adopted from step-1 of
example-18 (0.3 g, 95%). LCMS: m/z =621.85 (M+H).sup..
Step-2: Synthesis of
(1s,4s)-4-amino-N-(3-(((8-isopropyl-2(1-methylpiperidin-4-yl)oxy)pyrazolo-
[1.5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohcxane-1-carhoxamide.
##STR00283##
[0423] The process of this step was adopted from step-2 of
example-18 (0.2 g, 94.7). LCMS: m/z =521.45 (M+H).sup.-.
Step-3: Synthesis of
(1s,4-4-(((E)-4-(dimethylamino)-4-oxobut-2-en-1-yl)amino)-N-(3-(((8-isopr-
opyl-2((1-rnethylpiperidin-4-yl
)oxy)pyrazolo[1.5-a][1,3.5]triazin-4-yl)amino)methyl)
phenyl)cyclohexane-1-carboxamicle
##STR00284##
[0425] To a solution of
(1,4-cis)-4-amino-N-(3-(((8-isopropyl-2((1-methylpiperidin-4-yl)oxy)pyraz-
olo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl)cyclohexane-1-carhoxamid-
e (0.1 g,0.19 mmol) and (E)-4-bromo-N,N-dimethylbut-2-enamide
(0.044 g,0.23 mmol) in ACN (3 mL ) was added DIPEA (0.06mL, 0.384
mmol) at RT, then stirred for 12hr, After reaction completion,
quenched with water and diluted with ethyl acetate. The aqueous
layer was separated and extracted with ethyl acetate (2.times.15
mL). The combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by preparative HPLC (Method: A: 0.1% TFA in Water, B:ACN,
Column: EVO C18 (21.2mm.times.150 mm particle size 5 .mu.m)) to
afford desired title compound (0.030 g, 20%). .sup.1H NMR
(DMSO-d.sub.6, 400MHz): 9.84 (s, 1H), 7.97 (s, 1H), 7.85 (m, 1H),
7.55-7.53 (d, 1H), 7.48 (s, 1H), 7.25-7.21 (t, 1H), 7.05-6.96 (rn,
2H), 6.70-6.66 (m, 1H), 5.12 (s, 1H), 4.62 (s, 2H), 4.25-4.19 (m,
3H), 3.44-3.43 (d, 3.09-2.89 (m, 4H), 2.80-2.77 (rn, 3H), 2.66 (s,
1H), 2.32-2.35 (in, 3I 1), 2.11-2.10 (m, 2H), 1.86-1.84 (d, 2H),
1.74-1.68 (m, 4H), 1.62-1.57 (rn, 2H), 1.22 (s, 6H): LCMS: m/z
=632.80 (M+H).sup.-.
Example-28: Synthesis of Synthesis of
1-acryloyl-N-(3-(((8-isopropyl-2-methoxypyrazolo[1,5-a][1,3,5
]triazin-4-yl)amino)methyl)phenyl)piperidine-4-carboxamide
(Compound- 136). Step- 1 : Synthesis
of8-isopropyl-2-methoxy-N-(3-nitrobenzyl)pvrazolo[1.5-a][1,3.5]triazin-4--
amine
##STR00285##
[0427] NaH(0.08 g, 2.05 mmol) was added to DMSO (4 mL) under inert
atmosphere and stirred for 15 min. tert-butyl
(4-hydroxybutyl)carbamate (0.38 g, 2.05 mmol) was added to the
reaction mixture and continued stirring for 10 min.
8-isopropyl-2-(methylsulfonyl)-N-(3-nitrobenzyl)pyrazolo[1,5][1,3,5]triaz-
in-4-amine (0.2 g. 0.51 mmol) was added and stirred for 10 min.
Then the reaction mixture allowed to heat at 60.degree. C. for 1 h.
After completion of the reaction, cooled to room temperature and
was quenched with ice-water anti diluted with ethyl acetate. The
aqueous layer was separated anti extracted with ethyl acetate
(2.times.25mI,). combined organic phase was washed with brine,
dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue
was purified by 100-200 silica gel column chromatography to afford
desired title compound (0.1 g, 57%). LCMS: m/z=343.2
(M+H).sup..
[0428] Step-2: Synthesis of
N-(3-aminobenzyl)-8-isopropyl-2-methoxypyrazolo[1,5-a][1.3,5]triazin-4-am-
ine:
##STR00286##
[0429] To a solution of
8-isopropyl-2-methoxy-N-(3-nitrobenzyl)pyrazolo[1,5-a][1,3,5]amine
(0.1 g, 0.29 mmol) in THF:McOH 1:Water (3:2:1) were added zinc
(0.08 g, 1.46 mmol) and ammonium chloride (0.078 g, 1.46 mmol). The
reaction mixture was stirred at room temperature for 4 h. After
completion of reaction the reaction mixture filtered through
celiteand diluted with ethyl acetate. The aqueous layer was
separated and extracted with ethyl acetate (2.times.25 mL ). The
combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by 100-200 silica gel column chromatography to afford
desired title compound (0.06 g, Crude). LCMS: m/z=313.25
(M+H).sup.1 .
Step-3: Synthesis of 1
-acryloyl-N-(3-(((8-isopropyl-2-methoxypyrazolo[1 ,5-a][1 .3
,5]triazin-4-yl)amino)methyl)phenyl)piperidine-4-carboxamide
##STR00287##
[0431] The process of this step was adopted from example-16. The
obtained crude compound was purified by combiflash to afford the
title compound (0.05 g, 65.7%). .sup.1HNMR (DMSO-d.sub.6, 400 MHz):
.delta. 9.92 (s, 1H), 9.30-9.27 (t, 1H), 7.95 (s, 1H), 7.55-7.51
(rn, 2H), 7.25-7.21 (t, 1 H), 7.02-7.01 (d, 1H), 6.84-6.77 (m, 1H),
6.H-6.06 (d, 1H), 5.67-5.64 (d, 1H), 4.60-4.59 (d, 2H), 4.44-4.41
(d, 1H), 4.10-4.07 (d, 1H), 3.83 (s. 31 i), 3.59 (s, 11 i),
3.07-2.98 (in. 2H), 2.67-2.57 (m, 4H), 1.81-1.75 (t, 21 I),
1.49-1.46 (m,31 I), 1.27-1.25 (d, 6H): LCMS: m/z=475.3
(M+H).sup.1.
Example-29: Synthesis of
(E)-4-(dimethylamino)-1-(4-(2-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl-
)amino)pyrazolo[1,5a][1,3,5]triazin-4-yl)arnino)methyl)phenyl)piperazin
-1-yl)but-2-en-1-one (Compound-137)
-1: Synthesis of
tert-butyl4-(2-(((8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-
-4-yl)amino)methyl)phenyl)piperazine-1-carboxylate
##STR00288##
[0433] The process of this step was adopted from step-1 of
example-4 (0.8 g, 80%). LCMS: m/z =498.2 (M+H).sup..
Step-2: Synthesis of
4-(tert-butoxycarbonyl)-1-(2-(((8-isopropyl-2-(methylsulfonyl)pyrazolo
[1.5-a][1.3.5]triazin-4-yl)amino)rnethyl)phenyl)piperazine
1-oxide
##STR00289##
[0435] The process of this step was adopted from step-2 of
example-4 (0.7 g, crude). LCMS: m/z =546.3 (M+H).sup.-.
Step-3: Synthesis of
4-(tert-butoxylcarbonyl)-1-2-(((8-isopropyl-2((tetrahydro-2H-pyran-4-yl)a-
mino)pyrazolo[1.5-a][1,3,5]triazin-4-yl
)amino)methyl)phenyl)piperazine 1-oxide
##STR00290##
[0437] The process of this step was adopted from step-3 of
example-4 (0.6 g, crude). LCMS m/z =567.7 (M+H).sup..
Step-4: Synthesis of
8-isopropyl-N4-(2-(piperazin-1-yl)benzyl)-N2-(tetrahydro-2-pyran-4-yl)pyr-
azolo[1,5-a][1.3.5]triazine-2.4-diamine.
##STR00291##
[0439] The process of this step was adopted from step-4 of
example-4 (0.40 g, crude). LCMS: m/z=451 .3 (M+H).sup.1.
Step-6: Synthesis of
(E)-4-(dimethylatnino)-1-(4-(2-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-y-
l)amino)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)phenyl),piperazin--
1-yl)but-2-en-1-one
##STR00292##
[0441] To a cooled solution of (E)-4-(dimethylamino)but-2-enoic
acid (0.073 g, 0.44 mmol) in DMF (5 mI,) at 0.degree. C. was added
NATU (0.25 g, 0.666 mmol) followed by DIPEA (032 mL., 1.77 mmol)
and finally added 8-i
sopropyl-N4-(2-(piperazin-1-yl)benzyl)-N2-(tetrahydro-2H-pyran-4-yl)p-
yrazolo[1,5-a][1,3,5]triazine-2,4-diamine(0.2 g, 0.444 mmol).
reaction mixture was stirred for 1 h at room temperature.sub.-- The
reaction mixture was quenched with ice-water and diluted with ethyl
acetate. The aqueous layer was separated and extracted with ethyl
acetate (2.times.25m1). The combined organic phase was washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated the
crude residue was purified by preparative HPLC (Method: A: 0.01%
TFA in water, B: ACN: MeOH, Column: Kinetex Evo C 18:(150 mm*21.2
mm) to afford desired title compound as TFA salt. To removal TFA
passed the compound through vari pure basic resin column using McOH
as eluent to afford the title compound as free base (0.07 g, 38%).
.sup.1H NMR (DMSO-d.sub.6400MHz): .delta. 8.80-8.50 (d, 1 H), 7.72
(s, 1H), 7.23-7.16 (m, 3H), 7.06 (t, HI), 6.80 (s, 1H), 6.64-6.63
(m, 2H), 4.76-4.74 (d. 2H), 3.82-3.73 (m, 8H), 3.04-3.03 (d, 2H),
2.92-2.86 (m, 5H), 2.67 (s. 1H), 2.45 (s, 1H). 2.15 (s, 6H): 1.82
(s, 1H), 1.46 (s, 2H), 1.24-1.22 (d, 7H): LCMS: m/z=562.45
(M+H).sup.+.
Example-30: Synthesis of
(E)-1-(4-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1.5-a-
][1,3,5]triazin-4-yl)amino)methyl)piperidin-1-yl)-4-(pyrrolidin-1-yl)but-2-
-en-1-one (Compound-138)
Step-1: Synthesis of tert-butyl
4-(((8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)m-
ethyl)piperidine-1-carboxylate
##STR00293##
[0443] tert-Butyl 4-(aminomethyl)piperidine-1-carboxylate (0.619 g,
2.89 mmol) was added to a solution of
4-chloro-8-isopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine
(0.7 g, 2.89 mmol) in acetonitrile (50 mL) at 0.degree. C.. The
resulting reaction mixture was stirred at ambient temperature for
15 h. After completion of the reaction, the reaction mixture was
concentrated under vacuum and purified the residue in 100-200 mesh
silica column by eluting.sup., with 5% ethyl acetate-hexane to
afford the title compound (0.9 g, 74%). LCMS: m/z=421.4
(M+H).sup.-.
Step-2: Synthesis of tert-butyl
4-(((8-isopropyl-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl)ami-
no)methyl)piperidine-1-carboxylate
##STR00294##
[0445] The process of this step was adopted from step-2 of
example-4 (0.5 g, 58%) m/z =451.4 (M+H).sup.-.
Step-3: Synthesis of tert-butyl
4-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1,5-a][1,3,5-
]triazin-4-yl)amino)methyl)piperidine-1-carboxylate
##STR00295##
[0447] The process of this step was adopted from step-3 of
example-4 (0.32 g, 67.6%). LCMS: m/z=474.5 (M+H).sup.1.
[0448] Step-4: Synthesis of
8-isopropyl-N4-(piperidin-4-ylmethyl)-N2-(tetrahydro-2H-pyran-4-yl)pyrazo-
lo[1,5-a][1,3,5]triazine-2,4-diamine.
##STR00296##
[0449] The process of this step was adopted from step-4 of
example-4 (0.22 g, 93.6%). LCMS: m/z=374.1 (M+H).sup.+.
Step-4: Synthesis of
(E)-4-bronno-144-(8-isopropyl-2-((tetrahydro-2H-pyran-4-yl )amino)
pvrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)piperidin-1-yl)but-2-en-1-
-one
##STR00297##
[0451] To a cooled solution of (E)-4-bromobut-2-enoic acid (0.66 g,
0.8 mmol) in DMF (10 mL) at 0.degree. C. was added HATU (0.3 g, 0.8
mmol) followed by DIPEA (0.139 mL, 0.8 mmol) and finally added
8-isopropyl-N4-(piperidin-4-ylmethyl)-N2-(tetrahydro-2 H-pyran
-4-yl) pyrazolo[1,5-a][1,3,5]triazine-2,4-diamine (0.15 g, 0.4
mmol). The reaction mixture was stirred for 1 h at room
temperature. The reaction mixture was quenched with ice-water and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.25 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated. The residue was purified by 100-200 silica gel
column chromatography to afford desired title compound (0.135 g,
86.5%). LCMS: m/z=522.0 (M+H).sup.1.
Step-5: Synthesis of (E)-1-(4-(((8-i
sopropv1-2-((tetrahydro-2H-pyran-4-yl)amino)pyazolo[1,5-a][1,3,5]triazin--
4-yl)amino)methyl)piperidin-1-yl)-4-(pyrrolidin-1-yl)but-2-en-1-one
##STR00298##
[0453] K.sub.2CO.sub.3 (0.09 g, 0.65 mmol) and pyrrolidine (0.028
g, 0.39 mnol) was sequentially added to a solution of
(E)-4-promo-1-(4-(((8-isopropyl-2-((tetrahydro-2H-pyran-4-yl)amino)
pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)methyl)piperidin-1-yl)but-2-en-1-
-one (0.17 g, 0.33 mmol) in ACN (10 mL) and the resulting mix was
allowed to stir for 4 h at 60.degree. C. The reaction mixture was
quenched with water and diluted with ethyl acetate. The aqueous
layer was separated and extracted with ethyl acetate (2.times.25
mL). Thc combined organic phase was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by prep. HPLC (Method- A: 0.1% formic acid in water, 13:
Acetonitrile-Methanol, Column; KINETEXC18 (21.2 mm*150 mm, 5
.mu.m)) to afford desired title compound (0.02 g, 11.9%).
.sup.1HNMR (DMSO-d.sub.6, 400 MHz); .delta. 8.30 (d, 1H), 7.68 (s,
1H), 6.89 (d, 1H), 6.54-6.14 (m, 2H), 4.38-4.35 (d, 1 H), 4.02-3.99
(d, 1H), 4.87-4.84 (d, 2H), 140-3.29 (m, 2H), 3.16 (d, 2H),
3.014-2.86 (m, 2H), 2.58-2.41 (m, 8H), 2.02-1.97 (rn, 2H),
1.88-1.78 (m, 2H), 1.68-1.67 (rn, 5H), 1.53-1.45 (m, 2H), 1.23-1.2
(m, 6H),1.06-1.04 (m, 2H); LCMS: m/z=511.80 (M+H).sup.+
Example-31: Synthesis of
1-acryloyl-N-(3-(((3-isopropyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazol-
o[1,5-al]pyrimidin-7-yl)amino)methyl)phenyl)azetidine-2-carboxamide
(Compound-139)
Step-1: Synthesis of
5-chloro-3-isopropyl-N-(3-nitrohenzyl)pyrazolo[1,5-a]pyrimidin 7
amine
##STR00299##
[0455] The process of this step was adopted from step-1 of
example-4 (0.8 g, 99%). LCMS:m/z=346 (M+H).sub.+.
Step-2: Synthesis of tert-butyl (5-chloro-3-isopropylpyrazolo
[1,5-a]pyrimidin-7-yl)(3-nitrohenzy)carbamate
##STR00300##
[0457] To a solution of
5-chloro-3-isopropyl-N-(3nitrobenzyl)pyrazolo[1,5-a]pyrimidin
-7-amine(0.8 g, 2.3 I mmol) in CCl.sub.4 (20 mL) was added
(BOC).sub.2O (0.6 g, 2.78 mmol), followed by DMAP) (0.028 g, 0.23
mmol) and reaction mixture was stirred at room temperature for
After completion of the reaction, the reaction mixture was
concentrated to afford the title compound (1 g, 86%). LCMS: m/z=446
(M+H).sub..
Step-3: Synthesis of tert-butyl
(3-isopropyl-5-((tetrahydro-2H-pyran-4-yl)amino)
pyrazolo[1,5-a]pyrimidin-7-yl)(3-nitrohenzyl)carbamate
##STR00301##
[0459] To a solution of tert-butyl
(5-chloro-3-isopropylpyrazolo[1,5-a]pyrimidin-7-yl)(3-nitrohenzyl)carhatr-
iate (1 g, 2.24 mmol) and tetrahydro-2H-pyran-4-amine (0.68 g, 6.74
mmol) in toluene (20 mL) was added NaO'Bu (0.31 g, 3.28 mmol) and
degassed with N.sub.2 gas for 10min. Then added BINAP (0.2 g,
0.33mmol) followed by (dba).sub.3 (0.1g, 0.11 mmol), then reaction
mixture was heated to 100.degree. C. for 12h. After completion of
the reaction, the reaction mixture was cooled to room temperature
and filtered through celitc bed and washed with ethyl acetate
(2.times.20mL). The filtrate was concentrated under vacuum and
purified by column chromatography using 100-200 silica gel to
afford the title compound (0.8 g, 70%). LCMS: m/z=511
(M.times.H).sup.1.
Step-4: Synthesis of tert-butyl
(3-aminobenzyl)(3-isopropyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1-
,5-a]pyrimidin-7-yl)carbarnate.
##STR00302##
[0461] To a solution of tert-butyl
(3-isopropyl-5((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1
,5-a]pyrimidin-7-yl)(3-ni trobenzyl)carbamate (0.8 g, 1.56 mmol) in
THF:MeOH:Water (3:2:1 ratio, 20 mL) were added zinc (1.02 g, 15.68
mmol) and ammonium chloride (1.7 g, 31.36 mmol). The reaction
mixture was stirred at room temperature for 4h. After completion of
reaction, the reaction mixture was filtered through celite and
diluted with ethyl acetate. The aqueous layer was separated and
extracted with ethyl acetate (2.times.25 mL). The combined organic
phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated. The residue was purified by 100-200 silica gel
column chromatography to afford desired title compound (0.6 g,
87%). LCMS; m/z=480.8 (M+H).sup.+.
Step-5: Synthesis of ten-butyl (3-(1-acryloylazetidine-2-carhox ami
do)benzyl)(3-isopropyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazolo[1.5-a]-
pyrimidin-7-yl)carbamate
##STR00303##
[0463] The process of this step was adopted from example-1 (0.3 g,
crude). LCMS: m/z=617.9 (M+H).sup..
Step-6: Synthesis of 1 -acryloyl-N -(3-(((3-i
sopropyl-5-((tetruhydro-2H-pyran-4-yl)amino) pyrazolo[1,5-a
]pyrimidin-7-yl)amino)methyl)phenyl)azetidine-2-carboxamide
##STR00304##
[0465] The process of this step was adopted from step-2 of
example-2. The obtained crude compound was purified by preparative
HPLC (Method; A; 0.02% Ammonia in water, B; ACN:MeOH (1;1) Column;
X bridge C 18 (150 mm*21.2 mm) to afford desired title compound as
free base(0.1g, 10%). .sup.1HNMR (DMSO-d.sub.6, 400 MHz); .delta.
10.17 (s, 1 H), 7.83-7.81 (m, 1 H), 7.61 (s, 1 H), 7.58-7.52 (m,
2H), 7.29-7.25 (m, 1 H), 7_05-7.02 (t, 1 H), 6.57-6.55 (m, 2H),
6.36-6.29(m, 1 H), 6.13-6.05 (m, 2H), 5.72-5.69 (m, 1 H), 5.58-5.57
(m, 1 H), 5.01 (s, 2H), 4.43-4.42 (d, 2H), 4.21-4.15 (in, 1H),
3.88-3.79 (m, 2H), 3.37-3.34 (in, 2H), 2.95-2.92 (m, 1 H), 2.20 (m
1H). 1.84-1.81 (d, 2H), 1.38-1.33 (m, 2H), 1.24-1.22 (d, 6H); LCMS:
m/z=518.8 (M+H).sup.1.
[0466] Although the present application has been illustrated by
certain of the preceding examples, it is not to be construed as
being limited thereby but rather, the present application
encompasses the generic arca as hereinbefore disclosed. Various
modifications and embodiments can be made without departing from
the spirit and scope thereof. For example, the following compounds
which can be prepared by following similar procedure as described
above with suitable modifications known to the one ordinary skilled
in the art arc also included in the scope of the present
application.
##STR00305## ##STR00306## ##STR00307## ##STR00308## ##STR00309##
##STR00310## ##STR00311## ##STR00312##
Example-32: Biochemical Assay for CDK7:
[0467] The ability of compounds to inhibit CDK7 kinase activity was
tested in a TR-FRET assay using 5 nM of CDK7/CyeH/MNAT1 obtained
from lnvitrogen, USA. Test compounds were pre-incubated with the
kinase at room temperature for 60 min. After incubation, substrate
mix 1100 nM Ultra-light Ml3P (Perkin Ulmer, USA) and 1mM ATP
(Sigrna)I was added. The above reaction was stopped by the addition
of 40 mM EDTA after 60 minutes of kinase reaction. 1 nM Eu-labelled
antiphospho-MBP antibody [Perkin Elmer, USA] was added, mixed well
and the fluorescence emission at 615 nm and 665 nm [excitation at
340 nm] was measured. The final DMSO concentration in the assay was
1%. For IC.sub.50 determination, appropriate concentrations were
made by 1/3.sup.rd serial dilutions of 10 mM DMSO stock solution of
test compound. All the fluorescence measurements were made in a
Victor 3 Multilabcl Counter [Perkin Elmer, USA].
[0468] The IC.sub.50 was determined by fitting the dose response
data to sigmoidal cure fitting equation using GraphPad Prism
software V5. To identify compounds that inhibit CDK7 irreversibly,
time depended inhibition studies were carried by pre-incubating
compound with the enzyme at three time points (20, 60 and 180 min)
and carrying out assay as described above.
[0469] Selected compounds were screened in the above mentioned
assay procedure. The % inhibition of the selected compounds and the
IC.sub.50 values are summarised in the below table; wherein group
"A" refers to an IC.sub.50 value of less than or equal to 300 nM,
"B" refres to IC.sub.50 value in range of 300.01 to 1000 nM and
group "C" refres to IC.sub.50 value of greater than 1000 nM.
TABLE-US-00028 Compound % inhibition Compound % inhibition No. @ 10
.mu.M IC.sub.50 No. @ 10 .mu.M IC.sub.50 1. 88 B 2. 98 A 3. 96 A 4.
93 A 5. 94 A 6. 91 A 7. 91 C 8. 96 A 9. 85 C 10. 89 B 11. 89 B 12.
92 B 13. 93 B 14. 97 A 15. 98 A 16. -- A 17. 84 C 18. 101 A 19. 100
A 20. 100 A 21. 96 A 22. 98 A 23. 98 A 24. 98 A 25. 99 A 26. 95 B
27. 100 A 28. 99 A 29. 98 A 30. 98 A 31. 100 A 32. 91 A 33. 86 B
34. 73 C 35. 99 A 36. 91 B 37. 91 A 38. 95 A 39. 76 C 40. 99 A 41.
92 B 42. 99 A 43. 79 B 44. 94 A 45. 73 C 46. 98 A 47. 100 A 48. 98
A 49. 84 C 50. 97 A 51. 91 B 52. 96 -- 53. 99 A 54. 98 A 55. 99 A
56. 97 A 57. 84 C 58. 96 B 59. 98 A 60. 98 A 61. 76 C 62. 97 A 63.
94 B 64. 91 B 65. 86 C 66. 98 A 67. 91 B 68. 92 B 69. 98 A 70. 85 B
71. 92 B 72. 84 B 73. 101 A 74. 99 A 75. 97 A 76. 81 C 77. 99 A 78.
100 A 79. 96 A 80. 94 B 81. 98 A 82. 93 B 83. 96 A 84. 97 A 85. 99
A 86. 99 A 87. 32 -- 88. 97 A 89. 38 -- 90. 95 A 91. 94 A 92. 97 A
93. 96 A 94. 99 A 95. 93 A 96. 87 B 97. 87 B 98. 88 B 99. 82 C 100.
82 B 101. 96 A 102. 82 B 103. 80 C 104. 78 C 105. 92 A 106. 91 C
107. 95 A 108. 99 A 109. 98 A 110. 99 A 111. 95 A 112. 99 A 113. --
A 114. 97 A 115. 94 A 116. 92 A 117. 95 A 118. 93 B 119. 99 A 120.
93 A 121. 93 A 122. -- A 123. 96 A 124. 94 A 125. 86 A 126. 80 C
127. 95 A 128. 93 A 129. 94 A 130. 97 A 131. 86 B 132. 90 A 133. 97
A 134. 97 A 135. 97 A 136. 73 C 137. 99 A 138. 64 C 139. 98 A .sup.
40A. -- A .sup. 40B. -- A .sup. 40C. -- A .sup. 40D. -- B .sup.
48A. -- A
Example-33: In vivo Efficacy of CDK7 Inhibitor in the MV4- 11 Human
Acute Myeloid Leukemia (AML) Cancer Xenograft Model
[0470] The effect of the CDK7 inhibitor to inhibit the growth of
MV4-11 xenograft tumors implanted in mice was evaluated. 13riefly,
MV4-11 cells were grown in lscove's Modified Dulbccco's Medium
(Sigma Aldrich) medium supplemented with 10% FBS (Invitrogen) and
1% penicillin streptomycin (Invitorgen). To establish tumors,
15.times.10.sup.6 MV4-11 cells were injected subcutaneously in 200
.mu.l of 1:1 HBSS (Sigma: H4641) and ECM gel (Corning) into the
right hind flank of male anhymic nude mice (Invigo), Tumor volume
was measured thrice a week and body weight was monitored daily. To
estimate tumor volumes, the length (D) and width (d), of the
xenograft tumors were measured manually with calipers and the tumor
volume was calculated using the formula:Tumor
volume=(D.times.d2)/2. Treatment was initiated when the average
tumor size had reached approximately 250 mm3. The animals were
randomized based on tumor volumes into two groups of seven animals
each. To evaluate efficacy, compound- 115 was administered
intraperitoneally once per day (q24h/qd schedule). The treatment
period was for 14 days after which (fie overall efficacy was
evaluated based on tumor volume changes observed during the
treatment period. Tumor volumes were analyzed using one-way ANOVA
with Dunnett s multiple comparisons test for comparison of
treatment versus control group. Results arc shown in graph (FIG.
1).
Example-34: Inhibition of RNA Polymerase II CTD Phosphorylation in
Cell Western assay
[0471] 25000 cells (MDA-MB-231/NCI-H358) wcrc seeded in 96 well
black clear bottom plate incubated for overnight before addition of
selected compound. 3 fold dilutions of selected compound diluted in
DMSO starting from 10 .mu.M, added to cells and incubated for 4
hours at 37.degree. CO.sub.2incubator. Cells wcrc washed once with
100 .mu.l of phosphate buffered saline (sigma # P3813), then fixed
with 100 .mu.l/well of 4% Para formaldehyde for 60 min at room
temperature, in dark. The cells were washed 3 times with 100 .mu.l
of wash buffer (PBS with 0.1% Triton-X-100), later blocked for 2
hours at room temperature in blocking buffer (5%, 13SA in PBST).
Cells were stained overnight at 4.degree. C. with Phospho RNA Pol
11 (S5) (Millipore #04-1572, Abcam #5131) or Phospho RNA Pol II
(Ser-2), antibody (Bethyl labs #A300-654-A) in blocking buffer.
Post incubation cells were washed with Delifia wash buffer (Perkin
Elmer # 4010-0010). Cells were treated with LANCE secondary
antibodies (LANCE.RTM. Eu-W1024 .sub.Perkin elmer # AD)-0076 for
phospho Ser-5 CTD RNA pot-II and Delfia IM-N1 anti rabbit 1 g6
Perkin elmer AD0106 for phospho Ser-2 CTD RNA pol-II) for 2 hours
in assay buffer (Perkin Elmer #1244-1H), Cells wcrc washed 3 times
with Delfia wash buffer post incubation, Enhancement solution
(Perkin Elmer #1244-105) added and incubated for 20 mins. Europium
readings wcrc taken in Victor-3 instrument. Cell normalization was
done using IIoechst dye (0.5 .mu.g/ml).
[0472] The inhibition (EC.sub.50 in .mu.M) of RNA polymerase II CTD
phosphosphorylation for selected compounds was evaluated in below
table:
[0473] Selected compounds were screened in the above mentioned
assay procedure. The EC.sub.50 values are summarised in the below
(able; wherein group "A" refers to an EC.sub.50 value of less than
or equal to 1 .mu.M, group B refres to EC.sub.50 value in range of
1 to 10 .mu.M and group "C" refres to EC.sub.50 value of greater
than 10 .mu.M.
TABLE-US-00029 Compound No. EC.sub.50 Compound No. EC.sub.50 1 A 2
A 3 A 4 A 5 B 6 B 8 A 13 B 14 B 15 C 18 A 20 A 21 A 24 A 32 B 38 B
69 B 78 A 83 A 85 B 88 B 90 B 91 C 92 B 93 B 94 A 95 A 96 B 97 B
101 B 105 C 106 B 107 B 108 B 109 B 110 B 111 B 114 B 115 A 116 B
117 B 119 C 122 C 124 C 125 A 128 B 129 B 131 C 132 C 134 B
* * * * *