U.S. patent application number 17/570139 was filed with the patent office on 2022-04-28 for therapeutic compounds and uses thereof.
This patent application is currently assigned to GENENTECH, INC.. The applicant listed for this patent is CONSTELLATION PHARMACEUTICALS, INC., GENENTECH, INC.. Invention is credited to Sarah Bronner, Kevin X. Chen, Patrick Cyr, Kwong Wah Lai, Steven Magnuson, Jeremy M. Murray, F. Anthony Romero, Vickie Hsiao-Wei Tsui, John Wai, Fei Wang.
Application Number | 20220127265 17/570139 |
Document ID | / |
Family ID | 1000006075117 |
Filed Date | 2022-04-28 |
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United States Patent
Application |
20220127265 |
Kind Code |
A1 |
Cyr; Patrick ; et
al. |
April 28, 2022 |
THERAPEUTIC COMPOUNDS AND USES THEREOF
Abstract
The present invention relates to a compound formula (I):
##STR00001## and to salts thereof, wherein R.sup.1, R.sup.2 X, and
Y have any of the values defined herein, and compositions and uses
thereof. The compounds are useful as inhibitors of CBP and/or
EP300. Also included are pharmaceutical compositions comprising a
compound of formula (I) or a pharmaceutically acceptable salt
thereof, and methods of using such compounds and salts in the
treatment of various CBP and/or EP300-mediated disorders.
Inventors: |
Cyr; Patrick; (South San
Francisco, CA) ; Bronner; Sarah; (South San
Francisco, CA) ; Romero; F. Anthony; (South San
Francisco, CA) ; Magnuson; Steven; (South San
Francisco, CA) ; Tsui; Vickie Hsiao-Wei; (South San
Francisco, CA) ; Murray; Jeremy M.; (South San
Francisco, CA) ; Wai; John; (Shanghai, CN) ;
Lai; Kwong Wah; (Shanghai, CN) ; Wang; Fei;
(Shanghai, CN) ; Chen; Kevin X.; (Shanghai,
CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
GENENTECH, INC.
CONSTELLATION PHARMACEUTICALS, INC. |
South San Francisco
Cambridge |
CA
MA |
US
US |
|
|
Assignee: |
GENENTECH, INC.
South San Francisco
CA
CONSTELLATION PHARMACEUTICALS, INC.
Cambridge
MA
|
Family ID: |
1000006075117 |
Appl. No.: |
17/570139 |
Filed: |
January 6, 2022 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16198581 |
Nov 21, 2018 |
11247989 |
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17570139 |
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PCT/US2017/034326 |
May 24, 2017 |
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16198581 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 471/04 20130101;
C07D 519/00 20130101 |
International
Class: |
C07D 471/04 20060101
C07D471/04; C07D 519/00 20060101 C07D519/00 |
Foreign Application Data
Date |
Code |
Application Number |
May 24, 2016 |
CN |
PCT/CN2016/083118 |
Claims
1. A compound of formula (Ia): ##STR00691## or a salt thereof,
wherein: R.sup.1 is: C.sub.1-12alkyl, ##STR00692## R.sup.2 is
--C(O)--N(R.sup.e).sub.2, or --C(O)--R.sup.e; Y is selected from
the group consisting of: ##STR00693## ##STR00694## ##STR00695##
##STR00696## ##STR00697## ##STR00698## ##STR00699## ##STR00700##
##STR00701## ##STR00702## ##STR00703## ##STR00704## ##STR00705##
##STR00706## ##STR00707## ##STR00708## ##STR00709## and each
R.sup.e is independently selected from hydrogen, C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.2-4alkynyl, and C.sub.2-5cycloalkyl,
wherein each C.sub.1-4alkyl, C.sub.2-4alkenyl, C.sub.2-4alkynyl,
and C.sub.2-5cycloalkyl is optionally substituted with one or more
groups independently selected from oxo, halo, amino, hydroxyl,
C.sub.1-3alkoxy, and C.sub.1-C.sub.3 alkyl that is optionally
substituted with one or more groups independently selected from
halo.
2. The compound or salt of claim 1 wherein, R.sup.1 is selected
from the group consisting of: methyl, ##STR00710##
3. The compound or salt of any one of claim 1, wherein R.sup.2 is
C.sub.1-4alkanoyl.
4. The compound or salt thereof of claim 1 selected from the group
consisting of: ##STR00711## ##STR00712## ##STR00713## ##STR00714##
##STR00715## ##STR00716## or a salt thereof.
5. A compound or salt thereof selected from the group consisting
of: ##STR00717## or a salt thereof.
6. A composition comprising a compound as described in claim 1 or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable adjuvant, carrier, or vehicle.
7. A method for treating a CBP and/or EP300-mediated cancer in an
animal by selectively inhibiting a bromodomain of the CBP and/or
EP300 in the animal, comprising administering an effective amount
of a compound or a pharmaceutically acceptable salt thereof, as
described in claim 1, to the animal, wherein the animal has a
mutation in the CBP and/or EP300, and wherein the cancer is
selected from lung cancer, breast cancer, pancreatic cancer,
colorectal cancer, or melanoma.
8. A composition comprising a compound as described in claim 5 or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable adjuvant, carrier, or vehicle.
9. A method for treating a CBP and/or EP300-mediated cancer in an
animal by selectively inhibiting a bromodomain of the CBP and/or
EP300 in the animal, comprising administering an effective amount
of a compound or a pharmaceutically acceptable salt thereof, as
described in claim 5, to the animal, wherein the animal has a
mutation in the CBP and/or EP300, and wherein the cancer is
selected from lung cancer, breast cancer, pancreatic cancer,
colorectal cancer, or melanoma.
Description
PRIORITY OF INVENTION
[0001] This application is a continuation of U.S. application Ser.
No. 16/198,581, filed 21 Nov. 2018, which is a continuation of
International Application No. PCT/US2017/034326, filed 24 May 2017,
which claims priority to International Patent Application
PCT/CN2016/083118, filed 24 May 2016. The entire content of the
applications referenced above are hereby incorporated by
reference.
TECHNICAL FIELD OF THE INVENTION
[0002] The present invention relates to compounds useful as
inhibitors of CBP/EP300 and methods of treating cancer using such
inhibitors.
BACKGROUND OF THE INVENTION
[0003] Chromatin is a complex combination of DNA and protein that
makes up chromosomes. It is found inside the nuclei of eukaryotic
cells and is divided between heterochromatin (condensed) and
euchromatin (extended) forms. The major components of chromatin are
DNA and proteins. Histones are the chief protein components of
chromatin, acting as spools around which DNA winds. The functions
of chromatin are to package DNA into a smaller volume to fit in the
cell, to strengthen the DNA to allow mitosis and meiosis, and to
serve as a mechanism to control expression and DNA replication. The
chromatin structure is controlled by a series of post-translational
modifications to histone proteins, notably histones H3 and H4, and
most commonly within the "histone tails" which extend beyond the
core nucleosome structure. Histone tails tend to be free for
protein-protein interaction and are also the portion of the histone
most prone to post-translational modification. These modifications
include acetylation, methylation, phosphorylation,
ubiquitinylation, and SUMOylation. These epigenetic marks are
written and erased by specific enzymes that place the tags on
specific residues within the histone tail, thereby forming an
epigenetic code, which is then interpreted by the cell to allow
gene specific regulation of chromatin structure and thereby
transcription.
[0004] Of all classes of proteins, histones are amongst the most
susceptible to post-translational modification. Histone
modifications are dynamic, as they can be added or removed in
response to specific stimuli, and these modifications direct both
structural changes to chromatin and alterations in gene
transcription. Distinct classes of enzymes, namely histone
acetyltransferases (HATs) and histone deacetylases (HDACs),
acetylate or de-acetylate specific histone lysine residues (Struhl
K., Genes Dev., 1989, 12, 5, 599-606).
[0005] Bromodomains, which are approximately 110 amino acids long,
are found in a large number of chromatin-associated proteins and
have been identified in approximately 70 human proteins, often
adjacent to other protein motifs (Jeanmougin F., et al., Trends
Biochem. Sci., 1997, 22, 5, 151-153; and Tamkun J. W., et al.,
Cell, 1992, 7, 3, 561-572). Interactions between bromodomains and
modified histones may be an important mechanism underlying
chromatin structural changes and gene regulation.
Bromodomain-containing proteins have been implicated in disease
processes including cancer, inflammation and viral replication.
See, e.g., Prinjha et al., Trends Pharm. Sci., 33(3):146-153 (2012)
and Muller et al., Expert Rev., 13(29):1-20 (September 2011).
[0006] Cell-type specificity and proper tissue functionality
requires the tight control of distinct transcriptional programs
that are intimately influenced by their environment. Alterations to
this transcriptional homeostasis are directly associated with
numerous disease states, most notably cancer, immuno-inflammation,
neurological disorders, and metabolic diseases. Bromodomains reside
within key chromatin modifying complexes that serve to control
distinctive disease-associated transcriptional pathways. This is
highlighted by the observation that mutations in
bromodomain-containing proteins are linked to cancer, as well as
immune and neurologic dysfunction. Hence, the selective inhibition
of bromodomains across a specific family, such as the selective
inhibition of a bromodomain of CBP/EP300, creates varied
opportunities as novel therapeutic agents in human dysfunction.
[0007] There is a need for treatments for cancer, immunological
disorders, and other CBP/EP300 bromodomain related diseases.
SUMMARY OF THE INVENTION
Compounds of Formula (I)
[0008] One aspect is a compound of formula (I):
##STR00002##
or a salt thereof, wherein:
[0009] R.sup.1 is C.sub.1-12alkyl, C.sub.2-12alkenyl,
C.sub.2-12alkynyl, 3-12 membered carbocycle, and 3-12 membered
heterocycle, wherein each C.sub.1-12alkyl, C.sub.2-12alkenyl,
C.sub.2-12alkynyl, 3-12 membered carbocycle, and 3-12 membered
heterocycle of R.sup.1 is optionally substituted with one or more
groups Rd;
[0010] R.sup.2 is --C(O)--N(R.sup.e).sub.2,
--S(O)--N(R.sup.e).sub.2, --S(O).sub.2--N(R.sup.e).sub.2,
--C(O)--R.sup.e, --C(O)--O--R.sup.e, --S(O)--R.sup.e, or
--S(O).sub.2--R.sup.e,
[0011] X is absent, --C(.dbd.O)--, or C.sub.1-3alkyl; and Y is
phenyl, a 9-membered bicyclic carbocycle, a 10-membered bicyclic
carbocycle, a 9-membered bicyclic heterocycle, or a 10-membered
bicyclic heterocycle, wherein Y is optionally substituted with Ra
and wherein Y is further optionally substituted with one or more
groups R.sup.b;
[0012] or wherein wherein --X--Y taken together is selected from
the group consisting of:
##STR00003## ##STR00004## ##STR00005## ##STR00006## ##STR00007##
##STR00008##
[0013] each R.sup.a is independently selected from the group
consisting of a 5-membered carbocycle, a 6-membered carbocycle, a
5-membered heterocycle, and a 6-membered heterocycle, which a
5-membered carbocycle, 6-membered carbocycle, 5-membered
heterocycle, and 6-membered heterocycle are optionally substituted
with one or more groups R.sup.c;
[0014] each R.sup.b is independently selected from the group
consisting of halo, cyano, hydroxy, amino, C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4 alkoxy,
C.sub.1-4alkoxycarbonyl, C.sub.1-4alkanoyl,
--C(O)--N(R.sup.f).sub.2, --N(R.sup.f)C(O)--R.sup.f, and
C.sub.1-4alkanoyloxy, wherein each C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4 alkoxy,
C.sub.1-4alkoxycarbonyl, C.sub.1-4alkanoyl, and
C.sub.1-4alkanoyloxy, is optionally substituted with one or more
groups independently selected from oxo, halo, amino, hydroxy,
C.sub.1-3alkoxy, and C.sub.1-C.sub.3 alkyl that is optionally
substituted with one or more groups independently selected from
halo;
[0015] each R.sup.c is independently selected from the group
consisting of halo, cyano, hydroxy, amino, C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4 alkoxy,
C.sub.1-4alkoxycarbonyl, C.sub.1-4alkanoyl, and
C.sub.1-4alkanoyloxy, wherein each C.sub.1-4alkyl, C.sub.2-4
alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4alkoxy, C.sub.1-4
alkoxycarbonyl, C.sub.1-4alkanoyl, and C.sub.1-4alkanoyloxy, is
optionally substituted with one or more groups independently
selected from oxo, halo, amino, hydroxy, C.sub.1-3alkoxy, and
C.sub.1-C.sub.3 alkyl that is optionally substituted with one or
more groups independently selected from halo;
[0016] each R.sup.d is independently selected from the group
consisting of oxo, halo, cyano, hydroxy, amino, C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4alkoxy,
C.sub.1-4alkoxycarbonyl, C.sub.1-4alkanoyl, and
C.sub.1-4alkanoyloxy, wherein each C.sub.1-4alkyl, C.sub.2-4
alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4alkoxy, C.sub.1-4
alkoxycarbonyl, C.sub.1-4alkanoyl, and C.sub.1-4alkanoyloxy, is
optionally substituted with one or more groups independently
selected from oxo, halo, amino, hydroxy, C.sub.1-3alkoxy, and
C.sub.1-C.sub.3 alkyl that is optionally substituted with one or
more groups independently selected from halo;
[0017] each R.sup.e is independently selected from hydrogen,
C.sub.1-4alkyl, C.sub.2-4alkenyl, C.sub.2-4alkynyl, and
C.sub.2-5cycloalkyl, wherein each C.sub.1-4alkyl, C.sub.2-4alkenyl,
C.sub.2-4alkynyl, and C.sub.2-5cycloalkyl is optionally substituted
with one or more groups independently selected from oxo, halo,
amino, hydroxyl, C.sub.1-3alkoxy, and C.sub.1-C.sub.3 alkyl that is
optionally substituted with one or more groups independently
selected from halo; and
[0018] each R.sup.f is independently selected from hydrogen and
C.sub.1-4alkyl;
[0019] or a compound selected from the group consisting of:
##STR00009##
or a salt thereof.
[0020] Another aspect is a compound of formula (I):
##STR00010##
or a salt thereof, wherein:
[0021] R.sup.1 is C.sub.1-12alkyl, C.sub.2-12alkenyl,
C.sub.2-12alkynyl, 3-12 membered carbocycle, and 3-12 membered
heterocycle, wherein each C.sub.1-12alkyl, C.sub.2-12alkenyl,
C.sub.2-12alkynyl, 3-12 membered carbocycle, and 3-12 membered
heterocycle of R.sup.1 is optionally substituted with one or more
groups Rd;
[0022] R.sup.2 is --C(O)--N(R.sup.e).sub.2,
--S(O)--N(R.sup.e).sub.2, --S(O).sub.2--N(R.sup.e).sub.2,
--C(O)--R.sup.e, --C(O)--O--R.sup.e, --S(O)--R.sup.e, or
--S(O).sub.2--R.sup.e,
[0023] X is absent, --C(.dbd.O)--, or C.sub.1-3alkyl;
[0024] Y is phenyl, a 9-membered bicyclic carbocycle, a 10-membered
bicyclic carbocycle, a 9-membered bicyclic heterocycle, or a
10-membered bicyclic heterocycle, wherein Y is optionally
substituted with R.sup.a and wherein Y is further optionally
substituted with one or more groups R.sup.b;
[0025] each R.sup.a is independently selected from the group
consisting of a 5-membered carbocycle, a 6-membered carbocycle, a
5-membered heterocycle, and a 6-membered heterocycle, which a
5-membered carbocycle, 6-membered carbocycle, 5-membered
heterocycle, and 6-membered heterocycle are optionally substituted
with one or more groups R.sup.c;
[0026] each R.sup.b is independently selected from the group
consisting of halo, cyano, hydroxy, amino, C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4 alkoxy,
C.sub.1-4alkoxycarbonyl, C.sub.1-4alkanoyl,
--C(O)--N(R.sup.f).sub.2, --N(R.sup.f)C(O)--R.sup.f, and
C.sub.1-4alkanoyloxy, wherein each C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4 alkoxy,
C.sub.1-4alkoxycarbonyl, C.sub.1-4alkanoyl, and
C.sub.1-4alkanoyloxy, is optionally substituted with one or more
groups independently selected from oxo, halo, amino, hydroxy,
C.sub.1-3alkoxy, and C.sub.1-C.sub.3 alkyl that is optionally
substituted with one or more groups independently selected from
halo;
[0027] each R.sup.c is independently selected from the group
consisting of halo, cyano, hydroxy, amino, C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4 alkoxy,
C.sub.1-4alkoxycarbonyl, C.sub.1-4alkanoyl, and
C.sub.1-4alkanoyloxy, wherein each C.sub.1-4alkyl, C.sub.2-4
alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4alkoxy, C.sub.1-4
alkoxycarbonyl, C.sub.1-4alkanoyl, and C.sub.1-4alkanoyloxy, is
optionally substituted with one or more groups independently
selected from oxo, halo, amino, hydroxy, C.sub.1-3alkoxy, and
C.sub.1-C.sub.3 alkyl that is optionally substituted with one or
more groups independently selected from halo;
[0028] each R.sup.d is independently selected from the group
consisting of oxo, halo, cyano, hydroxy, amino, C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4alkoxy,
C.sub.1-4alkoxycarbonyl, C.sub.1-4alkanoyl, and
C.sub.1-4alkanoyloxy, wherein each C.sub.1-4alkyl, C.sub.2-4
alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4alkoxy, C.sub.1-4
alkoxycarbonyl, C.sub.1-4alkanoyl, and C.sub.1-4alkanoyloxy, is
optionally substituted with one or more groups independently
selected from oxo, halo, amino, hydroxy, C.sub.1-3alkoxy, and
C.sub.1-C.sub.3 alkyl that is optionally substituted with one or
more groups independently selected from halo; each R.sup.e is
independently selected from hydrogen, C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.2-4alkynyl, and C.sub.2-5cycloalkyl,
wherein each C.sub.1-4alkyl, C.sub.2-4alkenyl, C.sub.2-4alkynyl,
and C.sub.2-5cycloalkyl is optionally substituted with one or more
groups independently selected from oxo, halo, amino, hydroxyl,
C.sub.1-3alkoxy, and C.sub.1-C.sub.3 alkyl that is optionally
substituted with one or more groups independently selected from
halo; and each R is independently selected from hydrogen and
C.sub.1-4alkyl.
[0029] Another aspect includes a composition comprising a compound
of formula (I) or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable adjuvant, carrier, or vehicle.
[0030] Another aspect includes a method for treating a CBP and/or
EP300-mediated disorder in an animal comprising administering a
compound of formula (I) or a pharmaceutically acceptable salt
thereof to the animal.
[0031] Another aspect includes a method for treating a CBP and/or
EP300-mediated disorder in an animal, wherein the disorder is
cancer, comprising administering a compound of formula (I) or a
pharmaceutically acceptable salt thereof to the animal.
[0032] Another aspect includes a method for treating a CBP and/or
EP300-mediated disorder in an animal, wherein the disorder is a
fibrotic disease, comprising administering a compound of formula
(I) or a pharmaceutically acceptable salt thereof to the
animal.
[0033] Another aspect includes a method for treating a CBP and/or
EP300-mediated disorder in an animal, wherein the disorder is a
fibrotic lung disease, comprising administering a compound of
formula (I) or a pharmaceutically acceptable salt thereof to the
animal.
[0034] Another aspect includes a compound of formula (I) or a
pharmaceutically acceptable salt thereof for use in medical
therapy.
[0035] Another aspect includes a compound of formula (I) or a
pharmaceutically acceptable salt thereof for the prophylactic or
therapeutic treatment of a CBP and/or EP300-mediated disorder.
[0036] Another aspect includes the use of a compound of formula (I)
or a pharmaceutically acceptable salt thereof to prepare a
medicament for treating a CBP and/or EP300-mediated disorder in an
animal (e.g. a mammal such as a human).
[0037] Another aspect includes compounds for the study of CBP
and/or EP300.
[0038] Another aspect includes synthetic intermediates and
synthetic processes disclosed herein that are useful for preparing
a compound of formula (I) or a salt thereof.
DETAILED DESCRIPTION
Compounds and Definitions
[0039] Definitions and terms are described in more detail below.
Chemical elements are identified in accordance with the Periodic
Table of the Elements, CAS version, Handbook of Chemistry and
Physics, 75.sup.th Ed.
[0040] Unless otherwise stated, compounds of formula (I) include
enantiomeric, diastereomeric and geometric (or conformational)
isomeric forms of a given structure. For example, the R and S
configurations for each asymmetric center, Z and E double bond
isomers, Z and E conformational isomers, single stereochemical
isomers, as well as enantiomeric, diastereomeric, and geometric (or
conformational) mixtures are included. Unless otherwise stated, all
tautomeric forms of structures depicted herein are included.
Additionally, unless otherwise stated, structures depicted herein
are also meant to include compounds that differ only in the
presence of one or more isotopically enriched atoms. For example,
compounds of formula (I), wherein the independent replacement or
enrichment of one or more hydrogen by deuterium or tritium, carbon
by .sup.13C-- or .sup.14C carbon, nitrogen by a .sup.15N nitrogen,
sulfur by a .sup.33S, .sup.34S or .sup.36S sulfur, oxygen by a
.sup.17O or .sup.18O oxygen, or fluorine by a .sup.18F are
included. Such compounds are useful, for example, as analytical
tools, as probes in biological assays, or as therapeutic
agents.
[0041] Where a particular enantiomer is described, it may, in
certain embodiments be provided substantially free of the
corresponding enantiomer, and may also be referred to as "optically
enriched." "Optically-enriched," as used herein, means that the
mixture of enantiomers is made up of a significantly greater
proportion of one enantiomer, and may be described by enantiomeric
excess (ee %). In certain embodiments, the mixture of enantiomers
is made up of at least about 90% by weight of a given enantiomer
(about 90% ee). In other embodiments, the mixture of enantiomers is
made up of at least about 95%, 98% or 99% by weight of a given
enantiomer (about 95%, 98% or 99% ee). Enantiomers and
diastereomers may be isolated from racemic mixtures by any method
known to those skilled in the art, including recrystallization from
solvents in which one stereoisomer is more soluble than the other,
chiral high pressure liquid chromatography (HPLC), supercritical
fluid chromatography (SFC), the formation and crystallization of
chiral salts, which are then separated by any of the above methods,
or prepared by asymmetric syntheses and optionally further
enriched. See, for example, Jacques et al., Enantiomers, Racemates
and Resolutions (Wiley Interscience, New York, 1981); Wilen, et
al., Tetrahedron 33:2725 (1977); Eliel, E. L. Stereochemistry of
Carbon Compounds (McGraw-Hill, N Y, 1962); Wilen, S. H. Tables of
Resolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed.,
Univ. of Notre Dame Press, Notre Dame, Ind. 1972).
[0042] The term "heteroatom" means any atom independently selected
from an atom other than carbon or hydrogen, for example, one or
more of oxygen, sulfur, nitrogen, phosphorus or silicon (including
any oxidized form of nitrogen, sulfur, phosphorus or silicon; and
the quaternized form of any nitrogen).
[0043] The terms "halo" and "halogen" as used herein refer to an
atom selected from fluorine (fluoro, --F), chlorine (chloro, --Cl),
bromine (bromo, --Br) and iodine (iodo, --I).
[0044] The term "oxo" refers to .dbd.O.
[0045] The term "amino" includes --NH.sub.2,
--NH(C.sub.1-C.sub.3alkyl), and
--N(C.sub.1-C.sub.3alkyl).sub.2.
[0046] The term "unsaturated", as used herein, means that a moiety
has one or more units of unsaturation.
[0047] The term "carbocyclyl" used alone or as part of a larger
moiety, refers to a saturated, partially unsaturated, or aromatic
ring system having 3 to 20 carbon atoms. In one embodiment,
carbocyclyl includes 3 to 12 carbon atoms (C.sub.3-C.sub.12). In
another embodiment, carbocyclyl includes C.sub.3-C.sub.8,
C.sub.3-C.sub.10 or C.sub.5-C.sub.10. In other embodiment,
carbocyclyl, as a monocycle, includes C.sub.3-C.sub.8,
C.sub.3-C.sub.6 or C.sub.5-C.sub.6. In another embodiment,
carbocyclyl, as a bicycle, includes C.sub.7-C.sub.12. In another
embodiment, carbocyclyl, as a spiro system, includes
C.sub.5-C.sub.12. Examples of monocyclic carbocyclyls include
cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl,
1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl,
perdeuteriocyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl,
1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl,
cyclononyl, cyclodecyl, cycloundecyl, phenyl, and cyclododecyl;
bicyclic carbocyclyls having 7 to 12 ring atoms include [4,3],
[4,4], [4,5], [5,5], [5,6] or [6,6] ring systems, for example
bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, naphthalene, and
bicyclo[3.2.2]nonane; and spiro carbocyclyls include
spiro[2.2]pentane, spiro[2.3]hexane, spiro[2.4]heptane,
spiro[2.5]octane and spiro[4.5]decane. The term carbocyclyl
includes aryl ring systems as defined herein. The term carbocycyl
also includes cycloalkyl rings (e.g. saturated or partially
unsaturated mono-, bi-, or spiro-carbocycles).
[0048] The term "alkyl," as used herein, refers to a saturated
linear or branched-chain hydrocarbon radical. In one embodiment,
the alkyl radical is one to eighteen carbon atoms
(C.sub.1-C.sub.18). In other embodiments, the alkyl radical is
C.sub.0-C.sub.6, C.sub.0-C.sub.5, C.sub.0-C.sub.3,
C.sub.1-C.sub.12, C.sub.1-C.sub.10, C.sub.1-C.sub.8,
C.sub.1-C.sub.6, C.sub.1-C.sub.5, C.sub.1-C.sub.4 or
C.sub.1-C.sub.3. C.sub.0 alkyl refers to a bond. Examples of alkyl
groups include methyl (Me, --CH.sub.3), ethyl (Et,
--CH.sub.2CH.sub.3), 1-propyl (n-Pr, n-propyl,
--CH.sub.2CH.sub.2CH.sub.3), 2-propyl (i-Pr, i-propyl,
--CH(CH.sub.3).sub.2), 1-butyl (n-Bu, n-butyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-methyl-1-propyl (i-Bu,
i-butyl, --CH.sub.2CH(CH.sub.3).sub.2), 2-butyl (s-Bu, s-butyl,
--CH(CH.sub.3)CH.sub.2CH.sub.3), 2-methyl-2-propyl (t-Bu, t-butyl,
--C(CH.sub.3).sub.3), 1-pentyl (n-pentyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-pentyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.3), 3-pentyl
(--CH(CH.sub.2CH.sub.3).sub.2), 2-methyl-2-butyl
(--C(CH.sub.3).sub.2CH.sub.2CH.sub.3), 3-methyl-2-butyl
(--CH(CH.sub.3)CH(CH.sub.3).sub.2), 3-methyl-1-butyl
(--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2), 2-methyl-1-butyl
(--CH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.3), 1-hexyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-hexyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 3-hexyl
(--CH(CH.sub.2CH.sub.3)(CH.sub.2CH.sub.2CH.sub.3)),
2-methyl-2-pentyl (--C(CH.sub.3).sub.2CH.sub.2CH.sub.2CH.sub.3),
3-methyl-2-pentyl (--CH(CH.sub.3)CH(CH.sub.3)CH.sub.2CH.sub.3),
4-methyl-2-pentyl (--CH(CH.sub.3)CH.sub.2CH(CH.sub.3).sub.2),
3-methyl-3-pentyl (--C(CH.sub.3)(CH.sub.2CH.sub.3).sub.2),
2-methyl-3-pentyl (--CH(CH.sub.2CH.sub.3)CH(CH.sub.3).sub.2),
2,3-dimethyl-2-butyl (--C(CH.sub.3).sub.2CH(CH.sub.3).sub.2),
3,3-dimethyl-2-butyl (--CH(CH.sub.3)C(CH.sub.3).sub.3, heptyl,
octyl, nonyl, decyl, undecyl and dodecyl.
[0049] The term "alkenyl," as used herein, denotes a linear or
branched-chain hydrocarbon radical with at least one carbon-carbon
double bond. An alkenyl includes radicals having "cis" and "trans"
orientations, or alternatively, "E" and "Z" orientations. In one
example, the alkenyl radical is two to eighteen carbon atoms
(C.sub.2-C.sub.18). In other examples, the alkenyl radical is
C.sub.2-C.sub.12, C.sub.2-C.sub.10, C.sub.2-C.sub.8,
C.sub.2-C.sub.6 or C.sub.2-C.sub.3. Examples include, but are not
limited to, ethenyl or vinyl (--CH.dbd.CH.sub.2), prop-1-enyl
(--CH.dbd.CHCH.sub.3), prop-2-enyl (--CH.sub.2CH.dbd.CH.sub.2),
2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl,
buta-1,3-dienyl, 2-methylbuta-1,3-diene, hex-1-enyl, hex-2-enyl,
hex-3-enyl, hex-4-enyl and hexa-1,3-dienyl.
[0050] The term "alkynyl," as used herein, refers to a linear or
branched hydrocarbon radical with at least one carbon-carbon triple
bond. In one example, the alkynyl radical is two to eighteen carbon
atoms (C.sub.2-C.sub.18). In other examples, the alkynyl radical is
C.sub.2-C.sub.12, C.sub.2-C.sub.10, C.sub.2-C.sub.8,
C.sub.2-C.sub.6 or C.sub.2-C.sub.3. Examples include, but are not
limited to, ethynyl (--C.ident.CH), prop-1-ynyl
(--C.ident.CCH.sub.3), prop-2-ynyl (propargyl,
--CH.sub.2C.ident.CH), but-1-ynyl, but-2-ynyl and but-3-ynyl.
[0051] The term "alkoxy" refers to a linear or branched radical
represented by the formula --OR in which R is alkyl, alkenyl,
alkynyl or carbocycyl. Alkoxy groups include methoxy, ethoxy,
propoxy, isopropoxy, and cyclopropoxy.
[0052] The term "haloalkyl," as used herein, refers to an alkyl as
defined herein that is substituted with one or more (e.g. 1, 2, 3,
or 4) halo groups.
[0053] The term "aryl" used alone or as part of a larger moiety as
in "arylalkyl", "arylalkoxy", or "aryloxyalkyl", refers to a
monocyclic, bicyclic or tricyclic, carbon ring system, that
includes fused rings, wherein at least one ring in the system is
aromatic. The term "aryl" may be used interchangeably with the term
"aryl ring". In one embodiment, aryl includes groups having 6-20
carbon atoms (C.sub.6-C.sub.20 aryl). In another embodiment, aryl
includes groups having 6-10 carbon atoms (C.sub.6-C.sub.10 aryl).
Examples of aryl groups include phenyl, naphthyl, anthracyl,
biphenyl, phenanthrenyl, naphthacenyl,
1,2,3,4-tetrahydronaphthalenyl, 1H-indenyl, 2,3-dihydro-1H-indenyl,
and the like, which may be substituted or independently substituted
byone or more substituents described herein. A particular aryl is
phenyl. In another embodiment aryl includes an aryl ring fused to
one or more carbocyclic rings, such as indanyl, dihydrophenanthryl,
or tetrahydronaphthyl, and the like, where the radical or point of
attachment is on an aromatic ring.
[0054] The term "heteroaryl" used alone or as part of a larger
moiety, e.g., "heteroarylalkyl", or "heteroarylalkoxy", refers to a
monocyclic, bicyclic or tricyclic ring system having 5 to 14 ring
atoms, wherein at least one ring is aromatic and contains at least
one heteroatom. In one embodiment, heteroaryl includes 4-6 membered
monocyclic aromatic groups where one or more ring atoms is
nitrogen, sulfur or oxygen that is independently optionally
substituted. In another embodiment, heteroaryl includes 5-6
membered monocyclic aromatic groups where one or more ring atoms is
nitrogen, sulfur or oxygen that is independently optionally
substituted. In some embodiments, the heteroaryl group is a
C.sub.1-C.sub.20 heteroaryl group, where the heteroaryl ring
contains 1-20 carbon atoms and the remaining ring atoms include one
or more nitrogen, sulfur, or oxygen atoms. Example heteroaryl
groups include thienyl, furyl, imidazolyl, pyrazolyl, thiazolyl,
isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl,
oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl,
pyrimidyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl,
tetrazolo[1,5-b]pyridazinyl, imidazol[1,2-a]pyrimidinyl, purinyl,
benzoxazolyl, benzofuryl, benzothiazolyl, benzothiadiazolyl,
benzotriazolyl, benzoimidazolyl, indolyl, 1,3-thiazol-2-yl,
1,3,4-triazol-5-yl, 1,3-oxazol-2-yl, 1,3,4-oxadiazol-5-yl,
1,2,4-oxadiazol-5-yl, 1,3,4-thiadiazol-5-yl, 1H-tetrazol-5-yl,
1,2,3-triazol-5-yl, pyrid-2-yl N-oxide, and
pyrazolo[4,3-c]pyridinyl. The terms "heteroaryl" also includes
groups in which a heteroaryl is fused to one or more aryl,
carbocyclyl, or heterocyclyl rings, where the radical or point of
attachment is on the heteroaryl ring. Nonlimiting examples include
indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl,
indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl,
cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl,
4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl,
phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and
pyrido[2,3-b]-1,4-oxazin-3(4H)-one. A heteroaryl group may be
mono-, bi- or tri-cyclic.
[0055] As used herein, the term "heterocyclyl" or "heterocycle"
refers to a "carbocyclyl" as defined herein, wherein one or more
(e.g. 1, 2, 3, or 4) carbon atoms have been replaced with a
heteroatom (e.g. O, N, or S). In some embodiments, a heterocyclyl
or heterocycle refers to a saturated ring system, such as a 3 to 12
membered saturated heterocyclyl ring system. In some embodiments, a
heterocyclyl or heterocycle refers to a heteroaryl ring system,
such as a 5 to 14 membered heteroaryl ring system. A heterocyclyl
or heterocycle can optionally be substituted with one or more
substituents independently selected from those defined herein.
[0056] In one example, heterocyclyl or heterocycle includes 3-12
ring atoms and includes monocycles, bicycles, tricycles and spiro
ring systems, wherein the ring atoms are carbon, and one to five
ring atoms is a heteroatom selected from nitrogen, sulfur or
oxygen, which is independently optionally substituted by one or
more groups. In one example, heterocyclyl or heterocycle includes 1
to 4 heteroatoms. In another example, heterocyclyl or heterocycle
includes 3- to 7-membered monocycles having one or more heteroatoms
selected from nitrogen, sulfur or oxygen. In another example,
heterocyclyl or heterocycle includes 4- to 6-membered monocycles
having one or more heteroatoms selected from nitrogen, sulfur or
oxygen. In another example, heterocyclyl or heterocycle includes
3-membered monocycles. In another example, heterocyclyl or
heterocycle includes 4-membered monocycles. In another example,
heterocyclyl or heterocycle includes 5-6 membered monocycles. In
one example, the heterocyclyl or heterocycle group includes 0 to 3
double bonds. Any nitrogen or sulfur heteroatom may optionally be
oxidized (e.g. NO, SO, SO.sub.2), and any nitrogen heteroatom may
optionally be quaternized (e.g. [NR.sub.4].sup.+Cl.sup.-,
[NR.sub.4].sup.+OH.sup.-). Example heterocyclyls or heterocycles
include oxiranyl, aziridinyl, thiiranyl, azetidinyl, oxetanyl,
thietanyl, 1,2-dithietanyl, 1,3-dithietanyl, pyrrolidinyl,
dihydro-1H-pyrrolyl, dihydrofuranyl, tetrahydrofuranyl,
dihydrothienyl, tetrahydrothienyl, imidazolidinyl, piperidinyl,
piperazinyl, morpholinyl, thiomorpholinyl,
1,1-dioxo-thiomorpholinyl, dihydropyranyl, tetrahydropyranyl,
hexahydrothiopyranyl, hexahydropyrimidinyl, oxazinanyl,
thiazinanyl, thioxanyl, homopiperazinyl, homopiperidinyl, azepanyl,
oxepanyl, thiepanyl, oxazepinyl, oxazepanyl, diazepanyl,
1,4-diazepanyl, diazepinyl, thiazepinyl, thiazepanyl,
tetrahydrothiopyranyl, oxazolidinyl, thiazolidinyl,
isothiazolidinyl, 1,1-dioxoisothiazolidinonyl, oxazolidinonyl,
imidazolidinonyl, 4,5,6,7-tetrahydro[2H]indazolyl,
tetrahydrobenzoimidazolyl, 4,5,6,7-tetrahydrobenzo[d]imidazolyl,
1,6-dihydroimidazol[4,5-d]pyrrolo[2,3-b]pyridinyl, thiazinyl,
oxazinyl, thiadiazinyl, oxadiazinyl, dithiazinyl, dioxazinyl,
oxathiazinyl, thiatriazinyl, oxatriazinyl, dithiadiazinyl,
imidazolinyl, dihydropyrimidyl, tetrahydropyrimidyl, 1-pyrrolinyl,
2-pyrrolinyl, 3-pyrrolinyl, indolinyl, thiapyranyl, 2H-pyranyl,
4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, pyrazolidinyl,
dithianyl, dithiolanyl, pyrimidinonyl, pyrimidindionyl,
pyrimidin-2,4-dionyl, piperazinonyl, piperazindionyl,
pyrazolidinylimidazolinyl, 3-azabicyclo[3.1.0]hexanyl,
3,6-diazabicyclo[3.1.1]heptanyl, 6-azabicyclo[3.1.1]heptanyl,
3-azabicyclo[3.1.1]heptanyl, 3-azabicyclo[4.1.0]heptanyl,
azabicyclo[2.2.2]hexanyl, 2-azabicyclo[3.2.1]octanyl,
8-azabicyclo[3.2.1]octanyl, 2-azabicyclo[2.2.2]octanyl,
8-azabicyclo[2.2.2]octanyl, 7-oxabicyclo[2.2.1]heptane,
azaspiro[3.5]nonanyl, azaspiro[2.5]octanyl, azaspiro[4.5]decanyl,
1-azaspiro[4.5]decan-2-only, azaspiro[5.5]undecanyl,
tetrahydroindolyl, octahydroindolyl, tetrahydroisoindolyl,
tetrahydroindazolyl, 1,1-dioxohexahydrothiopyranyl. Examples of
5-membered heterocyclyls or heterocycles containing a sulfur or
oxygen atom and one to three nitrogen atoms are thiazolyl,
including thiazol-2-yl and thiazol-2-yl N-oxide, thiadiazolyl,
including 1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl,
oxazolyl, for example oxazol-2-yl, and oxadiazolyl, such as
1,3,4-oxadiazol-5-yl, and 1,2,4-oxadiazol-5-yl. Example 5-membered
ring heterocyclyls or heterocycles containing 2 to 4 nitrogen atoms
include imidazolyl, such as imidazol-2-yl; triazolyl, such as
1,3,4-triazol-5-yl; 1,2,3-triazol-5-yl, 1,2,4-triazol-5-yl, and
tetrazolyl, such as 1H-tetrazol-5-yl. Example benzo-fused
5-membered heterocyclyls or heterocycles are benzoxazol-2-yl,
benzthiazol-2-yl and benzimidazol-2-yl. Example 6-membered
heterocyclyls or heterocycles contain one to three nitrogen atoms
and optionally a sulfur or oxygen atom, for example pyridyl, such
as pyrid-2-yl, pyrid-3-yl, and pyrid-4-yl; pyrimidyl, such as
pyrimid-2-yl and pyrimid-4-yl; triazinyl, such as
1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl; pyridazinyl, in
particular pyridazin-3-yl, and pyrazinyl. The pyridine N-oxides and
pyridazine N-oxides and the pyridyl, pyrimid-2-yl, pyrimid-4-yl,
pyridazinyl and the 1,3,4-triazin-2-yl groups, are other example
heterocyclyl groups.
[0057] The term "heterocyclyl" or "heterocycle" also includes
groups in which a heterocyclyl is fused to one or more aryl,
carbocyclyl, or heterocyclyl rings, where the radical or point of
attachment is on the heterocyclyl ring. Nonlimiting examples
include tetrahydroquinolinyl and tetrahydroisoquinolinyl.
[0058] As used herein, the term "partially unsaturated" refers to a
ring moiety that includes at least one double or triple bond
between ring atoms but the ring moiety is not aromatic.
[0059] As used herein, the term "inhibitor" refers to a compound
that binds to and inhibits the bromodomain of CBP and/or EP300 with
measurable affinity and activity. In certain embodiments, an
inhibitor has an IC.sub.50 or binding constant of less about 20
.mu.M, less than about 1 .mu.M, less than about 500 nM, less than
about 100 nM, or less than about 10 nM.
[0060] The terms "measurable affinity" and "measurably inhibit," as
used herein, refer to a measurable reduction in activity (e.g.,
reduction in recognition of lysine acetyl recognition of chromatin)
of the bromodomain of CBP and/or EP300 between: (i) a sample
comprising a compound of formula (I) or composition thereof and
such bromodomain, and (ii) an equivalent sample comprising such
bromodomain, in the absence of said compound, or composition
thereof.
[0061] "Pharmaceutically acceptable salts" include both acid and
base addition salts. It is to be understood that when a compound or
Example herein is shown as a specific salt, the corresponding
free-base, as well as other salts of the corresponding free-base
(including pharmaceutically acceptable salts of the corresponding
free-base) are contemplated.
[0062] "Pharmaceutically acceptable acid addition salt" refers to
those salts which retain the biological effectiveness and
properties of the free bases and which are not biologically or
otherwise undesirable, formed with inorganic acids such as
hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,
carbonic acid, phosphoric acid and the like, and organic acids may
be selected from aliphatic, cycloaliphatic, aromatic, araliphatic,
heterocyclic, carboxylic, and sulfonic classes of organic acids
such as formic acid, acetic acid, propionic acid, glycolic acid,
gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid,
maleic acid, maloneic acid, succinic acid, fumaric acid, tartaric
acid, citric acid, aspartic acid, ascorbic acid, glutamic acid,
anthranilic acid, benzoic acid, cinnamic acid, mandelic acid,
embonic acid, phenylacetic acid, methanesulfonic acid,
ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid,
salicyclic acid and the like.
[0063] "Pharmaceutically acceptable base addition salts" include
those derived from inorganic bases such as sodium, potassium,
lithium, ammonium, calcium, magnesium, iron, zinc, copper,
manganese, aluminum salts and the like. Particularly base addition
salts are the ammonium, potassium, sodium, calcium and magnesium
salts. Salts derived from pharmaceutically acceptable organic
nontoxic bases includes salts of primary, secondary, and tertiary
amines, substituted amines including naturally occurring
substituted amines, cyclic amines and basic ion exchange resins,
such as isopropylamine, trimethylamine, diethylamine,
triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol,
tromethamine, dicyclohexylamine, lysine, arginine, histidine,
caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine,
glucosamine, methylglucamine, theobromine, purines, piperizine,
piperidine, N-ethylpiperidine, polyamine resins and the like.
Particular organic non-toxic bases are isopropylamine,
diethylamine, ethanolamine, tromethamine, dicyclohexylamine,
choline, and caffeine.
[0064] The term "tautomer" or "tautomeric form" refers to
structural isomers of different energies which are interconvertible
via a low energy barrier. For example, proton tautomers (also known
as prototropic tautomers) include interconversions via migration of
a proton, such as keto-enol and imine-enamine isomerizations.
Valence tautomers include interconversions by reorganization of
some of the bonding electrons.
[0065] A "solvate" refers to an association or complex of one or
more solvent molecules and a compound of the present invention.
Examples of solvents include water, isopropanol, ethanol, methanol,
DMSO, ethyl acetate, acetic acid and ethanolamine. The term
"hydrate" refers to the complex where the solvent molecule is
water.
[0066] "Therapeutically effective amount" refers to an amount of a
compound of the present invention that (i) treats the particular
disease, condition or disorder, (ii) attenuates, ameliorates or
eliminates one or more symptoms of the particular disease,
condition, or disorder, or (iii) delays the onset of one or more
symptoms of the particular disease, condition or disorder described
herein. In the case of cancer, the therapeutically effective amount
of the drug may reduce the number of cancer cells; reduce the tumor
size; inhibit (i.e., slow to some extent and preferably stop)
cancer cell infiltration into peripheral organs; inhibit (i.e.,
slow to some extent and preferably stop) tumor metastasis; inhibit,
to some extent, tumor growth; and/or relieve to some extent one or
more of the symptoms associated with the cancer. For cancer
therapy, efficacy can, for example, be measured by assessing the
time to disease progression (TTP) and/or determining the response
rate (RR). In the case of immunological disorders, the therapeutic
effective amount is an amount sufficient to decrease or alleviate
an allergic disorder, the symptoms of an autoimmune and/or
inflammatory disease, or the symptoms of an acute inflammatory
reaction (e.g. asthma).
[0067] "Treatment" (and variations such as "treat" or "treating")
refers to clinical intervention in an attempt to alter the natural
course of the individual or cell being treated, and can be
performed either for prophylaxis or during the course of clinical
pathology. Desirable effects of treatment include one or more of
preventing recurrence of disease, alleviation of symptoms,
diminishment of any direct or indirect pathological consequences of
the disease, stabilized (i.e., not worsening) state of disease,
preventing metastasis, decreasing the rate of disease progression,
amelioration or palliation of the disease state, prolonging
survival as compared to expected survival if not receiving
treatment and remission or improved prognosis. In certain
embodiments, a compound of formula (I) is used to delay development
of a disease or disorder or to slow the progression of a disease or
disorder. Those individuals in need of treatment include those
already with the condition or disorder as well as those prone to
have the condition or disorder, (for example, through a genetic
mutation or abberent expression of a gene or protein).
[0068] "CBP/EP300 bromodomain inhibitor" or "CBP and/or EP300
bromodomain inhibitor" refers to a compound that binds to the CBP
bromodomain and/or EP300 bromodomain and inhibits and/or reduces a
biological activity of CBP and/or EP300. In some embodiments,
CBP/EP300 bromodomain inhibitor binds to the CBP and/or EP300
primarily (e.g., solely) through contacts and/or interactions with
the CBP bromodomain and/or EP300 bromodomain. In some embodiments,
CBP/EP300 bromodomain inhibitor binds to the CBP and/or EP300
through contacts and/or interactions with the CBP bromodomain
and/or EP300 bromodomain as well as additional CBP and/or EP300
residues and/or domains. In some embodiments, CBP/EP300 bromodomain
inhibitor substantially or completely inhibits the biological
activity of the CBP and/or EP300. In some embodiments, the
biological activity is binding of the bromodomain of CBP and/or
EP300 to chromatin (e.g., histones associated with DNA) and/or
another acetylated protein. In certain embodiments, the CBP/EP300
bromodomain inhibitor blocks CBP/EP300 activity so as to restore a
functional response by T-cells (e.g., proliferation, cytokine
production, target cell killing) from a dysfunctional state to
antigen stimulation. In some embodiments, the CBP/EP300 bromodomain
inhibitor binds to and inhibits CBP bromodomain. In some
embodiments, the CBP/EP300 bromodomain inhibitor binds to and
inhibits EP300 bromodomain.
[0069] As used herein, "a" or "an" means one or more, unless
clearly indicated otherwise. As used herein, "another" means at
least a second or more.
Exemplary Values for Compounds of Formula (I)
[0070] In certain embodiments the compound is a compound of formula
(Id):
##STR00011##
or a salt thereof, wherein:
[0071] U is CH or N; V is CH; and W is CH or N; or
[0072] U is CH or N; V is N; and W is CH; and
[0073] one of R.sup.3, R.sup.4, and R.sup.5 is selected from the
group consisting of hydrogen and R.sup.a and the remainder of
R.sup.3, R.sup.4, and R.sup.5 R.sup.b are independently selected
from the group consisting of hydrogen, halo, cyano, hydroxy, amino,
C.sub.1-4alkyl, C.sub.2-4alkenyl, C.sub.2-4alkynyl,
C.sub.2-6cycloalkyl, (C.sub.2-6cycloalkyl) C.sub.1-4alkyl,
C.sub.1-4alkoxy, C.sub.1-4alkoxycarbonyl, C.sub.1-4alkanoyl, and
C.sub.1-4alkanoyloxy, wherein each C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.2-4alkynyl, C.sub.2-6cycloalkyl,
(C.sub.2-6cycloalkyl) C.sub.1-4alkyl, C.sub.1-4 alkoxy,
C.sub.1-4alkoxycarbonyl, C.sub.1-4alkanoyl, and
C.sub.1-4alkanoyloxy, is optionally substituted with one or more
groups independently selected from oxo, halo, amino, hydroxy,
C.sub.1-3alkoxy, and C.sub.1-C.sub.3 alkyl that is optionally
substituted with one or more groups independently selected from
halo.
[0074] In certain embodiments the compound is a compound of formula
(Ia):
##STR00012##
or a salt thereof.
[0075] In certain embodiments the compound is a compound of formula
(Ib):
##STR00013##
or a salt thereof.
[0076] In certain embodiments the compound is a compound of formula
(Ic):
##STR00014##
or a salt thereof.
[0077] In certain embodiments R.sup.1 is C.sub.1-12alkyl or 3-12
membered heterocycle, wherein each C.sub.1-12alkyl and 3-12
membered heterocycle of R.sup.1 is optionally substituted with one
or more groups R.sup.d.
[0078] In certain embodiments R.sup.1 is C.sub.1-3alkyl or 3-6
membered heterocycle, wherein each C.sub.1-3alkyl or 3-6 membered
heterocycle of R.sup.1 is optionally substituted with one or more
groups R.sup.d.
[0079] In certain embodiments R.sup.1 is C.sub.1-3alkyl or 3-6
membered heterocycle, wherein each C.sub.1-3alkyl or 3-6 membered
heterocycle of R.sup.1 is optionally substituted with one or more
groups independently selected from the group consisting of oxo,
halo, C.sub.2-6cycloalkyl, and C.sub.1-4alkoxy.
[0080] In certain embodiments R.sup.1 is selected from the group
consisting of: methyl,
##STR00015##
[0081] In certain embodiments R.sup.1 is:
##STR00016##
[0082] In certain embodiments R.sup.2 is C.sub.1-4alkanoyl
[0083] In certain embodiments R.sup.2 is selected from the group
consisting of:
##STR00017##
[0084] In certain embodiments R.sup.2 is --C(.dbd.O)CH.sub.3.
[0085] In certain embodiments Y is phenyl, wherein Y is optionally
substituted with R.sup.a and wherein Y is further optionally
substituted with one or more groups R.sup.b.
[0086] In certain embodiments Y is 9-membered bicyclic carbocycle
or a 10-membered bicyclic carbocycle, wherein Y is optionally
substituted with R.sup.a and wherein Y is further optionally
substituted with one or more groups R.sup.b.
[0087] In certain embodiments Y is a 9-membered bicyclic
heterocycle or a 10-membered bicyclic heterocycle, wherein Y is
optionally substituted with R.sup.a and wherein Y is further
optionally substituted with one or more groups R.sup.b.
[0088] In certain embodiments --X--Y is selected from the group
consisting of:
##STR00018## ##STR00019## ##STR00020## ##STR00021## ##STR00022##
##STR00023## ##STR00024## ##STR00025## ##STR00026## ##STR00027##
##STR00028## ##STR00029## ##STR00030## ##STR00031## ##STR00032##
##STR00033## ##STR00034##
[0089] In certain embodiments Y is selected from the group
consisting of:
##STR00035## ##STR00036## ##STR00037## ##STR00038## ##STR00039##
##STR00040## ##STR00041## ##STR00042## ##STR00043## ##STR00044##
##STR00045## ##STR00046## ##STR00047## ##STR00048## ##STR00049##
##STR00050##
[0090] In certain embodiments --X--Y is selected from the group
consisting of:
##STR00051## ##STR00052## ##STR00053## ##STR00054##
[0091] In certain embodiments --X--Y is selected from the group
consisting of:
##STR00055## ##STR00056## ##STR00057##
[0092] In certain embodiments --X--Y is selected from the group
consisting of:
##STR00058## ##STR00059## ##STR00060##
[0093] In certain embodiments Y is selected from the group
consisting of:
##STR00061##
[0094] In certain embodiments --X--Y is selected from the group
consisting of:
##STR00062##
[0095] In certain embodiments the compound or salt is selected from
the group consisting of:
##STR00063## ##STR00064## ##STR00065## ##STR00066## ##STR00067##
##STR00068## ##STR00069## ##STR00070## ##STR00071## ##STR00072##
##STR00073## ##STR00074## ##STR00075## ##STR00076## ##STR00077##
##STR00078## ##STR00079## ##STR00080## ##STR00081## ##STR00082##
##STR00083## ##STR00084## ##STR00085## ##STR00086##
##STR00087## ##STR00088## ##STR00089## ##STR00090## ##STR00091##
##STR00092##
and salts thereof.
Uses, Formulation and Administration of Compounds of Formula
(I)
Pharmaceutically Acceptable Compositions
[0096] Another aspect includes a pharmaceutical composition
comprising a compound of formula (I) or a pharmaceutically
acceptable salt thereof. In one embodiment, the composition further
comprises a pharmaceutically acceptable carrier, adjuvant, or
vehicle. In another embodiment, the composition further comprises
an amount of the compound effective to measurably inhibit a
bromodomain of CBP and/or EP300. In certain embodiments, the
composition is formulated for administration to a patient in need
thereof.
[0097] The term "patient" or "individual" as used herein, refers to
an animal, such as a mammal, such as a human. In one embodiment,
patient or individual refers to a human.
[0098] The term "pharmaceutically acceptable carrier, adjuvant, or
vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that
does not destroy the pharmacological activity of the compound with
which it is formulated. Pharmaceutically acceptable carriers,
adjuvants or vehicles that may be used in the compositions of this
invention include, but are not limited to, ion exchangers, alumina,
aluminum stearate, lecithin, serum proteins, such as human serum
albumin, buffer substances such as phosphates, glycine, sorbic
acid, potassium sorbate, partial glyceride mixtures of saturated
vegetable fatty acids, water, salts or electrolytes, such as
protamine sulfate, disodium hydrogen phosphate, potassium hydrogen
phosphate, sodium chloride, zinc salts, colloidal silica, magnesium
trisilicate, polyvinyl pyrrolidone, cellulose-based substances,
polyethylene glycol, sodium carboxymethylcellulose, polyacrylates,
waxes, polyethylene-polyoxypropylene-block polymers, polyethylene
glycol and wool fat.
[0099] Compositions comprising a compound of formula (I) or salt
thereof may be administered orally, parenterally, by inhalation
spray, topically, transdermally, rectally, nasally, buccally,
sublingually, vaginally, intraperitoneal, intrapulmonary,
intradermal, epidural or via an implanted reservoir. The term
"parenteral" as used herein includes subcutaneous, intravenous,
intramuscular, intra-articular, intra-synovial, intrasternal,
intrathecal, intrahepatic, intralesional and intracranial injection
or infusion techniques.
[0100] In one embodiment, the composition comprising a compound of
formula (I) or salt thereof is formulated as a solid dosage form
for oral administration. Solid dosage forms for oral administration
include capsules, tablets, pills, powders, and granules. In certain
embodiments, the solid oral dosage form comprising a compound of
formula (I) or a salt thereof further comprises one or more of (i)
an inert, pharmaceutically acceptable excipient or carrier, such as
sodium citrate or dicalcium phosphate, and (ii) filler or extender
such as starches, lactose, sucrose, glucose, mannitol, or silicic
acid, (iii) binders such as carboxymethylcellulose, alginates,
gelatin, polyvinylpyrrolidinone, sucrose or acacia, (iv) humectants
such as glycerol, (v) disintegrating agent such as agar, calcium
carbonate, potato or tapioca starch, alginic acid, certain
silicates or sodium carbonate, (vi) solution retarding agents such
as paraffin, (vii) absorption accelerators such as quaternary
ammonium salts, (viii) a wetting agent such as cetyl alcohol or
glycerol monostearate, (ix) absorbent such as kaolin or bentonite
clay, and (x) lubricant such as talc, calcium stearate, magnesium
stearate, polyethylene glycols or sodium lauryl sulfate. In certain
embodiments, the solid oral dosage form is formulated as capsules,
tablets or pills. In certain embodiments, the solid oral dosage
form further comprises buffering agents. In certain embodiments,
such compositions for solid oral dosage forms may be formulated as
fillers in soft and hard-filled gelatin capsules comprising one or
more excipients such as lactose or milk sugar, polyethylene glycols
and the like.
[0101] In certain embodiments, tablets, dragees, capsules, pills
and granules of the compositions comprising a compound of formula
(I) or salt thereof optionally comprise coatings or shells such as
enteric coatings. They may optionally comprise opacifying agents
and can also be of a composition that they release the active
ingredient(s) only, or preferentially, in a certain part of the
intestinal tract, optionally, in a delayed manner. Examples of
embedding compositions include polymeric substances and waxes,
which may also be employed as fillers in soft and hard-filled
gelatin capsules using such excipients as lactose or milk sugar as
well as high molecular weight polethylene glycols and the like.
[0102] In another embodiment, a composition comprises
micro-encapsulated compound of formula (I) or salt thereof, and
optionally, further comprises one or more excipients.
[0103] In another embodiment, compositions comprise liquid dosage
formulations comprising a compound of formula (I) or salt thereof
for oral administration, and optionally further comprise one or
more of pharmaceutically acceptable emulsions, microemulsions,
solutions, suspensions, syrups and elixirs. In certain embodiments,
the liquid dosage form optionally, further comprise one or more of
an inert diluent such as water or other solvent, a solubilizing
agent, and an emulsifier such as ethyl alcohol, isopropyl alcohol,
ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in
particular, cottonseed, groundnut, corn, germ, olive, castor, and
sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene
glycols or fatty acid esters of sorbitan, and mixtures thereof. In
certain embodiments, liquid oral compositions optionally further
comprise one or more adjuvant, such as a wetting agent, a
suspending agent, a sweetening agent, a flavoring agent and a
perfuming agent.
[0104] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution, suspension or emulsion in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, U.S.P.
and isotonic sodium chloride solution. In addition, sterile, fixed
oils are conventionally employed as a solvent or suspending medium.
For this purpose any bland fixed oil can be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid are used in the preparation of injectables.
[0105] Injectable formulations can be sterilized, for example, by
filtration through a bacterial-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use.
[0106] In order to prolong the effect of a compound of formula (I),
it is often desirable to slow the absorption of the compound from
subcutaneous or intramuscular injection. This may be accomplished
by the use of a liquid suspension of crystalline or amorphous
material with poor water solubility. The rate of absorption of the
compound then depends upon its rate of dissolution that, in turn,
may depend upon crystal size and crystalline form. Alternatively,
delayed absorption of a parenterally administered compound form is
accomplished by dissolving or suspending the compound in an oil
vehicle. Injectable depot forms are made by forming microencapsule
matrices of the compound in biodegradable polymers such as
polylactide-polyglycolide. Depending upon the ratio of compound to
polymer and the nature of the particular polymer employed, the rate
of compound release can be controlled. Examples of other
biodegradable polymers include poly(orthoesters) and
poly(anhydrides). Depot injectable formulations are also prepared
by entrapping the compound in liposomes or microemulsions that are
compatible with body tissues.
[0107] In certain embodiments, the composition for rectal or
vaginal administration are formulated as suppositories which can be
prepared by mixing a compound of formula (I) or a salt thereof with
suitable non-irritating excipients or carriers such as cocoa
butter, polyethylene glycol or a suppository wax, for example those
which are solid at ambient temperature but liquid at body
temperature and therefore melt in the rectum or vaginal cavity and
release the compound of formula (I).
[0108] Example dosage forms for topical or transdermal
administration of a compound of formula (I) include ointments,
pastes, creams, lotions, gels, powders, solutions, sprays,
inhalants or patches. The compound of formula (I) or a salt thereof
is admixed under sterile conditions with a pharmaceutically
acceptable carrier, and optionally preservatives or buffers.
Additional formulation examples include an ophthalmic formulation,
ear drops, eye drops, transdermal patches. Transdermal dosage forms
can be made by dissolving or dispensing the compound of formula (I)
or a salt thereof in medium, for example ethanol or
dimethylsulfoxide. Absorption enhancers can also be used to
increase the flux of the compound across the skin. The rate can be
controlled by either providing a rate controlling membrane or by
dispersing the compound in a polymer matrix or gel.
[0109] Nasal aerosol or inhalation formulations of a compound of
formula (I) or a salt thereof may be prepared as solutions in
saline, employing benzyl alcohol or other suitable preservatives,
absorption promotors to enhance bioavailability, fluorocarbons,
and/or other conventional solubilizing or dispersing agents.
[0110] In certain embodiments, pharmaceutical compositions may be
administered with or without food. In certain embodiments,
pharmaceutically acceptable compositions are administered without
food. In certain embodiments, pharmaceutically acceptable
compositions of this invention are administered with food.
[0111] Specific dosage and treatment regimen for any particular
patient will depend upon a variety of factors, including age, body
weight, general health, sex, diet, time of administration, rate of
excretion, drug combination, the judgment of the treating
physician, and the severity of the particular disease being
treated. The amount of a provided compound of formula (I) or salt
thereof in the composition will also depend upon the particular
compound in the composition.
[0112] In one embodiment, the therapeutically effective amount of
the compound of the invention administered parenterally per dose
will be in the range of about 0.01-100 mg/kg, alternatively about
0.1 to 20 mg/kg of patient body weight per day, with the typical
initial range of compound used being 0.3 to 15 mg/kg/day. In
another embodiment, oral unit dosage forms, such as tablets and
capsules, contain from about 5 to about 100 mg of the compound of
the invention.
[0113] An example tablet oral dosage form comprises about 2 mg, 5
mg, 25 mg, 50 mg, 100 mg, 250 mg or 500 mg of a compound of formula
(I) or salt thereof, and further comprises about 5-30 mg anhydrous
lactose, about 5-40 mg sodium croscarmellose, about 5-30 mg
polyvinylpyrrolidone (PVP) K30 and about 1-10 mg magnesium
stearate. The process of formulating the tablet comprises mixing
the powdered ingredients together and further mixing with a
solution of the PVP. The resulting composition can be dried,
granulated, mixed with the magnesium stearate and compressed to
tablet form using conventional equipment. An example of an aerosol
formulation can be prepared by dissolving about 2-500 mg of a
compound of formula (I) or salt thereof, in a suitable buffer
solution, e.g. a phosphate buffer, and adding a tonicifier, e.g. a
salt such sodium chloride, if desired. The solution may be
filtered, e.g. using a 0.2 micron filter, to remove impurities and
contaminants.
Uses of Compounds and Pharmaceutically Acceptable Compositions
[0114] Another aspect includes the use of a compound of formula (I)
or a salt thereof for the inhibition of a bromodomain (in vitro or
in vivo) (e.g., in vitro or in vivo inhibition of the bromodomain
of CBP/EP300).
[0115] Another embodiment includes a method for treating a
bromodomain-mediated disorder (e.g., CBP/EP300 bromodomain-mediated
disorder) in an animal comprising administering a compound of
formula (I), or a pharmaceutically acceptable salt thereof to the
animal. CBP/EP300-mediated disorders include, but are not limited
to those disorders described herein.
[0116] Another embodiment includes a method of increasing efficacy
of a cancer treatment comprising a cytotoxic agent in an animal
comprising administering to the animal an effective amount of a
compound of formula (I) or a pharmaceutically acceptable salt
thereof.
[0117] Another embodiment includes a method of extending the
duration of response to a cancer therapy in an animal, comprising
administering to an animal undergoing the cancer therapy a compound
of formula (I) or a pharmaceutically acceptable salt thereof,
wherein the duration of response to the cancer therapy when the
compound of formula (I) or the pharmaceutically acceptable salt
thereof is administered is extended over the duration of response
to the cancer therapy in the absence of the administration of the
compound of formula (I) or the pharmaceutically acceptable salt
thereof.
[0118] Another embodiment includes a method of treating cancer in
an individual comprising administering to the individual (a) a
compound of formula (I) or a pharmaceutically acceptable salt
thereof, and (b) a cytotoxic agent. In one embodiment the cytotoxic
agent is selected from anti-microtubule agents, platinum
coordination complexes, alkylating agents, antibiotic agents,
topoisomerase II inhibitors, antimetabolites, topoisomerase I
inhibitors, hormones and hormonal analogues, signal transduction
pathway inhibitors, non-receptor tyrosine kinase angiogenesis
inhibitors, immunotherapeutic agents, proapoptotic agents,
inhibitors of LDH-A, inhibitors of fatty acid biosynthesis, cell
cycle signaling inhibitors, HDAC inhibitors, proteasome inhibitors,
and inhibitors of cancer metabolism. In one embodiment the
cytotoxic agent is a taxane. In one embodiment the taxane is
paclitaxel or docetaxel. In one embodiment the cytotoxic agent is a
platinum agent. In one embodiment the cytotoxic agent is an
antagonist of EGFR. In one embodiment the antagonist of EGFR is
N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine or a
pharmaceutically acceptable salt thereof (e.g., erlotinib). In one
embodiment the cytotoxic agent is a RAF inhibitor. In one
embodiment the RAF inhibitor is a BRAF or CRAF inhibitor. In one
embodiment the RAF inhibitor is vemurafenib. In one embodiment the
cytotoxic agent is a PI3K inhibitor.
[0119] In certain embodiments, treatment may be administered after
one or more symptoms have developed. In other embodiments,
treatment may be administered in the absence of symptoms. For
example, treatment may be administered to a susceptible individual
prior to the onset of symptoms (e.g., in light of a history of
symptoms and/or in light of genetic or other susceptibility
factors). Treatment may also be continued after symptoms have
resolved, for example to prevent or delay their recurrence.
[0120] In some embodiments, the CBP/EP300 bromodomain inhibitor
interferes with the associating of CBP and/or EP300 with histones,
in particular acetylated lysines in histones. In some embodiments,
the CBP/EP300 bromodomain inhibitor inhibits binding of CBP and/or
EP300 to chromatin (e.g., histone associated DNA). In some
embodiments, the CBP/EP300 bromodomain inhibitor inhibits and/or
reduces binding of the CBP bromodomain and/or EP300 bromodomain to
chromatin (e.g., histone associated DNA). In some embodiments, the
CBP/EP300 bromodomain inhibitor does not affect association of
other domains of CBP and/or EP300 to chromatin. In some
embodiments, CBP/EP300 bromodomain inhibitor binds to the CBP
and/or EP300 primarily (e.g., solely) through contacts and/or
interactions with the CBP bromodomain and/or EP300 bromodomain. In
some embodiments, CBP/EP300 bromodomain inhibitor binds to the CBP
and/or EP300 through contacts and/or interactions with the CBP
bromodomain and/or EP300 bromodomain as well as additional CBP
and/or EP300 residues and/or domains. Methods of assaying
association with chromatin are known in the art and include, but
are not limited to, chromatin fractionation, BRET assay (Promega),
FRAP assay, Chromatin Immunoprecipitation (ChTP), biophysical
binding assay, and/or Histone Association Assay. See, e.g., Das et
al., BioTechniques 37:961-969 (2004).
[0121] In some embodiments, the CBP/EP300 bromodomain inhibitor
does not affect effector function in CD8 cells (i.e., effector
function is substantially the same in the presence and/or absence
of the CBP/EP300 bromodomain inhibitor). In some embodiments, the
CBP/EP300 bromodomain inhibitor does not affect expression levels
of perforin, granzyme, and/or EOMES (i.e., expression levels of one
or more perforin, granzyme, and/or EOMES are substantially the same
in the presence and/or absence of the CBP/EP300 bromodomain
inhibitor). In some embodiments, the CBP/EP300 bromodomain
inhibitor does not affect expression levels of effector cytokines
IFN-.gamma. and/or TNF.alpha. (i.e., expression levels of effector
cytokines IFN-.gamma. and/or TNF.alpha. are substantially the same
in the presence and/or absence of the CBP/EP300 bromodomain
inhibitor). In some embodiments, the CBP/EP300 bromodomain
inhibitor enhances naive T cell responsiveness to CD3/CD28
stimulation in the presence of Treg cells.
[0122] In some embodiments, the CBP/EP300 bromodomain inhibitor
does not substantially bind to (e.g., does not bind to) the HAT
domain of CBP and/or EP300. In some embodiments, the CBP/EP300
bromodomain inhibitor does not substantially bind to (e.g., does
not bind to) the HAT domain of CBP and/or EP300 as identified in
Delvecchio et al., Nat. Struct. & Mol. Biol. 20:1040-1046
(2013), which is incorporated by reference in its entirety. In some
embodiments, the CBP/EP300 bromodomain inhibitor does not
substantially bind to one or more residues of the amino acid
sequence ENKFSAKRLQTTR
LGNHLEDRVNKFLRRQNHPEAGEVFVRVVASSDKTVEVKPGMKSRFVDSGEMSESFPY
RTKALFAFEEIDGVDVCFFGMHVQEYGSDCPPPNTRRVYISYLDSIHFFRPRCLRTAVYH
EILIGYLEYVKKLGYVTGHIWACPPSEGDDYIFHCHPPDQKIPKPKRLQEWYKKMLDKA
FAERIIHDYKDIFKQATEDRLTSAKELPYFEGDFWPNVLEESIKELEQEEEERKKEESTAA
SETTEGSQGDSKNAKKKNNKKTNKNKSSISRANKKKPSMPNVSNDLSQKLYATMEKH
KEVFFVIHLHAGPVINTLPPIVDPDPLLSCDLMDGRDAFLTLARDKHWEFSSLRRSKWST
LCMLVELHTQGQD (amino acid residues 1321-1701 of UniProt No. Q92793
(SEQ ID NO:1)). In some embodiments, the CBP/EP300 bromodomain
inhibitor does not substantially bind to one or more residues of
the amino acid sequence
ENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHASDKTVEVKPGMKAR
FVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPPPNQRRVYISYLDSV
HFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDYIFHCHPPDQKIPKPK
RLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFEGDFWPNVLEESIKEL
EQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLSRGNKKKPGMPNVSN
DLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDLMDGRDAFLTLARDKH
LEFSSLRRAQWSTMCMLVELHTQSQD (amino acid residues 1285-1664 of
UniProt No. Q09472 (SEQ ID NO:2)). In some embodiments, the
CBP/EP300 bromodomain inhibitor does not inhibit the histone
acetyltransferase (HAT) catalytic activity of CBP and/or EP300.
[0123] Compounds that are CBP/EP300 bromodomain inhibitors are
expected to have improved and/or distinct properties over other
compounds, such as "HAT" inhibitor compounds. HAT inhibition is
expected to result in a global reduction in protein acetylation
(histone and non-histone), likely affecting cell viability in a
significant way. In some embodiments, CBP/EP300 bromodomain
inhibition preserves the HAT activity of these proteins while
resulting in the reduction of transcriptional activity of a
relatively small subset of target genes.
[0124] In some embodiments, provided are methods of enhancing
immune function in an individual having cancer comprising
administering an effective amount of any CBP/EP300 bromodomain
inhibitors disclosed herein. In some embodiments of any of the
methods, the CD8 T cells in the individual have enhanced priming,
activation, proliferation, and/or cytolytic activity relative to
prior to the administration of the CBP/EP300 bromodomain inhibitor.
In some embodiments, the number of CD8 T cells is elevated relative
to prior to administration of the CBP/EP300 bromodomain inhibitors.
In some embodiments, the CD8 T cells have reduced levels of
expression of one or more of the following biomarkers: IFNA17,
IGFI, FSCN1, SUMO2, CIorf129, EIF2S2, TDGF1, AIDA, CCR4, CD160,
MC4R, KRTAP2-2, MTIJP, OR4N2, KRTAP4-5, MTIL//MTIL, ILI3, LCEID,
KIR2DL2, LOC158696, LIF, IL28A, TAS2R13, CTLA4, and/or FOXP3
relative to prior to administration of the CBP/EP300 bromodomain
inhibitor. In some embodiments, the CD8 T cells have reduced levels
of expression of CD160 and/or KIR2DL2 relative to prior to
administration of the CBP/EP300 bromodomain inhibitor.
[0125] In some embodiments of the methods of enhancing immune
function, the enhanced immune function is characterized by Treg
cells in the individual (e.g., at the tumor site(s)) have reduced
levels of expression of one or more of the following markers:
IL28A, GPR87, ANKRD37, CABLES1, RAPGEF2, TRIM69, MT1L//MT1L,
FAM1138, FOXP3, CSF2, OCM2, GLIPR1, FGFBP2, CTLA4, CST7, GOLGA6L1,
IFIT3, FAM13A, APOD, AK2, CLDN1, HSD11B1, DNAJC12, PHEX, IL2,
FOXD4L3, GNA15, ZBTB32, RDH10, OR52E5, CYP2A6, GZMH, CCL20, ADM,
LOC100131541, RNF122, FAM36A, AMY2B, GPR183, MYOF, IL29, AIDA,
SPRYI, ENOPHI, IL1RN, SLAMF1, PGM2L1, SSBP3, MMP23B, HISTIH3J,
MYO1B, BEND5, S1PR1, CDK6, GPR56, ZC3HIZA, DOK5, DUSPI, CYB5R2,
KCNAB2, LAG3, KLF10, GK, SHC4, IL12RB2, CD109, HAVCR2 (TIM-3), LTA,
FAM40B, HMGCSI, HSPA1A, ZNF705A, CMAH, KIF3A, CHN1, KBTBD8, TNF,
MOP-1, RASGRP4, INSIG1, SLAMF7, OR10H4, LPL, HIST1H2BJ, LIF, IGF1,
IL18RAP, OR52N4, OR1D2, CCR4, CXCR5, IL1R1, MICAL2, NRN1, PICALM,
B3GNT5, IFI44L, CXCR3, ICOS, IFIT2, NCR3, HSPA1B, CD80, GNG2,
C7orf68, GPR171, RPS10P7, IL23A, LOC283174, PLK2, EMP1, FNBP1L,
CD226, RBMS3, IL23R, PTGER4, GZMB, F5, and/or HIST1H2BK relative to
prior to administration of CBP/EP300 bromodomain inhibitor. In some
embodiments, the Treg cell biomarker is one or more of LAG3, CTLA4,
and/or FOXP3. In some embodiments of the methods of enhancing
immune function, the enhanced immune function is characterized by
enhanced naive T cell responsiveness to CD3/CD28 stimulation in the
presence of Treg cells. In some embodiments, the CD8 T cell priming
is characterized by increased T cell proliferation and/or enhanced
cytolytic activity in CD8 T cells. In some embodiments, the CD8 T
cell activation is characterized by an elevated frequency of
T-IFN.sup.+ CD8 T cells. In some embodiments, the CD8 T cell is an
antigen-specific T-cell. In some embodiments, the immune evasion is
inhibited.
[0126] In some embodiments, the methods provided herein are useful
in treating conditions where enhanced immunogenicity is desired
such as increasing tumor immunogenicity for the treatment of
cancer. For example, provided herein are CBP/EP300 bromodomain
inhibitors for use to enhance T-cell function to upregulate
cell-mediated immune responses and for the treatment of T cell
dysfunctional disorders, tumor immunity. In some embodiments, the
CBP/EP300 bromodomain inhibitors promote anti-tumor immunity by
inhibiting the suppressive function of regulatory T (Treg) cells
and/or relieving T cell exhaustion on chronically stimulated
CD8.sup.+ T cells. CBP/EP300 bromodomain inhibitors are further
useful in reducing FOXP3 expression during extra-thymic Treg cell
differentiation. Continual FOXP3 expression is essential to
maintain suppressive activity in Treg cells. In some embodiments,
reduced FOXP3 expression through CBP/EP300 bromodomain inhibition
impairs Treg cells suppressive activity and promotes tumor
antiimmunity. Treg cells are highly enriched in tumors derived from
multiple cancer indications, including melanoma, NSCLC, renal,
overian, colon, pancreatic, hepatocellular, and breast cancer. In a
subset of these indications, increased intratumoral Treg cell
densities are associated with poor patient prognosis. These
indications include NSCLC, ovarian, pancreatic, hepatocellular, and
breat cancer. CBP/EP300 bromodomain inhibitors are predicted to
impair intrtumoral Treg cell function in these cancer indications
to enhance effector T cell activity. In other embodiments, the
CBP/EP300 bromodomain inhibitors may be used to treat infectious
diseases, where some pathogens may have evolved to manipulate
regulatory T (Treg) cells to immunosuppress the host to ensure
survival, such as in retrovial infections (e.g., HIV),
mycobacterial infections (e.g., tuberculosis), and parasitic
infections (e.g., Leishmania and malaria).
[0127] In some embodiments, the methods provided herein are useful
in treating a CBP and/or EP300-mediated disorder involving
fibrosis. In some embodiments, the CBP and/or EP300-mediated
disorder is a fibrotic disease. Certain fibrotic diseases may
include, for example, pulmonary fibrosis, silicosis, cystic
fibrosis, renal fibrosis, liver fibrosis, liver cirrhosis, primary
sclerosing cholangitis, primary biliary cirrhosis, endomyocardial
fibrosis, mediastinal fibrosis, myelofibrosis, retroperitoneal
fibrosis, progressive massive fibrosis, nephrogenic systemic
fibrosis, Crohn's disease, keloid, myocardial infarction, systemic
sclerosis or arthro fibrosis.
[0128] In other embodiments, the CBP and/or EP300-mediated disorder
is a fibrotic lung disease. Fibrotic lung diseases may include, for
example, idiopathic pulmonary fibrosis, fibrotic interstitial lung
disease, interstitial pneumonia, fibrotic variant of non-specific
interstitial pneumonia, cystic fibrosis, lung fibrosis, chronic
obstructive pulmonary lung disease (COPD), or pulmonary arterial
hypertension. In certain embodiments, the fibrotic lung disease is
idiopathic pulmonary fibrosis.
CBP and/or EP300-Mediated Disorders
[0129] A "CBP and/or EP300-mediated disorder" is characterized by
the participation of the bromodomains of CBP and/or EP300 in the
inception, manifestation of one or more symptoms or disease
markers, severity, or progression of a disorder. In one embodiment
the bromodomain-mediated disorder is a CBP bromodomain-mediated
disorder. In one embodiment the bromodomain-mediated disorder is an
EP300 bromodomain-mediated disorder.
[0130] CBP and/or EP300 bromodomain-mediated disorders include
cancers, including, but not limited to acoustic neuroma, acute
leukemia, acute lymphocytic leukemia, acute myelocytic leukemia
(monocytic, myeloblastic, adenocarcinoma, angiosarcoma,
astrocytoma, myelomonocytic and promyelocytic), acute T-cell
leukemia, basal cell carcinoma, bile duct carcinoma, bladder
cancer, brain cancer, breast cancer, bronchogenic carcinoma,
cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic
leukemia, chronic lymphocytic leukemia, chronic myelocytic
(granulocytic) leukemia, chronic myelogenous leukemia, colon
cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma,
diffuse large B-cell lymphoma, dysproliferative changes (dysplasias
and metaplasias), embryonal carcinoma, endometrial cancer,
endotheliosarcoma, ependymoma, epithelial carcinoma,
erythroleukemia, esophageal cancer, estrogen-receptor positive
breast cancer, essential thrombocythemia, Ewing's tumor,
fibrosarcoma, follicular lymphoma, germ cell testicular cancer,
glioma, glioblastoma, gliosarcoma, heavy chain disease,
hemangioblastoma, hepatoma, hepatocellular cancer, hormone
insensitive prostate cancer, leiomyosarcoma, leukemia, liposarcoma,
lung cancer, lymphagioendotheliosarcoma, lymphangiosarcoma,
lymphoblastic leukemia, lymphoma (Hodgkin's and non-Hodgkin's),
malignancies and hyperproliferative disorders of the bladder,
breast, colon, lung, ovaries, pancreas, prostate, skin and uterus,
lymphoid malignancies of T-cell or B-cell origin, medullary
carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma,
multiple myeloma, myelogenous leukemia, myeloma, myxosarcoma,
neuroblastoma, NUT midline carcinoma (NMC), non-small cell lung
cancer, oligodendroglioma, oral cancer, osteogenic sarcoma, ovarian
cancer, pancreatic cancer, papillary adenocarcinomas, papillary
carcinoma, pinealoma, polycythemia vera, prostate cancer, rectal
cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma,
sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, small
cell lung carcinoma, solid tumors (carcinomas and sarcomas), small
cell lung cancer, stomach cancer, squamous cell carcinoma,
synovioma, sweat gland carcinoma, thyroid cancer, Waldenstr6m's
macroglobulinemia, testicular tumors, uterine cancer and Wilms'
tumor.
[0131] In certain embodiments, the cancer is lung cancer, breast
cancer, pancreatic cancer, colorectal cancer, and/or melanoma. In
certain embodiments, the cancer is lung. In certain embodiments,
the lung cancer is NSCLC. In certain embodiments, the cancer is
breast cancer.
[0132] In certain embodiments, the cancer is melanoma.
[0133] CBP and/or EP300-mediated disorders also include
inflammatory diseases, inflammatory conditions, and autoimmune
diseases, including, but not limited to: Addison's disease, acute
gout, ankylosing spondylitis, asthma, atherosclerosis, Behcet's
disease, bullous skin diseases, chronic obstructive pulmonary
disease (COPD), Crohn's disease, dermatitis, eczema, giant cell
arteritis, glomerulonephritis, hepatitis, hypophysitis,
inflammatory bowel disease, Kawasaki disease, lupus nephritis,
multiple sclerosis, myocarditis, myositis, nephritis, organ
transplant rejection, osteoarthritis, pancreatitis, pericarditis,
Polyarteritis nodosa, pneumonitis, primary biliary cirrhosis,
psoriasis, psoriatic arthritis, rheumatoid arthritis, scleritis,
sclerosing cholangitis, sepsis, systemic lupus erythematosus,
Takayasu's Arteritis, toxic shock, thyroiditis, type I diabetes,
ulcerative colitis, uveitis, vitiligo, vasculitis, and Wegener's
granulomatosis.
[0134] CBP and/or EP300-mediated disorders also include AIDS;
chronic kidney diseases, including, but are not limited to diabetic
nephropathy, hypertensive nephropathy, HIV-associated nephropathy,
glomerulonephritis, lupus nephritis, IgA nephropathy, focal
segmental glomerulosclerosis, membranous glomerulonephritis,
minimal change disease, polycystic kidney disease and tubular
interstitial nephritis; acute kidney injury or disease or condition
including, but are not limited to ischemia-reperfusion induced,
cardiac and major surgery induced, percutaneous coronary
intervention induced, radio-contrast agent induced, sepsis induced,
pneumonia induced, and drug toxicity induced; obesity;
dyslipidemia; hypercholesterolemia; Alzheimer's disease; metabolic
syndrome; hepatic steatosis; type II diabetes; insulin resistance;
and diabetic retinopathy.
[0135] CBP and/or EP300 inhibitors may also be used to provide male
contraception.
[0136] CBP and/or EP300-mediated disorders also include fibrotic
diseases. Certain fibrotic diseases may include, for example,
pulmonary fibrosis, silicosis, cystic fibrosis, renal fibrosis,
liver fibrosis, liver cirrhosis, primary sclerosing cholangitis,
primary biliary cirrhosis, endomyocardial fibrosis, mediastinal
fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive
massive fibrosis, nephrogenic systemic fibrosis, Crohn's disease,
keloid, myocardial infarction, systemic sclerosis or arthro
fibrosis.
[0137] CBP and/or EP300-mediated disorders also include fibrotic
lung diseases. Fibrotic lung diseases may include, for example,
idiopathic pulmonary fibrosis, fibrotic interstitial lung disease,
interstitial pneumonia, fibrotic variant of non-specific
interstitial pneumonia, cystic fibrosis, lung fibrosis, chronic
obstructive pulmonary lung disease (COPD), or pulmonary arterial
hypertension. In certain embodiments, the fibrotic lung disease is
idiopathic pulmonary fibrosis.
Co-Administration of Compounds and Other Agents
[0138] The compounds of formula (I) or salts thereof may be
employed alone or in combination with other agents for treatment.
For example, the second agent of the pharmaceutical combination
formulation or dosing regimen may have complementary activities to
the compound of formula (I) such that they do not adversely affect
each other. The compounds may be administered together in a unitary
pharmaceutical composition or separately. In one embodiment a
compound or a pharmaceutically acceptable salt can be
co-administered with a cytotoxic agent to treat proliferative
diseases and cancer.
The term "co-administering" refers to either simultaneous
administration, or any manner of separate sequential
administration, of a compound of formula (I) or a salt thereof, and
a further active pharmaceutical ingredient or ingredients,
including cytotoxic agents and radiation treatment. If the
administration is not simultaneous, the compounds are administered
in a close time proximity to each other. Furthermore, it does not
matter if the compounds are administered in the same dosage form,
e.g. one compound may be administered topically and another
compound may be administered orally.
[0139] Those additional agents may be administered separately from
an inventive compound-containing composition, as part of a multiple
dosage regimen. Alternatively, those agents may be part of a single
dosage form, mixed together with a compound of this invention in a
single composition. If administered as part of a multiple dosage
regime, the two active agents may be submitted simultaneously,
sequentially or within a period of time from one another normally
within five hours from one another.
[0140] As used herein, the term "combination," "combined," and
related terms refers to the simultaneous or sequential
administration of therapeutic agents in accordance with this
invention. For example, a compound of the present invention may be
administered with another therapeutic agent simultaneously or
sequentially in separate unit dosage forms or together in a single
unit dosage form. Accordingly, the present invention provides a
single unit dosage form comprising a compound of formula (I), an
additional therapeutic agent, and a pharmaceutically acceptable
carrier, adjuvant, or vehicle.
[0141] The amount of both an inventive compound and additional
therapeutic agent (in those compositions which comprise an
additional therapeutic agent as described above) that may be
combined with the carrier materials to produce a single dosage form
will vary depending upon the host treated and the particular mode
of administration. In certain embodiments, compositions of this
invention are formulated such that a dosage of between 0.01-100
mg/kg body weight/day of an inventive can be administered.
[0142] Typically, any agent that has activity against a disease or
condition being treated may be co-administered. Examples of such
agents can be found in Cancer Principles and Practice of Oncology
by V. T. Devita and S. Hellman (editors), 6.sup.th edition (Feb.
15, 2001), Lippincott Williams & Wilkins Publishers. A person
of ordinary skill in the art would be able to discern which
combinations of agents would be useful based on the particular
characteristics of the drugs and the disease involved.
[0143] In one embodiment, the treatment method includes the
co-administration of a compound of formula (I) or a
pharmaceutically acceptable salt thereof and at least one cytotoxic
agent. The term "cytotoxic agent" as used herein refers to a
substance that inhibits or prevents a cellular function and/or
causes cell death or destruction. Cytotoxic agents include, but are
not limited to, radioactive isotopes (e.g., At.sup.211, I.sup.131,
I.sup.125, Y.sup.90, Re.sup.186, Re.sup.188, Sm.sup.153,
Bi.sup.212, P.sup.32, Pb.sup.212 and radioactive isotopes of Lu);
chemotherapeutic agents; growth inhibitory agents; enzymes and
fragments thereof such as nucleolytic enzymes; and toxins such as
small molecule toxins or enzymatically active toxins of bacterial,
fungal, plant or animal origin, including fragments and/or variants
thereof.
[0144] Exemplary cytotoxic agents can be selected from
anti-microtubule agents, platinum coordination complexes,
alkylating agents, antibiotic agents, topoisomerase II inhibitors,
antimetabolites, topoisomerase I inhibitors, hormones and hormonal
analogues, signal transduction pathway inhibitors, non-receptor
tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents,
proapoptotic agents, inhibitors of LDH-A; inhibitors of fatty acid
biosynthesis; cell cycle signaling inhibitors; HDAC inhibitors,
proteasome inhibitors; and inhibitors of cancer metabolism.
[0145] "Chemotherapeutic agent" includes chemical compounds useful
in the treatment of cancer. Examples of chemotherapeutic agents
include erlotinib (TARCEVA.RTM., Genentech/OSI Pharm.), bortezomib
(VELCADE.RTM., Millennium Pharm.), disulfiram, epigallocatechin
gallate, salinosporamide A, carfilzomib, 17-AAG(geldanamycin),
radicicol, lactate dehydrogenase A (LDH-A), fulvestrant
(FASLODEX.RTM., AstraZeneca), sunitib (SUTENT.RTM., Pfizer/Sugen),
letrozole (FEMARA.RTM., Novartis), imatinib mesylate (GLEEVEC.RTM.,
Novartis), finasunate (VATALANIB.RTM., Novartis), oxaliplatin
(ELOXATIN.RTM., Sanofi), 5-FU (5-fluorouracil), leucovorin,
Rapamycin (Sirolimus, RAPAMUNE.RTM., Wyeth), Lapatinib
(TYKERB.RTM., GSK572016, Glaxo Smith Kline), Lonafamib (SCH 66336),
sorafenib (NEXAVAR.RTM., Bayer Labs), gefitinib (IRESSA.RTM.,
AstraZeneca), AG1478, alkylating agents such as thiotepa and
CYTOXAN.RTM. cyclosphosphamide; alkyl sulfonates such as busulfan,
improsulfan and piposulfan; aziridines such as benzodopa,
carboquone, meturedopa, and uredopa; ethylenimines and
methylamelamines including altretamine, triethylenemelamine,
triethylenephosphoramide, triethylenethiophosphoramide and
trimethylomelamine; acetogenins (especially bullatacin and
bullatacinone); a camptothecin (including topotecan and
irinotecan); bryostatin; callystatin; CC-1065 (including its
adozelesin, carzelesin and bizelesin synthetic analogs);
cryptophycins (particularly cryptophycin 1 and cryptophycin 8);
adrenocorticosteroids (including prednisone and prednisolone);
cyproterone acetate; 5.alpha.-reductases including finasteride and
dutasteride); vorinostat, romidepsin, panobinostat, valproic acid,
mocetinostat dolastatin; aldesleukin, talc duocarmycin (including
the synthetic analogs, KW-2189 and CB1-TM1); eleutherobin;
pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards
such as chlorambucil, chlomaphazine, chlorophosphamide,
estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide
hydrochloride, melphalan, novembichin, phenesterine, prednimustine,
trofosfamide, uracil mustard; nitrosoureas such as carmustine,
chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine;
antibiotics such as the enediyne antibiotics (e.g., calicheamicin,
especially calicheamicin .gamma.1I and calicheamicin .omega.1I
(Angew Chem. Intl. Ed. Engl. 1994 33:183-186); dynemicin, including
dynemicin A; bisphosphonates, such as clodronate; an esperamicin;
as well as neocarzinostatin chromophore and related chromoprotein
enediyne antibiotic chromophores), aclacinomysins, actinomycin,
authramycin, azaserine, bleomycins, cactinomycin, carabicin,
caminomycin, carzinophilin, chromomycinis, dactinomycin,
daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine,
ADRIAMYCIN.RTM. (doxorubicin), morpholino-doxorubicin,
cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and
deoxydoxorubicin), epirubicin, esorubicin, idarubicin,
marcellomycin, mitomycins such as mitomycin C, mycophenolic acid,
nogalamycin, olivomycins, peplomycin, porfiromycin, puromycin,
quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin,
ubenimex, zinostatin, zorubicin; anti-metabolites such as
methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as
denopterin, methotrexate, pteropterin, trimetrexate; purine analogs
such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine;
pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine,
carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine,
floxuridine; androgens such as calusterone, dromostanolone
propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals
such as aminoglutethimide, mitotane, trilostane; folic acid
replenisher such as frolinic acid; aceglatone; aldophosphamide
glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil;
bisantrene; edatraxate; defofamine; demecolcine; diaziquone;
elfomithine; elliptinium acetate; an epothilone; etoglucid; gallium
nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as
maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidamnol;
nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone;
podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK.RTM.
polysaccharide complex (JHS Natural Products, Eugene, Oreg.);
razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid;
triaziquone; 2,2',2''-trichlorotriethylamine; trichothecenes
(especially T-2 toxin, verracurin A, roridin A and anguidine);
urethan; vindesine; dacarbazine; mannomustine; mitobronitol;
mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C");
cyclophosphamide; thiotepa; taxoids, e.g., TAXOL (paclitaxel;
Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANE.RTM.
(Cremophor-free), albumin-engineered nanoparticle formulations of
paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.),
and TAXOTERE.RTM. (docetaxel, doxetaxel; Sanofi-Aventis);
chloranmbucil; GEMZAR.RTM. (gemcitabine); 6-thioguanine;
mercaptopurine; methotrexate; platinum analogs such as cisplatin
and carboplatin; vinblastine; etoposide (VP-16); ifosfamide;
mitoxantrone; vincristine; NAVELBINE.RTM. (vinorelbine);
novantrone; teniposide; edatrexate; daunomycin; aminopterin;
capecitabine (XELODA.RTM.); ibandronate; CPT-11; topoisomerase
inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such
as retinoic acid; and pharmaceutically acceptable salts, acids and
derivatives of any of the above.
[0146] Chemotherapeutic agent also includes (i) anti-hormonal
agents that act to regulate or inhibit hormone action on tumors
such as anti-estrogens and selective estrogen receptor modulators
(SERMs), including, for example, tamoxifen (including
NOLVADEX.RTM.; tamoxifen citrate), raloxifene, droloxifene,
iodoxyfene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018,
onapristone, and FARESTON.RTM. (toremifine citrate); (ii) aromatase
inhibitors that inhibit the enzyme aromatase, which regulates
estrogen production in the adrenal glands, such as, for example,
4(5)-imidazoles, aminoglutethimide, MEGASE.RTM. (megestrol
acetate), AROMASIN.RTM. (exemestane; Pfizer), formestanie,
fadrozole, RIVISOR.RTM. (vorozole), FEMARA.RTM. (letrozole;
Novartis), and ARIMIDEX.RTM. (anastrozole; AstraZeneca); (iii)
anti-androgens such as flutamide, nilutamide, bicalutamide,
leuprolide and goserelin; buserelin, tripterelin,
medroxyprogesterone acetate, diethylstilbestrol, premarin,
fluoxymesterone, all transretionic acid, fenretinide, as well as
troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); (iv)
protein kinase inhibitors; (v) lipid kinase inhibitors; (vi)
antisense oligonucleotides, particularly those which inhibit
expression of genes in signaling pathways implicated in aberrant
cell proliferation, such as, for example, PKC-alpha, Ralf and
H-Ras; (vii) ribozymes such as VEGF expression inhibitors (e.g.,
ANGIOZYME.RTM.) and HER2 expression inhibitors; (viii) vaccines
such as gene therapy vaccines, for example, ALLOVECTIN.RTM.,
LEUVECTIN.RTM., and VAXID.RTM.; PROLEUKIN.RTM., rIL-2; a
topoisomerase 1 inhibitor such as LURTOTECAN.RTM.; ABARELIX.RTM.
rmRH; and (ix) pharmaceutically acceptable salts, acids and
derivatives of any of the above.
[0147] Chemotherapeutic agent also includes antibodies such as
alemtuzumab (Campath), bevacizumab (AVASTIN.RTM., Genentech);
cetuximab (ERBITUX.RTM., Imclone); panitumumab (VECTIBIX.RTM.,
Amgen), rituximab (RITUXAN.RTM., Genentech/Biogen Idec), pertuzumab
(OMNITARG.RTM., 2C4, Genentech), trastuzumab (HERCEPTIN.RTM.,
Genentech), tositumomab (Bexxar, Corixia), and the antibody drug
conjugate, gemtuzumab ozogamicin (MYLOTARG.RTM., Wyeth). Additional
humanized monoclonal antibodies with therapeutic potential as
agents in combination with the compounds of the invention include:
apolizumab, aselizumab, atlizumab, bapineuzumab, bivatuzumab
mertansine, cantuzumab mertansine, cedelizumab, certolizumab pegol,
cidfusituzumab, cidtuzumab, daclizumab, eculizumab, efalizumab,
epratuzumab, erlizumab, felvizumab, fontolizumab, gemtuzumab
ozogamicin, inotuzumab ozogamicin, ipilimumab, labetuzumab,
lintuzumab, matuzumab, mepolizumab, motavizumab, motovizumab,
natalizumab, nimotuzumab, nolovizumab, numavizumab, ocrelizumab,
omalizumab, palivizumab, pascolizumab, pecfusituzumab, pectuzumab,
pexelizumab, ralivizumab, ranibizumab, reslivizumab, reslizumab,
resyvizumab, rovelizumab, ruplizumab, sibrotuzumab, siplizumab,
sontuzumab, tacatuzumab tetraxetan, tadocizumab, talizumab,
tefibazumab, tocilizumab, toralizumab, tucotuzumab celmoleukin,
tucusituzumab, umavizumab, urtoxazumab, ustekinumab, visilizumab,
and the anti-interleukin-12 (ABT-874/J695, Wyeth Research and
Abbott Laboratories) which is a recombinant exclusively
human-sequence, full-length IgG.sub.1 .lamda. antibody genetically
modified to recognize interleukin-12 p40 protein.
[0148] Chemotherapeutic agent also includes "EGFR inhibitors,"
which refers to compounds that bind to or otherwise interact
directly with EGFR and prevent or reduce its signaling activity,
and is alternatively referred to as an "EGFR antagonist." Examples
of such agents include antibodies and small molecules that bind to
EGFR. Examples of antibodies which bind to EGFR include MAb 579
(ATCC CRL HB 8506), MAb 455 (ATCC CRL HB8507), MAb 225 (ATCC CRL
8508), MAb 528 (ATCC CRL 8509) (see, U.S. Pat. No. 4,943,533,
Mendelsohn et al.) and variants thereof, such as chimerized 225
(C225 or Cetuximab; ERBUTIX.RTM.) and reshaped human 225 (H225)
(see, WO 96/40210, Imclone Systems Inc.); IMC-11F8, a fully human,
EGFR-targeted antibody (Imclone); antibodies that bind type II
mutant EGFR (U.S. Pat. No. 5,212,290); humanized and chimeric
antibodies that bind EGFR as described in U.S. Pat. No. 5,891,996;
and human antibodies that bind EGFR, such as ABX-EGF or Panitumumab
(see WO98/50433, Abgenix/Amgen); EMD 55900 (Stragliotto et al. Eur.
J. Cancer 32A:636-640 (1996)); EMD7200 (matuzumab) a humanized EGFR
antibody directed against EGFR that competes with both EGF and
TGF-alpha for EGFR binding (EMD/Merck); human EGFR antibody,
HuMax-EGFR (GenMab); fully human antibodies known as E1.1, E2.4,
E2.5, E6.2, E6.4, E2.11, E6. 3 and E7.6. 3 and described in U.S.
Pat. No. 6,235,883; MDX-447 (Medarex Inc); and mAb 806 or humanized
mAb 806 (Johns et al., J. Biol. Chem. 279(29):30375-30384 (2004)).
The anti-EGFR antibody may be conjugated with a cytotoxic agent,
thus generating an immunoconjugate (see, e.g., EP659,439A2, Merck
Patent GmbH). EGFR antagonists include small molecules such as
compounds described in U.S. Pat. Nos. 5,616,582, 5,457,105,
5,475,001, 5,654,307, 5,679,683, 6,084,095, 6,265,410, 6,455,534,
6,521,620, 6,596,726, 6,713,484, 5,770,599, 6,140,332, 5,866,572,
6,399,602, 6,344,459, 6,602,863, 6,391,874, 6,344,455, 5,760,041,
6,002,008, and 5,747,498, as well as the following PCT
publications: WO98/14451, WO98/50038, WO99/09016, and WO99/24037.
Particular small molecule EGFR antagonists include OSI-774
(CP-358774, erlotinib, TARCEVA.RTM. Genentech/OSI Pharmaceuticals);
PD 183805 (CI 1033, 2-propenamide,
N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quin-
azolinyl]-, dihydrochloride, Pfizer Inc.); ZD1839, gefitinib
(IRESSA.RTM.)
4-(3'-Chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoli-
ne, AstraZeneca); ZM 105180
((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-1382
(N8-(3-chloro-4-fluoro-phenyl)-N2-(1-methyl-piperidin-4-yl)-pyrimido[5,4--
d]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166
((R)-4-[4-[(1-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol)-
;
(R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimi-
dine); CL-387785
(N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide); EKB-569
(N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6-quinolinyl]-4-(-
dimethylamino)-2-butenamide) (Wyeth); AG1478 (Pfizer); AG1571 (SU
5271; Pfizer); dual EGFR/HER2 tyrosine kinase inhibitors such as
lapatinib (TYKERB.RTM., GSK572016 or N-[3-chloro-4-[(3
fluorophenyl)methoxy]phenyl]-6[5[[[2methylsulfonyl)ethyl]amino]methyl]-2--
furanyl]-4-quinazolinamine).
[0149] Chemotherapeutic agents also include "tyrosine kinase
inhibitors" including the EGFR-targeted drugs noted in the
preceding paragraph; small molecule HER2 tyrosine kinase inhibitor
such as TAK165 available from Takeda; CP-724,714, an oral selective
inhibitor of the ErbB2 receptor tyrosine kinase (Pfizer and OSI);
dual-HER inhibitors such as EKB-569 (available from Wyeth) which
preferentially binds EGFR but inhibits both HER2 and
EGFR-overexpressing cells; lapatinib (GSK572016; available from
Glaxo-SmithKline), an oral HER2 and EGFR tyrosine kinase inhibitor;
PKI-166 (available from Novartis); pan-HER inhibitors such as
canertinib (CI-1033; Pharmacia); Raf-1 inhibitors such as antisense
agent ISIS-5132 available from ISIS Pharmaceuticals which inhibit
Raf-1 signaling; non-HER targeted TK inhibitors such as imatinib
mesylate (GLEEVEC.RTM., available from Glaxo SmithKline);
multi-targeted tyrosine kinase inhibitors such as sunitinib
(SUTENT.RTM., available from Pfizer); VEGF receptor tyrosine kinase
inhibitors such as vatalanib (PTK787/ZK222584, available from
Novartis/Schering AG); MAPK extracellular regulated kinase I
inhibitor CI-1040 (available from Pharmacia); quinazolines, such as
PD 153035,4-(3-chloroanilino) quinazoline; pyridopyrimidines;
pyrimidopyrimidines; pyrrolopyrimidines, such as CGP 59326, CGP
60261 and CGP 62706; pyrazolopyrimidines,
4-(phenylamino)-7H-pyrrolo[2,3-d] pyrimidines; curcumin (diferuloyl
methane, 4,5-bis (4-fluoroanilino)phthalimide); tyrphostines
containing nitrothiophene moieties; PD-0183805 (Warner-Lamber);
antisense molecules (e.g. those that bind to HER-encoding nucleic
acid); quinoxalines (U.S. Pat. No. 5,804,396); tryphostins (U.S.
Pat. No. 5,804,396); ZD6474 (Astra Zeneca); PTK-787
(Novartis/Schering AG); pan-HER inhibitors such as CI-1033
(Pfizer); Affinitac (ISIS 3521; Isis/Lilly); imatinib mesylate
(GLEEVEC.RTM.); PKI 166 (Novartis); GW2016 (Glaxo SmithKline);
CI-1033 (Pfizer); EKB-569 (Wyeth); Semaxinib (Pfizer); ZD6474
(AstraZeneca); PTK-787 (Novartis/Schering AG); INC-1C11 (Imclone),
rapamycin (sirolimus, RAPAMUNE.RTM.); or as described in any of the
following patent publications: U.S. Pat. No. 5,804,396; WO
1999/09016 (American Cyanamid); WO 1998/43960 (American Cyanamid);
WO 1997/38983 (Warner Lambert); WO 1999/06378 (Warner Lambert); WO
1999/06396 (Warner Lambert); WO 1996/30347 (Pfizer, Inc); WO
1996/33978 (Zeneca); WO 1996/3397 (Zeneca) and WO 1996/33980
(Zeneca).
[0150] Chemotherapeutic agents also include dexamethasone,
interferons, colchicine, metoprine, cyclosporine, amphotericin,
metronidazole, alemtuzumab, alitretinoin, allopurinol, amifostine,
arsenic trioxide, asparaginase, BCG live, bevacuzimab, bexarotene,
cladribine, clofarabine, darbepoetin alfa, denileukin, dexrazoxane,
epoetin alfa, elotinib, filgrastim, histrelin acetate, ibritumomab,
interferon alfa-2a, interferon alfa-2b, lenalidomide, levamisole,
mesna, methoxsalen, nandrolone, nelarabine, nofetumomab,
oprelvekin, palifermin, pamidronate, pegademase, pegaspargase,
pegfilgrastim, pemetrexed disodium, plicamycin, porfimer sodium,
quinacrine, rasburicase, sargramostim, temozolomide, VM-26, 6-TG,
toremifene, tretinoin, ATRA, valrubicin, zoledronate, and
zoledronic acid, and pharmaceutically acceptable salts thereof.
[0151] Chemotherapeutic agents also include hydrocortisone,
hydrocortisone acetate, cortisone acetate, tixocortol pivalate,
triamcinolone acetonide, triamcinolone alcohol, mometasone,
amcinonide, budesonide, desonide, fluocinonide, fluocinolone
acetonide, betamethasone, betamethasone sodium phosphate,
dexamethasone, dexamethasone sodium phosphate, fluocortolone,
hydrocortisone-17-butyrate, hydrocortisone-17-valerate,
aclometasone dipropionate, betamethasone valerate, betamethasone
dipropionate, prednicarbate, clobetasone-17-butyrate,
clobetasol-17-propionate, fluocortolone caproate, fluocortolone
pivalate and fluprednidene acetate; immune selective
anti-inflammatory peptides (ImSAIDs) such as
phenylalanine-glutamine-glycine (FEG) and its D-isomeric form (feG)
(IMULAN BioTherapeutics, LLC); anti-rheumatic drugs such as
azathioprine, ciclosporin (cyclosporine A), D-penicillamine, gold
salts, hydroxychloroquine, leflunomideminocycline, sulfasalazine,
tumor necrosis factor alpha (TNF.alpha.) blockers such as
etanercept (Enbrel), infliximab (Remicade), adalimumab (Humira),
certolizumab pegol (Cimzia), golimumab (Simponi), Interleukin 1
(IL-1) blockers such as anakinra (Kineret), T cell costimulation
blockers such as abatacept (Orencia), Interleukin 6 (IL-6) blockers
such as tocilizumab (ACTEMERA.RTM.); Interleukin 13 (IL-13)
blockers such as lebrikizumab; Interferon alpha (IFN) blockers such
as Rontalizumab; Beta 7 integrin blockers such as rhuMAb Beta7; IgE
pathway blockers such as Anti-MI prime; Secreted homotrimeric LTa3
and membrane bound heterotrimer LTa1/.beta.2 blockers such as
Anti-lymphotoxin alpha (LTa); radioactive isotopes (e.g.,
At.sup.211, I.sup.131, I.sup.125, Y.sup.90, Re.sup.186, Re.sup.188,
Sm.sup.153, Bi.sup.212, P.sup.32, Pb.sup.212 and radioactive
isotopes of Lu); miscellaneous investigational agents such as
thioplatin, PS-341, phenylbutyrate, ET-18-OCH.sub.3, or farnesyl
transferase inhibitors (L-739749, L-744832); polyphenols such as
quercetin, resveratrol, piceatannol, epigallocatechine gallate,
theaflavins, flavanols, procyanidins, betulinic acid and
derivatives thereof; autophagy inhibitors such as chloroquine;
delta-9-tetrahydrocannabinol (dronabinol, MARINOL.RTM.);
beta-lapachone; lapachol; colchicines; betulinic acid;
acetylcamptothecin, scopolectin, and 9-aminocamptothecin);
podophyllotoxin; tegafur (UFTORAL.RTM.); bexarotene
(TARGRETIN.RTM.); bisphosphonates such as clodronate (for example,
BONEFOS.RTM. or OSTAC.RTM.), etidronate (DIDROCAL.RTM.), NE-58095,
zoledronic acid/zoledronate (ZOMETA.RTM.), alendronate
(FOSAMAX.RTM.), pamidronate (AREDIA.RTM.), tiludronate
(SKELID.RTM.), or risedronate (ACTONEL.RTM.); and epidermal growth
factor receptor (EGF-R); vaccines such as THERATOPE.RTM. vaccine;
perifosine, COX-2 inhibitor (e.g. celecoxib or etoricoxib),
proteosome inhibitor (e.g. PS341); CCI-779; tipifarnib (R11577);
orafenib, ABT510; Bcl-2 inhibitor such as oblimersen sodium
(GENASENSE.RTM.); pixantrone; farnesyltransferase inhibitors such
as lonafarnib (SCH 6636, SARASAR.TM.); and pharmaceutically
acceptable salts, acids or derivatives of any of the above; as well
as combinations of two or more of the above such as CHOP, an
abbreviation for a combined therapy of cyclophosphamide,
doxorubicin, vincristine, and prednisolone; and FOLFOX, an
abbreviation for a treatment regimen with oxaliplatin
(ELOXATIN.TM.) combined with 5-FU and leucovorin.
[0152] Chemotherapeutic agents also include non-steroidal
anti-inflammatory drugs with analgesic, antipyretic and
anti-inflammatory effects. NSAIDs include non-selective inhibitors
of the enzyme cyclooxygenase. Specific examples of NSAIDs include
aspirin, propionic acid derivatives such as ibuprofen, fenoprofen,
ketoprofen, flurbiprofen, oxaprozin and naproxen, acetic acid
derivatives such as indomethacin, sulindac, etodolac, diclofenac,
enolic acid derivatives such as piroxicam, meloxicam, tenoxicam,
droxicam, lornoxicam and isoxicam, fenamic acid derivatives such as
mefenamic acid, meclofenamic acid, flufenamic acid, tolfenamic
acid, and COX-2 inhibitors such as celecoxib, etoricoxib,
lumiracoxib, parecoxib, rofecoxib, rofecoxib, and valdecoxib.
NSAIDs can be indicated for the symptomatic relief of conditions
such as rheumatoid arthritis, osteoarthritis, inflammatory
arthropathies, ankylosing spondylitis, psoriatic arthritis,
Reiter's syndrome, acute gout, dysmenorrhoea, metastatic bone pain,
headache and migraine, postoperative pain, mild-to-moderate pain
due to inflammation and tissue injury, pyrexia, ileus, and renal
colic.
[0153] In certain embodiments, chemotherapeutic agents include, but
are not limited to, doxorubicin, dexamethasone, vincristine,
cyclophosphamide, fluorouracil, topotecan, interferons, platinum
derivatives, taxanes (e.g., paclitaxel, docetaxel), vinca alkaloids
(e.g., vinblastine), anthracyclines (e.g., doxorubicin),
epipodophyllotoxins (e.g., etoposide), cisplatin, an mTOR inhibitor
(e.g., a rapamycin), methotrexate, actinomycin D, dolastatin 10,
colchicine, trimetrexate, metoprine, cyclosporine, daunorubicin,
teniposide, amphotericin, alkylating agents (e.g., chlorambucil),
5-fluorouracil, campthothecin, cisplatin, metronidazole, and
imatinib mesylate, among others. In other embodiments, a compound
of the present invention is administered in combination with a
biologic agent, such as bevacizumab or panitumumab.
[0154] In certain embodiments, compounds of the present invention,
or a pharmaceutically acceptable composition thereof, are
administered in combination with an antiproliferative or
chemotherapeutic agent selected from any one or more of abarelix,
aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine,
amifostine, anastrozole, arsenic trioxide, asparaginase,
azacitidine, BCG live, bevacuzimab, fluorouracil, bexarotene,
bleomycin, bortezomib, busulfan, calusterone, capecitabine,
camptothecin, carboplatin, carmustine, cetuximab, chlorambucil,
cladribine, clofarabine, cyclophosphamide, cytarabine,
dactinomycin, darbepoetin alfa, daunorubicin, denileukin,
dexrazoxane, docetaxel, doxorubicin (neutral), doxorubicin
hydrochloride, dromostanolone propionate, epirubicin, epoetin alfa,
elotinib, estramustine, etoposide phosphate, etoposide, exemestane,
filgrastim, floxuridine, fludarabine, fulvestrant, gefitinib,
gemcitabine, gemtuzumab, goserelin acetate, histrelin acetate,
hydroxyurea, ibritumomab, idarubicin, ifosfamide, imatinib
mesylate, interferon alfa-2a, interferon alfa-2b, irinotecan,
lenalidomide, letrozole, leucovorin, leuprolide acetate,
levamisole, lomustine, megestrol acetate, melphalan,
mercaptopurine, 6-MP, mesna, methotrexate, methoxsalen, mitomycin
C, mitotane, mitoxantrone, nandrolone, nelarabine, nofetumomab,
oprelvekin, oxaliplatin, paclitaxel, palifermin, pamidronate,
pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium,
pentostatin, pipobroman, plicamycin, porfimer sodium, procarbazine,
quinacrine, rasburicase, rituximab, sargramostim, sorafenib,
streptozocin, sunitinib maleate, talc, tamoxifen, temozolomide,
teniposide, VM-26, testolactone, thioguanine, 6-TG, thiotepa,
topotecan, toremifene, tositumomab, trastuzumab, tretinoin, ATRA,
uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine,
zoledronate, or zoledronic acid.
[0155] Chemotherapeutic agents also include treatments for
Alzheimer's Disease such as donepezil hydrochloride and
rivastigmine; treatments for Parkinson's Disease such as
L-DOPA/carbidopa, entacapone, ropinrole, pramipexole,
bromocriptine, pergolide, trihexephendyl, and amantadine; agents
for treating multiple sclerosis (MS) such as beta interferon (e.g.,
Avonex.RTM. and Rebif.RTM.), glatiramer acetate, and mitoxantrone;
treatments for asthma such as albuterol and montelukast sodium;
agents for treating schizophrenia such as zyprexa, risperdal,
seroquel, and haloperidol; anti-inflammatory agents such as
corticosteroids, TNF blockers, IL-1 RA, azathioprine,
cyclophosphamide, and sulfasalazine; immunomodulatory and
immunosuppressive agents such as cyclosporin, tacrolimus,
rapamycin, mycophenolate mofetil, interferons, corticosteroids,
cyclophophamide, azathioprine, and sulfasalazine; neurotrophic
factors such as acetylcholinesterase inhibitors, MAO inhibitors,
interferons, anti-convulsants, ion channel blockers, riluzole, and
anti-Parkinsonian agents; agents for treating cardiovascular
disease such as beta-blockers, ACE inhibitors, diuretics, nitrates,
calcium channel blockers, and statins; agents for treating liver
disease such as corticosteroids, cholestyramine, interferons, and
anti-viral agents; agents for treating blood disorders such as
corticosteroids, anti-leukemic agents, and growth factors; and
agents for treating immunodeficiency disorders such as gamma
globulin.
Additionally, chemotherapeutic agents include pharmaceutically
acceptable salts, acids or derivatives of any of chemotherapeutic
agents, described herein, as well as combinations of two or more of
them.
[0156] In another embodiment, provided are methods of using
CBP/EP300 bromodomain inhibitors to treat and/or delay progression
of cancer in combination with a PD-1 axis binding antagonist.
Further provided herein are methods of enhancing immune function in
an individual having cancer comprising administering to the
individual an effective amount of a CBP/EP300 bromodomain inhibitor
and an effective amount of a PD-1 axis binding antagonist. A PD-1
axis binding antagonist includes a PD-1 binding antagonist, a PD-L1
binding antagonist and a PD-L2 binding antagonist.
[0157] The term "PD-1 axis binding antagonist" is a molecule that
inhibits the interaction of a PD-1 axis binding partner with either
one or more of its binding partner, so as to remove T-cell
dysfunction resulting from signaling on the PD-1 signaling
axis--with a result being to restore or enhance T-cell function
(e.g., proliferation, cytokine production, target cell killing). As
used herein, a PD-1 axis binding antagonist includes a PD-1 binding
antagonist, a PD-L1 binding antagonist and a PD-L2 binding
antagonist.
[0158] The term "PD-1 binding antagonists" is a molecule that
decreases, blocks, inhibits, abrogates or interferes with signal
transduction resulting from the interaction of PD-1 with one or
more of its binding partners, such as PDL1, PDL2. In some
embodiments, the PD-1 binding antagonist is a molecule that
inhibits the binding of PD-1 to its binding partners. In a specific
aspect, the PD-1 binding antagonist inhibits the binding of PD-1 to
PDL1 and/or PDL2. For example, PD-1 binding antagonists include
anti-PD-1 antibodies, antigen binding fragments thereof,
immunoadhesins, fusion proteins, oligopeptides and other molecules
that decrease, block, inhibit, abrogate or interfere with signal
transduction resulting from the interaction of PD-1 with PDL1
and/or PDL2. In one embodiment, a PD-1 binding antagonist reduces
the negative co-stimulatory signal mediated by or through cell
surface proteins expressed on T lymphocytes mediated signaling
through PD-1 so as render a dysfunctional T-cell less dysfunctional
(e.g., enhancing effector responses to antigen recognition). In
some embodiments, the PD-1 binding antagonist is an anti-PD-1
antibody. In a specific aspect, a PD-1 binding antagonist is
nivolumab described herein (also known as MDX-1106-04, MDX-1106,
ONO-4538, BMS-936558, and OPDIVO.RTM.). In another specific aspect,
a PD-1 binding antagonist is pembrolizumab described herein (also
known as MK-3475, Merck 3475, KEYTRUDA.RTM., and SCH-900475). In
another specific aspect, a PD-1 binding antagonist is CT-011
described herein (also known as hBAT or hBAT-1). In yet another
specific aspect, a PD-1 binding antagonist is AMP-224 (also known
as B7-DCIg) described herein.
[0159] The term "PDL1 binding antagonists" is a molecule that
decreases, blocks, inhibits, abrogates or interferes with signal
transduction resulting from the interaction of PDL1 with either one
or more of its binding partners, such as PD-1, B7-1. In some
embodiments, a PDL1 binding antagonist is a molecule that inhibits
the binding of PDL1 to its binding partners. In a specific aspect,
the PDL1 binding antagonist inhibits binding of PDL1 to PD-1 and/or
B7-1. In some embodiments, the PDL1 binding antagonists include
anti-PDL1 antibodies, antigen binding fragments thereof,
immunoadhesins, fusion proteins, oligopeptides and other molecules
that decrease, block, inhibit, abrogate or interfere with signal
transduction resulting from the interaction of PDL1 with one or
more of its binding partners, such as PD-1, B7-1. In one
embodiment, a PDL1 binding antagonist reduces the negative
co-stimulatory signal mediated by or through cell surface proteins
expressed on T lymphocytes mediated signaling through PDL1 so as to
render a dysfunctional T-cell less dysfunctional (e.g., enhancing
effector responses to antigen recognition). In some embodiments, a
PDL1 binding antagonist is an anti-PDL1 antibody. In a specific
aspect, an anti-PDL1 antibody is YW243.55.S70 described herein. In
another specific aspect, an anti-PDL1 antibody is MDX-1105
described herein (also known as BMS-936559). In still another
specific aspect, an anti-PDL1 antibody is MPDL3280A described
herein. In still another specific aspect, an anti-PDL1 antibody is
MEDI4736 described herein.
[0160] The term "PDL2 binding antagonists" is a molecule that
decreases, blocks, inhibits, abrogates or interferes with signal
transduction resulting from the interaction of PD-L2 with either
one or more of its binding partners, such as PD-1. In some
embodiments, a PD-L2 binding antagonist is a molecule that inhibits
the binding of PD-L2 to its binding partners. In a specific aspect,
the PD-L2 binding antagonist inhibits binding of PD-L2 to PD-1. In
some embodiments, the PD-L2 antagonists include anti-PD-L2
antibodies, antigen binding fragments thereof, immunoadhesins,
fusion proteins, oligopeptides and other molecules that decrease,
block, inhibit, abrogate or interfere with signal transduction
resulting from the interaction of PD-L2 with either one or more of
its binding partners, such as PD-1. In one embodiment, a PD-L2
binding antagonist reduces the negative co-stimulatory signal
mediated by or through cell surface proteins expressed on T
lymphocytes mediated signaling through PD-L2 so as render a
dysfunctional T-cell less dysfunctional (e.g., enhancing effector
responses to antigen recognition). In some embodiments, a PD-L2
binding antagonist is an immunoadhesin.
[0161] Alternative names for "PD-1" include CD279 and SLEB2.
Alternative names for "PD-L1" include B7-H 1, B7-4, CD274, and
B7-H. Alternative names for "PD-L2" include B7-DC, Btdc, and CD273.
In some embodiments, PD-1, PD-L1, and PD-L2 are human PD-1, PD-L1
and PD-L2. In some embodiments, the PD-1 binding antagonist is a
molecule that inhibits the binding of PD-1 to its ligand binding
partners. In a specific aspect the PD-1 ligand binding partners are
PD-L1 and/or PD-L2. In another embodiment, a PD-L1 binding
antagonist is a molecule that inhibits the binding of PD-L1 to its
binding partners. In a specific aspect, PD-L1 binding partners are
PD-1 and/or B7-1. In another embodiment, the PD-L2 binding
antagonist is a molecule that inhibits the binding of PD-L2 to its
binding partners. In a specific aspect, a PD-L2 binding partner is
PD-1. The antagonist may be an antibody, an antigen binding
fragment thereof, an immunoadhesin, a fusion protein, or
oligopeptide. In some embodiment, the PD-1 binding antagonist is an
anti-PD-1 antibody (e.g., a human antibody, a humanized antibody,
or a chimeric antibody). In some embodiments, the anti-PD-1
antibody is selected from the group consisting of MDX-1 106, Merck
3475 (also known as: pembrolizumab, lambrolizumab, or MK-3475),
nivolumab (BMS-936558), CT-011, and MPDL3280A. In some embodiments,
the PD-1 binding antagonist is an immunoadhesin (e.g., an
immunoadhesin comprising an extracellular or PD-1 binding portion
of PD-L1 or PD-L2 fused to a constant region (e.g., an Fc region of
an immunoglobulin sequence). In some embodiments, the PD-1 binding
antagonist is AMP-224. In some embodiments, the PD-L1 binding
antagonist is anti-PD-L1 antibody. In some embodiments, the
anti-PD-L1 binding antagonist is selected from the group consisting
of YW243.55.S70, MPDL3280A and MDX-1 105. MDX-1 105, also known as
BMS-936559, is an anti-PD-L1 antibody described in WO2007/005874.
Antibody YW243.55.S70 (heavy and light chain variable region
sequences shown in SEQ ID Nos. 20 and 21, respectively) is an
anti-PD-L1 described in WO 2010/077634 A1. MDX-1 106, also known as
MDX-1 106-04, ONO-4538 or BMS-936558, is an anti-PD-1 antibody
described in WO2006/121168. Merck 3745, also known as MK-3475 or
SCH-900475, is an anti-PD-1 antibody described in WO2009/114335.
CT-011, also known as hBAT or hBAT-1, is an anti-PD-1 antibody
described in WO2009/101611. AMP-224, also known as B7-DCIg, is a
PD-L2-Fc fusion soluble receptor described in WO2010/027827 and
WO2011/066342. In some embodiments, the anti-PD-1 antibody is MDX-1
106. Alternative names for "MDX-1106" include MDX-1 106-04,
ONO-4538, BMS-936558 or Nivolumab. In some embodiments, the
anti-PD-1 antibody is Nivolumab (CAS Registry Number: 946414-94-4).
In some embodiments, the cancer is melanoma, NSCLC, and renal cell
carcinoma.
[0162] For treating an inflammatory disease or an autoimmune
disease, a compound of formula (I) or a pharmaceutically acceptable
salt thereof may be co-administered with methotrexate, tofacitinib,
6-mercaptopurine, azathioprine sulphasalazine, mesalazine,
olsalazine chloroquinine/hydroxychloroquinine, penicillamine,
aurothiomalate (intramuscular and oral), azathioprine, cochicine,
corticosteroids (oral, inhaled, and local injection), a beta-2
adrenoreceptor agonist (salbutamol, terbutaline, salmeteral), a
xanthine (theophylline, aminophylline), cromoglycate, nedocromil,
ketotifen, ipratropium and oxitropium, cyclosporin, FK506,
rapamycin, mycophenolate mofetil, leflunomide, an NSAID (e.g.
ibuprofen), a corticosteroid (e.g. prednisolone), a
phosphodiesterase inhibitor, an adensosine agonist, an
antithrombotic agent, a complement inhibitor, an adrenergic agent,
an agent that interferes with signalling by proinflammatory
cytokines such as TNF or IL-1 (e.g., a NIK, TKK, p38 or MAP kinase
inhibitor), an IL-1 converting enzyme inhibitor, a T-cell
signalling inhibitor (e.g. a kinase inhibitor), a metalloproteinase
inhibitor, sulfasalazine, a 6-mercaptopurine, an angiotensin
converting enzyme inhibitor, a soluble cytokine receptor (e.g.
soluble p55 or p75 TNF receptors and the derivatives p75TNFRigG
(etanercept) and p55TNFRigG (Lenercept), siL-IRI, siL-IRII,
siL-6R), an antiinflammatory cytokine (e.g. IL-4, IL-10, IL-11,
IL-13 and TGF), celecoxib, folic acid, hydroxychloroquine sulfate,
rofecoxib, etanercept, infliximab, adalimumab, certolizumab,
tocilizumab, abatacept, naproxen, valdecoxib, sulfasalazine,
methylprednisolone, meloxicam, methylprednisolone acetate, gold
sodium thiomalate, aspirin, triamcinolone acetonide, propoxyphene
napsylate/apap, folate, nabumetone, diclofenac, piroxicam,
etodolac, diclofenac sodium, oxaprozin, oxycodone HCl, hydrocodone
bitartrate/apap, diclofenac sodium/misoprostol, fentanyl, anakinra,
tramadol HCl, salsalate, sulindac, cyanocobalamin/fa/pyridoxine,
acetaminophen, alendronate sodium, prednisolone, cortisone,
betamethasone, morphine sulfate, lidocaine hydrochloride,
indomethacin, glucosamine sulf/chondroitin, amitriptyline HCl,
sulfadiazine, oxycodone HCVacetaminophen, olopatadine HCl
misoprostol, naproxen sodium, omeprazole, cyclophosphamide,
rituximab, IL-1 TRAP, MRA, CTLA4-IG, IL-18 BP, anti-IL-12,
Anti-IL1S, BIRB-796, SCIO-469, VX-702, AMG-548, VX-740,
Roflumilast, IC-485, CDC-801, SIPI agonists (such as FTY720), a PKC
family inhibitor (e.g. Ruboxistaurin or AEB-071) or Mesopram. In
certain embodiments, a compound of formula (I) or a
pharmaceutically acceptable salt thereof may be co-administered
with methotrexate or leflunomide. In moderate or severe Rheumatoid
arthritis cases, a compound of formula (I) or a pharmaceutically
acceptable salt thereof may be co-administered with cyclosporine
and anti-TNF antibodies as noted above. A compound of formula (I)
or a pharmaceutically acceptable salt thereof may also be
co-administered with: budenoside; epidermal growth factor; a
corticosteroid; cyclosporin, sulfasalazine; an aminosalicylate;
6-mercaptopurine; azathioprine; metronidazole; a lipoxygenase
inhibitor; mesalamine; olsalazine; balsalazide; an antioxidant; a
thromboxane inhibitor; an IL-1 receptor antagonist; an anti-IL-1
monoclonal antibody; an anti-IL-6 monoclonal antibody; a growth
factor; an elastase inhibitor; a pyridinyl-imidazole compound; an
antibody to or antagonist of other human cytokines or growth
factors (e.g. TNF, LT, IL-1, IL-2, IL-6, IL-7, IL-8, IL-12, IL-15,
IL-16, IL-23, EMAP-II, GM-CSF, FGF, and PDGF); a cell surface
molecule (e.g. CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45,
CD69, or CD90 or their ligands); methotrexate; cyclosporine; FK506;
rapamycin; mycophenolate mofetil; leflunomide; an NSAID (e.g.
ibuprofen); a corticosteroid (e.g. prednisolone); a
phosphodiesterase inhibitor; an adenosine agonist; an
antithrombotic agent; a complement inhibitor; an adrenergic agent;
an agent that interferes with signalling by proinflammatory
cytokines such as TNF 5 or IL-1 (e.g. a NIK, IKK, or MAP kinase
inhibitor); an IL-1 converting enzyme inhibitor; a TNF converting
enzyme inhibitor; a T-cell signalling inhibitor such as kinase
inhibitors; a metalloproteinase inhibitor; sulfasalazine;
azathioprine; a 6-mercaptopurine; an angiotensin converting enzyme
inhibitor; a soluble cytokine receptor (e.g. soluble p55 or p75 TNF
receptors, siL-IRI, siL-IRII, siL-6R), and an antiinflammatory
cytokine (e.g. IL-4, IL-10, IL-11, IL-13 or TGF).
[0163] For treating Crohn's disease, a compound of formula (I) or a
pharmaceutically acceptable salt thereof may be co-administered
with a TNF antagonist (e.g. an anti-TNF antibody), D2E7
(adalimumab), CA2 (infliximab), CDP 571, a TNFR-Ig construct,
(p75TNFRigG (etanercept)), a p55TNFRigG (LENERCEPT.TM.) inhibitor,
or a PDE4 inhibitor.
[0164] For treating inflammatory bowel disease, a compound of
formula (I) or a pharmaceutically acceptable salt thereof may be
co-administered with a corticosteroid (e.g. budenoside or
dexamethasone); sulfasalazine, 5-aminosalicylic acid; olsalazine;
an agent that interferes with synthesis or action of
proinflammatory cytokines such as IL-1 (e.g. an IL-1 converting
enzyme inhibitor or IL-lra); a T cell signaling inhibitor (e.g. a
tyrosine kinase inhibitor); 6-mercaptopurine; IL-11; mesalamine;
prednisone; azathioprine; mercaptopurine; infliximab;
methylprednisolone sodium succinate; diphenoxylate/atrop sulfate;
loperamide hydrochloride; methotrexate; omeprazole; folate;
ciprofloxacin/dextrose-water; hydrocodone bitartrate/apap;
tetracycline hydrochloride; fluocinonide; metronidazole;
thimerosal/boric acid; cholestyramine/sucrose; ciprofloxacin
hydrochloride; hyoscyamine sulfate; meperidine hydrochloride;
midazolam hydrochloride; oxycodone HCl/acetaminophen; promethazine
hydrochloride; sodium phosphate; sulfamethoxazole/trimethoprim;
celecoxib; polycarbophil; propoxyphene napsylate; hydrocortisone;
multivitamins; balsalazide disodium; codeine phosphate/apap;
colesevelam HCl; cyanocobalamin; folic acid; levofloxacin;
methylprednisolone; natalizumab or interferon-gamma.
[0165] For treating multiple sclerosis, a compound of formula (I)
or a pharmaceutically acceptable salt thereof may be
co-administered with a corticosteroid; prednisolone;
methylprednisolone; azathioprine; cyclophosphamide; cyclosporine;
methotrexate; 4-aminopyridine; tizanidine; interferon-la
(AVONEX.RTM.; Biogen); interferon-1b (BETASERON.RTM.;
Chiron/Berlex); interferon-n3) (Interferon Sciences/Fujimoto),
interferon-(Alfa Wassermann/J&J), interferon 1A-IF
(Serono/Inhale Therapeutics), Peginterferon 2b
(Enzon/Schering-Plough), Copolymer 1 (Cop-1; COPAXONE.RTM.; Teva
Pharmaceutical Industries, Inc.); hyperbaric oxygen; intravenous
immunoglobulin; cladribine; an antibody to or antagonist of other
human cytokines or growth factors and their receptors (e.g. TNF,
LT, IL-1, IL-2, IL-6, IL-7, IL-8, IL-12, IL-23, IL-15, IL-16,
EMAP-II, GM-CSF, FGF, or PDGF).
[0166] For treating AIDS a compound of formula (I) or a
pharmaceutically acceptable salt thereof may be co-administered
with antibodies to cell surface molecules such as CD2, CD3, CD4,
CD8, CD19, CD20, CD25, CD28, CD30, CD40, CD45, CD69, CD80, CD86,
CD90 or their ligands. A compound of formula (I) or a
pharmaceutically acceptable salt thereof may also be
co-administered with methotrexate, cyclosporine, FK506, rapamycin,
mycophenolate mofetil, leflunomide, an S1P1 agonist, an NSAID (e.g.
ibuprofen), a corticosteroid (e.g. prednisolone), a
phosphodiesterase inhibitor, an adensosine agonist, an
antithrombotic agent, a complement inhibitor, an adrenergic agent,
an agent that interferes with signalling by proinflammatory
cytokines such as TNF or IL-1 (e.g., a NIK, TKK, p38 or MAP kinase
inhibitor), an IL-1 converting enzyme inhibitor, a TACE inhibitor,
a T-cell signaling inhibitor (e.g. a kinase inhibitor), a
metalloproteinase inhibitor, sulfasalazine, azathioprine, a
6-mercaptopurine, an angiotensin converting enzyme inhibitor, a
soluble cytokine receptor (e. g. soluble p55 or p75 TNF receptors,
siL-IRI, siL-IRII, or siL-6R), or an antiinflammatory cytokine
(e.g. IL-4, IL-10, IL-13 or TGF).
[0167] A compound of formula (I) or a pharmaceutically acceptable
salt thereof may also be co-administered with agents, such as
alemtuzumab, dronabinol, daclizumab, mitoxantrone, xaliproden
hydrochloride, fampridine, glatiramer acetate, natalizumab,
sinnabidol, immunokine NNS03, ABR-215062, AnergiX.MS, chemokine
receptor antagonists, BBR-2778, calagualine, CPI-1189, LEM
(liposome encapsulated mitoxantrone), THC.CBD (cannabinoid
agonist), MBP-8298, mesopram (PDE4 inhibitor), MNA-715, an
anti-IL-6 receptor antibody, neurovax, pirfenidone allotrap 1258
(RDP-1258), sTNF-R1, talampanel, teriflunomide, TGF-beta2,
tiplimotide, a VLA-4 antagonist (e.g. TR-14035, VLA4 Ultrahaler, or
Antegran-ELAN/Biogen), an interferon gamma antagonist, or an IL-4
agonist.
[0168] For treating ankylosing spondylitis a compound of formula
(I) or a pharmaceutically acceptable salt thereof may be
co-administered with ibuprofen, diclofenac, misoprostol, naproxen,
meloxicam, indomethacin, diclofenac, celecoxib, rofecoxib,
sulfasalazine, methotrexate, azathioprine, minocyclin, prednisone,
an anti-TNF antibody, D2E7 (HUMIRA.RTM.), CA2 (infliximab), CDP
571, a TNFR-Ig construct, (p75TNFRigG (ENBREL.RTM.), or p55TNFRigG
(LENERCEPT.RTM.).
[0169] For treating asthma a compound of formula (I) or a
pharmaceutically acceptable salt thereof may be co-administered
with albuterol, salmeterol/fluticasone, montelukast sodium,
fluticasone propionate, budesonide, prednisone, salmeterol
xinafoate, levalbuterol HCl, albuterol sulfate/ipratropium,
prednisolone sodium phosphate, triamcinolone acetonide,
beclomethasone dipropionate, ipratropium bromide, azithromycin,
pirbuterol acetate, prednisolone, theophylline anhydrous,
methylprednisolone sodium succinate, clarithromycin, zafirlukast,
formoterol fumarate, influenza virus vaccine, amoxicillin
trihydrate, flunisolide, cromolyn sodium, fexofenadine
hydrochloride, flunisolide/menthol, amoxicillin/clavulanate,
levofloxacin, guaifenesin, dexamethasone sodium phosphate,
moxifloxacin HCl, doxycycline hyclate, guaifenesin/d-methorphan,
p-ephedrine/cod/-chlorphenir, gatifloxacin, cetirizine
hydrochloride, mometasone furoate, salmeterol xinafoate,
benzonatate, cephalexin, pe/hydrocodone/chlorphenir, cetirizine
HCl/pseudoephed, phenylephrine/cod/promethazine,
codeine/promethazine, cefprozil, dexamethasone,
guaifenesin/pseudoephedrine, chlorpheniramine/hydrocodone,
nedocromil sodium, terbutaline sulfate, epinephrine,
methylprednisolone, an anti-IL-13 antibody, or metaproterenol
sulfate.
[0170] For treating COPD a compound of formula (I) or a
pharmaceutically acceptable salt thereof may be co-administered
with albuterol sulfate/ipratropium, ipratropium bromide,
salmeterol/fluticasone, albuterol, salmeterol xinafoate,
fluticasone propionate, prednisone, theophylline anhydrous,
methylprednisolone sodium succinate, montelukast sodium,
budesonide, formoterol fumarate, triamcinolone acetonide,
levofloxacin, guaifenesin, azithromycin, beclomethasone
dipropionate, levalbuterol HCl, flunisolide, ceftriaxone sodium,
amoxicillin trihydrate, gatifloxacin, zafirlukast,
amoxicillin/clavulanate, flunisolide/menthol,
chlorpheniramine/hydrocodone, metaproterenol sulfate,
methylprednisolone, mometasone furoate,
p-ephedrine/cod/chlorphenir, pirbuterol acetate,
p-ephedrine/loratadine, terbutaline sulfate, tiotropium bromide,
(R,R)-formoterol, TgAAT, cilomilast, or roflumilast.
[0171] For treating psoriasis, a compound of formula (I) or a
pharmaceutically acceptable salt thereof may be co-administered
with calcipotriene, clobetasol propionate, triamcinolone acetonide,
halobetasol propionate, tazarotene, methotrexate, fluocinonide,
betamethasone diprop augmented, fluocinolone acetonide, acitretin,
tar shampoo, betamethasone valerate, mometasone furoate,
ketoconazole, pramoxine/fluocinolone, hydrocortisone valerate,
flurandrenolide, urea, betamethasone, clobetasol propionate/emoll,
fluticasone propionate, azithromycin, hydrocortisone, moisturizing
formula, folic acid, desonide, pimecrolimus, coal tar, diflorasone
diacetate, etanercept folate, lactic acid, methoxsalen, he/bismuth
subgal/znox/resor, methylprednisolone acetate, prednisone,
sunscreen, halcinonide, salicylic acid, anthralin, clocortolone
pivalate, coal extract, coal tar/salicylic acid, coal tar/salicylic
acid/sulfur, desoximetasone, diazepam, emollient,
fluocinonide/emollient, mineral oil/castor oil/na lact, mineral
oil/peanut oil, petroleum/isopropyl myristate, psoralen, salicylic
acid, soap/tribromsalan, thimerosal/boric acid, celecoxib,
infliximab, cyclosporine, alefacept, efalizumab, tacrolimus,
pimecrolimus, PUVA, UVB, sulfasalazine, ABT-874 or ustekinamab.
[0172] For treating psoriatic arthritis, a compound of formula (I)
or a pharmaceutically acceptable salt thereof may be
co-administered with methotrexate, etanercept, rofecoxib,
celecoxib, folic acid, sulfasalazine, naproxen, leflunomide,
methylprednisolone acetate, indomethacin, hydroxychloroquine
sulfate, prednisone, sulindac, betamethasone diprop augmented,
infliximab, methotrexate, folate, triamcinolone acetonide,
diclofenac, dimethylsulfoxide, piroxicam, diclofenac sodium,
ketoprofen, meloxicam, methylprednisolone, nabumetone, tolmetin
sodium, calcipotriene, cyclosporine, diclofenac sodium/misoprostol,
fluocinonide, glucosamine sulfate, gold sodium thiomalate,
hydrocodone bitartrate/apap, ibuprofen, risedronate sodium,
sulfadiazine, thioguanine, valdecoxib, alefacept, D2E7
(adalimumab), or efalizumab.
[0173] For treating lupus, a compound of formula (I) or a
pharmaceutically acceptable salt thereof may be co-administered
with an NSAID (e.g. diclofenac, naproxen, ibuprofen, piroxicam, or
indomethacin); a COX2 inhibitor (e.g. celecoxib, rofecoxib, or
valdecoxib); an anti-malarial (e.g. hydroxychloroquine); a steroid
(e.g. prednisone, prednisolone, budenoside, or dexamethasone); a
cytotoxic (e.g. azathioprine, cyclophosphamide, mycophenolate
mofetil, or methotrexate); an inhibitor of PDE4, or a purine
synthesis inhibitor (e.g. Cellcept.RTM.). For example, a compound
of formula (I) or a pharmaceutically acceptable salt thereof may be
co-administered with sulfasalazine, 5-aminosalicylic acid,
olsalazine, Imuran.RTM., an agent that interferes with the
synthesis, production, or action of a proinflammatory cytokine
(e.g. IL-1), or a caspase inhibitor (e.g. a IL-1 converting enzyme
inhibitor or IL-1ra).
[0174] A compound of formula (I) or a pharmaceutically acceptable
salt thereof may also be co-administered with a T cell signaling
inhibitor (e.g. a tyrosine kinase inhibitor), or a molecule that
targets T cell activation (e.g. CTLA-4-IgG, an anti-B7 family
antibody, or an anti-PD-1 family antibody).
[0175] A compound of formula (I) or a pharmaceutically acceptable
salt thereof can also be co-administered with an IL-11 antibody, an
anti-cytokine antibody (e.g. fonotolizumab (anti-IFNg antibody)),
or an anti-receptor receptor antibodies (e.g. an anti-IL-6 receptor
antibody or an antibody to a B-cell surface molecule).
[0176] A compound of formula (I) or a pharmaceutically acceptable
salt thereof can also be co-administered with LJP 394 (abetimus),
an agent that depletes or inactivates B-cells (e.g. Rituximab
(anti-CD20 antibody) or lymphostat-B (anti-BlyS antibody)), a TNF
antagonist (e.g. an anti-TNF antibody), D2E7 (adalimumab), CA2
(infliximab), CDP 571, a TNFR-Ig construct, (p75TNFRigG
(etanercept), or p55TNFRigG (LENERCEPT.TM.).
[0177] A compound of formula (I) or a pharmaceutically acceptable
salt thereof can also be co-administered with one or more agents
used in the prevention or treatment of AIDS: an HIV reverse
transcriptase inhibitor, an HIV protease inhibitor, an
immunomodulator, or another retroviral drug. Examples of reverse
transcriptase inhibitors include, but are not limited to, abacavir,
adefovir, didanosine, dipivoxil delavirdine, efavirenz,
emtricitabine, lamivudine, nevirapine, rilpivirine, stavudine,
tenofovir, zalcitabine, and zidovudine. Examples of protease
inhibitors include, but are not limited to, amprenavir, atazanavir,
darunavir, indinavir, fosamprenavir, lopinavir, nelfinavir,
ritonavir, saquinavir, and tipranavir. Examples of other retroviral
drugs include, but are not limited to, elvitegravir, enfuvirtide,
maraviroc and raltegravir.
[0178] For treating type II diabetes, hepatic steatosis, insulin
resistance, metabolic syndrome or a related disorder, a compound of
formula (I) or a pharmaceutically acceptable salt thereof may be
co-administered with insulin or insulins that have been modified to
improve the duration of action in the body; agents that stimulate
insulin secretion such as acetohexamide, chlorpropamide, glyburide,
glimepiride, glipizide, glicazide, glycopyramide, gliquidone,
rapaglinide, nataglinide, tolazamide or tolbutamide; agents that
are glucagon-like peptide agonists such as exanatide, liraglutide
or taspoglutide; agents that inhibit dipeptidyl-peptidase IV such
as vildagliptin, sitagliptin, saxagliptin, linagliptin, allogliptin
or septagliptin; agents that bind to the peroxisome
proliferator-activated receptor gamma such as rosiglitazone or
pioglitazone; agents that decrease insulin resistance such as
metformin; or agents that reduce glucose absorbance in the small
intestine such as acarbose, miglitol or voglibose.
[0179] For treating acute kidney disorders or a chronic kidney
disease, a compound of formula (I) or a pharmaceutically acceptable
salt thereof may be co-administered with dopamine, a diuretic (e.g.
furosemide), bumetanide, thiazide, mannitol, calcium gluconate,
sodium bicarbonate, albuterol, paricalcitol, doxercalciferol,
cinacalcet, or bardoxalone methyl.
[0180] The amount of both the compound of formula (I) or salt
thereof and additional agent (in those compositions which comprise
an additional therapeutic agent as described above) that may be
combined with the carrier materials to produce a single dosage form
will vary depending upon the host treated and the particular mode
of administration. In certain embodiments, compositions of this
invention are formulated such that a dosage of between 0.01-100
mg/kg body weight/day of an inventive can be administered.
[0181] The additional therapeutic agent and the compound of formula
(I) may act synergistically. Therefore, the amount of additional
therapeutic agent in such compositions may be less than that
required in a monotherapy utilizing only that therapeutic agent, or
there may be fewer side effects for the patient given that a lower
dose is used. In certain embodiments, in such compositions a dosage
of between 0.01-1,000 .mu.g/kg body weight/day of the additional
therapeutic agent can be administered.
[0182] Provided herein are methods of extending the duration of
response to a cytotoxic agent in an individual with cancer
comprising administering to the individual (a) an effective amount
of a compound of formula (I) or a pharmaceutically acceptable salt
thereof and (b) an effective amount of the cytotoxic agent.
[0183] In certain embodiments of any of the methods, the cytotoxic
agent is a targeted therapy. In certain embodiments, the targeted
therapy is one or more of an EGFR antagonist, RAF inhibitor, and/or
PI3K inhibitor.
[0184] In certain embodiments of any of the methods, the targeted
therapy is an EGFR antagonist. In certain embodiments of any of the
methods, the EGFR antagonist is
N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine
and/or a pharmaceutical acceptable salt thereof. In certain
embodiments, the EGFR antagonist is
N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine. In
certain embodiments, the EGFR antagonist is
N-(4-(3-fluorobenzyloxy)-3-chlorophenyl)-6-(5-((2-(methylsulfonyl)ethylam-
ino)methyl)furan-2-yl)quinazolin-4-amine,di4-methylbenzenesulfonate
or a pharmaceutically acceptable salt thereof (e.g.,
lapatinib).
[0185] In certain embodiments of any of the methods, targeted
therapy is a RAF inhibitor. In certain embodiments, the RAF
inhibitor is a BRAF inhibitor. In certain embodiments, the RAF
inhibitor is a CRAF inhibitor.
[0186] In certain embodiments, the BRAF inhibitor is vemurafenib.
In certain embodiments, the RAF inhibitor is
3-(2-cyanopropan-2-yl)-N-(4-methyl-3-(3-methyl-4-oxo-3,4-dihydroquinazoli-
n-6-ylamino)phenyl)benzamide or a pharmaceutically acceptable salt
thereof (e.g., AZ628 (CAS #878739-06-1)).
[0187] In certain embodiments of any of the methods, the targeted
therapy is a PI3K inhibitor.
[0188] In certain embodiments of any of the methods, the cytotoxic
agent is chemotherapy. In certain embodiments of any of the
methods, the chemotherapy is a taxane. In certain embodiments, the
taxane is paclitaxel. In certain embodiments, the taxane is
docetaxel.
[0189] In certain embodiments of any of the methods, the cytotoxic
agent is a platinum agent. In certain embodiments, the platinum
agent is carboplatin. In certain embodiments, the platinum agent is
cisplatin. In certain embodiments of any of the methods, the
cytotoxic agent is a taxane and a platinum agent. In certain
embodiments, the taxane is paclitaxel. In certain embodiments, the
taxane is docetaxel. In certain embodiments, the platinum agent is
carboplatin. In certain embodiments, the platinum agent is
cisplatin.
[0190] In certain embodiments of any of the methods, the cytotoxic
agent is a vinca alkyloid. In certain embodiments, the vinca
alkyloid is vinorelbine. In certain embodiments of any of the
methods, the chemotherapy is a nucleoside analog. In certain
embodiments, the nucleoside analog is gemcitabine.
[0191] In certain embodiments of any of the methods, the cytotoxic
agent is radiotherapy.
[0192] In certain embodiments of any of the methods, the compound
of formula (I) or a pharmaceutically acceptable salt thereof is
concomitantly administered with the cytotoxic agent (e.g., targeted
therapy, chemotherapy, and/or radiotherapy). In certain
embodiments, the compound of formula (I) or a pharmaceutically
acceptable salt thereof is administered prior to and/or
concurrently with the cytotoxic agent (e.g., targeted therapy,
chemotherapy, and/or radiotherapy).
Exemplification of Compounds of Formula (I)
[0193] As depicted in the Examples of Compounds of Formula (I),
below, in certain exemplary embodiments, compounds of Formula (I)
are prepared according to the following general procedures. It will
be appreciated that, although the general methods depict the
synthesis of certain compounds of the present invention, the
following general methods, and other methods known to one of
ordinary skill in the art, can be applied to all compounds and
subclasses and species of each of these compounds, as described
herein.
##STR00093##
[0194] Compounds of formula (4) may be prepared by general
synthetic methods as shown in Scheme 1.
[0195] Reaction between cyano-ketone (1) and hydrazine in a
suitable solvent such as ethanol at a temperature ranging from
about room temperature to reflux and for a time varying from about
30 minutes to about 2 hours, can readily produce bicycle-pyrazole
(2). The bromo pyrazole (3) can be formed by converting the amino
pyrazole (2) using a nitrite such as, but not limited to,
isoamylnitrite, sodium nitrite, or tert-butyl nitrite and a copper
salt such as, but not limited to, copper(II) bromide in organic an
solvent such as, but not limited to, acetonitrile at a temperature
of about 20.degree. C. to about 60.degree. C. for a time of about 5
hours. The alkylation of pyrazole N.sup.1 nitrogen of (2) can be
carried out using an alkyl iodide/bromide/mesylate/triflate in the
presence of an inorganic base such as, but not limited to, sodium
hydride or cesium carbonate in a suitable organiv solvent such as,
but not limited to, N,N-dimethylformamide (DMF) or tetrahydrofuran
(THF) at a temperature ranging from about 0.degree. C. to
120.degree. C. and for a time varying from about 30 minutes to
about 16 hours to form compounds of formula (4).
##STR00094##
[0196] Compounds of formula (8) and (11) may be prepared by general
synthetic methods as shown in Scheme 2.
[0197] Deprotection of N-tert-butoxycarbonyl (Boc) group using a
protic acid such as, but not limited to, trifluoroacetic acid or
hydrochloric acid, and subsequent N-acetylation using acetic
anhydride in the presence of a base such as, but not limited to,
triethylamine (TEA) can readily afford compounds of formula (5).
Compounds of formula (8) can be prepared from the bromide (5) upon
treatment with aryl, heteroaryl or heterocyclic boronic acids or
boronate esters (7) under palladium catalyst conditions such as,
but not limited to, [1,1'-bis(diphenylphosphino)ferrocene]
dichloropalladium(II) in the presence of water and an inorganic
base such as, but not limited to, sodium carbonate, potassium
carbonate, or potassium phosphate in an organic solvent such as,
but not limited to, 1,4-dioxane at an elevated temperature.
Alternatively, reaction between bromide (5) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) in the
presence of base such as, but not limited to, potassium acetate or
potassium 2-ethyl heanoate, under palladium catalyst conditions
such as, but not limited to, [1,1'-bis(diphenylphosphino)ferrocene]
dichloropalladium(II) in the presence of an organic solvent such
as, but not limited to, 1,4-dioxane or dimethylformamide, can
produce the corresponding boronate ester that upon treatment with
aryl, heteroaryl or heterocyclic halides under the analogous
palladium catalyst conditions can also yield compounds of formula
(8). To access compounds of formula (11), Boc-protected (4) can be
directly elaborated to compounds of formula (9) using either
coupling procedure described above. Subjection of piperidine (9) to
the protic conditions described above, followed by treatment with a
base such as, but not limited to, triethylamine (TEA), and urea
(10) provides compounds of formula (11).
##STR00095##
[0198] Compounds of formula (8a) and (11a) may be prepared by
general synthetic methods as shown in Scheme 3.
[0199] Deprotection of N-tert-butoxycarbonyl (Boc) group using a
protic acid such as, but not limited to, trifluoroacetic acid or
hydrochloric acid, and subsequent N-acetylation using acetic
anhydride in the presence of a base such as, but not limited to,
triethylamine (TEA) can readily afford compounds of formula (5).
Compounds of formula (8a) can be prepared from the bromide (5) upon
treatment with aryl, heteroaryl or heterocyclic boronic acids or
boronate esters (7a) under palladium catalyst conditions such as,
but not limited to, [1,1'-bis(diphenylphosphino)ferrocene]
dichloropalladium(II) in the presence of water and an inorganic
base such as, but not limited to, sodium carbonate, potassium
carbonate, or potassium phosphate in an organic solvent such as,
but not limited to, 1,4-dioxane at an elevated temperature.
Alternatively, reaction between bromide (5) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) in the
presence of base such as, but not limited to, potassium acetate or
potassium 2-ethyl heanoate, under palladium catalyst conditions
such as, but not limited to, [1,1'-bis(diphenylphosphino)ferrocene]
dichloropalladium(II) in the presence of an organic solvent such
as, but not limited to, 1,4-dioxane or dimethylformamide, can
produce the corresponding boronate ester that upon treatment with
aryl, heteroaryl or heterocyclic halides under the analogous
palladium catalyst conditions can also yield compounds of formula
(8a). To access compounds of formula (11a), Boc-protected (4) can
be directly elaborated to compounds of formula (9a) using either
coupling procedure described above. Subjection of piperidine (9a)
to the protic conditions described above, followed by treatment
with a base such as, but not limited to, triethylamine (TEA), and
urea (10) provides compounds of formula (11a).
##STR00096##
[0200] Compounds of formula (15) may be prepared by general
synthetic methods as shown in Scheme 4.
[0201] Compounds of formula (15) can be prepared from the bromide
(13) upon treatment with aryl, heteroaryl or heterocyclic boronic
acids or boronate esters under palladium catalyst conditions such
as, but not limited to,
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) or
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-
-amino-1,1'-biphenyl-2-yl) palladium(II) in the presence of water
and an inorganic base such as, but not limited to, sodium carbonate
in an organic solvent such as, but not limited to, 1,4-dioxane at
an elevated temperature. Alternatively, reaction between triflate
(12) under the analogous palladium catalyst conditions can also
yield compounds of formula (15).
##STR00097##
[0202] Compounds of formula (18) may be prepared by general
synthetic methods as shown in Scheme 5.
[0203] Compounds of formula (18) can be prepared from the triflate
(16) upon treatment with amines (17) in the presence of an organic
solvent such as, but not limited to, dimethyl sulfoxide (DMSO)
under microwave irradiation. When the salt form of amines (17) were
utilized, a base such as, but not limited to, triethylamine was
added to the reaction conditions.
##STR00098##
[0204] Compounds of formulas (21) and (23) may be prepared by
general synthetic methods as shown in Scheme 6.
[0205] Compounds of formula (23) can be prepared from the bromide
(5) upon treatment with indole or azaindole boronic acids or
boronate esters (22) under palladium catalyst conditions such as,
but not limited to, [1,1'-bis(diphenylphosphino)ferrocene]
dichloropalladium(II) in the presence of water and an inorganic
base such as, but not limited to, sodium carbonate, potassium
carbonate, or potassium phosphate in an organic solvent such as,
but not limited to, 1,4-dioxane at an elevated temperature.
Alternatively, reaction between bromide (5) and indole or azaindole
boronic acids or boronate esters (19) under the analogous palladium
catalyst conditions, followed by deprotection of
N-tert-butoxycarbonyl (Boc) group using a protic acid such as, but
not limited to, trifluoroacetic acid or hydrochloric acid yields
compounds of formula (21).
##STR00099##
[0206] Compounds of formula (26) may be prepared by general
synthetic methods as shown in Scheme 7.
[0207] Compounds of formula (26) can be prepared from (24) upon
treatment with aryl, heteroaryl, or heterocyclic iodides or
bromides (25) under copper catalyst conditions such as, but not
limited to, copper (I) iodide in the presence of an inorganic base
such as, but not limited to, potassium carbonate or potassium
phosphate and a ligand such as, but not limited to,
N,N'-dimethylethylenediame or (1R,2R)-cyclohexane-1,2-diamine in an
organic solvent such as, but not limited to, 1,4-dioxane at an
elevated temperature.
##STR00100##
[0208] Compounds of formula (28) may be prepared by general
synthetic methods as shown in Scheme 8.
[0209] Compounds of formula (28) can be prepared from the bromide
(5) upon treatment with indoles or azaindoles (27) under copper
catalyst conditions such as, but not limited to, copper (I) iodide
in the presence of an inorganic base such as, but not limited to,
potassium phosphate and a ligand such as, but not limited to,
(1R,2R)-cyclohexane-1,2-diamine in an organic solvent such as, but
not limited to, toluene at an elevated temperature.
##STR00101##
[0210] Compounds of formula (34) may be prepared by general
synthetic methods as shown in Scheme 9.
[0211] Compounds of formula (32) can be prepared from the bromide
(5) upon treatment with boronic esters or boronate acids (31) under
palladium catalyst conditions such as, but not limited to,
[1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium(II) in the
presence of water and an inorganic base such as, but not limited
to, sodium carbonate, in an organic solvent such as, but not
limited to, 1,4-dioxane at an elevated temperature. Treatment of
the ester (32) with hydroxide sources such as, but not limited to,
lithium hydroxide in the presence of water and organic solvents
such as, but not limited to, methanol and/or tetrahydrofuran yields
carboxylic acids of formula (33). Reaction of carboxylic acids (33)
with a coupling reagent such as, but not limited to,
1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium
3-oxide hexafluorophosphate, a base such as, but not limited to,
diisopropylethylamine, and an amine (34) provides amides of formula
(35).
General Procedure for Intermediates A & B
##STR00102## ##STR00103##
[0212] Step 1: ethyl
3-((tert-butoxycarbonyl)(2-cyanoethyl)amino)propanoate
##STR00104##
[0214] To a solution of ethyl 3-aminopropanoate hydrochloride
(366.5 g, 2.39 mol) in MeOH (1.2 L) at room temperature was added
NaOH (95.6 g, 2.39 mol) in portions. The mixture was heated to
70.degree. C., acrylonitrile (158 g, 2.98 mol) was added dropwise
and the reaction mixture stirred for 6 h. The solution was cooled
to 0.degree. C. before (Boc).sub.2O (521 g, 2.39 mol) was added.
The reaction was stirred at room temperature for 6 h, filtered, and
washed with MeOH (200 mL). The filtrate was concentrated in vacuo
to give a yellow oil residue that was re-dissolved in EtOAc and
water (500 mL). The aqueous layer was extracted with EtOAc (800
mL). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (638 g) as light yellow oil that required no further
purification. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.17 (q,
J=7.2 Hz, 2H), 3.68-3.62 (m, 4H), 2.57-2.53 (m, 4H), 1.49 (s, 9H),
1.29 (t, J=7.2 Hz, 3H).
Step 2: tert-butyl 3-cyano-4-oxopiperidine-1-carboxylate
##STR00105##
[0216] To toluene (2.7 L) at 25.degree. C. was added NaH (80 g, 2.0
mol) portion-wise and the suspension was heated to 80.degree. C.
Ethyl 3-((tert-butoxycarbonyl)(2-cyanoethyl)amino)propanoate (270
g, crude) in anhydrous toluene (270 mL) was added dropwise. The
mixture was heated to 100.degree. C. and stirred for 5 hours. The
mixture was cooled to room temperature, quenched with sat. aq.
ammonium chloride (800 mL) and washed with hexanes (800 mL). The
aqueous phase was acidified with HCl (2 N) to pH 6 and then
extracted with EtOAc (1 L.times.2). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo to give the title compound (310 g) as yellow
oil that required no further purification. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 4.17-4.14 (m, 1H), 3.59-3.56 (m, 2H), 3.43-3.41
(m, 2H), 2.70-2.66 (m, 2H), 1.51 (s, 9H).
Step 3: tert-butyl
3-amino-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00106##
[0218] A mixture of tert-butyl
3-cyano-4-oxopiperidine-1-carboxylate (310 g, 1.38 mol) and
hydrazine mono-hydrate (140 mL, 2.08 mol) in EtOH (1.5 L) was
heated to 60.degree. C. for 2 h. The mixture was concentrated in
vacuo to give the crude product that was dissolved in EtOAc (1 L)
and washed with water (1 L.times.2). The organic layer was dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo
to afford the title compound (230 g, 70%) as a colorless solid.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 4.28 (s, 2H), 3.66-3.63
(m, 2H), 2.62-2.59 (m, 2H), 1.49 (s, 9H).
Step 4: tert-butyl
3-bromo-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00107##
[0220] To a stirred mixture of tert-butyl
3-amino-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(120 g, 503.6 mmol), CuBr.sub.2 (112.5 g, 503.6 mmol) and MeCN (1.2
L) at 0.degree. C. was added isopentyl nitrite (76.7 g, 654.7 mmol)
and the reaction mixture stirred for 20 min. The temperature was
raised to 60.degree. C. and the reaction mixture was stirred for an
additional 5 h. After cooling the reaction to room temperature, the
reaction mixture was quenched with water (1 L) and the mixture was
extracted with EtOAc (1 L.times.2). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (petroleum ether/EtOAc=4:1) to afford the title
compound (Intermediate A, 52 g, 34%) as light yellow solid. LCMS
M/Z (M+H) 302.
Step 5: tert-butyl
3-bromo-1-methyl-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00108##
[0222] To a stirred solution of tert-butyl
3-bromo-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(Intermediate A, 32 g, 105.9 mmol) in THE at 0.degree. C. (350 mL)
was added NaH (5.08 g, 127.1 mmol) and the mixture was stirred for
30 min. Methyliodide (18.05 g, 127.1 mmol) was added dropwise and
the mixture stirred for an additional 2 h. The mixture was quenched
with water and extracted with EtOAc (300 mL.times.2). The combined
organic layers were dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by silica
gel chromatography (petroleum ether/EtOAc=8:1) to afford the title
compound (16 g, 48%) as a colorless oil. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 4.24 (s, 2H), 3.70 (s, 3H), 3.69-3.67 (m, 2H),
2.70-2.67 (m, 2H), 1.47 (s, 9H).
Step 6:
1-(3-bromo-1-methyl-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl-
)ethanone
##STR00109##
[0224] A mixture of tert-butyl
3-bromo-1-methyl-6,7-dihydro-1H-pyrazolo[4,3-c]
pyridine-5(4H)-carboxylate (12 g, 38.0 mmol) and trifluoroacetic
acid (40 mL) in DCM (80 mL) was stirred at room temperature for 2
h. The mixture was concentrated in vacuo and the residue was
re-dissolved in DCM (120 mL). The mixture was cooled to 0.degree.
C. before TEA (12.1 g, 120 mmol) and acetic anhydride (5.3 g, 52
mmol) were added dropwise. The mixture stirred at room temperature
for an additional 2 h before water (100 mL) was added. The organic
layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (DCM/MeOH=20:1) to afford the title compound
(Intermediate B, 8.5 g, 87%) as white solid. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 4.40-4.39 (m, 2H), 3.88-3.78 (m, 2H), 3.72 (s,
3H), 2.83-2.70 (m, 2H), 2.20-2.17 (m, 3H).
General Procedure for Intermediate C
##STR00110##
[0225] Step 1: tert-butyl
3-bromo-1-(cyclopropylmethyl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H-
)-carboxylate
##STR00111##
[0227] To a stirred solution of tert-butyl
3-bromo-6,7-dihydro-1H-pyrazolo[4,3-c] pyridine-5(4H)-carboxylate
(Intermediate A, 6.0 g, 19.8 mmol) in DMF (40 mL) was added
Cs.sub.2CO.sub.3 (9.70 g, 29.8 mmol) and (bromomethyl)cyclopropane
(4.0 g, 29.8 mmole). The reaction mixture was heated to 80.degree.
C. for 12 h. The mixture was diluted with EtOAc (200 mL), washed
with brine (100 mL.times.3), dried over Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by silica
gel chromatography (eluent gradient from petroleum ether to
petroleum ether/tert-butyl methyl ether/THF=10:1:1) to give the
title compound (3.0 g, 42%) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 4.29 (s, 2H), 3.85 (d, J=3.4 Hz, 2H), 3.71 (t,
J=5.2 Hz, 2H), 2.67 (t, J=5.2 Hz, 2H), 1.49 (s, 9H), 1.25-1.18 (m,
1H), 0.61-0.55 (m, 2H), 0.35-0.31 (m, 2H).
Step 2:
1-(3-bromo-1-(cyclopropylmethyl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone
##STR00112##
[0229] A mixture of tert-butyl
3-bromo-1-(cyclopropylmethyl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H-
)-carboxylate (3.0 g, 8.4 mmol) and trifluoroacetic acid (30 mL) in
DCM (30 mL) was stirred at room temperature for 2 h. The solvent
was concentrated in vacuo and the crude product was re-dissolved in
DCM (120 mL). The solution was cooled to 0.degree. C. before TEA
(2.49 g, 24.6 mmol) and acetic anhydride (1.26 g, 12.3 mmol) were
added dropwise. The reaction mixture was stirred at room
temperature for additional 2 h before it was quenched with water.
The organic layer was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude residue was purified
by silica gel chromatography (DCM/MeOH=20:1) to afford the title
compound (2.40 g, 96%) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 4.49-4.33 (m, 3H), 3.90-3.70 (m, 4H), 2.77-2.67
(m, 2H), 2.23-2.19 (m, 3H), 1.28-1.18 (m, 1H), 0.63-0.58 (m, 2H),
0.36-0.32 (m, 2H).
General Procedure for Intermediate D
##STR00113##
[0230] Step 1: tert-butyl
3-bromo-1-(oxetan-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carb-
oxylate
##STR00114##
[0232] To a solution of tert-butyl
3-bromo-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(Intermediate A, 40.0 g, 132 mmol) in DMF (500 mL) was added
Cs.sub.2CO.sub.3 (87 g, 264 mmol) and 3-iodooxetane (27 g, 146
mmol). The mixture was heated to 60.degree. C. for 12 h before
3-iodooxetane (5 g, 27.0 mmol) was added and the mixture was
stirred at 60.degree. C. for an additional 6 h. After cooling the
reaction to room temperature, the mixture was filtered, washed with
EtOAc (500 mL) and concentrated in vacuo. The crude residue was
purified by silica gel chromatography (petroleum ether:tert-butyl
methyl ether:THF=from 100:1:1 to 5:1:1) to give the title compound
(30 g, 64%) as a white solid. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 5.30-5.25 (m 1H), 5.18-5.14 (m, 2H), 4.95-4.91 (m, 2H),
4.28 (s, 2H), 3.73-3.66 (m, 2H), 2.64 (t, J=5.6 Hz, 2H), 1.48 (s,
9H).
Step 2:
1-(3-bromo-1-(oxetan-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5-
(4H)-yl)ethanone
##STR00115##
[0234] To a solution of tert-butyl
3-bromo-1-(oxetan-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carb-
oxylate (25.0 g, 70.0 mmol) in DCM (50 mL) was added
trifluoroacetic acid (50 mL) dropwise at 0.degree. C. The mixture
was stirred at room temperature for 2 h. The mixture was
concentrated in vacuo and the residue was re-dissolved in DCM (500
mL). The mixture was cooled to 0.degree. C. before triethylamine
(48.8 mL, 350 mmol) and acetic anhydride (7.2 g, 70.0 mmol) were
added dropwise. The mixture was stirred at room temperature for
additional 2 h. The reaction was quenched with water. The organic
layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (DCM/MeOH=80:1) to give the title compound
(Intermediate D, 17.0 g, 81%) as a light yellow solid. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 5.32-5.27 (m 1H), 5.16-5.13 (m, 2H),
4.95-4.91 (m, 2H), 4.47-4.31 (m, 2H), 3.88-3.70 (m, 2H), 2.75-2.63
(m, 2H), 2.17 (s, 3H).
General Procedure for Intermediate E
##STR00116##
[0235] Step 1: (R)-tetrahydrofuran-3-yl Methanesulfonate
##STR00117##
[0237] To a solution of (R)-tetrahydrofuran-3-ol (25 g, 253.7 mmol)
in DCM (250 mL) at 0.degree. C. was added triethylamine (119 mL,
851.2 mmol) and mesyl chloride (39 g, 340.48 mmol) dropwise. The
mixture was stirred at room temperature for 12 h. The reaction was
quenched with water (100 mL) and extracted with DCM (100
mL.times.2). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (47 g, 99%) as a brown oil. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 5.35-5.27 (m, 1H), 4.05-3.83 (m, 4H), 3.04 (s,
3H), 2.28-2.20 (m, 2H).
Step 2: (S)-tert-butyl
3-bromo-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5-
(4H)-carboxylate
##STR00118##
[0239] To a solution of tert-butyl
3-bromo-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(Intermediate A, 24.8 g, 82 mmol) in DMF (200 mL) was added
Cs.sub.2CO.sub.3 (79 g, 246 mmol) and (R)-tetrahydrofuran-3-yl
methanesulfonate (17.4 g, 98 mmol). The mixture was heated to
80.degree. C. for 12 h. After cooling the reaction to room
temperature, the mixture was concentrated in vacuo. The crude
residue was purified by silica gel chromatography (petroleum
ether/EtOAc=from 10:1 to 3:1) to give the title compound (50 g,
71%) as a yellow oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
4.97-4.78 (m, 1H), 4.13 (s, 2H), 3.98-3.86 (m, 2H), 3.81-3.67 (m,
2H), 3.56 (t, J=5.6 Hz, 2H), 2.68 (t, J=5.6 Hz, 2H), 2.33-2.08 (m,
2H), 1.38 (s, 9H).
Step 3:
(S)-1-(3-bromo-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,-
3-c]pyridin-5(4H)-yl)ethanone
##STR00119##
[0241] To a solution of (S)-tert-butyl
3-bromo-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5-
(4H)-carboxylate (29 g, 78 mmol) in DCM (300 mL) was added
trifluroacetic acid (70 mL) dropwise. The mixture was stirred at
room temperature for 2 h. The solvent was concentrated in vacuo and
the crude residue was re-dissolved in DMF (100 mL). The mixture was
cooled to 0.degree. C. before triethylamine (22 mL, 156 mmol) and
acetic anhydride (8.7 g, 86 mmol) were added dropwise. The mixture
was stirred at room temperature for an additional 2 h. The reaction
was quenched with water (200 mL) at 0.degree. C. and extracted with
EtOAc (150 mL.times.3). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography
(DCM/MeOH=30:1) to give the title compound (Intermediate E, 21.3 g,
87%) as a white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
4.78-4.67 (m, 1H), 4.45-4.29 (m, 2H), 4.15-4.06 (m, 2H), 3.96-3.92
(m, 2H), 3.88-3.70 (m, 2H), 2.71-2.67 (m, 2H), 2.38-2.34 (m, 2H),
2.16 (s, 3H).
General procedure for Intermediate F
##STR00120##
Step 1: tetrahydrofuran-3-yl Methanesulfonate
##STR00121##
[0243] To a solution of tetrahydrofuran-3-ol (10 g, 113.5 mmol) in
DCM (150 mL) was added MsCl (15.6 g, 136.2 mmol) and TEA (23 g, 227
mmol). The reaction mixture was stirred at room temperature for 18
h. Water (100 mL) was added and the mixture was extracted with DCM
(100 mL.times.2). The combined organic layers was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to
give the title compound (16 g, 85%) as a brown oil. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 5.27-5.25 (m, 1H), 4.00-3.83 (m, 4H),
3.01 (s, 3H), 2.23-2.18 (m, 2H).
Step 2: tert-butyl
3-bromo-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5-
(4H)-carboxylate
##STR00122##
[0245] To a solution of tert-butyl
3-bromo-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(Intermediate A, 20.0 g, 66.0 mmol) in DMF (100 mL) was added
Cs.sub.2CO.sub.3 (40.0 g, 123 mmol) and tetrahydrofuran-3-yl
methanesulfonate (16.0 g, 98.0 mmol). The mixture was heated to
80.degree. C. for 12 h. The solution was concentrated in vacuo and
the crude residue was purified by silica gel chromatography (eluent
from petroleum ether/EtOAc=10:1 to 3:1) to give the title compound
(17 g, 69%) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 4.78-4.69 (m, 1H), 4.26 (s, 2H), 4.18-3.86 (m, 4H), 3.72
(s, 2H), 2.72-2.62 (m, 2H), 2.44-2.22 (m, 2H), 1.48 (s, 9H).
Step 3:
1-(3-bromo-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]-
pyridin-5(4H)-yl)ethanone
##STR00123##
[0247] To a solution of tert-butyl
3-bromo-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo
[4,3-c]pyridine-5(4H)-carboxylate (17.0 g, 45.0 mmol) in DCM (60
mL) was added TFA (30 mL) dropwise. The reaction solution was
stirred at room temperature for 2 h. The solvent was removed by
evaporation and the crude product was re-dissolved in DMF (50 mL).
The mixture was cooled to 0.degree. C. before TEA (41.0 g, 40.5
mmol) and acetic anhydride (7.0 g, 68.0 mmol) were added dropwise.
The ice bath was removed and the reaction was stirred at room
temperature for additional 2 h. Water (50 mL) was added and the
solution was extracted with EtOAc (150 mL.times.3). The combined
organic layers were dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by silica
gel chromatography (DCM/MeOH=30:1) to give the title compound
(Intermediate F, 12.0 g, 82%) as a white solid. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 4.96-4.92 (m 1H), 4.28 (s, 2H),
3.99-3.95 (m, 2H), 3.80-3.68 (m, 4H), 2.82-2.70 (m, 2H), 2.29-2.19
(m, 2H), 2.10-2.08 (m, 3H).
General procedure for Intermediates G & H
##STR00124##
Step 1: tetrahydro-2H-pyran-4-yl Methanesulfonate
##STR00125##
[0249] To a solution of tetrahydro-2H-pyran-4-ol (5 g, 49.0 mmol)
and triethylamine (8.2 mL, 58.7 mmol) in DCM (100 mL) was added
mesyl chloride (16.8 g, 146.9 mmol) dropwise at 0.degree. C. under
a nitrogen atmosphere. The mixture was stirred at room temperature
for 5 h. Water (100 mL) was added and extracted with DCM (100
mL.times.2). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (4 g, 45%) as a yellow solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 4.85-4.81 (m 1H), 3.90-3.87 (m, 2H), 3.52-3.46
(m, 2H), 2.99 (s, 3H), 2.01-1.97 (m, 2H), 1.83-1.80 (m, 2H).
Step 2: tert-butyl
3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
ne-5(4H)-carboxylate
##STR00126##
[0251] To a solution of tert-butyl
3-bromo-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(Intermediate A, 6 g, 19.8 mmol) in DMF (40 mL) was added
Cs.sub.2CO.sub.3 (19.5 g, 59.6 mmol) and tetrahydro-2H-pyran-4-yl
methanesulfonate (3.9 g, 21.8 mmol). The mixture was heated to
80.degree. C. for 12 h under a nitrogen atmosphere. After cooling
the reaction to room temperature, the mixture was filtered. The
mixture was diluted with EtOAc (100 mL) and washed with brine (100
mL.times.2). The organic layer was concentrated in vacuo. The crude
residue was purified by silica gel chromatography (petroleum
ether:tert-butyl methyl ether:THF=from 10:1:1 to 2:1:1) to give the
title compound (Intermediate G, 3.2 g, 47%) as a clear oil. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 4.35-4.25 (m, 1H), 4.17 (s,
2H), 3.95-3.93 (m, 2H), 3.62-3.57 (m, 2H), 3.42 (t, J=11.2 Hz, 2H),
2.74-2.73 (m, 2H), 1.98-1.89 (m, 2H), 1.80-1.77 (m, 2H), 1.41 (s,
9H).
Step 3:
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,-
3-c]pyridin-5 (4H)-yl)ethanone
##STR00127##
[0253] To a solution of tert-butyl
3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
ne-5(4H)-carboxylate (Intermediate G, 3.2 g, 8.3 mmol) in DCM (20
mL) was added trifluoroacetic acid (20 mL) dropwise at 0.degree. C.
The mixture was stirred at room temperature for 2 h. The mixture
was concentrated in vacuo and the residue was re-dissolved in DCM
(30 mL). The mixture was cooled to 0.degree. C. before
triethylamine (2.9 mL, 21 mmol) and acetic anhydride (0.93 g, 9.1
mmol) were added dropwise. The mixture was stirred at room
temperature for an additional 0.5 h. The reaction was quenched with
water (60 mL). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (DCM/MeOH=50:1)
to give the title compound (Intermediate H, 2.1 g, 77%) as a light
yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.33-4.29
(m, 1H), 4.28 (s 2H), 3.95-3.92 (m, 2H), 3.70-3.67 (m, 2H),
3.43-3.36 (m, 2H), 2.84-2.69 (m, 2H), 2.09-2.08 (m, 3H), 1.96-1.91
(m, 2H), 1.80-1.76 (m, 2H).
General Procedure for Intermediate I
##STR00128## ##STR00129##
[0254] Step 1: N-(2-chlorobenzyl)-2, 2-dimethoxyacetamide
##STR00130##
[0256] To a solution of 2-chlorobenzylamine (30.0 g, 212 mmol) in
MeOH (200 mL) was added triethylamine (36.7 mL, 265 mmol) and
methyl dimethoxyacetate (31.0 g, 233 mmol). The reaction was heated
to 80.degree. C. for 20 h. After cooling to room temperature, the
reaction was concentrated in vacuo. EtOAc (300 ml) was added,
washed with 1N HCl (300 mL.times.2) and sat. aq. NaHCO.sub.3 (300
mL.times.2). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (40.0 g, crude) as colorless oil that required no
further purification. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.39-7.33 (m, 2H), 7.24-7.21 (m, 2H), 4.73 (s, 1H), 4.56 (d, J=6.0
Hz, 2H), 3.38 (s, 6H).
Step 2: 8-chloroisoquinolin-3(2H)-one
##STR00131##
[0258] To a solution of concentrated sulfuric acid (100 mL) at
0.degree. C. was added N-(2-chlorobenzyl)-2, 2-dimethoxyacetamide
(40.0 g, 164 mmol). The reaction was stirred at room temperature
for 16 h. The reaction was poured into ice water and the mixture
was basified with ammonium hydroxide to pH 8. The yellow
precipitate was filtered off, washed with water, and dried in vacuo
to give the title compound (20.0 g, crude) as a yellow solid that
required no further purification. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 9.11 (s, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.56-7.52 (m, 1H),
7.45 (d, J=7.6 Hz, 1H), 7.00 (s, 1H).
Step 3: 8-chloro-3-((triisopropylsilyl)oxy)isoquinoline
##STR00132##
[0260] To a solution of 8-chloroisoquinolin-3(2H)-one (12.0 g, 66.8
mmol) in DMF (15 mL) at 0.degree. C. was added imidazole (13.6 g,
200.5 mmol) and chlorotriisopropylsilane (17.2 mL, 80.2 mmol). The
reaction was stirred at room temperature for 12 h. The reaction
mixture was concentrated in vacuo. The crude residue was dissolved
in EtOAc (70 mL) and washed with H.sub.2O (40 mL.times.2). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by silica
gel column chromatography (petroleum ether) to give the title
compound (13.0 g, 62%) as colourless oil. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.16 (s, 1H), 7.79 (d, J=8.4 Hz, 1H),
7.65-7.56 (m, 1H), 7.55-7.47 (m, 1H), 7.23 (s, 1H), 1.46-1.30 (m,
3H), 1.06 (d, J=7.6 Hz, 18H).
Step 4:
8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-((triisopropylsi-
lyl)-oxy)isoquinoline
##STR00133##
[0262] To a solution of
8-chloro-3-((triisopropylsilyl)oxy)isoquinoline (13.0 g, 38.7 mmol)
in 1,4-dioxane (10 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (913
mg, 1.2 mmol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (19.6
g, 77.4 mmol) and KOAc (11.4 g, 116.1 mmol). The mixture was heated
to 80.degree. C. for 2 h under a nitrogen atmosphere. After cooling
the reaction to room temperature, the reaction mixture was used to
the next step directly without further purification. LCMS M/Z
(M-TIPS+H) 272.
Step 5:
1-(1-(tetrahydro-2H-pyran-4-yl)-3-(3-((triisopropylsilyl)oxy)isoqu-
inolin-8-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00134##
[0264] To the above step cooled solution was added
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 8.9 g, 27.0 mmol),
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (1.2 g,
1.5 mmol) and K.sub.3PO.sub.4 (20.5 g, 96.5 mmol), 1,4-dioxane (5
mL) and water (3 mL). The reaction mixture was heated to 90.degree.
C. for 4 h under a nitrogen atmosphere. After cooling to room
temperature, the mixture was filtered and concentrated in vacuo.
DCM (60 mL) was added and washed with water (50 mL.times.2). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by silica
gel chromatography (DCM/MeOH=20:1) to give the title compound (6.0
g, 28%) as a brown solid. LCMS M/Z (M-TIPS+H) 393.
Step 6:
1-(3-(3-hydroxyisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7--
dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00135##
[0266] To a solution of
1-(1-(tetrahydro-2H-pyran-4-yl)-3-(3-((triisopropylsilyl)oxy)isoquinolin--
8-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (6.0
g, 10.9 mmol) in THE (10 mL) at 20.degree. C. was added TBAF (54.7
ml, 54.7 mmol, 1 M in THF). The reaction was stirred at room
temperature for 12 h. The reaction mixture was concentrated in
vacuo. The crude residue was dissolved in DCM (50 mL) and washed
with H.sub.2O (150 mL.times.2). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography
(DCM/MeOH=10:1) to give the title compound (5.0 g, 86% purity) as a
brown solid. LCMS M/Z (M+H) 393.
Step 7:
8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyr-
azolo[4,3-c]pyridin-3-yl)isoquinolin-3-yl
trifluoromethanesulfonate
##STR00136##
[0268] To a solution of
1-(3-(3-hydroxyisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-
-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (5.0 g, 12.7 mmol) in
DCM (10 mL) at 0.degree. C. was added triethylamine (5.3 mL, 38.2
mmol) and trifluoromethanesulfonic anhydride (3.2 mL, 19.1 mmol).
The reaction was stirred at room temperature for 1 h. DCM (100 mL)
was added and washed with water (80 mL.times.2). The organic layer
was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (EtOAc) to give the title compound (Intermediate I,
1.7 g, 23%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.84-9.74 (m, 1H), 8.21 (s, 1H), 8.16 (d, J=8.4 Hz, 1H),
8.05-7.95 (m, 1H), 7.84-7.72 (m, 1H), 4.59 (s, 2H), 4.50-4.45 (m,
1H), 4.05-3.98 (m, 2H), 3.89-3.74 (m, 2H), 3.51 (d, J=12.0 Hz, 1H),
3.01-2.80 (m, 2H), 2.20-2.00 (m, 5H), 1.99-1.88 (m, 2H).
Example 1 (Procedure A)
1-(3-(1H-indol-3-yl)-1-methyl-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)--
yl)ethanone
##STR00137##
[0269] Step 1: tert-butyl
3-(5-acetyl-1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-1-
H-indole-1-carboxylate
##STR00138##
[0271] To a solution of
1-(3-bromo-1-methyl-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethano-
ne (Intermediate B, 500 mg, 1.92 mmol) in 1,4-dioxane (8 mL) and
water (2 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (146
mg, 0.2 mmol), Na.sub.2CO.sub.3 (408 mg, 3.84 mmol) and
(1-(tert-butoxycarbonyl)-1H-indol-3-yl)boronic acid (552 mg, 2.1
mmol). The mixture was heated to 110.degree. C. for 18 h under a
nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
The crude residue was purified by silica gel chromatography
(DCM/MeOH=20:1) to give the title compound (400 mg, 53%) as a brown
solid.
Step 2:
1-(3-(1H-indol-3-yl)-1-methyl-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
n-5(4H)-yl)ethanone
##STR00139##
[0273] To a solution of tert-butyl
3-(5-acetyl-1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-1-
H-indole-1-carboxylate (200 mg, 0.51 mmol) in DCM (2 mL) at
0.degree. C. was added trifluoroacetic acid (2 mL, 3.4 mmol). The
mixture was stirred at room temperature for 2 h and concentrated in
vacuo. The crude residue was purified by reverse phase
chromatography (acetonitrile 30-60%/0.1% NH.sub.4OH in water) to
give the title compound (15 mg, 10%) as a white solid. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.23 (s, 1H), 8.26-8.21 (m, 1H),
7.52-7.40 (m, 2H), 7.15-7.06 (m, 2H), 4.58 (s, 2H), 3.85-3.68 (m,
5H), 2.80-2.68 (m, 2H), 2.12 (s, 3H). LCMS M/Z (M+H) 295.
Example 2 (Procedure B)
1-(1-methyl-3-(1-methyl-1H-indol-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyrid-
in-5(4H)-yl)ethanone
##STR00140##
[0275] To a solution of
1-(3-(1H-indol-3-yl)-1-methyl-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-
-yl)ethanone (Example 1, 150 mg, 0.51 mmol) in DMF (2 mL) at
0.degree. C. was added sodium hydride (60%, 41 mg, 1.02 mmol) by
portionwise. The mixture was stirred at room temperature for 0.5 h.
Iodomethane (0.073 mL, 1.02 mmol) was added dropwise and the
mixture stirred at room temperature for an additional 1 h. The
mixture was quenched with water (20 mL) and extracted with EtOAc
(20 mL.times.3). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by reverse phase chromatography
(acetonitrile 36-66%/0.1% NH.sub.4OH in water) to give the title
compound (47 mg, 29%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.28-8.24 (m, 1H), 7.55-7.42 (m, 2H),
7.22-7.18 (m, 1H), 7.12-7.08 (m, 1H), 4.61-4.54 (m, 2H), 3.85 (s,
1H), 3.79-3.72 (m, 5H), 2.82-2.67 (m, 2H), 2.13-2.07 (m, 3H). LCMS
M/Z (M+H) 309.
Example 3 (Procedure C)
1-(1-methyl-3-(6-(1-methyl-1H-pyrazol-4-yl)-1H-indol-3-yl)-6,7-dihydro-1H--
pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00141##
[0276] Step 1: tert-butyl 6-bromo-1H-indole-1-carboxylate
##STR00142##
[0278] To a solution of 6-bromo-1H-indole (10.0 g, 51.0 mmol) in
DCM (100 mL) was added di-tert-butyl dicarbonate (13.4 g, 61.2
mmol), DMAP (623 mg, 5.1 mmol) and triethylamine (21.2 mL, 153.0
mmol). The mixture was stirred at 20.degree. C. for 16 h under a
nitrogen atmosphere. The mixture was concentrated in vacuo. The
crude residue was purified by silica gel chromatography (petroleum
ether/EtOAc=10:1) to give the title compound (14.5 g, 96%) as a
yellow solid.
Step 2: tert-butyl
6-(1-methyl-1H-pyrazol-4-yl)-1H-indole-1-carboxylate
##STR00143##
[0280] To a solution of tert-butyl 6-bromo-1H-indole-1-carboxylate
(10.0 g, 33.8 mmol) in 1,4-dioxane (90 mL) and water (30 mL) was
added [1,1'-is(diphenylphosphino)ferrocene]-dichloropalladium (II)
(2.0 g, 3.4 mmol), Na.sub.2CO.sub.3 (10.7 g, 101.3 mmol) and
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(8.4 g, 40.5 mmol). The mixture was heated to 110.degree. C. for 16
h under a nitrogen atmosphere. After cooling the reaction to room
temperature, water (100 mL) was added and extracted with EtOAc (100
mL.times.3). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (petroleum
ether/EtOAc=2:1) to give the title compound (9.0 g, 90%) as a light
brown solid.
Step 3: tert-butyl
3-bromo-6-(1-methyl-1H-pyrazol-4-yl)-1H-indole-1-carboxylate
##STR00144##
[0282] To a solution of tert-butyl
6-(1-methyl-1H-pyrazol-4-yl)-1H-indole-1-carboxylate (7.5 g, 25.2
mmol) in DCM (80 mL) was added N-bromosuccinimide (4.49 g, 25.2
mmol). The mixture was stirred at 40.degree. C. for 16 h under a
nitrogen atmosphere. Water (120 mL) was added and the mixture was
extracted with DCM (120 mL.times.2). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo to give the title compound (7.0 g, 74%) as
brown oil that required no further purification.
Step 4: tert-butyl
6-(1-methyl-1H-pyrazol-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)-1H-indole-1-carboxylate
##STR00145##
[0284] To a solution of tert-butyl
3-bromo-6-(1-methyl-1H-pyrazol-4-yl)-1H-indole-1-carboxylate (7.0
g, 18.6 mmol) in DMF (40 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (1.4 g,
1.9 mmol), KOAc (5.5 g, 55.8 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (7.0 g,
27.9 mmol). The mixture was heated to 70.degree. C. for 12 h under
a nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was concentrated in vacuo. The crude
residue was purified by silica gel chromatography (petroleum
ether/EtOAc=5:1) to give the title compound (3.0 g, 38%) as a
yellow oil.
Step 5: tert-butyl
3-(5-acetyl-1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-6-
-(1-methyl-1H-pyrazol-4-yl)-1H-indole-1-carboxylate
##STR00146##
[0286] To a solution of
1-(3-bromo-1-methyl-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethano-
ne (Intermediate B, 838 mg, 3.25 mmol) in 1,4-dioxane (15 mL) and
water (5 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (238
mg, 0.33 mmol), K.sub.2CO.sub.3 (898 mg, 6.50 mmol) and tert-butyl
6-(1-methyl-1H-pyrazol-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)-1H-indole-1-carboxylate (2.5 g, 3.25 mmol). The mixture was
heated to 70.degree. C. for 16 h under a nitrogen atmosphere. After
cooling the reaction to room temperature, water (50 mL) was added
and extracted with EtOAc (60 mL.times.3). The combined organic
layers were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (DCM/MeOH=20:1) to give the title compound (1.0 g,
65%) as a brown solid.
Step 6:
1-(1-methyl-3-(6-(1-methyl-1H-pyrazol-4-yl)-1H-indol-3-yl)-6,7-dih-
ydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00147##
[0288] To a solution of tert-butyl
3-(5-acetyl-1-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-6-
-(1-methyl-1H-pyrazol-4-yl)-1H-indole-1-carboxylate (700 mg, 1.48
mmol) in DCM (7 mL) was added trifluoroacetic acid (7 mL). The
mixture was stirred at 20.degree. C. for 3 h under a nitrogen
atmosphere. The mixture was concentrated in vacuo to give the crude
residue that was dissolved in EtOAc (50 mL), washed with sat. aq.
NaHCO.sub.3 (50 mL.times.2) and brine (50 mL.times.2), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by recrystallization to give the title
compound (202 mg, 36%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.21 (s, 1H), 8.21-8.15 (m, 1H), 8.08 (s,
1H), 7.83 (s, 1H), 7.52 (s, 1H), 7.47-7.37 (m, 1H), 7.28-7.26 (m,
1H), 4.57 (s, 2H), 3.86 (s, 3H), 3.80-3.68 (m, 5H), 2.80-2.67 (m,
2H), 2.11 (s, 3H). LCMS M/Z (M+H) 375.
Example 4 (Procedure D)
1-(3-(6-methylisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1-
H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00148##
[0289] Step 1:
N-(2-bromo-4-chlorobenzylidene)-2,2-dimethoxyethanamine
##STR00149##
[0291] To a solution of 2-bromo-4-chlorobenzaldehyde (40.0 g,
182.27 mmol) in toluene (100 mL) was added 2,2-dimethoxyethanamine
(19.16 g, 182.27 mmol). The mixture was heated to 110.degree. C.
for 16 h under a nitrogen atmosphere. After cooling the reaction to
room temperature, the mixture was concentrated in vacuo to give the
title compound (59.0 g, crude) as colorless oil that required no
further purification. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
8.57 (s, 1H), 7.96 (d, J=8.4 Hz, 1H), 7.57 (s, 1H), 7.31-7.28 (m,
1H), 4.67 (t, J=5.2 Hz, 1H), 3.80 (d, J=4.4 Hz, 2H), 3.41 (s,
6H).
Step 2: N-(2-bromo-4-chlorobenzyl)-2,2-dimethoxyethanamine
##STR00150##
[0293] To a solution of
N-(2-bromo-4-chlorobenzylidene)-2,2-dimethoxyethanamine (59.0 g,
192.45 mmol) in MeOH (200 mL) at 0.degree. C. was added sodium
borohydride (5.82 g, 153.96 mmol) portionwise. The mixture was
stirred at 28.degree. C. for 2 h under a nitrogen atmosphere and
concentrated in vacuo. Water (200 mL) was added and extracted with
DCM (200 mL.times.3). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to
give the title compound (60.0 g, crude) as colorless oil that
required no further purification. LCMS M/Z (M+H) 308.
Step 3:
N-(2-bromo-4-chlorobenzyl)-N-(2,2-dimethoxyethyl)-4-methylbenzenes-
ulfonamide
##STR00151##
[0295] To a solution of
N-(2-bromo-4-chlorobenzyl)-2,2-dimethoxyethanamine (60.0 g, 194.43
mmol) in DCM (300 mL) at 0.degree. C. was added
4-methylbenzene-1-sulfonyl chloride (37.0 g, 194.43 mmol),
N,N-dimethylpyridin-4-amine (1.19 g, 9.72 mmol) and triethylamine
(53.9 mL, 388.85 mmol). The mixture was stirred at 28.degree. C.
for 10 h under a nitrogen atmosphere. Water (300 mL) was added and
extracted with DCM (300 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (petroleum ether/EtOAc=5:1) to give the title
compound (70 g, 78%) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.72 (d, J=7.6 Hz, 1H), 7.52 (s, 1H), 7.45 (d,
J=8.4 Hz, 1H), 7.35-7.27 (m, 3H), 4.48 (s, 2H), 4.36 (t, J=5.2 Hz,
1H), 3.29 (d, J=5.2 Hz, 2H), 3.23 (s, 6H), 2.45 (s, 3H). LCMS M/Z
(M+H) 462.
Step 4: 8-bromo-6-chloroisoquinoline
##STR00152##
[0297] To a solution of aluminum trichloride (43.22 g, 324.12 mmol)
in 1,2-dichloroethane (200 mL) at 0.degree. C. was added
N-(2-bromo-4-chlorobenzyl)-N-(2,2-dimethoxyethyl)-4-methylbenzenesulfonam-
ide (30.0 g, 64.82 mmol) in 1,2-dichloroethane (200 mL) dropwise.
The mixture was stirred at 28.degree. C. for 16 h under a nitrogen
atmosphere and concentrated in vacuo. The crude residue was
purified by silica gel chromatography (petroleum ether/EtOAc=10:1)
to give the title compound (6.2 g, 39%) as a yellow solid. LCMS M/Z
(M+H) 242.
Step 5:
6-chloro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoli-
ne
##STR00153##
[0299] To a solution of 8-bromo-6-chloroisoquinoline (3.0 g, 12.37
mmol) in 1,4-dioxane (50 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (905
mg, 1.24 mmol), potassium 2-ethylhexanoate (3.38 g, 18.56 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (4.71
g, 18.56 mmol). The mixture was heated to 80.degree. C. for 1 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 290.
Step 6:
1-(3-(6-chloroisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-d-
ihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00154##
[0301] To the above step cooled solution was added
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 6.12 g, 18.65 mmol),
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (758
mg, 1.04 mmol) and K.sub.3PO.sub.4 (4.40 g, 20.72 mmol),
1,4-dioxane (30 mL) and water (20 mL). The reaction mixture was
heated to 80.degree. C. for 8 h under nitrogen atmosphere. After
cooling to room temperature, the mixture was filtered and
concentrated in vacuo. EtOAc (200 mL) was added and washed with
water (120 mL.times.2). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (DCM/MeOH=30:1)
to give the title compound (1.6 g, 31%) as a brown solid. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 9.78-9.70 (m, 1H), 8.58 (d,
J=5.6 Hz, 1H), 8.15 (s, 1H), 7.88 (d, J=5.6 Hz, 1H), 7.68-7.55 (m,
1H), 4.55 (s, 2H), 4.54-4.42 (m, 1H), 4.06-3.96 (m, 2H), 3.87-3.77
(m, 2H), 3.51 (t, J=12.0 Hz, 2H), 3.01-2.80 (m, 2H), 2.15-2.00 (m,
5H), 1.98-1.87 (m, 2H). LCMS M/Z (M+H) 411.
Step 7:
1-(3-(6-methylisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-d-
ihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00155##
[0303] To a solution of
1-(3-(6-chloroisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro--
1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (200 mg, 0.49 mmol) in
toluene (10 mL) and water (2 mL) was added potassium
methyltrifluoroborate (59 mg, 0.49 mmol), butyl
di-1-adamantylphosphine (175 mg, 0.49 mmol), palladium(II) acetate
(109 mg, 0.49 mmol) and Cs.sub.2CO.sub.3 (159 mg, 0.49 mmol). The
mixture was heated to 80.degree. C. for 16 h under a nitrogen
atmosphere. After cooling the reaction to room temperature, EtOAc
(100 mL) was added and washed with water (100 mL.times.3) and brine
(100 mL). The organic phase was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by reverse phase chromatography (acetonitrile
30-60%/0.05% NH.sub.4OH in water) to give the title compound (15
mg, 8%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.66 (s, 1H), 8.48 (d, J=6.0 Hz, 1H), 7.81-7.73 (m, 2H),
7.54-7.42 (m, 1H), 4.52 (s, 2H), 4.51-4.40 (s, 1H), 4.08-3.95 (m,
2H), 3.87-3.76 (m, 2H), 3.52 (t, J=12.0 Hz, 2H), 3.00-2.81 (m, 2H),
2.56 (s, 3H), 2.13-2.03 (m, 5H), 2.01-1.94 (m, 2H). LCMS M/Z (M+H)
391.
Example 5 (Procedure E)
1-(3-(6-methoxyisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro--
1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00156##
[0305] To a solution of
1-(3-(6-chloroisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro--
1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (200 mg, 0.49 mmol) in
1,4-dioxane (6 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (46 mg,
0.10 mmol),
methanesulfonato(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1-
'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (82 mg, 0.10
mmol), t-BuONa (65 mg, 0.68 mmol) and MeOH (0.1 mL, 2.43 mmol). The
mixture was heated to 50.degree. C. for 20 h under a nitrogen
atmosphere. After cooling the reaction to room temperature, the
mixture was filtered and concentrated in vacuo. The crude residue
was purified by reverse phase chromatography (acetonitrile
15-45%/0.05% NH.sub.40H in water) to give the title compound (30
mg, 90% purity) which was further separated by using chiral SFC
(SFC 80; Chiralpak AS 250.times.30 mm I.D, 5 um; Supercritical
CO.sub.2/MeOH+NH.sub.3.H.sub.2O=30/30; 60 ml/min) to give the title
compound (18 mg, 9%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.66-9.59 (m, 1H), 8.41 (d, J=5.6 Hz, 1H),
7.75 (d, J=5.6 Hz, 1H), 7.36 (d, J=2.4 Hz, 1H), 7.24-7.14 (m, 1H),
4.51 (s, 2H), 4.50-4.39 (m, 1H), 4.05-3.95 (m, 2H), 3.94 (s, 3H),
3.86-3.76 (m, 2H), 3.51 (t, J=12.0 Hz, 2H), 3.00-2.79 (m, 2H),
2.16-2.00 (m, 5H), 1.99-1.87 (m, 2H). LCMS M/Z (M+H) 407.
Example 6 (Procedure F)
1-(1-(tetrahydro-2H-pyran-4-yl)-3-(6-vinylisoquinolin-8-yl)-6,7-dihydro-1H-
-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00157##
[0307] To a solution of
1-(3-(6-chloroisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro--
1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (3.0 g, 7.3 mmol) in
THE (30 mL) and water (6 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (348 mg,
0.73 mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphe-
nyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (574 mg, 0.73
mmol), Na.sub.2CO.sub.3 (1.55 g, 14.6 mmol) and
4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (1.46 g, 9.49
mmol). The mixture was heated to 60.degree. C. for 16 h under a
nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
The crude residue was purified by silica gel chromatography
(DCM/MeOH=30:1) to give the title compound (1.5 g, 51%) as a yellow
solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.70-9.64 (m,
1H), 8.50 (d, J=5.6 Hz, 1H), 7.96 (s, 1H), 7.83 (d, J=6.0 Hz, 1H),
7.74 (s, 1H), 7.04-6.92 (m, 1H), 6.12 (d, J=17.6 Hz, 1H), 5.51 (d,
J=11.2 Hz, 1H), 4.53 (s, 2H), 4.48-4.38 (m, 1H), 4.04-3.95 (m, 2H),
3.87-3.76 (m, 2H), 3.50 (t, J=11.6 Hz, 2H), 3.00-2.79 (m, 2H),
2.13-1.98 (m, 5H), 1.97-1.92 (m, 2H). LCMS M/Z (M+H) 403.
Example 7 (Procedure G)
1-(3-(6-ethylisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-
-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00158##
[0309] To a solution of
1-(1-(tetrahydro-2H-pyran-4-yl)-3-(6-vinylisoquinolin-8-yl)-6,7-dihydro-1-
H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (Example 6, 200 mg, 0.50
mmol) in MeOH (5 mL) was added 10% Pd/C (53 mg). The mixture was
stirred at room temperature for 24 h under a hydrogen atmosphere
(15 Psi). The mixture was filtered and the filtrate was
concentrated in vacuo. The crude residue was purified by reverse
phase chromatography (acetonitrile 15-45%/0.05% NH.sub.4OH in
water) to give the title compound (14 mg, 7%) as a white solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.67 (s, 1H), 8.48 (d,
J=6.0 Hz, 1H), 7.80 (d, J=5.6 Hz, 1H), 7.77 (s, 1H), 7.59-7.45 (m,
1H), 4.52 (s, 2H), 4.50-4.39 (m, 1H), 4.05-3.93 (m, 2H), 3.88-3.74
(m, 2H), 3.51 (t, J=11.6 Hz, 2H), 2.99-2.77 (m, 2H), 2.86 (q, J=7.6
Hz, 2H), 2.19-2.00 (m, 5H), 1.99-1.88 (m, 2H), 1.31 (t, J=7.6 Hz,
2H). LCMS M/Z (M+1) 405.
Example 8 (Procedure H)
1-(3-(6-(hydroxymethyl)isoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7--
dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00159##
[0310] Step 1:
8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4-
,3-c]pyridin-3-yl)isoquinoline-6-carbaldehyde
##STR00160##
[0312] To a solution of
1-(1-(tetrahydro-2H-pyran-4-yl)-3-(6-vinylisoquinolin-8-yl)-6,7-dihydro-1-
H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (Example 6, 300 mg, 0.75
mmol) in THE (3 mL) and water (3 mL) was added osmium(VIII) oxide
(500 mg, 1.97 mmol). The mixture was stirred at room temperature
for 0.5 h. Sodium periodate (294 mg, 1.37 mmol) was added and the
mixture stirred for an additional 2 h. Water (30 mL) was added and
the mixture was extracted with EtOAc (30 mL.times.3). The combined
organic layers were dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo to give the title compound (300 mg,
crude) as a yellow solid that required no further purification.
LCMS M/Z (M+H) 405.
Step 2:
1-(3-(6-(hydroxymethyl)isoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4--
yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00161##
[0314] To a solution of
8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4-
,3-c]pyridin-3-yl)isoquinoline-6-carbaldehyde (300 mg, 0.74 mmol)
in MeOH (5 mL) at 0.degree. C. was added sodium borohydride (84 mg,
2.23 mmol). The mixture was stirred at room temperature for 1 h
under a nitrogen atmosphere. The reaction was concentrated in
vacuo. Water (30 mL) was added and extracted with EtOAc (30
mL.times.3). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo.--The crude
residue was purified by reverse phase chromatography (acetonitrile
13-43%/0.05% NH.sub.4OH in water) to give the title compound (13
mg, 4%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.70 (s, 1H), 8.51 (d, J=5.6 Hz, 1H), 7.91-7.82 (m, 2H),
7.60-7.53 (m, 1H), 5.53 (t, J=5.6 Hz, 2H), 7.75 (d, J=5.2 Hz, 1H),
4.61-4.41 (m, 1H), 4.51 (s, 2H), 4.06-3.95 (m, 2H), 3.88-3.76 (m,
2H), 3.52 (t, J=12.0 Hz, 2H), 3.03-2.79 (m, 2H), 2.19-2.00 (m, 5H),
1.99-1.95 (m, 2H). LCMS M/Z (M+1) 407.
Example 9 (Procedure I)
1-(3-(5-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-
-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00162##
[0315] Step 1: (2-chloro-5-methylphenyl)methanamine
##STR00163##
[0317] To a solution of 2-chloro-5-methyl-benzonitrile (9.0 g, 59.4
mmol) in anhydrous THE (90 mL) at 0.degree. C. was added
BH.sub.3-THF (1.0 M, 178 mL, 178 mmol) dropwise under a nitrogen
atmosphere. The mixture was stirred at room temperature for 12 h.
The reaction was quenched with 2N HCl (110 mL) at 0.degree. C. and
then heated to 70.degree. C. for 1 h. After cooling the reaction to
room temperature, the solution was washed with DCM (150 mL). The
aqueous phase was basified with 1N NaOH to pH 8 and then extracted
with DCM (150 mL.times.3). The combined organic layers were dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo
to give the title compound (5.8 g, 63%) as yellow oil. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.22 (d, J=8.0 Hz, 1H), 7.17 (s, 1H),
6.99 (d, J=8.0 Hz, 1H), 3.88 (s, 2H), 2.32 (s, 3H).
Step 2: N-(2-chloro-5-methylbenzyl)-2,2-dimethoxyacetamide
##STR00164##
[0319] To a solution of (2-chloro-5-methylphenyl)methanamine (4.6
g, 29.6 mmol) in MeOH (40 mL) was added triethylamine (5.2 mL, 37.2
mmol) and methyl dimethoxyacetate (4.4 g, 33.1 mmol). The mixture
was heated to 80.degree. C. for 16 h in sealed tube. After cooling
the reaction to room temperature, the mixture was concentrated in
vacuo. EtOAc (100 mL) was added and washed with 1N HCl (50 mL),
WATER (50 mL), brine (50 mL). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to
give the title compound (4.1 g, 54%) as colorless oil. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 8.47-8.37 (m, 1H), 7.28 (d, J=8.4
Hz, 1H), 7.09-7.05 (s, 2H), 4.74 (s, 1H), 4.31 (d, J=6.4 Hz, 2H),
3.32 (s, 6H), 2.25 (s, 3H).
Step 3: 8-chloro-5-methylisoquinolin-3(2H)-one
##STR00165##
[0321] To a solution of sulfuric acid (40 mL) at 0.degree. C. was
added N-(2-chloro-5-methylbenzyl)-2,2-dimethoxyacetamide (5.0 g,
19.4 mmol). The reaction was stirred at room temperature for 16 h.
The reaction was poured into ice water (100 mL) and the mixture was
basified with ammonium hydroxide to pH 8. The yellow precipitate
was filtered off, washed with MeOH (10 mL), Et.sub.2O (10 mL), and
dried in vacuo to give the title compound (3.8 g, crude) as a
yellow solid that required no further purification. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.07 (s, 1H), 7.37-7.30 (m, 2H),
6.93 (s, 1H), 2.48 (s, 3H).
Step 4: 8-chloro-5-methylisoquinolin-3-yl
trifluoromethanesulfonate
##STR00166##
[0323] To a solution of 8-chloro-5-methylisoquinolin-3(2H)-one (1
g, 5.16 mmol) in DCM (10 mL) at 0.degree. C. was added
triethylamine (4.3 mL, 31 mmol) and trifluoromethanesulfonic
anhydride (1.3 mL, 7.75 mmol). The reaction was stirred at room
temperature for 1 h. DCM (50 mL) was added and washed with ice
water (40 mL.times.2). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (2 g, crude) as dark brown oil that required no
further purification.
Step 5:
8-chloro-5-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline
##STR00167##
[0325] To a solution of 8-chloro-5-methylisoquinolin-3-yl
trifluoromethanesulfonate (2 g, crude) in 1,4-dioxane (10 mL) and
water (2 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (449
mg, 0.6 mmol), Na.sub.2CO.sub.3 (1.95 g, 18.4 mmol) and
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(1.4 g, 6.7 mmol). The mixture was irradiated in a microwave at
90.degree. C. for 0.5 h. EtOAc (100 mL) was added and washed with
water (60 mL.times.2), brine (60 mL). The organic layer was dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The crude residue was purified by silica gel chromatography
(petroleum ether/EtOAc=1:1) to give the title compound (770 mg) as
a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.48 (s,
1H), 8.45 (s, 1H), 8.19 (s, 1H), 8.10 (s, 1H), 7.59-7.53 (m. 2H),
3.92 (s, 3H), 2.66 (s, 3H).
Step 6:
1-(1-(tetrahydro-2H-pyran-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxa-
borolan-2-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00168##
[0327] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 200 mg, 0.6 mmol) in
1,4-dioxane (4 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (17 mg,
0.04 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (14 mg, 0.02 mmol), KOAc (179
mg, 1.8 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (309
mg, 1.2 mmol). The mixture was heated to 80.degree. C. for 2 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 376.
Step 7:
1-(3-(5-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(te-
trahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)eth-
anone
##STR00169##
[0329] To the above step cooled solution was added
8-chloro-5-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline (94 mg,
0.36 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl-
)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (19 mg, 0.02 mmol),
K.sub.3PO.sub.4 (322 mg, 1.5 mmol), 1,4-dioxane (1 mL) and water (1
mL). The reaction mixture was heated to 90.degree. C. for 1 h under
a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. DCM (50 mL) was
added and washed with water (40 mL.times.2). The organic layer was
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The crude residue was purified by reverse phase
chromatography (acetonitrile 20-50%/0.2% formic acid in water) to
give the title compound (50 mg, 17%) as a yellow solid. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 9.66-9.62 (m, 1H), 8.08-8.06 (m, 2H),
7.92-7.90 (m, 1H), 7.55-7.50 (m, 1H), 7.44-7.32 (m, 1H), 4.62-4.45
(m, 2H), 4.34-4.21 (m, 1H), 4.19-4.14 (m, 2H), 4.09-3.77 (m, 2H),
4.00 (s, 3H), 3.59-3.53 (m, 2H), 2.96-2.81 (m, 2H), 2.76-2.70 (m,
3H), 2.48-2.37 (m, 2H), 2.24-2.02 (m, 3H), 2.00-1.88 (m, 2H). LCMS
M/Z (M+H) 471.
Example 10 (Procedure J)
N-methyl-3-(5-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetr-
ahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxa-
mide
##STR00170##
[0330] Step 1: tert-butyl
1-(tetrahydro-2H-pyran-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00171##
[0332] To a solution of tert-butyl
3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
ne-5(4H)-carboxylate (Intermediate G, 400 mg, 1.04 mmol) in
1,4-dioxane (10 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (30 mg,
0.06 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (25 mg, 0.03 mmol), KOAc (300
mg, 3.12 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (526
mg, 2.08 mmol). The mixture was heated to 80.degree. C. for 2 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 434.
Step 2: tert-butyl
3-(5-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-2-
H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00172##
[0334] To the above step cooled solution was added
8-chloro-5-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline (160 mg,
0.62 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl-
)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (25 mg, 0.03 mmol),
K.sub.3PO.sub.4 (661 mg, 3.12 mmol), 1,4-dioxane (5 mL) and water
(3 mL). The reaction mixture was heated to 90.degree. C. for 12 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. DCM (20 mL) was
added and washed with water (15 mL.times.2), brine (20 mL). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by silica
gel chromatography (DCM/MeOH=20:1) to give the title compound (180
mg, 37%) as a yellow solid. LCMS M/Z (M+H) 529.
Step 3:
5-methyl-3-(1-methyl-1H-pyrazol-4-yl)-8-(1-(tetrahydro-2H-pyran-4--
yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)isoquinoline
##STR00173##
[0336] To a solution of tert-butyl
3-(5-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-2-
H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(180 mg, 0.34 mmol) in DCM (3 mL) was added trifluoroacetic acid
(0.27 mL, 3.6 mmol). The mixture was stirred at room temperature
for 1 h and concentrated in vacuo to give the title compound (100
mg, crude) as yellow oil that required no further purification.
LCMS M/Z (M+H) 429.
Step 4:
N-methyl-3-(5-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-
-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-
-carboxamide
##STR00174##
[0338] To a solution of
5-methyl-3-(1-methyl-1H-pyrazol-4-yl)-8-(1-(tetrahydro-2H-pyran-4-yl)-4,5-
,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)isoquinoline (100
mg, 0.23 mmol) in DCM (2 mL) was added triethylamine (0.1 mL, 0.70
mmol) and N-methyl-1H-imidazole-1-carboxamide (58 mg, 0.47 mmol).
The reaction was stirred at room temperature for 12 h and
concentrated in vacuo. The crude residue was purified by reverse
phase chromatography (acetonitrile 23-53%/0.05% NH.sub.4OH in
water) to give the title compound (12 mg, 10%) as a light yellow
solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.58 (s, 1H),
8.08-8.05 (m, 2H), 7.91 (s, 1H), 7.56-7.49 (m, 1H), 7.41-7.36 (m,
1H), 4.47-4.39 (m, 1H), 4.35-4.21 (m, 3H), 4.20-4.12 (m, 2H), 4.00
(s, 3H), 3.90-3.84 (m, 2H), 3.62-3.51 (m, 2H), 2.89-2.83 (m, 2H),
2.78 (d, J=4.4 Hz, 3H), 2.73 (s, 3H), 2.50-2.36 (m, 2H), 2.00-1.91
(m, 2H). LCMS M/Z (M+H) 486.
Example 11 (Procedure K)
4-(8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo-
[4,3-c]pyridin-3-yl)isoquinolin-3-yl)-1-methyl-1H-pyrazole-3-carbonitrile
##STR00175##
[0339] Step 1:
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-3-ca-
rbonitrile
##STR00176##
[0341] To a solution of 4-bromo-1-methyl-1H-pyrazole-3-carbonitrile
(150 mg, 0.81 mmol) in 1,4-dioxane (4 mL) was added
(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (38 mg, 0.08
mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2-
'-amino-1,1'-biphenyl-2-yl) palladium(II) (63 mg, 0.08 mmol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (225
mg, 0.88 mmol) and KOAc (237 mg, 2.42 mmol). The mixture was heated
to 90.degree. C. for 12 h under a nitrogen atmosphere. After
cooling the reaction to room temperature, the reaction mixture was
used to the next step directly without further purification. LCMS
M/Z (M+H) 234.
Step 2:
4-(8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H--
pyrazolo[4,3-c]pyridin-3-yl)isoquinolin-3-yl)-1-methyl-1H-pyrazole-3-carbo-
nitrile
##STR00177##
[0343] To the above step cooled solution was added
8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4-
,3-c]pyridin-3-yl)isoquinolin-3-yl trifluoromethanesulfonate
(Intermediate I, 100 mg, 0.19 mmol),
(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (9 mg, 0.02
mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2-
'-amino-1,1'-biphenyl-2-yl) palladium(II) (15 mg, 0.02 mmol) and
Na.sub.2CO.sub.3 (40 mg, 0.38 mmol), 1,4-dioxane (1 mL) and water
(1 mL). The reaction mixture was heated to 60.degree. C. for 16 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. The crude residue
was purified by reverse phase chromatography (acetonitrile
25-55%/0.2% formic acid in water) to give the title compound (25
mg, 6%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.86 (s, 1H), 8.68 (s, 1H), 8.22 (s, 1H), 7.98 (d, J=8.0
Hz, 1H), 7.91-7.83 (m, 1H), 7.71-7.60 (m, 1H), 4.55 (s, 2H),
4.52-4.42 (m, 1H), 4.07-3.97 (m, 2H), 4.03 (s, 1H), 3.88-3.78 (m,
2H), 3.52 (d, J=12.0 Hz, 2H), 3.02-2.82 (m, 2H), 2.17-2.01 (m, 5H),
2.00-1.89 (m, 2H). LCMS M/Z (M+H) 482.
Example 12 (Procedure L)
5-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,-
3-c]pyridin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)-3,4-dihydroisoquinolin-1(2H-
)-one
##STR00178##
[0344] Step 1:
1-(1-(tetrahydro-2H-pyran-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-
-2-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00179##
[0346] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 600 mg, 1.83 mmol) in
1,4-dioxane (4 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (52 mg,
0.11 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (43 mg, 0.05 mmol), KOAc (538
mg, 5.48 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (928
mg, 3.66 mmol). The mixture was heated to 80.degree. C. for 3 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 376.
Step 2:
5-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyr-
azolo[4,3-c]pyridin-3-yl)-3,4-dihydroisoquinolin-1(2H)-one
##STR00180##
[0348] To the above step cooled solution was added
5-bromo-3,4-dihydroisoquinolin-1(2H)-one (238 mg, 1.06 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (47 mg, 0.06 mmol),
K.sub.3PO.sub.4 (509 mg, 2.4 mmol), 1,4-dioxane (1 mL) and water (1
mL). The reaction mixture was heated to 90.degree. C. for 1 h under
a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. Water (100 mL) was
added and extracted with EtOAc (50 mL.times.3). The combined
organic layers were washed with brine (50 mL.times.2), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography
(DCM/MeOH=30:1) to give the title compound (250 mg, 35%) as a white
solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.15-8.11 (m, 1H),
7.46-7.39 (m, 2H), 6.29-6.22 (m, 1H), 4.58-4.42 (m, 2H), 4.26-4.10
(m, 3H), 3.97-3.80 (m, 2H), 3.61-3.45 (m, 4H), 3.20-3.09 (m, 2H),
2.86-2.80 (m, 2H), 2.38-2.27 (m, 2H), 2.20-2.11 (m, 3H), 1.94-1.85
(m, 2H). LCMS M/Z (M+H) 395.
Step 3:
5-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyr-
azolo[4,3-c]pyridin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)-3,4-dihydroisoquino-
lin-1(2H)-one
##STR00181##
[0350] To a solution of
5-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4-
,3-c]pyridin-3-yl)-3,4-dihydroisoquinolin-1(2H)-one (100 mg, 0.25
mmol) in 1,4-dioxane (10 mL) was added copper(I) iodide (2 mg, 0.01
mmol), (1R,2R)-cyclohexane-1,2-diamine (6 mg, 0.05 mmol),
K.sub.3PO.sub.4 (161 mg, 0.76 mmol) and 4-iodo-1-methyl-1H-pyrazole
(79 mg, 0.38 mmol). The reaction mixture was heated to 100.degree.
C. for 12 h under a nitrogen atmosphere. After cooling to room
temperature, the mixture was filtered and concentrated in vacuo.
Water (50 mL) was added and extracted with DCM (30 mL.times.3). The
combined organic layers were dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude residue was purified
by reverse phase chromatography (acetonitrile 26-56%/0.2% formic
acid in water) to give the title compound (37 mg, 31%) as a yellow
solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.20-8.10 (m, 2H),
7.54 (d, J=5.2 Hz, 1H), 7.49-7.36 (m, 2H), 4.61-4.45 (m, 2H),
4.27-4.11 (m, 3H), 4.02-3.78 (m, 4H), 3.93 (s, 3H), 3.61-3.50 (m,
2H), 3.36-3.28 (m, 2H), 2.90-2.77 (m, 2H), 2.40-2.27 (m, 2H),
2.20-2.11 (m, 3H), 1.95-1.86 (m, 2H). LCMS M/Z (M+H) 475.
Example 13 (Procedure M)
1-(3-(2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c]pyridin-1-yl)-
-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)--
yl)ethanone
##STR00182##
[0351] Step 1:
2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c]pyridine
##STR00183##
[0353] To a solution of 5-chloro-2-methyl-1H-pyrrolo[2,3-c]pyridine
(900 mg, 5.4 mmol) in 1,4-dioxane (50 mL) and water (10 mL) was
added 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (258
mg, 0.54 mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphe-
nyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (425 mg, 0.54
mmol), Na.sub.2CO.sub.3 (1.72 g, 16.2 mmol) and
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(1.35 g, 6.48 mmol). The mixture was heated to 100.degree. C. for
16 h under a nitrogen atmosphere. After cooling the reaction to
room temperature, the mixture was filtered and concentrated in
vacuo. The crude residue was purified by silica gel chromatography
(DCM/MeOH=10:1) to give the title compound (700 mg, 61%) as a white
solid. LCMS M/Z (M+H) 213.
Step 2:
1-(3-(2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c]pyrid-
in-1-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
n-5(4H)-yl)ethanone
##STR00184##
[0355] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 250 mg, 0.76 mmol) in
toluene (10 mL) was added copper(I) iodide (7 mg, 0.04 mmol),
K.sub.3PO.sub.4 (646 mg, 3.05 mmol),
(1R,2R)-cyclohexane-1,2-diamine (17 mg, 0.15 mmol) and
2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c]pyridine
(162 mg, 0.76 mmol). The mixture was heated to 120.degree. C. for
12 h under a nitrogen atmosphere. After cooling the reaction to
room temperature, the mixture was filtered and concentrated in
vacuo. Water (100 mL) was added and extracted with EtOAc (50
mL.times.3). The combined organic layers were washed with brine (30
mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by reverse
phase chromatography (acetonitrile 20-50%/0.05% NH.sub.4OH in
water) to give the title compound (6 mg, 2%) as a white solid.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.38-8.34 (m, 1H), 7.95
(s, 1H), 7.89-7.86 (m, 1H), 7.64-7.60 (m, 1H), 6.40-6.36 (m, 1H),
4.40-4.19 (m, 3H), 4.17-4.12 (m, 2H), 4.06-3.81 (m, 2H), 3.96 (s,
3H), 3.56 (t, J=12.0 Hz, 2H), 2.95-2.81 (m, 2H), 2.42-2.27 (m, 5H),
2.19-2.04 (m, 3H), 1.95-1.92 (m, 2H). LCMS M/Z (M+H) 460.
Examples 14 & 15 (Procedure N)
(S)-1-[8-(5-acetyl-1-tetrahydropyran-4-yl-6,7-dihydro-4H-pyrazolo[4,3-c]py-
ridin-3-yl)-3-isoquinolyl]-N-methyl-pyrrolidine-3-carboxamide
and
(R)-1-[8-(5-acetyl-1-tetrahydropyran-4-yl-6,7-dihydro-4H-pyrazolo[4,3-c]py-
ridin-3-yl)-3-isoquinolyl]-N-methyl-pyrrolidine-3-carboxamide
##STR00185##
[0357] To a microwave vial was added
[8-(5-acetyl-1-tetrahydropyran-4-yl-6,7-dihydro-4H-pyrazolo[4,3-c]pyridin-
-3-yl)-3-isoquinolyl] trifluoromethanesulfonate (Intermediate I,
40.0 mg, 0.0763 mmol), N-methylpyrrolidine-3-carboxamide (39.1 mg,
0.305 mmol), and then DMSO (1.0 mL). The mixture was heated to
130.degree. C. for 0.5 h under microwave conditions. After cooling
to room temperature, the crude mixture was purified by reverse
phase preparative HPLC (acetonitrile 5-50%/0.1% ammonium hydroxide
in water) to give racemic
1-[8-(5-acetyl-1-tetrahydropyran-4-yl-6,7-dihydro-4H-pyrazolo[4,3-c]pyrid-
in-3-yl)-3-isoquinolyl]-N-methyl-pyrrolidine-3-carboxamide (18.6
mg, 49%) as a yellow solid that was separated by chiral SFC
(Chiralpak AD 150.times.21.2 mm I.D., 5 p1n); Supercritical
CO.sub.2/MeOH (0.1% NH.sub.3H.sub.2O)=60:40 at 70 mL/min) to give
(S)-1-[8-(5-acetyl-1-tetrahydropyran-4-yl-6,7-dihydro-4H-pyrazolo[4,3-c]p-
yridin-3-yl)-3-isoquinolyl]-N-methyl-pyrrolidine-3-carboxamide (6.7
mg, first peak) and
(R)-1-[8-(5-acetyl-1-tetrahydropyran-4-yl-6,7-dihydro-4H-pyrazolo[4,3-c]p-
yridin-3-yl)-3-isoquinolyl]-N-methyl-pyrrolidine-3-carboxamide (7.4
mg, seconds peak). Absolute configuration was arbitrarily assigned
to each diastereomer. Example 14: .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.44 (t, J=0.9 Hz, 1H), 7.95 (d, J=4.8 Hz,
1H), 7.61 (dt, J=8.6, 1.0 Hz, 1H), 7.53 (ddd, J=8.7, 7.0, 2.1 Hz,
1H), 7.21-7.07 (m, 1H), 6.63 (d, J=1.1 Hz, 1H), 4.50 (s, 2H),
4.48-4.37 (m, 1H), 4.07-3.95 (m, 2H), 3.87-3.75 (m, 2H), 3.75-3.67
(m, 1H), 3.65-3.60 (m, 1H), 3.56-3.42 (m, 4H), 3.12-3.02 (m, 1H),
2.99-2.78 (m, 2H), 2.62 (d, J=4.6 Hz, 3H), 2.21-2.00 (m, 7H),
1.98-1.88 (m, 2H). LCMS M/Z (M+H) 503.3. Example 15: .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.44 (d, J=1.0 Hz, 1H), 7.95 (d,
J=4.8 Hz, 1H), 7.61 (dt, J=8.5, 1.1 Hz, 1H), 7.53 (ddd, J=8.9, 7.0,
2.1 Hz, 1H), 7.19-7.08 (m, 1H), 6.67-6.59 (m, 1H), 4.50 (s, 2H),
4.46-4.47 (m, 1H), 4.01 (dd, J=10.9, 4.8 Hz, 2H), 3.87-3.77 (m,
2H), 3.73-3.66 (m, 1H), 3.65-3.58 (m, 1H), 3.54-3.42 (m, 4H),
3.11-3.03 (m, 1H), 2.94 (t, J=5.6 Hz, 2H), 2.62 (d, J=4.6 Hz, 3H),
2.18-2.00 (m, 7H), 1.96-1.88 (m, 2H). LCMS M/Z (M+H) 503.3.
Example 16 (Procedure O)
1-(3-(isoquinolin-4-yl)-1-(tetrahydro-2H-pyran-4-yl)-1,4,6,7-tetrahydro-5H-
-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one
##STR00186##
[0359] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 35 mg, 0.11 mmol) and
4-isoquinolylboronic acid (37 mg, 0.21 mmol) in dioxane (0.7 mL)
and water (0.2 mL) was added K.sub.3PO.sub.4.H.sub.2O (63 mg, 0.27
mmol),
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-
-1,1'-biphenyl)]palladium(II) methanesulfonate (4.8 mg, 0.0053
mmol) and 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl
(2.6 mg, 0.0053 mmol). The mixture was stirred at 100.degree. C.
for 5 h under a nitrogen atmosphere. The reaction mixture was then
cooled to room temperature and concentrated in vacuo. The crude
residue was dissolved in dichloromethane (5 mL), dried over
anhydrous MgSO.sub.4, filtered through celite and concentrated in
vacuo. The mixture obtained was purified by reverse phase
chromatography (acetonitrile 5-50%/0.1% ammonium hydroxide in
water) to give the title compound (38.2 mg, 90%) as a white solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.32 (s, 1H), 8.50-8.42
(m, 2H), 8.20 (d, J=8.0 Hz, 1H), 7.89-7.80 (m, 1H), 7.77-7.71 (m,
1H), 4.56-4.49 (m, 2H), 4.49-4.38 (m, 1H), 4.06-3.97 (m, 2H),
3.88-3.77 (m, 2H), 3.58-3.46 (m, 2H), 3.00-2.78 (m, 2H), 2.20-2.05
(m, 4H), 2.01 (s, 1H), 1.98-1.88 (m, 2H). LCMS M/Z (M+H) 377.
Example 17 (Procedure P)
5-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,-
3-c]pyridin-3-yl)-2-methyl-1,4-dihydroisoquinolin-3(2H)-one
##STR00187##
[0361] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 80 mg, 0.24 mmol) and
bis(pinacolato)diboron (124 mg, 0.488 mmol) in dioxane (1.2 mL) was
added KOAc (72 mg, 0.73 mmol),
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-
-1,1'-biphenyl)]palladium(II) methanesulfonate (5.9 mg, 0.0073
mmol) and 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl
(7.1 mg, 0.015 mmol). The mixture was stirred at 80.degree. C. for
16 h under a nitrogen atmosphere. The reaction mixture was cooled
to room temperature and
5-chloro-2-methyl-1,4-dihydroisoquinolin-3-one (26 mg, 0.13 mmol),
K.sub.3PO.sub.4.H.sub.2O (78 mg, 0.33 mmol), water (0.3 mL) and
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-
-1,1'-biphenyl)]palladium(II) methanesulfonate (5.9 mg, 0.0073
mmol) were added. The reaction mixture was stirred at 90.degree. C.
for 4 h under a nitrogen atmosphere. The reaction mixture was then
cooled to room temperature and concentrated in vacuo. The crude
residue was dissolved in dichloromethane (5 mL), dried over
anhydrous MgSO.sub.4, filtered through celite and concentrated in
vacuo. The mixture obtained was purified by SFC (Supercritical
CO.sub.2 5-40%/MeOH (0.1% NH.sub.3H.sub.2O)) to give the title
compound (28.3 mg, 28%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.37-7.19 (m, 3H), 4.55 (s, 2H), 4.42 (s,
2H), 4.40-4.31 (m, 1H), 4.02-3.94 (m, 2H), 3.78 (dt, J=14.7, 5.8
Hz, 2H), 3.65 (d, J=3.9 Hz, 2H), 3.54-3.43 (m, 2H), 2.96 (s, 3H),
2.93-2.73 (m, 2H), 2.13-1.98 (m, 5H), 1.87 (dd, J=12.7, 5.2 Hz,
2H). LCMS M/Z (M+H) 409.
Example 18 (Procedure Q)
7-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,-
3-c]pyridin-3-yl)-2-(4-methoxyphenyl)isoindolin-1-one
##STR00188##
[0362] Step 1:
7-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4-
,3-c]pyridin-3-yl)isoindolin-1-one
##STR00189##
[0364] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 350 mg, 1.07 mmol) and
bis(pinacolato)diboron (542 mg, 2.13 mmol) in dioxane (5.3 mL) was
added KOAc (314 mg, 3.20 mmol),
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-
-1,1'-biphenyl)]palladium(II) methanesulfonate (26 mg, 0.032 mmol)
and 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (31 mg,
0.064 mmol). The mixture was stirred at 80.degree. C. for 16 h
under a nitrogen atmosphere. The reaction mixture was cooled to
room temperature and 7-bromoisoindolin-1-one (113 mg, 0.533 mmol),
K.sub.3PO.sub.4.H.sub.2O (313 mg, 1.33 mmol), water (1.3 mL) and
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-
-1,1'-biphenyl)]palladium(II) methanesulfonate (26 mg, 0.032 mmol)
were added. The reaction mixture was stirred at 90.degree. C. for 4
h under a nitrogen atmosphere. The reaction mixture was then cooled
to room temperature and concentrated in vacuo. The crude residue
was dissolved in dichloromethane (20 mL), dried over anhydrous
MgSO.sub.4, filtered through celite and concentrated in vacuo. The
mixture obtained was purified by silica gel chromatography
(MeOH/iPrOAc=1:10 to 1:3) to give the title compound (144 mg, 36%)
as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6, 23/24 H)
.delta. 7.66-7.40 (m, 3H), 6.06-5.94 (m, 1H), 4.48 (d, J=9.5 Hz,
2H), 4.62-4.41 (m, 1H), 4.18 (dd, J=36.7, 11.6 Hz, 2H), 3.99-3.77
(m, 2H), 3.59-3.47 (m, 3H), 2.82 (dt, J=24.3, 5.9 Hz, 2H),
2.43-2.30 (m, 2H), 2.17-2.06 (m, 3H), 1.93 (d, J=13.2 Hz, 2H). LCMS
M/Z (M+H) 381.
Step 2:
7-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyr-
azolo[4,3-c]pyridin-3-yl)-2-(4-methoxyphenyl)isoindolin-1-one
##STR00190##
[0366] To a solution of
7-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4-
,3-c]pyridin-3-yl)isoindolin-1-one (30 mg, 0.079 mmol) in dioxane
(0.46 mL) was added 4-bromoanisole (14 mg, 0.075 mmol),
N,N'-dimethylethylenediame (1.0 .mu.L, 0.0095 mmol), copper(I)
iodide (1.8 mg, 0.0095 mmol) and K.sub.2CO.sub.3 (44 mg, 0.32
mmol). The mixture was stirred at 101.degree. C. for 24 h under a
nitrogen atmosphere. The reaction mixture was then cooled to room
temperature, diluted with dichloromethane (5 mL), filtered through
celite and concentrated in vacuo. The mixture obtained was purified
by reverse phase chromatography (acetonitrile 20-60%/0.1% formic
acid in water) to give the title compound (32.1 mg, 84%) as a white
solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.80-7.70 (m,
2H), 7.72-7.61 (m, 2H), 7.43 (ddd, J=11.2, 6.7, 1.9 Hz, 1H),
7.04-6.96 (m, 2H), 5.00 (s, 2H), 4.40 (s, 1H), 4.38-4.31 (m, 2H),
4.02-3.94 (m, 2H), 3.77 (s, 3H), 3.73 (t, J=6.1 Hz, 2H), 3.48 (dd,
J=13.7, 10.4 Hz, 2H), 2.94-2.72 (m, 2H), 2.16-2.03 (m, 4H), 1.98
(s, 1H), 1.88 (d, J=12.9 Hz, 2H). LCMS M/Z (M+H) 487.
Example 19 (Procedure R)
1-(3-(6-(4-methoxyphenyl)-1H-indol-1-yl)-1-(tetrahydro-2H-pyran-4-yl)-1,4,-
6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one
##STR00191##
[0367] Step 1: 6-(4-methoxyphenyl)-1H-indole
##STR00192##
[0369] To a solution of 6-bromo-1H-indole (150 mg, 0.765 mmol) and
(4-methoxyphenyl)boronic acid (151 mg, 0.995 mmol) in dioxane (2.6
mL) and water (0.6 mL) was added K.sub.3PO.sub.4.H.sub.2O (545 mg,
2.30 mmol),
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2-
'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate (25 mg, 0.031
mmol) and 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl
(15 mg, 0.031 mmol). The reaction mixture was stirred at 85.degree.
C. for 16 h under a nitrogen atmosphere. The reaction mixture was
then cooled to room temperature and concentrated in vacuo. The
crude residue was dissolved in dichloromethane (10 mL), dried over
anhydrous MgSO.sub.4, filtered through celite and concentrated in
vacuo. The mixture obtained was purified by silica gel
chromatography (iPrOAc/Heptane=1:19 to 1:9) to give the title
compound (134 mg, 78%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.17 (s, 1H), 7.67 (d, J=8.2 Hz, 1H),
7.62-7.48 (m, 3H), 7.34 (dd, J=8.2, 1.6 Hz, 1H), 7.22 (dd, J=3.2,
2.4 Hz, 1H), 7.03-6.94 (m, 2H), 6.56 (ddd, J=3.2, 2.1, 1.0 Hz, 1H),
3.86 (s, 3H). LCMS M/Z (M+H) 224.
Step 2:
1-(3-(6-(4-methoxyphenyl)-1H-indol-1-yl)-1-(tetrahydro-2H-pyran-4--
yl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one
##STR00193##
[0371] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 50 mg, 0.15 mmol) in
toluene (0.76 mL) was added 6-(4-methoxyphenyl)-1H-indole (51 mg,
0.23 mmol), K.sub.3PO.sub.4 (77 mg, 0.35 mmol), copper(I) iodide
(29 mg, 0.15 mmol) and (trans)-1,2-diaminocyclohexane (18 mg, 0.15
mmol). The mixture was stirred at 110.degree. C. for 16 h under a
nitrogen atmosphere. The reaction mixture was then cooled to room
temperature, diluted with dichloromethane (5 mL), filtered through
celite and concentrated in vacuo. The mixture obtained was purified
by reverse phase chromatography (acetonitrile 30-70%/0.1% ammonium
hydroxide in water) to give the title compound (49.6 mg, 69%) as a
white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.05-7.90
(m, 1H), 7.69 (dd, J=8.2, 2.1 Hz, 1H), 7.62-7.54 (m, 2H), 7.58-7.47
(m, 1H), 7.40 (dd, J=8.2, 1.6 Hz, 1H), 7.08-6.98 (m, 2H), 6.73-6.67
(m, 1H), 4.52 (d, J=11.4 Hz, 2H), 4.41 (tt, J=11.0, 4.5 Hz, 1H),
4.00 (ddd, J=9.3, 4.8, 2.5 Hz, 2H), 3.86-3.77 (m, 5H), 3.50 (td,
J=12.0, 2.4 Hz, 2H), 2.95-2.75 (m, 2H), 2.12 (s, 2H), 2.10-1.99 (m,
3H), 1.93 (dd, J=12.8, 4.2 Hz, 2H). LCMS M/Z (M+H) 471.
Example 20 (Procedure S)
1-(3-(2-(2,4-dimethylphenyl)imidazo[1,2-a]pyridin-8-yl)-1-(tetrahydro-2H--
pyran-4-yl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one
##STR00194##
[0372] Step
1:8-bromo-2-(2,4-dimethylphenyl)imidazo[1,2-a]pyridine
##STR00195##
[0374] To a solution of 3-bromopyridin-2-amine (200 mg, 1.16 mmol)
in 2-propanol (2.3 mL) was added
2-bromo-1-(2,4-dimethylphenyl)ethanone (315 mg, 1.39 mmol). The
mixture was stirred at 75.degree. C. for 72 h. The reaction mixture
was then cooled to room temperature and concentrated in vacuo. The
crude residue was partitioned between DCM (50 mL) and saturated
aqueous NaHCO.sub.3 solution (50 mL) and the two phases were
separated. The aqueous layer was extracted with DCM (2.times.50
mL). The combined organic layers were dried over anhydrous
MgSO.sub.4, filtered and concentrated in vacuo. The mixture
obtained was purified by silica gel chromatography
(iPrOAc/Heptane=1:9) to give the title compound (242 mg, 70%) as a
pale yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.12
(dd, J=6.7, 1.1 Hz, 1H), 7.84-7.77 (m, 1H), 7.74 (s, 1H), 7.43 (dd,
J=7.3, 1.0 Hz, 1H), 7.13-7.06 (m, 2H), 6.66 (dd, J=7.3, 6.7 Hz,
1H), 2.52 (s, 3H), 2.36 (s, 3H). LCMS M/Z (M+H) 301.
Step 2:
1-(3-(2-(2,4-dimethylphenyl)imidazo[1,2-a]pyridin-8-yl)-1-(tetrahy-
dro-2H-pyran-4-yl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-
-1-one
##STR00196##
[0376] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 50 mg, 0.15 mmol) and
bis(pinacolato)diboron (77 mg, 0.30 mmol) in dioxane (0.76 mL) was
added KOAc (45 mg, 0.46 mmol),
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-
-1,1'-biphenyl)]palladium(II) methanesulfonate (3.7 mg, 0.0046
mmol) and 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl
(4.4 mg, 0.0091 mmol). The mixture was stirred at 80.degree. C. for
16 h under a nitrogen atmosphere. The reaction mixture was cooled
to room temperature and
8-bromo-2-(2,4-dimethylphenyl)imidazo[1,2-a]pyridine (23 mg, 0.076
mmol), K.sub.3PO.sub.4.H.sub.2O (38 mg, 0.17 mmol), water (0.3 mL)
and
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-
-1,1'-biphenyl)]palladium(II) methanesulfonate (3.7 mg, 0.0046
mmol) were added. The reaction mixture was stirred at 90.degree. C.
for 4 h under a nitrogen atmosphere. The reaction mixture was then
cooled to room temperature and concentrated in vacuo. The crude
residue was dissolved in dichloromethane (5 mL), dried over
anhydrous MgSO.sub.4, filtered through celite and concentrated in
vacuo. The mixture obtained was purified by reverse phase
chromatography (acetonitrile 20-60%/0.1% ammonium hydroxide in
water) to give the title compound (21.3 mg, 30%) as a white solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.58 (ddd, J=6.7, 3.2,
1.2 Hz, 1H), 8.25 (s, 1H), 8.06-7.81 (m, 1H), 7.50-7.36 (m, 1H),
7.14-7.05 (m, 2H), 6.97 (q, J=7.0 Hz, 1H), 4.87 (s, 1H), 4.76 (s,
1H), 4.39 (tq, J=10.7, 4.6 Hz, 1H), 4.00 (dd, J=10.3, 4.6 Hz, 2H),
3.79 (dt, J=8.9, 5.7 Hz, 2H), 3.50 (tt, J=11.8, 1.8 Hz, 2H),
2.95-2.75 (m, 2H), 2.52 (s, 3H), 2.31 (s, 3H), 2.19-2.03 (m, 4H),
1.96-1.84 (m, 3H). LCMS M/Z (M+H) 470.
Example 21 (Procedure T)
1-(3-(2-(4-methoxyphenyl)benzo[d]oxazol-4-yl)-1-(tetrahydro-2H-pyran-4-yl)-
-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one
##STR00197##
[0377] Step 1: 4-bromo-2-(4-methoxyphenyl)benzo[d]oxazole
##STR00198##
[0379] To a solution of 3-bromo-2-nitro-phenol (120 mg, 0.550 mmol)
in toluene (1.4 mL) was added (4-methoxyphenyl)methanol (190 mg,
1.38 mmol) and 1,1'-bis(diphenylphosphino)ferrocene (9.2 mg, 0.016
mmol). The mixture was stirred at 150.degree. C. for 24 h in a
sealed vial under a nitrogen atmosphere. The reaction mixture was
then cooled to room temperature, diluted with dichloromethane (10
mL), filtered through celite and concentrated in vacuo. The mixture
obtained was purified by silica gel chromatography
(acetone/heptane=1:19 to 1:9) to give the title compound (69.2 mg,
41%) as a pink solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
8.29-8.20 (m, 2H), 7.50 (d, J=8.0 Hz, 2H), 7.19 (t, J=8.0 Hz, 1H),
7.07-6.98 (m, 2H), 3.90 (s, 3H). LCMS M/Z (M+H) 305.
Step 2:
1-(3-(2-(4-methoxyphenyl)benzo[d]oxazol-4-yl)-1-(tetrahydro-2H-pyr-
an-4-yl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one
##STR00199##
[0381] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 50 mg, 0.15 mmol) and
bis(pinacolato)diboron (77 mg, 0.30 mmol) in dioxane (0.76 mL) was
added KOAc (45 mg, 0.46 mmol),
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-
-1,1'-biphenyl)]palladium(II) methanesulfonate (3.7 mg, 0.0046
mmol) and 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl
(4.4 mg, 0.0091 mmol). The mixture was stirred at 80.degree. C. for
16 h under a nitrogen atmosphere. The reaction mixture was cooled
to room temperature and 4-bromo-2-(4-methoxyphenyl)-1,3-benzoxazole
(27 mg, 0.088 mmol), K.sub.3PO.sub.4.H.sub.2O (47 mg, 0.20 mmol),
water (0.3 mL) and
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-
-1,1'-biphenyl)]palladium(II) methanesulfonate (3.7 mg, 0.0046
mmol) were added. The reaction mixture was stirred at 90.degree. C.
for 4 h under a nitrogen atmosphere. The reaction mixture was then
cooled to room temperature and concentrated in vacuo. The crude
residue was dissolved in dichloromethane (5 mL), dried over
anhydrous MgSO.sub.4, filtered through celite and concentrated in
vacuo. The mixture obtained was purified by reverse phase
chromatography (acetonitrile 30-70%/0.1% formic acid in water) to
give the title compound (28.9 mg, 40%) as a white solid. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 8.29-8.15 (m, 2H), 7.73 (ddd,
J=8.1, 4.8, 1.1 Hz, 1H), 7.63 (td, J=7.9, 1.0 Hz, 1H), 7.44 (td,
J=8.0, 4.4 Hz, 1H), 7.22-7.12 (m, 2H), 4.86 (d, J=10.1 Hz, 2H),
4.40 (td, J=11.2, 4.5 Hz, 1H), 4.05-3.96 (m, 2H), 3.88 (d, J=1.3
Hz, 3H), 3.83 (dt, J=12.0, 5.7 Hz, 2H), 3.50 (tt, J=11.5, 1.8 Hz,
2H), 2.95-2.75 (m, 2H), 2.20-2.03 (m, 5H), 1.94-1.85 (m, 2H). LCMS
M/Z (M+H) 473.
Example 22 (Procedure U)
3-(5-acetyl-1-tetrahydrofuran-3-yl-6,7-dihydro-4H-pyrazolo[4,3-c]pyridin-3-
-yl)-N-(3-cyanophenyl)benzamide
##STR00200##
[0382] Step 1: methyl
3-(5-acetyl-1-tetrahydrofuran-3-yl-6,7-dihydro-4H-pyrazolo[4,3-c]pyridin--
3-yl)benzoate
##STR00201##
[0384] A solution of
1-(3-bromo-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-
-5(4H)-yl)ethanone (Intermediate F, 2.3 g, 7.3 mmol) and
(3-(methoxycarbonyl)phenyl)boronic acid (2.0 g, 11 mmol) in dioxane
(20 mL) was charged with aqueous sodium carbonate solution (1.0 M,
20 mL) and heated at 120.degree. C. for 20 minutes. The mixture was
then diluted with ethyl acetate and water and the layers were
partitioned and separated. The organic layer was washed once with
water, dried over anhydrous magnesium sulfate, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (dichloromethane/methanol=100:1 to 10:1) to give the
title compound (2.6 g, 7.0 mmol, 96% yield). LCMS M/Z (M+H)
370.
Step 2:
3-(5-acetyl-1-tetrahydrofuran-3-yl-6,7-dihydro-4H-pyrazolo[4,3-c]p-
yridin-3-yl)benzoic Acid
##STR00202##
[0386] A solution of methyl
3-(5-acetyl-1-tetrahydrofuran-3-yl-6,7-dihydro-4H-pyrazolo[4,3-c]pyridin--
3-yl)benzoate (3.2 g, 8.7 mmol) in THE (20 mL) and MeOH (10 mL) was
charged with aqueous lithium hydroxide solution (2.0 M, 10 mL) and
stirred at room temperature overnight. The mixture was then
concentrated in vacuo to remove the organic solvents. The pH of the
resulting aqueous mixture was adjusted to pH=4 by titration with
aqueous hydrochloric acid solution (1.0 M). The precipitate was
then collected by filtration, washed with water, and dried under
vacuum to afford the title compound (2.9 g, 8.2 mmol, 92% yield).
LCMS M/Z (M+H) 356.
Step 3:
3-(5-acetyl-1-tetrahydrofuran-3-yl-6,7-dihydro-4H-pyrazolo[4,3-c]p-
yridin-3-yl)-N-(3-cyanophenyl)benzamide
##STR00203##
[0388] A solution of
3-(5-acetyl-1-tetrahydrofuran-3-yl-6,7-dihydro-4H-pyrazolo[4,3-c]pyridin--
3-yl)benzoic acid (20 mg, 0.056 mmol) in DMF (0.5 mL) was charged
with
1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium
3-oxide hexafluorophosphate (44 mg, 0.11 mmol) and
diisopropylethylamine (15 mg, 0.11 mmol), then stirred at room
temperature for 5 min. To the mixture was added 3-aminobenzonitrile
(13 mg, 0.11 mmol). After stirring at room temperature for an
additional 1 h, the reaction mixture was purified by reverse phase
preparative HPLC (acetonitrile 5-50%/0.1% ammonium hydroxide in
water) to give the title compound (23 mug, 0.055 mmol, 90%).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.65 (s, 1H), 8.27
(ddd, J=2.8, 2.3, 1.1 Hz, 1H), 8.21-8.13 (m, 1H), 8.11-8.02 (m,
1H), 7.95-7.87 (m, 1H), 7.86-7.79 (m, 1H), 7.68-7.55 (m, 3H),
5.07-4.93 (m, 1H), 4.71 (d, J=2.6 Hz, 2H), 4.16-3.99 (m, 2H),
3.97-3.71 (m, 4H), 2.95-2.84 (m, 1H), 2.77 (d, J=6.0 Hz, 1H),
2.42-2.30 (m, 2H), 2.17-2.00 (m, 3H). LCMS M/Z (M+H) 456.2.
Example 23 (Procedure V)
5-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,-
3-c]pyridin-3-yl)-2-cyclopentyl-1,2-dihydroisoquinolin-3(4H)-one
##STR00204##
[0389] Step 1: 2-(2-chlorophenyl)-N-cyclopentylacetamide
##STR00205##
[0391] To a solution of 2-(2-chlorophenyl)acetic acid (17.5 g,
102.59 mmol) in DMF (100 mL) was added HATU (58.51 g, 153.88 mmol),
N,N-diisopropylethylamine (50.86 mL, 307.76 mmol) and
cyclopentylamine (12.06 mL, 123.1 mmol). The reaction mixture was
stirred at room temperature for 16 h under a nitrogen atmosphere.
The reaction solution was poured into water (200 mL) and extracted
with EtOAc (150 mL.times.3). The combined organic layers were
washed with brine (200 mL.times.2), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (petroleum
ether/EtOAc=3:1) to give the title compound (20 g, 82%) as a white
solid.
Step 2: 5-chloro-2-cyclopentyl-1,2-dihydroisoquinolin-3(4H)-one
##STR00206##
[0393] A mixture of 2-(2-chlorophenyl)-N-cyclopentylacetamide (3 g,
12.62 mmol), paraformaldehyde (1.89 g, 63.1 mmol), methanesulfonic
acid (25 mL, 384.98 mmol) and phosphorus pentoxide (3.0 g, 21.14
mmol) was heated to 80.degree. C. for 2 h. The reaction mixture was
poured into ice water (150 mL), neutralized with Na.sub.2CO.sub.3,
and then extracted with EtOAc (100 mL.times.3). The combined
organic layers were washed with brine (100 mL.times.3), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography (petroleum
ether/EtOAc=4:1) to give the title compound (600 mg, 19%) as a
green solid. LCMS M/Z (M+H) 250.
Step 3:
1-(1-(tetrahydro-2H-pyran-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxa-
borolan-2-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00207##
[0395] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 300 mg, 0.91 mmol) in
1,4-dioxane (4 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (26 mg,
0.05 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (22 mg, 0.03 mmol), KOAc (269
mg, 2.74 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (464
mg, 1.83 mmol). The mixture was heated to 80.degree. C. for 3 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 376.
Step 4:
5-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyr-
azolo[4,3-c]pyridin-3-yl)-2-cyclopentyl-1,2-dihydroisoquinolin-3(4H)-one
##STR00208##
[0397] To the above step cooled solution was added
5-chloro-2-cyclopentyl-1,2-dihydroisoquinolin-3(4H)-one (113 mg,
0.45 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl-
)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (23 mg, 0.03 mmol),
K.sub.3PO.sub.4 (240 mg, 1.13 mmol), 1,4-dioxane (1 mL) and water
(1 mL). The reaction mixture was heated to 90.degree. C. for 1 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. Water (100 mL) was
added and extracted with EtOAc (50 mL.times.3). The combined
organic layers were washed with brine (50 mL.times.2), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by reverse phase chromatography
(acetonitrile 30-60%/0.05% NH.sub.4OH in water) to give the title
compound (86 mg, 20%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.40 (d, J=8.0 Hz, 1H), 7.34-7.29 (m, 1H),
7.24 (d, J=6.8 Hz, 1H), 4.88-4.78 (m, 1H), 4.47-4.31 (m, 5H),
4.04-3.95 (m, 2H), 3.83-3.73 (m, 2H), 3.71-3.65 (m, 2H), 3.49 (t,
J=12.0 Hz, 2H), 2.94-2.75 (m, 2H), 2.12-2.01 (m, 5H), 1.92-1.83 (m,
2H), 1.80-1.64 (m, 4H), 1.62-1.52 (m, 4H). LCMS M/Z (M+H) 463.
Example 24
5-(8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo-
[4,3-c]pyridin-3-yl)-4-fluoroisoquinolin-3-yl)-N-methylpicolinamide
##STR00209##
[0398] Step 1: 8-chloro-4-fluoroisoquinolin-3(2H1)-one
##STR00210##
[0400] To a solution of 8-chloroisoquinolin-3(2H)-one (600 mg, 3.34
mmol) in THE (10 mL) was added
1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane
bis(tetrafluoroborate) (1.36 g, 3.84 mmol). The reaction was
stirred at room temperature for 16 h. DCM (80 mL) was added and
washed with water (20 mL.times.2). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to
give the title compound (650 mg, crude) as a yellow solid that
required no further purification. LCMS M/Z (M+H) 198.
Step 2: 8-chloro-4-fluoroisoquinolin-3-yl
trifluoromethanesulfonate
##STR00211##
[0402] To a solution of 8-chloro-4-fluoroisoquinolin-3(2H)-one (200
mg, 1.01 mmol) in DCM (4 mL) at 0.degree. C. was added
triethylamine (0.42 mL, 3.04 mmol) and trifluoromethanesulfonic
anhydride (0.42 mL, 2.53 mmol). The reaction was stirred at room
temperature for 12 h and concentrated in vacuo. The crude residue
was purified by silica gel chromatography (petroleum
ether/EtOAc=10:1) to give the title compound (100 mg, 30%) as a
yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.34 (s,
1H), 8.28 (d, J=8.4 Hz, 1H), 8.13-8.07 (m, 1H), 8.06-8.00 (m, 1H).
LCMS M/Z (M+H) 330.
Step 3:
5-(8-chloro-4-fluoroisoquinolin-3-yl)-N-methylpicolinamide
##STR00212##
[0404] To a solution of 8-chloro-4-fluoroisoquinolin-3-yl
trifluoromethanesulfonate (500 mg, 1.52 mmol) in 1,4-dioxane (4 mL)
and water (1 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (111
mg, 0.15 mmol), Na.sub.2CO.sub.3 (482 mg, 4.55 mmol) and
N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamide
(398 mg, 1.52 mmol). The mixture was irradiated in a microwave at
70.degree. C. for 0.5 h. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
The crude residue was purified by silica gel chromatography
(petroleum ether/EtOAc=3:1) to give the title compound (300 mg,
63%) as a yellow solid. LCMS M/Z (M+H) 316.
Step 4:
5-(4-fluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquin-
olin-3-yl)-N-methylpicolinamide
##STR00213##
[0406] To a solution of
5-(8-chloro-4-fluoroisoquinolin-3-yl)-N-methylpicolinamide (150 mg,
0.48 mmol) in 1,4-dioxane (3 mL) was added
(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (23 mg, 0.05
mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2-
'-amino-1,1'-biphenyl-2-yl) palladium(II) (37 mg, 0.05 mmol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (181
mg, 0.71 mmol) and KOAc (93 mg, 0.95 mmol). The mixture was heated
to 80.degree. C. for 1 h under a nitrogen atmosphere. After cooling
the reaction to room temperature, the reaction mixture was used to
the next step directly without further purification. LCMS M/Z (M+H)
408.
Step 5:
5-(8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H--
pyrazolo[4,3-c]pyridin-3-yl)-4-fluoroisoquinolin-3-yl)-N-methylpicolinamid-
e
##STR00214##
[0408] To the above step cooled solution was added
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 118 mg, 0.36 mmol),
(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (17 mg, 0.04
mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2-
'-amino-1,1'-biphenyl-2-yl) palladium(II) (28 mg, 0.04 mmol) and
Na.sub.2CO.sub.3 (76 mg, 0.72 mmol), 1,4-dioxane (2 mL) and water
(1 mL). The reaction mixture was heated to 60.degree. C. for 16 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. The crude residue
was purified by reverse phase chromatography (acetonitrile
29-59%/0.2% formic acid in water) to give the title compound (28
mg, 11%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.89 (s, 1H), 9.29 (s, 1H), 8.91 (d, J=4.8 Hz, 1H), 8.65
(d, J=8.0 Hz, 1H), 8.27-8.18 (m, 2H), 8.10-7.99 (m, 1H), 7.90-7.76
(m, 1H), 4.60 (s, 2H), 4.54-4.40 (m, 1H), 4.09-3.95 (m, 2H),
3.92-3.75 (m, 2H), 3.53 (t, J=11.6 Hz, 2H), 3.03-2.79 (m, 2H), 2.87
(d, J=4.4 Hz, 3H), 2.17-2.03 (m, 5H), 2.02-1.90 (m, 2H). LCMS M/Z
(M+H) 529.
Example 25
3-(6-(difluoromethyl)-7-(1-methyl-1H-pyrazol-4-yl)isoquinolin-4-yl)-N-meth-
yl-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4-
H)-carboxamide
##STR00215##
[0409] Step 1:
N-(3-bromo-4-chlorobenzylidene)-2,2-dimethoxyethanamine
##STR00216##
[0411] To a solution of 3-bromo-4-chlorobenzaldehyde (18.0 g, 82.02
mmol) in toluene (200 mL) was added 2,2-dimethoxyethanamine (10.35
g, 98.42 mmol). The mixture was heated to 110.degree. C. for 16 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was concentrated in vacuo to give the
title compound (25.0 g, crude) as yellow oil that required no
further purification.
Step 2: N-(3-bromo-4-chlorobenzyl)-2,2-dimethoxyethanamine
##STR00217##
[0413] To a solution of
N-(2-bromo-4-chlorobenzylidene)-2,2-dimethoxyethanamine (25.0 g,
81.54 mmol) in MeOH (150 mL) at 0.degree. C. was added sodium
borohydride (2.47 g, 65.24 mmol) portionwise. The mixture was
stirred at 28.degree. C. for 2 h under a nitrogen atmosphere and
concentrated in vacuo. Water (200 mL) was added and extracted with
DCM (200 mL.times.3). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to
give the title compound (25.0 g, crude) as colorless oil that
required no further purification. LCMS M/Z (M+H) 308.
Step 3:
N-(3-bromo-4-chlorobenzyl)-N-(2,2-dimethoxyethyl)-4-methylbenzenes-
ulfonamide
##STR00218##
[0415] To a solution of
N-(3-bromo-4-chlorobenzyl)-2,2-dimethoxyethanamine (27.8 g, 90.08
mmol) in DCM (200 mL) at 0.degree. C. was added
4-methylbenzene-1-sulfonyl chloride (17.17 g, 90.08 mmol),
N,N-dimethylpyridin-4-amine (550 mg, 4.5 mmol) and triethylamine
(24.97 mL, 180.17 mmol). The mixture was stirred at 28.degree. C.
for 16 h under a nitrogen atmosphere. Water (150 mL) was added and
extracted with DCM (150 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (petroleum ether/EtOAc=20:1) to give the title
compound (35 g, 84%) as colorless oil. LCMS M/Z (M+H) 462.
Step 4: 7-bromo-6-chloroisoquinoline
##STR00219##
[0417] To a solution of aluminum trichloride (44.66 g, 334.93 mmol)
in 1,2-dichloroethane (250 mL) at 0.degree. C. was added
N-(3-bromo-4-chlorobenzyl)-N-(2,2-dimethoxyethyl)-4-methylbenzenesulfonam-
ide (31.0 g, 66.99 mmol) in 1,2-dichloroethane (250 mL) dropwise.
The mixture was stirred at 28.degree. C. for 16 h under a nitrogen
atmosphere. The mixture was quenched with ice-water (130 mL) and
extracted with DCM (130 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (petroleum ether/EtOAc=10:1) to give the title
compound (11.0 g, 68%) as a white solid. LCMS M/Z (M+H) 242.
Step 5: 6-chloro-7-(1-methyl-1H-pyrazol-4-yl)isoquinoline
##STR00220##
[0419] To a solution of 7-bromo-6-chloroisoquinoline (12.1 g, 49.9
mmol) in 1,4-dioxane (100 mL) and water (10 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (3.65
g, 4.99 mmol), Na.sub.2CO.sub.3 (13.22 g, 124.74 mmol) and
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(15.57 g, 74.85 mmol). The mixture was heated to 70.degree. C. for
3 h under a nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
The crude residue was purified by silica gel chromatography
(petroleum ether/EtOAc=1:1) to give the title compound (11.5 g,
95%) as a brown solid. LCMS M/Z (M+H) 244.
Step 6: 7-(1-methyl-1H-pyrazol-4-yl)isoquinoline-6-carbaldehyde
##STR00221##
[0421] To a solution of
6-chloro-7-(1-methyl-1H-pyrazol-4-yl)isoquinoline (2.0 g, 8.21
mmol) in DMSO (15 mL) was added palladium(II) acetate (92 mg, 0.41
mmol), 1,2-bis(diphenylphosphino)ethane (327 mg, 0.82 mmol),
potassium formate (1.38 g, 16.41 mmol) and tert-butyl isocyanide
(1.11 mL, 9.85 mmol). The mixture was heated to 120.degree. C. for
6 h under a nitrogen atmosphere. After cooling the reaction to room
temperature, sat. aq. NaHCO.sub.3 (50 mL) was added and the mixture
stirred for an additional 30 min. The solution was extracted with
DCM (50 mL.times.3). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography (petroleum
ether/EtOAc=1:1) to give the title compound (1.0 g, 51%) as a white
solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.31 (s, 1H),
9.41 (s, 1H), 8.57 (d, J=6.0 Hz, 1H), 8.51 (s, 1H), 8.26 (s, 1H),
8.13 (s, 1H), 8.04 (d, J=5.6 Hz, 1H), 7.82 (s, 1H), 3.94 (s, 3H).
LCMS M/Z (M+H) 238.
Step 7:
6-(difluoromethyl)-7-(1-methyl-1H-pyrazol-4-yl)isoquinoline
##STR00222##
[0423] To a solution of
7-(1-methyl-1H-pyrazol-4-yl)isoquinoline-6-carbaldehyde (450 mg,
1.9 mmol) in DCM (10 mL) at 0.degree. C. was added
diethylaminosulfurtrifluoride (0.75 mL, 5.69 mmol). The mixture was
stirred at room temperature for 21 h. The mixture was poured into
sat. aq. NaHCO.sub.3 (10 mL) at 0.degree. C. and extracted with
EtOAc (20 mL.times.3). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography (petroleum
ether/EtOAc=2:3) to give the title compound (160 mg, 33%) as yellow
oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.31 (s, 1H), 8.60
(d, J=6.0 Hz, 1H), 8.25 (s, 1H), 7.99 (s, 1H), 7.76 (d, J=5.6 Hz,
1H), 7.72 (s, 1H), 7.63 (s, 1H), 6.79 (d, J=54.8 Hz, 1H), 4.03 (s,
3H).
Step 8:
4-bromo-6-(difluoromethyl)-7-(1-methyl-1H-pyrazol-4-yl)isoquinolin-
e
##STR00223##
[0425] To a solution of
6-(difluoromethyl)-7-(1-methyl-1H-pyrazol-4-yl)isoquinoline (160
mg, 0.6 mmol) in AcOH (1 mL) was added N-bromosuccinimide (109 mg,
0.6 mmol). The mixture was heated to 90.degree. C. for 2 h under a
nitrogen atmosphere. After cooling to room temperature, the mixture
was concentrated in vacuo. DCM (10 mL) was added and washed with
water (10 mL.times.3) and brine (10 mL). The organic layer was
dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo.
The crude residue was purified by silica gel chromatography
(DCM/MeOH=100:1) to give the title compound (95 mg, 45%) as a light
yellow solid. LCMS M/Z (M+H) 338.
Step 9: tert-butyl
1-(tetrahydro-2H-pyran-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00224##
[0427] To a solution of tert-butyl
3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
ne-5(4H)-carboxylate (Intermediate G, 500 mg, 1.29 mmol) in
1,4-dioxane (5 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (62 mg,
0.13 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (102 mg, 0.13 mmol), KOAc
(381 mg, 3.88 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (657
mg, 2.59 mmol). The mixture was heated to 80.degree. C. for 3 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 434.
Step 10: tert-butyl
3-(6-(difluoromethyl)-7-(1-methyl-1H-pyrazol-4-yl)isoquinolin-4-yl)-1-(te-
trahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carbo-
xylate
##STR00225##
[0429] To the above step cooled solution was added
4-bromo-6-(difluoromethyl)-7-(1-methyl-1H-pyrazol-4-yl)isoquinoline
(95 mg, 0.28 mmol),
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (13 mg,
0.028 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (22 mg, 0.028 mmol),
Na.sub.2CO.sub.3 (89 mg, 0.84 mmol), 1,4-dioxane (5 mL) and water
(2 mL). The reaction mixture was heated to 60.degree. C. for 16 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. DCM (50 mL) was
added and washed with water (30 mL.times.3) and brine (20 mL). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by silica
gel chromatography (DCM/MeOH=100:1) to give the title compound (70
mg, 10%) as a yellow solid. LCMS M/Z (M+H) 565.
Step 11:
6-(difluoromethyl)-7-(1-methyl-1H-pyrazol-4-yl)-4-(1-(tetrahydro--
2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)isoquinol-
ine
##STR00226##
[0431] To a solution of tert-butyl
3-(6-(difluoromethyl)-7-(1-methyl-1H-pyrazol-4-yl)isoquinolin-4-yl)-1-(te-
trahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carbo-
xylate (70 mg, 0.12 mmol) in DCM (1 mL) at 0.degree. C. was added
trifluoroacetic acid (1 mL, 13.42 mmol). The mixture was stirred at
room temperature for 12 h and concentrated in vacuo to give the
title compound (50 mg, crude) as brown oil that required no further
purification. LCMS M/Z (M+H) 465.
Step 12:
3-(6-(difluoromethyl)-7-(1-methyl-1H-pyrazol-4-yl)isoquinolin-4-y-
l)-N-methyl-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idine-5(4H)-carboxamide
##STR00227##
[0433] To a solution of
6-(difluoromethyl)-7-(1-methyl-1H-pyrazol-4-yl)-4-(1-(tetrahydro-2H-pyran-
-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)isoquinoline
(50 mg, 0.11 mmol) in DCM (5 mL) was added triethylamine (0.04 mL,
0.32 mmol) and N-methyl-1H-imidazole-1-carboxamide (20 mg, 0.16
mmol). The reaction was stirred at room temperature for 1 h. DCM
(50 mL) was added and washed with water (50 mL.times.3) and brine
(20 mL). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by reverse phase chromatography (acetonitrile
30-60%/0.05% NH.sub.4OH in water) to give the title compound (32
mg, 57%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.37 (s, 1H), 9.12 (s, 1H), 8.59 (s, 1H), 8.32 (s, 1H),
8.08 (s, 1H), 7.81 (s, 1H), 7.27 (t, J=56.0 Hz, 1H), 6.66-6.60 (m,
1H), 4.51-4.44 (m, 3H), 4.05-3.98 (m, 2H), 3.95 (s, 3H), 3.75-3.67
(m, 2H), 3.57-3.48 (m, 2H), 2.86-2.80 (m, 2H), 2.54 (d, J=4.4 Hz,
3H), 2.20-2.07 (m, 2H), 1.97-1.92 (m, 2H). LCMS M/Z (M+H) 522.
Examples 26 & 27
(S)-1-(3-(3-(1-methyl-1H-pyrazol-4-yl)quinolin-5-yl)-1-(tetrahydrofuran-3--
yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone and
(S)-1-(3-(2-(1-methyl-1H-pyrazol-4-yl)quinolin-5-yl)-1-(tetrahydrofuran-3-
-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00228##
[0434] Step 1:
(S)-1-(3-(quinolin-5-yl)-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo-
[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00229##
[0436] To a solution of
(S)-1-(3-bromo-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate E, 500 mg, 1.6 mmol) in
1,4-dioxane (10 mL) and water (2 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (116
mg, 0.2 mmol), K.sub.2CO.sub.3 (446 mg, 3.2 mmol) and
5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinoline (609 mg,
1.9 mmol). The mixture was heated to 120.degree. C. for 12 h under
a nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
DCM (50 mL) was added and washed with water (50 mL.times.2) and
brine (50 mL). The organic phase was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (DCM/MeOH=20:1)
to give the title compound (600 mg, 78%) as a yellow solid. LCMS
M/Z (M+H) 363.
Step 2:
(S)-5-(5-acetyl-1-(tetrahydrofuran-3-yl)-4,5,6,7-tetrahydro-1H-pyr-
azolo[4,3-c]pyridin-3-yl)quinoline 1-oxide
##STR00230##
[0438] To a solution of
(S)-1-(3-(quinolin-5-yl)-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo-
[4,3-c]pyridin-5(4H)-yl)ethanone (624 mg, 1.3 mmol) in DCM (10 mL)
was added 3-chlorobenzenecarboperoxoic acid (334 mg, 1.9 mmol). The
mixture was stirred at 26.degree. C. for 4 h. DCM (80 mL) was added
and washed with sat. aq. Na.sub.2S.sub.2O.sub.3 (50 mL.times.3),
water (50 mL) and brine (50 mL). The organic phase was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography
(DCM/MeOH=20:1) to give the title compound (230 mg, 45%) as a
yellow solid. LCMS M/Z (M+H) 379.
Step 3:
(S)-1-(3-(3-bromoquinolin-5-yl)-1-(tetrahydrofuran-3-yl)-6,7-dihyd-
ro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone and
(S)-1-(3-(2-bromoquinolin-5-yl)-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-p-
yrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00231##
[0440] To a solution of
(S)-5-(5-acetyl-1-(tetrahydrofuran-3-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4-
,3-c]pyridin-3-yl)quinoline 1-oxide (145 mg, 0.4 mmol) and 4 .ANG.
molecular sieves in DCM (13 mL) was added tetrabutylammonium
bromide (185 mg, 0.6 mmol). The mixture was stirred at room
temperature for 10 min and added 4-methylbenzenesulfonic anhydride
(187 mg, 0.6 mmol). The mixture was stirred at room temperature for
an additional 12 h. The mixture was filtered and concentrated in
vacuo. The crude residue was purified by silica gel chromatography
(DCM/MeOH=20:1) to give the mixture of title compounds (85 mg, 42%)
as a yellow solid. LCMS M/Z (M+H) 441.
Step 4:
(S)-1-(3-(3-(1-methyl-1H-pyrazol-4-yl)quinolin-5-yl)-1-(tetrahydro-
furan-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
and
(S)-1-(3-(2-(1-methyl-1H-pyrazol-4-yl)quinolin-5-yl)-1-(tetrahydrofuran-3-
-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00232##
[0442] To a solution of
(S)-1-(3-(3-bromoquinolin-5-yl)-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-p-
yrazolo[4,3-c]pyridin-5(4H)-yl)ethanone and
(S)-1-(3-(2-bromoquinolin-5-yl)-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-p-
yrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (80 mg, 0.15 mmol) in THE
(5 mL) and water (1 mL) was added
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(31 mg, 0.15 mmol),
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (7 mg, 0.01
mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (12 mg, 0.01 mmol),
Na.sub.2CO.sub.3 (32 mg, 0.3 mmol). The mixture was heated to
60.degree. C. for 16 h under a nitrogen atmosphere. After cooling
the reaction to room temperature, the mixture was filtered and
concentrated in vacuo. The crude residue was purified by reverse
phase chromatography (acetonitrile 30-60%/0.1% NH.sub.4OH in water)
to give the mixture of title compounds (50 mg, 62%) as a white
solid which was separated by using chiral SFC (Chiralpak AD
250.times.30 mm I.D., 5 um; Supercritical
CO.sub.2/MEOH+NH.sub.3.H.sub.2O=45/55; 50 ml/min) to give
(S)-1-(3-(3-(1-methyl-1H-pyrazol-4-yl)quinolin-5-yl)-1-(tetrahydrofuran-3-
-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)etanone (5 mg,
first peak) and
(S)-1-(3-(2-(1-methyl-1H-pyrazol-4-yl)quinolin-5-yl)-1-(tetrahy-
drofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
(21 mg, second peak). Example 26: .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.21 (s, 1H), 9.10 (s, 1H), 8.36 (s, 1H),
8.02-7.95 (m, 1H), 7.99 (s, 1H), 7.78-7.73 (m, 1H), 7.63-7.55 (m,
1H), 5.12-5.07 (m, 1H), 4.62-4.50 (m, 2H), 4.12-4.00 (m, 3H),
3.92-3.76 (m, 3H), 3.90 (s, 1H), 2.97-2.84 (m, 2H), 2.44-2.39 (m,
2H), 2.11-2.00 (m, 3H). LCMS M/Z (M+H) 443. Example 27: .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 8.79 (d, J=8.8 Hz, 1H), 8.49 (s,
1H), 8.18 (s, 1H), 7.96-7.91 (m, 1H), 7.86 (d, J=8.8 Hz, 1H),
7.80-7.75 (m, 1H), 7.56-7.46 (m, 1H), 5.09-5.03 (m, 1H), 4.50 (s,
2H), 4.12-4.01 (m, 2H), 3.97-3.79 (m, 4H), 3.93 (s, 3H), 2.95-2.82
(m, 2H), 2.42-2.35 (m, 2H), 2.10-2.00 (m, 3H). LCMS M/Z (M+H)
443.
Example 28
1-(3-(4-chloroisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1-
H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00233##
[0443] Step 1: 8-bromo-4-chloroisoquinoline
##STR00234##
[0445] To a solution of 8-bromoisoquinoline (12.2 g, 58.64 mmol) in
AcOH (150 mL) was added 1-chloropyrrolidine-2,5-dione (8.61 g, 64.5
mmol) portionwise. The mixture was heated to 117.degree. C. for 2 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, water (150 mL) was added and extracted with EtOAc (150
mL.times.3). The combined organic layers were washed with sat. aq.
NaHCO.sub.3 (150 mL.times.3), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (10.0 g, crude) as a yellow solid that required no
further purification. LCMS M/Z (M+H) 242.
Step 2:
4-chloro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoli-
ne
##STR00235##
[0447] To a solution of 8-bromo-4-chloroisoquinoline (500 mg, 2.06
mmol) in 1,4-dioxane (8 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (151
mg, 0.21 mmol), KOAc (607 mg, 6.19 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (785
mg, 3.09 mmol). The mixture was heated to 80.degree. C. for 4 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 290.
Step 3:
1-(3-(4-chloroisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-d-
ihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00236##
[0449] To the above step cooled solution was added
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 541 mg, 1.65 mmol),
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (75 mg,
0.1 mmol) and Na.sub.2CO.sub.3 (656 mg, 6.19 mmol), 1,4-dioxane (2
mL) and water (2 mL). The reaction mixture was heated to 65.degree.
C. for 2 h under nitrogen atmosphere. After cooling to room
temperature, the mixture was filtered and concentrated in vacuo.
The crude residue was purified by reverse phase chromatography
(acetonitrile 20-50%/0.2% formic acid in water) to give the title
compound (65 mg, 7%) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 9.62-9.58 (m, 1H), 8.64-8.60 (m, 1H), 8.29-8.25
(m, 1H), 7.90-7.85 (m, 1H), 7.68-7.63 (m, 1H), 4.62-4.45 (m, 2H),
4.35-4.24 (m, 1H), 4.23-4.13 (m, 2H), 4.06-3.81 (m, 2H), 3.57 (t,
J=12.0 Hz, 2H), 2.95-2.85 (m, 2H), 2.44-2.41 (m, 2H), 2.21-2.08 (m,
3H), 1.98-1.94 (m, 2H). LCMS M/Z M+H) 411.
Example 29
1-(3-(5-fluoroisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1-
H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00237##
[0450] Step 1: 5-(benzyloxy)isoquinoline
##STR00238##
[0452] To a stirred solution of isoquinolin-5-ol (1.0 g, 6.89 mmol)
in DMF (30 mL) at 0.degree. C. was added NaH (60%, 303 mg, 7.58
mmol) and the mixture was stirred for 30 min. Benzyl bromide (1.0
g, 5.86 mmol) was added dropwise and the mixture stirred for an
additional 1 h. The mixture was quenched with water (100 mL) and
extracted with EtOAc (50 mL.times.3). The combined organic layers
were washed with brine (50 mL.times.3), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (900 mg, crude) as yellow oil that required no
further purification. LCMS M/Z (M+H) 236.
Step 2: 5-(benzyloxy)-8-bromoisoquinoline
##STR00239##
[0454] To a solution of 5-(benzyloxy)isoquinoline (1.3 g, 5.53
mmol) and NaOAc (906 mg, 11.05 mmol) in AcOH (50 mL) at 26.degree.
C. was added bromine (0.28 mL, 5.53 mmol) dropwise. The mixture was
stirred at 26.degree. C. for 16 h. Water (100 mL) was added and
extracted with EtOAc (70 mL.times.3). The combined organic layers
were washed with sat. aq. NaHCO.sub.3 (100 mL.times.2), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography (petroleum
ether/EtOAc=4:1) to give the title compound (300 mg, 17%) as a
white solid. LCMS M/Z (M+H) 314.
Step 3:
1-(1-(tetrahydro-2H-pyran-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxa-
borolan-2-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00240##
[0456] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 2.3 g, 7.0 mmol) in
1,4-dioxane (40 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (200 mg,
0.42 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (165 mg, 0.21 mmol), KOAc
(2.0 g, 21.0 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (3.56
g, 14.0 mmol). The mixture was heated to 80.degree. C. for 3 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 376.
Step 4:
1-(3-(5-(benzyloxy)isoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)--
6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00241##
[0458] To the above step cooled solution was added
5-(benzyloxy)-8-bromoisoquinoline (1.1 g, 3.5 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (185 mg, 0.24 mmol),
K.sub.3PO.sub.4 (1.98 g, 9.3 mmol), 1,4-dioxane (10 mL) and water
(10 mL). The reaction mixture was heated to 90.degree. C. for 1 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. Water (100 mL) was
added and extracted with EtOAc (50 mL.times.3). The combined
organic layers were washed with brine (50 mL.times.3), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography
(DCM/MeOH=20:1) to give the title compound (1 g, 30%) as a brown
solid. LCMS M/Z (M+H) 483.
Step 5:
1-(3-(5-hydroxyisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7--
dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00242##
[0460] To a solution of
1-(3-(5-(benzyloxy)isoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dih-
ydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (1.0 g, 2.07 mmol)
in MeOH (20 mL) was added Pd(OH).sub.2 (100 mg) and AcOH (2 drops).
The mixture was stirred at 26.degree. C. for 5 h under a hydrogen
atmosphere (15 Psi). The mixture was filtered and concentrated in
vacuo. The crude residue was purified by Prep-TLC (DCM/MeOH=20:1)
to give the title compound (400 mg, 49%) as a yellow solid. LCMS
M/Z (M+H) 393.
Step 6:
1-(3-(5-fluoroisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-d-
ihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00243##
[0462] To a solution of
1-(3-(5-hydroxyisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-
-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (260 mg, 0.66 mmol) in
toluene (7 mL) was added
1,3-bis(2,6-di-i-propylphenyl)-2-chloroimidazolium chloride/cesium
fluoride admixture (1.69 g, 2.76 mmol). The mixture was stirred at
26.degree. C. for 0.5 h and then heated to 110.degree. C. for 24 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. The crude residue
was purified by Prep-TLC (DCM/MeOH=20:1) to give the title compound
(3 mg, 1%) as a yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 9.71-9.67 (m, 1H), 8.67-8.53 (m, 1H), 7.96-7.94 (m, 1H),
7.54-7.47 (m, 1H), 7.46-7.35 (m, 1H), 4.61-4.44 (m, 2H), 4.33-4.22
(m, 1H), 4.21-4.12 (m, 2H), 4.02-3.83 (m, 2H), 3.60-3.53 (m, 2H),
2.93-2.84 (m, 2H), 2.44-2.40 (m, 2H), 2.21-2.07 (m, 3H), 1.98-1.94
(m, 2H). LCMS M/Z (M+H) 395.
Example 30
1-(3-(6-chloro-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-
-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00244##
[0463] Step 1: (2-bromo-4-chlorophenyl)methanamine
##STR00245##
[0465] To a stirred solution of 2-bromo-4-chlorobenzonitrile (20.0
g, 138.6 mmol) in anhydrous THE (200 mL) at 0.degree. C. was added
borane (277 mL, 277.2 mmol, 1.0 M) in THE dropwise under a nitrogen
atmosphere. The resulting mixture was stirred at 22.degree. C. for
1 h and refluxed for 3 h. The reaction was quenched with 2N HCl
(300 mL) at 0.degree. C. and then stirred at 70.degree. C. for 1 h.
After cooling to room temperature, the solution was extracted with
DCM (400 mL) and the aqueous phase was adjusted to pH=8 by using 2N
NaOH. The mixture was extracted with DCM (300 mL.times.3). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4 and
concentrated in vacuo to give the title compound (12 g, 59%) as
yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.55-7.53 (m,
1H), 7.34-7.29 (m, 1H), 7.28-7.23 (m, 1H), 3.86 (s, 2H).
Step 2: N-(2-bromo-4-chlorobenzyl)-2,2-dimethoxyacetamide
##STR00246##
[0467] To a solution of (2-bromo-4-chloro-phenyl)methanamine (12 g,
54.4 mmol) in MeOH (80 mL) was added triethylamine (9.5 mL, 68.0
mmol) and methyl dimethoxyacetate (8.0 g, 49.9 mmol). The mixture
was heated to 80.degree. C. for 20 h. After cooling to room
temperature, the mixture was concentrated in vacuo. The crude
residue was dissolved in EtOAc (150 mL), washed with 1N HCl (150
mL), H.sub.2O (150 mL), brine (150 mL). The organic phase was dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo
to give the title compound (12 g, 68%) as a light yellow solid.
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 7.58 (d, J=1.6 Hz, 1H),
7.33 (d, J=8.4 Hz, 1H), 7.29-7.27 (m, 1H), 7.03 (s, 1H), 4.74 (s,
1H), 4.52 (d, J=6.4 Hz, 2H), 3.41 (s, 6H).
Step 3: 8-bromo-6-chloroisoquinolin-3(2H)-one
##STR00247##
[0469] To a solution of sulfuric acid (100 mL) at 0.degree. C. was
added N-(2-bromo-4-chlorobenzyl)-2,2-dimethoxyacetamide (12.0 g,
37.2 mmol). The reaction was heated to 50.degree. C. for 16 h. The
reaction was poured into ice water (150 mL) and the mixture was
basified with ammonium hydroxide to pH 8. The yellow precipitate
was filtered off, washed with water, and dried in vacuo. The crude
residue was purified by silica gel chromatography (DCM/MeOH=10:1)
to give the title compound (3.7 g, 39%) as a yellow solid.
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 9.10 (s, 1H), 7.93 (s,
1H), 7.69 (d, J=2.0 Hz, 1H), 6.94 (s, 1H).
Step 4: 8-bromo-6-chloro-3-((triisopropylsilyl)oxy)isoquinoline
##STR00248##
[0471] To a solution of 8-bromo-6-chloroisoquinolin-3(2H)-one (10
g, 38.7 mmol) in DMF (30 mL) at 0.degree. C. was added imidazole
(7.9 g, 116.1 mmol) and chlorotriisopropylsilane (12.4 mL, 58.0
mmol). The reaction was stirred at room temperature for 12 h. The
reaction mixture was concentrated in vacuo. The crude residue was
dissolved in DCM (50 mL) and washed with H.sub.2O (50 mL). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by silica
gel chromatography (petroleum ether) to give the title compound (10
g, 62%) as yellow oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
9.15 (s, 1H), 7.59 (s, 1H), 7.54 (s, 1H), 6.88 (s, 1H), 1.51-1.40
(m, 3H), 1.12 (d, J=7.2 Hz, 18H).
Step 5:
6-chloro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-((triis-
opropylsilyl)oxy)isoquinoline
##STR00249##
[0473] To a solution of
8-bromo-6-chloro-3-((triisopropylsilyl)oxy)isoquinoline (9.0 g,
21.7 mmol) in 1,4-dioxane (9 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) (1.6
g, 2.2 mmol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (8.3 g,
32.5 mmol) and 2-ethylhexanoyloxypotassium (11.9 g, 65.1 mmol). The
mixture was heated to 70.degree. C. for 1 h under a nitrogen
atmosphere. After cooling the reaction to room temperature, the
reaction mixture was used to the next step directly without further
purification. LCMS M/Z (M-TIPS+H) 306.
Step 6:
1-(3-(6-chloro-3-((triisopropylsilyl)oxy)isoquinolin-8-yl)-1-(tetr-
ahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethan-
one
##STR00250##
[0475] To the above step cooled solution in 1,4-dioxane (60 mL) and
water (12 mL) was added
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 6.39 g, 19.49 mmol),
[1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) (1.58
g, 2.16 mmol) and Na.sub.2CO.sub.3 (6.8 g, 64.9 mmol). The reaction
mixture was heated to 70.degree. C. for 4 h under nitrogen
atmosphere. After cooling to room temperature, the mixture was
filtered and concentrated in vacuo. EtOAc (200 mL) was added and
washed with water (120 mL.times.2). The organic layer was dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The crude residue was purified by silica gel chromatography
(DCM/MeOH=10:1) to give the title compound (2.6 g, 38% purity) as a
brown solid. LCMS M/Z (M-TIPS+H) 427.
Step 7:
1-(3-(6-chloro-3-hydroxyisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-
-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00251##
[0477] To a solution of
1-(3-(6-chloro-3-((triisopropylsilyl)oxy)isoquinolin-8-yl)-1-(tetrahydro--
2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
(2.6 g, 38% purity) in THE (10 mL) at 20.degree. C. was added TBAF
(22.3 ml, 22.3 mmol, 1 M in THF). The reaction was stirred at room
temperature for 12 h. The reaction mixture was concentrated in
vacuo. The crude residue was dissolved in DCM (50 mL) and washed
with H.sub.2O (150 mL.times.2). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by reverse phase chromatography
(acetonitrile 10-40%/0.225% formic acid in water) to give the title
compound (120 mg). LCMS M/Z (M+H) 427.
Step 8:
8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyr-
azolo[4,3-c]pyridin-3-yl)-6-chloroisoquinolin-3-yl
trifluoromethanesulfonate
##STR00252##
[0479] To a solution of
1-(3-(6-chloro-3-hydroxyisoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,-
7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (120 mg, 0.28
mmol) in DCM (3 mL) at 0.degree. C. was added triethylamine (0.12
mL, 0.84 mmol) and trifluoromethanesulfonic anhydride (0.06 mL,
0.37 mmol). The reaction was stirred at room temperature for 16 h.
DCM (30 mL) was added and washed with water (30 mL.times.2). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo to give the title compound (157 mg,
crude) as a brown solid that required no further purification. LCMS
M/Z (M+H) 559.
Step 9:
1-(3-(6-chloro-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(te-
trahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)eth-
anone
##STR00253##
[0481] To a solution of
8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4-
,3-c]pyridin-3-yl)-6-chloroisoquinolin-3-yl
trifluoromethanesulfonate (600 mg, crude) in 1,4-dioxane (10 mL)
and water (2 mL) was added
[1,1'-bis(diphenylphosphino)-ferrocene]dichloropalladium(II) (79
mg, 0.11 mmol), Na.sub.2CO.sub.3 (341 mg, 3.21 mmol) and
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(223 mg, 1.07 mmol). The mixture was irradiated in a microwave at
60.degree. C. for 0.5 h. EtOAc (50 mL) was added and washed with
water (40 mL), brine (40 mL). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by reverse phase chromatography
(acetonitrile 20-50%/0.05% NH.sub.4OH in water) to give the title
compound (10 mg) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 9.60 (s, 1H), 8.13-7.93 (m, 2H), 7.79-7.73 (m,
2H), 7.43-7.40 (m, 1H), 4.65-4.49 (m, 2H), 4.33-4.23 (m, 1H),
4.18-4.16 (m, 2H), 4.01-3.83 (m, 2H), 4.00 (s, 3H), 3.60-3.54 (m,
2H), 2.93-2.86 (m, 2H), 2.44-2.41 (m, 2H), 2.21-2.10 (m, 3H),
1.97-1.94 (m, 2H). LCMS M/Z (M+H) 491.
Example 31
1-(3-(6-ethyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro--
2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00254##
[0482] Step 1:
1-(3-(3-(1-methyl-1H-pyrazol-4-yl)-6-vinylisoquinolin-8-yl)-1-(tetrahydro-
-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00255##
[0484] To a solution of
1-(3-(6-chloro-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydr-
o-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
(Example 30, 110 mg, 0.22 mmol) in THE (3 mL) and water (0.6 mL)
was added 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl
(10 mg, 0.02 mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (16 mg, 0.02 mmol),
Na.sub.2CO.sub.3 (71 mg, 0.67 mmol) and
4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (345 mg, 2.24
mmol). The mixture was irradiated in a microwave at 60.degree. C.
for 0.5 h. EtOAc (50 mL) was added and extracted with water (40
mL), brine (40 mL). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (DCM/MeOH=10:1)
to give the title compound (70 mg, 65%) as a brown solid. LCMS M/Z
(M+H) 483.
Step 2:
1-(3-(6-ethyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tet-
rahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)etha-
none
##STR00256##
[0486] To a solution of
1-(3-(3-(1-methyl-1H-pyrazol-4-yl)-6-vinylisoquinolin-8-yl)-1-(tetrahydro-
-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
(70 mg, 0.15 mmol) in MeOH (5 mL) was added 10% Pd/C (50 mg). The
mixture was stirred at room temperature for 1 h under a hydrogen
atmosphere (15 Psi). The mixture was filtered and the filtrate was
concentrated in vacuo. The crude residue was purified by Prep-TLC
(DCM/MeOH=10:1) to give the title compound (6 mg, 9%) as a yellow
solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.54-9.50 (m, 1H),
8.05-8.02 (m, 2H), 7.78-7.75 (m, 1H), 7.60-7.56 (m, 1H), 7.36-7.34
(m, 1H), 4.66-4.43 (m, 2H), 4.33-4.23 (m, 1H), 4.21-4.12 (m, 2H),
4.04-3.81 (m, 2H), 4.00 (s, 3H), 3.60-3.54 (m, 2H), 2.94-2.81 (m,
4H), 2.53-2.38 (m, 2H), 2.22-2.05 (m, 3H), 1.97-1.94 (m, 2H),
1.40-1.34 (m, 3H). LCMS M/Z (M+H) 485.
Example 32
8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,-
3-c]pyridin-3-yl)-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline-4-carbonitrile
##STR00257##
[0487] Step 1:
2-chloro-6-((1-methyl-1H-pyrazol-4-yl)ethynyl)benzaldehyde
##STR00258##
[0488] a solution of 2-bromo-6-chlorobenzaldehyde (13 g, 59.2 mmol)
in triethylamine (650 mL) was added
bis(triphenylphosphine)palladium(II) dichloride (2.1 g, 3.0 mmol)
and 4-ethynyl-1-methyl-1H-pyrazole (10.4 g, 98 mmol) and copper(I)
iodide (650 mg, 3.4 mmol). The mixture was heated to 80.degree. C.
for 16 h under a nitrogen atmosphere. After cooling the reaction to
room temperature, the mixture was filtered and concentrated in
vacuo. The crude residue was purified by silica gel chromatography
(petroleum ether/EtOAc=5:1) to give the title compound (6.9 g, 48%)
as a yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.52
(s, 1H), 7.58 (s, 1H), 7.53 (s, 1H), 7.40 (d, J=7.2 Hz, 1H),
7.35-7.24 (m, 2H), 3.82 (s, 3H). LCMS M/Z (M+H) 245.
Step 2:
(2-chloro-6-((1-methyl-1H-pyrazol-4-yl)ethynyl)phenyl)methanol
##STR00259##
[0490] To a solution of
2-chloro-6-((1-methyl-1H-pyrazol-4-yl)ethynyl)benzaldehyde (6.9 g,
28.2 mmol) in MeOH (60 mL) at 0.degree. C. was added sodium
borohydride (1.7 g, 43.9 mmol) portionwise. The mixture was stirred
at room temperature for 2 h under a nitrogen atmosphere. The
reaction was concentrated in vacuo. Water (50 mL) was added and
extracted with EtOAc (50 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo to give the title compound (8.2 g, crude) as
a yellow solid that required no further purification. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.65 (s, 1H), 7.58 (s, 1H), 7.41 (d,
J=7.2 Hz, 1H), 7.34 (d, J=8.0 Hz, 1H), 7.23-7.19 (m, 1H), 5.01 (d,
J=6.0 Hz, 2H), 3.90 (s, 3H), 2.51 (t, J=6.4 Hz, 1H). LCMS M/Z (M+H)
247.
Step 3:
4-((2-(azidomethyl)-3-chlorophenyl)ethynyl)-1-methyl-1H-pyrazole
##STR00260##
[0492] To a solution of
(2-chloro-6-((1-methyl-1H-pyrazol-4-yl)ethynyl)phenyl)methanol (7.2
g, 29 mmol) in toluene (80 mL) was added diphenylphosphoryl azide
(9.6 g, 34.8 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (5.7 g,
37.6 mmol). The mixture was stirred at room temperature for 16 h
under a nitrogen atmosphere. DCM (30 mL) was added and washed with
water (30 mL.times.2). The organic phase was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (9 g, crude) as yellow oil that required no further
purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.13 (s,
1H), 7.74 (s, 1H), 7.62-7.52 (m, 1H), 7.46-7.38 (m, 1H), 7.28-7.10
(m, 1H), 4.73 (s, 2H), 3.87 (s, 3H). LCMS M/Z (M+H) 272.
Step 4:
8-chloro-4-iodo-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline
##STR00261##
[0494] To a solution of
4-((2-(azidomethyl)-3-chlorophenyl)ethynyl)-1-methyl-1H-pyrazole
(7.9 g, 28.9 mmol) in DCM (500 mL) was added iodine (36.8 g, 144.9
mmol) and K.sub.3PO.sub.4 (30.8 g, 145 mmol). The mixture was
stirred at room temperature for 24 h under a nitrogen atmosphere
and washed with water (400 mL.times.2). The organic phase was dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The crude residue was purified by silica gel chromatography
(petroleum ether/EtOAc=1:1) to give the title compound (10 g, 94%)
as a brown solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.43
(s, 1H), 8.47 (s, 1H), 8.18-8.15 (m, 2H), 7.90-1.86 (m, 2H), 3.95
(s, 3H). LCMS M/Z (M+H) 370.
Step 5:
8-chloro-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline-4-carbonitrile
##STR00262##
[0496] To a solution of
8-chloro-4-iodo-3-(1-methylpyrazol-4-yl)isoquinoline (500 mg, 1.35
mmol) in DMF (10 mL) was added copper(I) cyanide (133 mg, 1.49
mmol) and tetrakis(triphenylphosphine)palladium(0) (156 mg, 0.14
mmol). The mixture was heated to 120.degree. C. for 16 h under a
nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
DCM (50 mL) was added and washed with water (40 mL) and brine (40
mL). The organic phase was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude residue was purified
by silica gel chromatography (petroleum ether/EtOAc=1:1) to give
the title compound (300 mg, 83%) as a brown solid. LCMS M/Z (M+H)
269.
Step 6:
3-(1-methyl-1H-pyrazol-4-yl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxabor-
olan-2-yl)isoquinoline-4-carbonitrile
##STR00263##
[0498] To a solution of
8-chloro-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline-4-carbonitrile
(300 mg, 1.12 mmol) in 1,4-dioxane (5 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (52 mg,
0.11 mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphe-
nyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (87 mg, 0.11 mmol),
KOAc (328 mg, 3.35 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (850
mg, 3.35 mmol). The mixture was heated to 90.degree. C. for 12 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification.
Step 7:
8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyr-
azolo[4,3-c]pyridin-3-yl)-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline-4-carbo-
nitrile
##STR00264##
[0500] To the above reaction mixture was added
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 300 mg, 0.92 mmol),
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (38 mg,
0.08 mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphe-
nyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (63 mg, 0.08 mmol),
Na.sub.2CO.sub.3 (266 mg, 2.5 mmol) and water (1 mL). The mixture
was heated to 60.degree. C. for 12 h under a nitrogen atmosphere.
After cooling the reaction to room temperature, DCM (50 mL) was
added and washed with water (40 mL), brine (40 mL). The organic
phase was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by reverse
phase chromatography (acetonitrile 23-53%/0.2% formic acid in
water) to give the title compound (11 mg, 3%) as a white solid.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.90 (d, J=6.0 Hz, 1H),
8.52-8.38 (m, 2H), 8.22-8.17 (m, 1H), 7.96-7.84 (m, 1H), 7.66-7.54
(m, 1H), 4.68-4.51 (m, 2H), 4.32-4.27 (m, 1H), 4.19-4.16 (m, 2H),
4.04 (s, 3H), 4.02-3.84 (m, 2H), 3.61-3.55 (m, 2H), 2.99-2.81 (m,
2H), 2.48-2.39 (m, 2H), 2.27-2.06 (m, 3H), 1.98-1.95 (m, 2H). LCMS
M/Z (M+H) 482.
Example 33
1-(3-(4-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-
-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00265##
[0501] Step 1:
8-chloro-4-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline
##STR00266##
[0503] To a solution of
8-chloro-4-iodo-3-(1-methylpyrazol-4-yl)isoquinoline (300 mg, 0.81
mmol) in DMF (15 mL) and water (3 mL) was added potassium
methyltrifluoroborate (495 mg, 4.1 mmol), butyl
di-1-adamantylphosphine (30 mg, 0.08 mmol), palladium(II) acetate
(18 mg, 0.08 mmol) and Cs.sub.2CO.sub.3 (795 mg, 2.4 mmol). The
mixture was heated to 80.degree. C. for 20 h under a nitrogen
atmosphere. After cooling the reaction to room temperature, EtOAc
(50 mL) was added and washed with water (40 mL.times.2) and brine
(40 mL). The organic phase was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (petroleum
ether/EtOAc=1:1) to give the title compound (200 mg, 95%) as a
yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.57 (s,
1H), 7.98 (d, J=8.0 Hz, 1H), 7.95-7.90 (m, 2H), 7.67-7.57 (m, 2H),
4.02 (s, 3H), 2.79 (s, 3H). LCMS M/Z (M+H) 258.
Step 2:
4-methyl-3-(1-methyl-1H-pyrazol-4-yl)-8-(4,4,5,5-tetramethyl-1,3,2-
-dioxaborolan-2-yl)isoquinoline
##STR00267##
[0505] To a solution of
8-chloro-4-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline (200 mg,
0.78 mmol) in 1,4-dioxane (5 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (38 mg,
0.08 mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphe-
nyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (64 mg, 0.08 mmol),
KOAc (228 mg, 2.33 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (591
mg, 2.33 mmol). The mixture was heated to 90.degree. C. for 16 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
EtOAc (50 mL) was added and washed with water (40 mL.times.2) and
brine (40 mL). The organic phase was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (petroleum
ether/EtOAc=3:1) to give the title compound (100 mg, 37%) as a
brown solid. LCMS M/Z (M+H) 350.
Step 3:
1-(3-(4-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(te-
trahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)eth-
anone
##STR00268##
[0507] To a solution of
4-methyl-3-(1-methyl-1H-pyrazol-4-yl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxab-
orolan-2-yl)isoquinoline (100 mg, 0.29 mmol) in THE (3 mL) and
water (0.6 mL) was added
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 103 mg, 0.31 mmol),
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (14 mg,
0.03 mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphe-
nyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (24 mg, 0.03 mmol),
Na.sub.2CO.sub.3 (91 mg, 0.86 mmol). The mixture was heated to
60.degree. C. for 16 h under a nitrogen atmosphere. After cooling
the reaction to room temperature, EtOAc (50 mL) was added and
washed with water (40 mL), brine (40 mL). The organic phase was
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The crude residue was purified by reverse phase
chromatography (acetonitrile 20-50%/0.05% NH.sub.4OH in water) to
give the title compound (14 mg, 10%) as a white solid. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 9.61-9.48 (m, 1H), 8.14-8.04 (m, 1H),
7.95-7.90 (m, 2H), 7.84-7.74 (m, 1H), 7.61-7.50 (m, 1H), 4.63-4.44
(m, 2H), 4.31-4.25 (m, 1H), 4.20-4.12 (m, 2H), 4.01 (s, 3H),
4.00-3.83 (m, 2H), 3.59-3.54 (m, 2H), 2.96-2.82 (m, 2H), 2.81 (s,
3H), 2.48-2.39 (m, 2H), 2.21-2.05 (m, 3H), 1.98-1.95 (m, 2H). LCMS
M/Z (M+H) 471.
Example 34
N-methyl-3-(4-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetr-
ahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxa-
mide
##STR00269##
[0508] Step 1: tert-butyl
3-(4-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-2-
H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00270##
[0510] To a solution of
4-methyl-3-(1-methyl-1H-pyrazol-4-yl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxab-
orolan-2-yl)isoquinoline (300 mg, 0.86 mmol) in THE (10 mL) and
water (2 mL) was added tert-butyl
3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
ne-5(4H)-carboxylate (Intermediate G, 398 mg, 1.03 mmol),
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (43 mg,
0.09 mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphe-
nyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (71 mg, 0.09 mmol),
Na.sub.2CO.sub.3 (273 mg, 2.58 mmol). The mixture was heated to
60.degree. C. for 16 h under a nitrogen atmosphere. After cooling
the reaction to room temperature, DCM (70 mL) was added and washed
with water (50 mL), brine (50 mL). The organic phase was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography
(DCM/MeOH=50:1) to give the title compound (200 mg, 44%) as a brown
solid. LCMS M/Z (M+H) 529.
Step 2:
4-methyl-3-(1-methyl-1H-pyrazol-4-yl)-8-(1-(tetrahydro-2H-pyran-4--
yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)isoquinoline
##STR00271##
[0512] To a solution of tert-butyl
3-(4-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-2-
H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(200 mg, 0.38 mmol) in DCM (3 mL) at 0.degree. C. was added
trifluoroacetic acid (0.28 mL, 3.78 mmol). The mixture was stirred
at room temperature for 12 h. DCM (20 mL) was added and washed with
sat. aq. NaHCO.sub.3 (10 mL.times.3), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (180 mg, crude) as brown oil that required no
further purification. LCMS M/Z (M+H) 429.
Step 3:
N-methyl-3-(4-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-
-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-
-carboxamide
##STR00272##
[0514] To a solution of
4-methyl-3-(1-methyl-1H-pyrazol-4-yl)-8-(1-(tetrahydro-2H-pyran-4-yl)-4,5-
,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)isoquinoline (180
mg, 0.42 mmol) in DCM (5 mL) was added triethylamine (0.23 mL, 1.68
mmol) and N-methyl-1H-imidazole-1-carboxamide (105 mg, 0.84 mmol).
The mixture was stirred at room temperature for 12 h. DCM (40 mL)
was added, washed with water (40 mL.times.2) and brine (40 mL). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by
reverse phase chromatography (acetonitrile 20-50%/0.05% NH.sub.4OH
in water) to give the title compound (53 mg, 24%) as a white solid.
.sup.1H NMR (400 MHz, CDCl.sub.3) 9.44 (s, 1H), 8.01 (d, J=8.4 Hz,
1H), 7.86 (s, 1H), 7.80 (s, 1H), 7.72-7.68 (m, 1H), 7.49 (d, J=7.2
Hz, 1H), 4.85-4.72 (m, 1H), 4.35-4.21 (m, 3H), 4.13-4.10 (m, 2H),
3.96 (s, 3H), 3.85-3.82 (m, 2H), 3.56-3.50 (m, 2H), 2.84-2.81 (m,
2H), 2.78-2.67 (m, 6H), 2.48-2.32 (m, 2H), 1.96-1.87 (m, 2H). LCMS
M/Z (M+H) 486.
Example 35
1-(3-(1-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-
-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00273##
[0515] Step 1:
8-chloro-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline
##STR00274##
[0517] To a solution of 3-bromo-8-chloroisoquinoline (500 mg, 2.1
mmol) in 1,4-dioxane (10 mL) and water (2 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (75 mg,
0.1 mmol), Na.sub.2CO.sub.3 (437 mg, 4.1 mmol) and
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(472 mg, 2.3 mmol). The mixture was heated to 90.degree. C. for 16
h under a nitrogen atmosphere. After cooling the reaction to room
temperature, EtOAc (30 mL) was added and washed with water (20
mL.times.2). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (DCM/MeOH=20:1)
to give the title compound (430 mg, 85%) as a yellow solid. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 9.49 (s, 1H), 8.37 (s, 1H),
8.14 (s, 1H), 8.10 (s, 1H), 7.89-7.86 (m, 1H), 7.72-7.70 (m, 2H),
3.92 (s, 3H).
Step 2: 8-chloro-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline
2-oxide
##STR00275##
[0519] To a solution of
8-chloro-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline (1.38 g, 5.66
mmol) in DCM (100 mL) was added 3-chloroperoxybenzoic acid (3.45 g,
16.99 mmol). The mixture was stirred at 16.degree. C. for 3 h. DCM
(200 mL) was added and washed with sat. aq. NaHCO.sub.3 (100
mL.times.3). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by Prep-TLC (DCM/MeOH=20:1) to give the title
compound (600 mg, 41%) as a white solid.
Step 3:
8-chloro-1-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline
##STR00276##
[0521] To a solution of
8-chloro-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline 2-oxide (750 mg,
2.89 mmol), magnesium chloride (825 mg, 8.66 mmol) and copper(I)
chloride (14 mg, 0.15 mmol) in diethyl ether (50 mL) at 0.degree.
C. was added methylmagnesium bromide (3 M, 3.85 mL, 11.55 mmol).
The mixture was stirred at room temperature for 16 h. Sat. aq.
NH.sub.4Cl (100 mL) was added and extracted with DCM (50
mL.times.3). The combined organic layers were washed with brine (50
mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by Prep-TLC
(DCM/MeOH=50:1) to give the title compound (40 mg, 5%) as a yellow
solid. LCMS M/Z (M+H) 258.
Step 4:
1-(1-(tetrahydro-2H-pyran-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxa-
borolan-2-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00277##
[0523] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 200 mg, 0.61 mmol) in
1,4-dioxane (4 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (17 mg,
0.04 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (14 mg, 0.02 mmol), KOAc (179
mg, 1.83 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (309
mg, 1.22 mmol). The mixture was heated to 80.degree. C. for 3 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 376.
Step 5:
1-(3-(1-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(te-
trahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)eth-
anone
##STR00278##
[0525] To the above step cooled solution was added
8-chloro-1-methyl-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline (40 mg,
0.16 mmol), 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl
(8 mg, 0.02 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (6 mg, 0.01 mmol),
K.sub.3PO.sub.4 (117 mg, 0.55 mmol), 1,4-dioxane (1 mL) and water
(1 mL). The reaction mixture was heated to 90.degree. C. for 2 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. Water (50 mL) was
added and extracted with EtOAc (30 mL.times.3). The combined
organic layers were washed with brine (30 mL.times.2), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by reverse phase chromatography
(acetonitrile 18-48%/0.05% NH.sub.4OH in water) to give the title
compound (1 mg, 1%) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.03-8.00 (m, 2H), 7.84-7.82 (m, 1H), 7.69-7.61
(m, 2H), 7.42-7.40 (m, 1H), 4.45-4.35 (m, 1H), 4.26-4.22 (m, 2H),
4.15-4.12 (m, 2H), 4.00-3.75 (m, 2H), 3.97 (s, 3H), 3.58-3.52 (m,
2H), 2.93-2.86 (m, 2H), 2.39-2.36 (m, 5H), 2.21-2.04 (m, 3H),
1.94-1.91 (m, 2H). LCMS M/Z (M+H) 471.
Example 36
(S)-1-(3-(6-(1-methyl-1H-pyrazol-4-yl)naphthalen-1-yl)-1-(tetrahydrofuran--
3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00279##
[0526] Step 1: 6-(1-methyl-1H-pyrazol-4-yl)naphthalen-1-amine
##STR00280##
[0528] To a solution of 6-bromonaphthalen-1-amine (470 mg, 2.12
mmol) in 1,4-dioxane (25 mL) and water (5 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (155
mg, 0.21 mmol), Na.sub.2CO.sub.3 (449 mg, 4.24 mmol) and
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(528 mg, 2.54 mmol). The mixture was heated to 120.degree. C. for 3
h under a nitrogen atmosphere. After cooling the reaction to room
temperature, water (100 mL) was added and extracted with EtOAc (50
mL.times.3). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (DCM/MeOH=30:1)
to give the title compound (300 mg, 63%) as yellow solid. LCMS M/Z
(M+H) 224.
Step 2: 4-(5-bromonaphthalen-2-yl)-1-methyl-1H-pyrazole
##STR00281##
[0530] To a solution of
6-(1-methyl-1H-pyrazol-4-yl)naphthalen-1-amine (150 mg, 0.67 mmol)
in HCl (2.0 M, 6 mL) at 0.degree. C. was added sodium nitrite (51
mg, 0.74 mmol) in water (4 mL) slowly. The mixture was stirred at
0.degree. C. for 1 h. Copper (I) bromide (350 mg, 2.44 mmol) in
water (5 mL) at 0.degree. C. was added by dropwise. The mixture was
stirred at 0.degree. C. for an additional 1 h. Water (50 mL) was
added and extracted with EtOAc (30 mL.times.3). The combined
organic layers were dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by
Prep-TLC (DCM/MeOH=20:1) to give the title compound (40 mg, 17%) as
a yellow solid. LCMS M/Z (M+H) 287.
Step 3:
1-methyl-4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphtha-
len-2-yl)-1H-pyrazole
##STR00282##
[0532] To a solution of
4-(5-bromonaphthalen-2-yl)-1-methyl-1H-pyrazole (40 mg, 0.14 mmol),
KOAc (27 mg, 0.28 mmol) and
4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-
-dioxaborolane (70 mg, 0.28 mmol) in 1,4-dioxane (10 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (11 mg,
0.01 mmol). The mixture was heated to 80.degree. C. for 16 h under
a nitrogen atmosphere. After cooling the reaction to room
temperature, water (30 mL) was added and extracted with EtOAc (20
mL.times.3). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by Prep-TLC (petroleum ether/EtOAc=3:1) to
give the title compound (30 mg, 64%) as a yellow solid.
Step 4:
(S)-1-(3-(6-(1-methyl-1H-pyrazol-4-yl)naphthalen-1-yl)-1-(tetrahyd-
rofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00283##
[0534] To a solution of
(S)-1-(3-bromo-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate E, 28 mg, 0.09 mmol) in THE (5
mL) and water (1 mL) was added
1-methyl-4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-y-
l)-1H-pyrazole (30 mg, 0.09 mmol),
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (4 mg, 0.01
mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2-
'-amino-1,1'-biphenyl-2-yl) palladium(II) (7 mg, 0.01 mmol),
Na.sub.2CO.sub.3 (19 mg, 0.18 mmol). The mixture was heated to
60.degree. C. for 16 h under a nitrogen atmosphere. After cooling
the reaction to room temperature, water (50 mL) was added and
extracted with EtOAc (30 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by reverse
phase chromatography (acetonitrile 23-53%/0.05% NH.sub.4OH in
water) to give the title compounds (4 mg, 11%) as a white solid.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.16-8.14 (m, 1H),
7.98-7.85 (m, 3H), 7.76-7.74 (m, 1H), 7.66-7.57 (m, 1H), 7.55-7.47
(m, 1H), 7.45-7.40 (m, 1H), 4.93-4.91 (m, 1H), 4.56-4.39 (m, 2H),
4.23-4.18 (m, 2H), 4.16-4.01 (m, 2H), 4.00 (s, 3H), 3.99-3.82 (m,
2H), 2.91-2.85 (m, 2H), 2.61-2.43 (m, 2H), 2.20-2.04 (m, 3H). LCMS
M/Z (M+H) 442.
Example 37
N-methyl-3-(2-(1-methyl-1H-pyrazol-4-yl)-3-oxo-1,2,3,4-tetrahydroisoquinol-
in-5-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
ne-5(4H)-carboxamide
##STR00284##
[0535] Step 1: 5-chloro-1,2-dihydroisoquinolin-3(4H)-one
##STR00285##
[0537] A mixture of 2-(2-chlorophenyl)acetonitrile (5 g, 32.98
mmol), paraformaldehyde (1.09 g, 36.28 mmol) and pyrophosphoric
acid (29.35 g, 164.92 mmol) was heated to 180.degree. C. for 15
min. The reaction mixture was poured into ice water (200 mL),
neutralized with Na.sub.2CO.sub.3, and then extracted with DCM (100
mL.times.3). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (DCM/MeOH=20:1)
to give the title compound (400 mg, 7%) as a yellow solid. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.35 (d, J=8.0 Hz, 1H), 7.24-7.18
(m, 1H), 7.09 (d, J=8.0 Hz, 1H), 4.57 (s, 2H), 3.70 (s, 2H). LCMS
M/Z (M+H) 182.
Step 2:
5-chloro-2-(1-methyl-1H-pyrazol-4-yl)-1,2-dihydroisoquinolin-3(4H)-
-one
##STR00286##
[0539] To a solution of 5-chloro-1,2-dihydroisoquinolin-3(4H)-one
(500 mg, 2.75 mmol) in 1,4-dioxane (10 mL) was added copper(I)
iodide (26 mg, 0.14 mmol), (1R,2R)-cyclohexane-1,2-diamine (63 mg,
0.55 mmol), K.sub.3PO.sub.4 (1.75 g, 8.26 mmol) and
4-iodo-1-methyl-1H-pyrazole (859 mg, 4.13 mmol). The reaction
mixture was heated to 120.degree. C. for 12 h under a nitrogen
atmosphere. After cooling to room temperature, the mixture was
filtered and concentrated in vacuo. Water (100 mL) was added and
extracted with EtOAc (50 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (DCM/MeOH=20:1) to give the title compound (80 mg,
11%) as a yellow solid. LCMS M/Z (M+H) 262.
Step 3: tert-butyl
1-(tetrahydro-2H-pyran-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00287##
[0541] To a solution of tert-butyl
3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
ne-5(4H)-carboxylate (Intermediate G, 400 mg, 1.04 mmol) in
1,4-dioxane (8 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (30 mg,
0.06 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (25 mg, 0.03 mmol), KOAc (300
mg, 3.12 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (526
mg, 2.08 mmol). The mixture was heated to 80.degree. C. for 2 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 434.
Step 4: tert-butyl
3-(2-(1-methyl-1H-pyrazol-4-yl)-3-oxo-1,2,3,4-tetrahydroisoquinolin-5-yl)-
-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-
-carboxylate
##STR00288##
[0543] To the above step cooled solution was added
5-chloro-2-(1-methyl-1H-pyrazol-4-yl)-1,2-dihydroisoquinolin-3(4H)-one
(100 mg, 0.38 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (25 mg, 0.03 mmol),
K.sub.3PO.sub.4 (244 mg, 1.15 mmol), 1,4-dioxane (2 mL) and water
(2 mL). The reaction mixture was heated to 90.degree. C. for 1 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. Water (100 mL) was
added and extracted with EtOAc (50 mL.times.3). The combined
organic layers were washed with brine (50 mL.times.2), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography
(DCM/MeOH=20:1) to give the title compound (60 mg, 12%) as a yellow
solid. LCMS M/Z (M+H) 533.
Step 5:
2-(1-methyl-1H-pyrazol-4-yl)-5-(1-(tetrahydro-2H-pyran-4-yl)-4,5,6-
,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-1,2-dihydroisoquinolin-3(4H)-
-one
##STR00289##
[0545] To a solution of tert-butyl
3-(2-(1-methyl-1H-pyrazol-4-yl)-3-oxo-1,2,3,4-tetrahydroisoquinolin-5-yl)-
-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-
-carboxylate (60 mg, 0.11 mmol) in DCM (10 mL) at 0.degree. C. was
added trifluoroacetic acid (3 mL, 40 mmol). The mixture was stirred
at 0.degree. C. for 2 h and concentrated in vacuo to give the title
compound (50 mg, crude) as brown oil that required no further
purification. LCMS M/Z (M+H) 433.
Step 6:
N-methyl-3-(2-(1-methyl-1H-pyrazol-4-yl)-3-oxo-1,2,3,4-tetrahydroi-
soquinolin-5-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3--
c]pyridine-5(4H)-carboxamide
##STR00290##
[0547] To a solution of
2-(1-methyl-1H-pyrazol-4-yl)-5-(1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetr-
ahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-1,2-dihydroisoquinolin-3(4H)-one
(50 mg, 0.12 mmol) in DCM (15 mL) was added triethylamine (0.10 mL,
0.70 mmol) and N-methyl-1H-imidazole-1-carboxamide (43 mg, 0.35
mmol). The reaction was stirred at room temperature for 16 h. DCM
(40 mL) was added and washed with brine (30 mL.times.3). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by
reverse phase chromatography (acetonitrile 23-53%/0.2% formic acid
in water) to give the title compound (14 mg, 24%) as a white solid.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.98 (s, 2H), 7.58 (s,
1H), 7.35-7.27 (m, 3H), 4.88 (s, 2H), 4.87-4.80 (m, 1H), 4.26 (s,
2H), 4.25-4.17 (m, 1H), 4.16-4.10 (m, 2H), 3.91 (s, 3H), 3.87-3.80
(m, 4H), 3.55 (t, J=11.6 Hz, 2H), 2.85-2.77 (m, 5H), 2.40-2.29 (m,
2H), 1.95-1.87 (m, 2H). LCMS M/Z (M+H) 490.
Example 38
N-methyl-3-(3-methyl-2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahydroisoqui-
nolin-5-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idine-5(4H)-carboxamide
##STR00291##
[0548] Step 1:
N-(1-(2-chlorophenyl)propan-2-yl)-1-methyl-1H-pyrazol-4-amine
##STR00292##
[0550] To a solution of 1-(2-chlorophenyl)propan-2-one (4.0 g, 23.7
mmol) in MeOH (50 mL) was added 1-methyl-1H-pyrazol-4-amine (3.49
g, 26.09 mmol) and NaHCO.sub.3 (3.99 g, 47.44 mmol). The reaction
mixture was heated to 50.degree. C. for 2 h under a nitrogen
atmosphere. After cooling to 0.degree. C., sodium borohydride (916
mg, 24.22 mmol) was added portionwise. The mixture was stirred at
room temperature for 2 h under a nitrogen atmosphere. The reaction
was concentrated in vacuo. Water (100 mL) was added and extracted
with EtOAc (50 mL.times.3). The combined organic layers were dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The crude residue was purified by silica gel chromatography
(DCM/MeOH=20:1) to give the title compound (2.2 g, 55%) as brown
oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.38-7.35 (m, 1H),
7.25-7.15 (m, 3H), 7.14 (s, 1H), 6.95 (s, 1H), 3.81 (s, 3H),
3.51-3.41 (m, 1H), 3.14-3.07 (m, 1H), 2.77-2.69 (m, 1H), 1.16 (d,
J=6.4 Hz, 3H). LCMS M/Z (M+H) 250.
Step 2:
5-chloro-3-methyl-2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahydroi-
soquinoline
##STR00293##
[0552] To a solution of
N-(1-(2-chlorophenyl)propan-2-yl)-1-methyl-1H-pyrazol-4-amine (1.0
g, 4 mmol) in AcOH (1 mL) was added paraformaldehyde (240 mg, 8.01
mmol). The mixture was stirred at room temperature for 5 min under
a nitrogen atmosphere. After cooling to 0.degree. C.,
H.sub.2SO.sub.4 (3.2 mL 1) was added. The mixture was stirred at
room temperature for 16 h under a nitrogen atmosphere. The reaction
mixture was poured into water (100 mL), basified with solid
NaHCO.sub.3 to pH 8 and then extracted with EtOAc (50 mL.times.3).
The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (DCM/MeOH=20:1)
to give the title compound (300 mg, 29%) as yellow oil. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.25-7.09 (m, 4H), 7.05-6.97 (m, 1H),
4.22-4.02 (m, 2H), 3.85-3.81 (m, 3H), 3.76-3.66 (m, 1H), 3.08-3.00
(m, 1H), 2.78-2.72 (m, 1H), 1.08 (d, J=6.4 Hz, 3H). LCMS M/Z (M+H)
262.
Step 3: tert-butyl
1-(tetrahydro-2H-pyran-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00294##
[0554] To a solution of tert-butyl
3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
ne-5(4H)-carboxylate (Intermediate G, 1.0 g, 2.59 mmol) in
1,4-dioxane (20 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (74 mg,
0.16 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (61 mg, 0.08 mmol), KOAc (762
mg, 7.77 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.32
g, 5.18 mmol). The mixture was heated to 80.degree. C. for 5 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 434.
Step 4: tert-butyl
3-(3-methyl-2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahydroisoquinolin-5--
yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(-
4H)-carboxylate
##STR00295##
[0556] To the above step cooled solution was added
5-chloro-3-methyl-2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahydroisoquino-
line (300 mg, 1.15 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (57 mg, 0.07 mmol),
K.sub.3PO.sub.4 (612 mg, 2.88 mmol), 1,4-dioxane (4 mL) and water
(6 mL). The reaction mixture was heated to 90.degree. C. for 1 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. Water (50 mL) was
added and extracted with EtOAc (30 mL.times.3). The combined
organic layers were washed with brine (30 mL.times.3), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography
(DCM/MeOH=20:1) to give the title compound (70 mg, 11%) as a yellow
solid. LCMS M/Z (M+H) 533.
Step 5:
3-methyl-2-(1-methyl-1H-pyrazol-4-yl)-5-(1-(tetrahydro-2H-pyran-4--
yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-1,2,3,4-tetrahydroi-
soquinoline
##STR00296##
[0558] To a solution of tert-butyl
3-(3-methyl-2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahydroisoquinolin-5--
yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(-
4H)-carboxylate (70 mg, 0.12 mmol) in DCM (10 mL) at 0.degree. C.
was added trifluoroacetic acid (0.11 mL, 1.2 mmol). The mixture was
stirred at 0.degree. C. for 2 h and concentrated in vacuo to give
the title compound (50 mg, crude) as brown oil that required no
further purification. LCMS M/Z (M+H) 433.
Step 6:
N-methyl-3-(3-methyl-2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahyd-
roisoquinolin-5-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4-
,3-c]pyridine-5(4H)-carboxamide
##STR00297##
[0560] To a solution of
3-methyl-2-(1-methyl-1H-pyrazol-4-yl)-5-(1-(tetrahydro-2H-pyran-4-yl)-4,5-
,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-1,2,3,4-tetrahydroisoquino-
line (50 mg, 0.12 mmol) in DCM (10 mL) was added triethylamine
(0.10 mL, 0.70 mmol) and N-methyl-1H-imidazole-1-carboxamide (43
mg, 0.35 mmol). The reaction was stirred at room temperature for 16
h. DCM (40 mL) was added and washed with brine (30 mL.times.3). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by
reverse phase chromatography (acetonitrile 23-53%/0.2% formic acid
in water) to give the title compound (2 mg, 3%) as a white solid.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.24-7.08 (m, 4H), 7.02
(s, 1H), 4.48-4.39 (m, 1H), 4.30-4.07 (m, 7H), 3.97-3.88 (m, 1H),
3.86 (s, 3H), 3.80-3.65 (m, 3H), 3.55 (t, J=11.6 Hz, 2H), 3.20-3.12
(m, 1H), 2.80 (d, J=4.4 Hz, 3H), 2.75-2.62 (m, 1H), 2.39-2.29 (m,
2H), 1.95-1.86 (m, 2H), 1.03 (d, J=6.4 Hz, 3H). LCMS M/Z (M+H)
490.
Example 39
N-methyl-3-(2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahydroisoquinolin-5-y-
l)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4-
H)-carboxamide
##STR00298##
[0561] Step 1:
5-bromo-2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahydroisoquinoline
##STR00299##
[0563] To a solution of 5-bromo-1,2,3,4-tetrahydroisoquinoline (1.0
g, 4.72 mmol) in ethylene glycol (15 mL) was added copper(I) iodide
(898 mg, 4.72 mmol), K.sub.3PO.sub.4 (3.0 g, 14.15 mmol) and
4-iodo-1-methyl-1H-pyrazole (1.96 g, 9.43 mmol). The reaction
mixture was heated to 120.degree. C. for 3 h under a nitrogen
atmosphere. After cooling to room temperature, the mixture was
filtered and concentrated in vacuo. Water (50 mL) was added and
extracted with DCM (30 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (petroleum ether/EtOAc=1:1) to give the title
compound (0.28 g, 42% purity) as a brown solid which was further
purified by reverse phase chromatography (acetonitrile 5-35/0.05%
HCl in water) to give the title compound (0.05 g, 4%, HCl salt) as
a light yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
9.77 (s, 1H), 8.65 (s, 1H), 8.26 (s, 1H), 8.11 (d, J=8.0 Hz, 1H),
7.99 (d, J=7.2 Hz, 1H), 7.60-7.54 (m, 1H), 4.55 (t, J=8.0 Hz, 2H),
3.95 (s, 3H), 3.38 (t, J=8.0 Hz, 2H), 2.54 (s, 2H). LCMS M/Z (M+H)
292.
Step 2: tert-butyl
1-(tetrahydro-2H-pyran-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00300##
[0565] To a solution of tert-butyl
3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
ne-5(4H)-carboxylate (Intermediate G, 100 mg, 0.26 mmol) in
1,4-dioxane (4 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (14 mg,
0.03 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (24 mg, 0.03 mmol), KOAc (76
mg, 0.78 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (131
mg, 0.52 mmol). The mixture was heated to 80.degree. C. for 2 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 434.
Step 3: tert-butyl
3-(2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahydroisoquinolin-5-yl)-1-(te-
trahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carbo-
xylate
##STR00301##
[0567] To the above step cooled solution was added
5-bromo-2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahydroisoquinoline
(25 mg, 0.09 mmol),
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (5 mg, 0.01
mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (8 mg, 0.01 mmol),
K.sub.3PO.sub.4 (59 mg, 0.28 mmol), 1,4-dioxane (1 mL) and water (1
mL). The reaction mixture was heated to 90.degree. C. for 1 h under
a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. DCM (50 mL) was
added and washed with water (40 mL), brine (40 mL). The organic
layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (DCM/MeOH=20:1) to give the title compound (50 mg,
52%) as brown oil. LCMS M/Z (M+H) 519.
Step 4:
2-(1-methyl-1H-pyrazol-4-yl)-5-(1-(tetrahydro-2H-pyran-4-yl)-4,5,6-
,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-1,2,3,4-tetrahydroisoquinoli-
ne
##STR00302##
[0569] To a solution of tert-butyl
3-(2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahydroisoquinolin-5-yl)-1-(te-
trahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carbo-
xylate (50 mg, 0.1 mmol) in DCM (1 mL) at 0.degree. C. was added
trifluoroacetic acid (0.07 mL, 0.96 mmol). The mixture was stirred
at room temperature for 1 h and concentrated in vacuo to give the
title compound (43 mg, crude) as brown oil that required no further
purification. LCMS M/Z (M+H) 419.
Step 5:
N-methyl-3-(2-(1-methyl-1H-pyrazol-4-yl)-1,2,3,4-tetrahydroisoquin-
olin-5-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyri-
dine-5(4H)-carboxamide
##STR00303##
[0571] To a solution of
2-(1-methyl-1H-pyrazol-4-yl)-5-(1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetr-
ahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-1,2,3,4-tetrahydroisoquinoline
(43 mg, 0.10 mmol) in DCM (3 mL) was added triethylamine (0.07 mL,
0.5 mmol) and N-methyl-1H-imidazole-1-carboxamide (25 mg, 0.2
mmol). The reaction was stirred at room temperature for 12 h. DCM
(50 mL) was added and washed with water (40 mL), brine (40 mL). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by
reverse phase chromatography (acetonitrile 15-45%/0.05% NH.sub.4OH
in water) to give the title compound (7 mg, 14%) as a white solid.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.30 (s, 1H), 7.22 (d,
J=7.6 Hz, 1H), 7.17-7.11 (m, 2H), 7.04 (s, 1H), 4.46-4.37 (m, 1H),
4.25-4.09 (m, 6H), 3.90-3.80 (m, 4H), 3.58-3.45 (m, 4H), 3.26-3.21
(m, 2H), 3.04-3.01 (m, 2H), 2.85-2.77 (m, 4H), 2.41-2.25 (m, 2H),
1.91-1.87 (m, 2H). LCMS M/Z (M+H) 476.
Example 40
N-methyl-3-(2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c]pyridin-
-1-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-
-5(4H)-carboxamide
##STR00304##
[0572] Step 1: tert-butyl
3-(2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c]pyridin-1-yl)-1-
-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-c-
arboxylate
##STR00305##
[0574] To a solution of tert-butyl
3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
ne-5(4H)-carboxylate (Intermediate G, 1.0 g, 2.59 mmol) in toluene
(20 mL) was added copper(I) iodide (25 mg, 0.13 mmol),
K.sub.3PO.sub.4 (2.2 g, 10.36 mmol),
(1R,2R)-cyclohexane-1,2-diamine (59 mg, 0.52 mmol) and
2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c]pyridine
(500 mg, 2.36 mmol). The mixture was heated to 120.degree. C. for
12 h under a nitrogen atmosphere. After cooling the reaction to
room temperature, the mixture was filtered and concentrated in
vacuo. Water (100 mL) was added and extracted with EtOAc (50
mL.times.3). The combined organic layers were washed with brine (50
mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (DCM/MeOH=20:1) to give the title compound (90 mg,
7%) as a yellow solid. LCMS M/Z (M+H) 518.
Step 2:
3-(2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c]pyridin--
1-yl)-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]py-
ridine
##STR00306##
[0576] To a solution of tert-butyl
3-(2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c]pyridin-1-yl)-1-
-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-c-
arboxylate (90 mg, 0.17 mmol) in DCM (10 mL) at 0.degree. C. was
added trifluoroacetic acid (0.06 mL, 0.87 mmol). The mixture was
stirred at 0.degree. C. for 2 h and concentrated in vacuo to give
the title compound (60 mg, crude) as a yellow solid that required
no further purification. LCMS M/Z (M+H) 418.
Step 3:
N-methyl-3-(2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c-
]pyridin-1-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]-
pyridine-5(4H)-carboxamide
##STR00307##
[0578] To a solution of
3-(2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-c]pyridin-1-yl)-1-
-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine
(60 mg, 0.14 mmol) in DCM (10 mL) at 0.degree. C. was added
triethylamine (0.04 mL, 0.29 mmol) and
N-methyl-1H-imidazole-1-carboxamide (36 mg, 0.29 mmol). The
reaction was stirred at room temperature for 16 h. DCM (40 mL) was
added and washed with brine (30 mL.times.3). The organic layer was
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The crude residue was purified by reverse phase
chromatography (acetonitrile 28-58%/0.2% formic acid in water) to
give the title compound (9 mg, 13%) as a yellow solid. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.59 (s, 1H), 8.08 (s, 1H), 7.95 (s,
1H), 7.68 (s, 1H), 6.46 (s, 1H), 5.10-5.02 (m, 1H), 4.31-4.18 (m,
1H), 4.16-4.11 (m, 4H), 3.97 (s, 3H), 3.86 (t, J=5.6 Hz, 2H), 3.56
(t, J=11.6 Hz, 2H), 2.91-2.80 (m, 2H), 2.77 (d, J=3.6 Hz, 3H), 2.42
(s, 3H), 2.36-2.25 (m, 2H), 1.98-1.87 (m, 2H). LCMS M/Z (M+H)
475.
Example 41
1-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,-
3-c]pyridin-3-yl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-indole-6-carbon-
itrile
##STR00308##
[0579] Step 1: 5-bromo-6-chloro-1-(phenylsulfonyl)-1H-indole
##STR00309##
[0581] To a stirred solution of 5-bromo-6-chloro-1H-indole (5.0 g,
21.69 mmol) in DMF (50 mL) at 0.degree. C. was added NaH (60%, 1.3
g, 32.54 mmol) and the mixture was stirred at room temperature for
30 min. Benzenesulfonyl chloride (3.33 mL, 26.03 mmol) was added
dropwise and the mixture stirred for an additional 2 h. The mixture
was quenched with sat. aq. NH.sub.4Cl (100 mL) and extracted with
EtOAc (100 mL.times.3). The combined organic layers were washed
with brine (50 mL.times.3), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude residue was
recrystalized with DCM (20 mL) and TBME (200 mL) to give the title
compound (6.0 g, 75%) as a yellow solid. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 8.13 (s, 1H), 7.98-7.93 (m, 2H), 7.90 (s, 1H),
7.74 (d, J=4.0 Hz, 1H), 7.69-7.63 (m, 1H), 7.60-7.51 (m, 2H), 6.73
(d, J=4.0 Hz, 1H).
Step 2: 5-bromo-6-chloro-2-methyl-1-(phenylsulfonyl)-1H-indole
##STR00310##
[0583] To a solution of
5-bromo-6-chloro-1-(phenylsulfonyl)-1H-indole (1.0 g, 2.56 mmol) in
THE (10 mL) was added LDA (1.92 mL, 3.84 mmol, 2 M in THF). The
reaction was stirred at -78.degree. C. for 2 h under a nitrogen
atmosphere. Iodomethane (0.24 mL, 3.84 mmol) was added drop wise,
the cooling bath was removed and the mixture was stirred at room
temperature for 1 h. The mixture was quenched with sat. aq.
NH.sub.4Cl (20 nL) and extracted with EtOAc (100 mL.times.3). The
combined organic layers were dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo to give the title compound (0.8
g, crude) as a yellow solid that required no further purification.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.33 (s, 1H), 7.81-7.75
(m, 2H), 7.65 (s, 1H), 7.64-7.57 (m, 1H), 7.54-7.46 (m, 2H), 6.27
(s, 1H), 2.57 (s, 3H).
Step 3:
6-chloro-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1-(phenylsulfonyl)--
1H-indole
##STR00311##
[0585] To a solution of
5-bromo-6-chloro-2-methyl-1-(phenylsulfonyl)-1H-indole (11.76 g, 26
mmol) in 1,4-dioxane (100 mL) and water (25 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (1.9 g,
2.6 mmol), Na.sub.2CO.sub.3 (5.51 g, 51.99 mmol) and
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(6.49 g, 31.19 mmol). The mixture was heated to 90.degree. C. for
12 h under a nitrogen atmosphere. After cooling the reaction to
room temperature, water (200 mL) was added and extracted with EtOAc
(200 mL.times.3). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography (petroleum
ether/EtOAc=2:1) to give the title compound (4.6 g, 46%) as a
yellow solid. LCMS M/Z (M+H) 386.
Step 4:
6-chloro-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-indole
##STR00312##
[0587] To a solution of
6-chloro-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1-(phenylsulfonyl)-1H-indo-
le (1.8 g, 4.66 mmol) in THE (30 mL) was added TBAF (9.33 ml, 9.33
mmol, 1 M in THF). The mixture was heated to 65.degree. C. for 2 h
under a nitrogen atmosphere. The reaction mixture was concentrated
in vacuo. The crude residue was dissolved in EtOAc (100 mL) and
washed with water (100 mL.times.3). The organic layer was dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The crude residue was purified by silica gel chromatography
(petroleum ether/EtOAc=6:1) to give the title compound (1.1 g, 86%)
as a yellow solid. LCMS M/Z (M+H) 246.
Step 5:
1-(3-(6-chloro-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-indol-1-yl-
)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-
-yl)ethanone
##STR00313##
[0589] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 1.1 g, 3.35 mmol) in
toluene (8 mL) was added copper(I) iodide (32 mg, 0.17 mmol),
K.sub.3PO.sub.4 (2.85 g, 13.41 mmol),
(1R,2R)-cyclohexane-1,2-diamine (77 mg, 0.67 mmol) and
6-chloro-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-indole (906 mg,
3.69 mmol). The mixture was heated to 120.degree. C. for 12 h under
a nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
The crude residue was dissolved in EtOAc (100 mL) and washed with
water (100 mL.times.3). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (DCM/MeOH=100:1)
to give the title compound (1.0 g, 55%) as a yellow solid. LCMS M/Z
(M+H) 493.
Step 6:
1-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyr-
azolo[4,3-c]pyridin-3-yl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-indole--
6-carbonitrile
##STR00314##
[0591] To a solution of
1-(3-(6-chloro-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-indol-1-yl)-1-(te-
trahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)eth-
anone (154 mg, 0.31 mmol) in 1,4-dioxane (1.5 mL) and water (1.5
mL) was added potassium hexacyanoferrate(II) trihydrate (92 mg,
0.16 mmol), KOAc (4 mg, 0.04 mmol),
methanesulfonato(2-di-t-butylphosphino-2',4',6'-tri-i-propyl-1,1'-bipheny-
l)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (25 mg, 0.03 mmol) and
2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (13 mg, 0.03
mmol). The mixture was heated to 120.degree. C. for 12 h under a
nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
The crude residue was purified by reverse phase chromatography
(acetonitrile 18-48%/0.2% formic acid in water) to give the title
compound (15 mg, 10%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.13 (s, 1H), 7.84 (s, 1H), 7.77 (s, 1H),
7.53-7.43 (m, 1H), 6.58 (s, 1H), 4.51-4.42 (m, 1H), 4.28-4.18 (m,
2H), 4.02-3.96 (m, 2H), 3.91 (s, 3H), 3.87-3.78 (m, 2H), 3.56-3.45
(m, 2H), 3.03-2.87 (m, 2H), 2.34-2.30 (m, 3H), 2.10-1.87 (m, 7H).
LCMS M/Z (M+H) 484.
Example 42
1-(3-(6-(difluoromethyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-indol-1--
yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4-
H)-yl)ethanone
##STR00315##
[0592] Step 1:
2,2-difluoro-2-(2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1-(phenylsulfonyl)--
1H-indol-6-yl)-1-phenylethanone
##STR00316##
[0594] To a solution of
6-chloro-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1-(phenylsulfonyl)-1H-indo-
le (4.0 g, 9.85 mmol) in toluene (40 mL) was added
2,2-difluoro-1-phenyl-ethanone (3.08 g, 19.7 mmol),
chloro[(tricyclohexylphosphine)-2-(2'-aminobiphenyl)]palladium(II)
(505 mg, 0.98 mmol) and K.sub.3PO.sub.4 (8.36 g, 39.39 mmol). The
reaction mixture was heated to 120.degree. C. for 16 h under a
nitrogen atmosphere. After cooling the reaction to room
temperature, water (100 mL) was added and extracted with DCM (100
mL.times.2). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (3 g, crude) as a yellow solid that required no
further purification. LCMS M/Z (M+H) 506.
Step 2:
6-(difluoromethyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1-(phenyl-
sulfonyl)-1H-indole
##STR00317##
[0596] To a solution of
2,2-difluoro-2-(2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1-(phenylsulfonyl)--
1H-indol-6-yl)-1-phenylethanone (0.5 g, 0.30 mmol) in toluene (5
mL) and water (0.08 mL) was added KOH (50 mg, 0.89 mmol). The
reaction mixture was heated to 100.degree. C. for 6 h. After
cooling the reaction to room temperature, EtOAc (100 mL) was added
and washed with water (50 mL.times.3). The combined organic layers
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (petroleum ether/EtOAc=2/1) to give the title
compound (0.2 g, 50%) as a yellow solid. LCMS M/Z (M+H) 402.
Step 3:
6-(difluoromethyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-indole
##STR00318##
[0598] To a solution of
6-(difluoromethyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1-(phenylsulfony-
l)-1H-indole (400 mg, 1.0 mmol) in THE (4 mL) was added TBAF (2.0
ml, 2.0 mmol, 1 M in THF). The mixture was heated to 65.degree. C.
for 8 h under a nitrogen atmosphere. The reaction mixture was
concentrated in vacuo. The crude residue was dissolved in EtOAc
(100 mL) and washed with water (100 mL.times.3). The organic layer
was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (petroleum ether/EtOAc=2:1) to give the title
compound (150 mg, 58%) as a yellow solid. LCMS M/Z (M+H) 262.
Step 4:
1-(3-(6-(difluoromethyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H--
indol-1-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone
##STR00319##
[0600] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 130 mg, 0.40 mmol) in
toluene (2 mL) was added copper(I) iodide (4 mg, 0.02 mmol),
K.sub.3PO.sub.4 (336 mg, 1.58 mmol),
(1R,2R)-cyclohexane-1,2-diamine (9 mg, 0.08 mmol) and
6-(difluoromethyl)-2-methyl-5-(1-methyl-1H-pyrazol-4-yl)-1H-indole
(114 mg, 0.44 mmol). The mixture was heated to 120.degree. C. for
12 h under a nitrogen atmosphere. After cooling the reaction to
room temperature, the mixture was filtered and concentrated in
vacuo. The crude residue was purified by reverse phase
chromatography (acetonitrile 25-55%/0.2% formic acid in water) to
give the title compound (31 mg, 15%) as a white solid. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.84 (s, 1H), 7.58 (s, 1H), 7.57
(s, 1H), 7.30-7.26 (m, 1H), 6.96 (t, J=55.2 Hz, 1H), 6.51 (s, 1H),
4.51-4.42 (m, 1H), 4.25-4.18 (m, 2H), 4.02-3.96 (m, 2H), 3.90 (s,
3H), 3.87-3.78 (m, 2H), 3.56-3.45 (m, 2H), 3.03-2.87 (m, 2H),
2.34-2.30 (m, 3H), 2.10-1.95 (m, 7H). LCMS M/Z (M+H) 509.
Example 43
1-(1-methyl-3-(4-(1-methyl-1H-pyrazol-4-yl)benzoyl)-6,7-dihydro-1H-pyrazol-
o[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00320##
[0601] Step 1: methyl 4-(1-methyl-1H-pyrazol-4-yl)benzoate
##STR00321##
[0603] To a solution of methyl 4-bromobenzoate (10 g, 46.5 mmol) in
1,4-dioxane (75 mL) and water (25 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (3.4 g,
4.65 mmol), Na.sub.2CO.sub.3 (14.8 g, 139.5 mmol) and
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(14.5 g, 69.8 mmol). The mixture was heated to 120.degree. C. for
12 h under a nitrogen atmosphere. After cooling the reaction to
room temperature, the mixture was filtered and concentrated in
vacuo. The crude residue was purified by silica gel chromatography
(petroleum ether/EtOAc=5:1) to give the title compound (4.6 g, 46%)
as yellow oil.
Step 2: 4-(1-methyl-1H-pyrazol-4-yl)benzoic Acid
##STR00322##
[0605] To a solution of methyl 4-(1-methyl-1H-pyrazol-4-yl)benzoate
(4.6 g, 21.3 mmol) in MeOH (50 mL) was added sodium hydroxide (5M,
42.6 mL). The mixture was heated to 50.degree. C. for 5 h. After
cooling the reaction to room temperature, the mixture was washed
with EtOAc (100 mL). The aqueous layer was acidified with HCl (2 N)
to pH 2-3 and then extracted with EtOAc (100 mL.times.3). The
combined organic layers were dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo to give the title compound (3.5
g, crude) as a white solid that required no further
purification.
Step 3:
N-methoxy-N-methyl-4-(1-methyl-1H-pyrazol-4-yl)benzamide
##STR00323##
[0607] To a solution of 4-(1-methyl-1H-pyrazol-4-yl)benzoic acid
(3.5 g, 17.3 mmol) in DCM (100 mL) was added
N,O-dimethylhydroxylamine hydrochloride (2.0 g, 20.8 mmol),
(dimethylamino)-N,N-dimethyl(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy)-me-
thaniminium hexafluorophosphate (7.9 g, 20.8 mmol) and
N,N-diisopropylethylamine (8.58 mL, 51.9 mmol). The mixture was
stirred at room temperature for 8 h. The reaction mixture was
washed with 1 N HCl (100 mL.times.2), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (petroleum
ether/EtOAc=3:1) to give the title compound (3.2 g, 75%) as a
yellow solid.
Step 4: tert-butyl
1-methyl-3-(4-(1-methyl-1H-pyrazol-4-yl)benzoyl)-6,7-dihydro-1H-pyrazolo[-
4,3-c]pyridine-5(4H)-carboxylate
##STR00324##
[0609] To a solution of tert-butyl
3-bromo-1-methyl-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(300 mg, 0.95 mmol) in THE (6 mL) at -78.degree. C. was slowly
added n-BuLi (2.5 M, 0.46 mL, 1.14 mmol) under a nitrogen
atmosphere. After stirred at -78.degree. C. for 30 min, the mixture
was added a solution of
N-methoxy-N-methyl-4-(1-methyl-1H-pyrazol-4-yl)benzamide (279 mg,
1.14 mmol) in THE (0.5 mL) by dropwise. The mixture was stirred at
-78.degree. C. for an additional 2 h. The reaction mixture was
quenched with water (20 mL) and extracted with EtOAc (20
mL.times.2). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by reverse phase chromatography (acetonitrile
45-75%/0.2% formic acid in water) to give the title compound (50
mg, 13%) as a white solid.
Step 5:
(4-(1-methyl-1H-pyrazol-4-yl)phenyl)(1-methyl-4,5,6,7-tetrahydro-1-
H-pyrazolo[4,3-c]pyridin-3-yl)methanone
##STR00325##
[0611] A mixture of tert-butyl
1-methyl-3-(4-(1-methyl-1H-pyrazol-4-yl)benzoyl)-6,7-dihydro-1H-pyrazolo[-
4,3-c]pyridine-5(4H)-carboxylate (50 mg, 0.12 mmol) and
trifluoroacetic acid (2 mL) in DCM (2 mL) was stirred at room
temperature for 3 h. The mixture was concentrated in vacuo to give
the title compound (30 mg, crude) as brown oil that required no
further purification.
Step 6:
1-(1-methyl-3-(4-(1-methyl-1H-pyrazol-4-yl)benzoyl)-6,7-dihydro-1H-
-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00326##
[0613] To a solution of
(4-(1-methyl-1H-pyrazol-4-yl)phenyl)(1-methyl-4,5,6,7-tetrahydro-1H-pyraz-
olo[4,3-c]pyridin-3-yl)methanone (30 mg, 0.09 mmol) in DCM (1 mL)
was added triethylamine (0.026 mL, 0.19 mmol) and acetic anhydride
(0.018 mL, 0.19 mmol). The mixture was stirred at room temperature
for 2 h. DCM (5 mL) was added and washed with water (5 mL.times.2).
The organic layer was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude residue was purified
by reverse phase chromatography (acetonitrile 32-62%/0.2% formic
acid in water) to give the title compound (6 mg, 18%) as a white
solid. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33-8.28 (m, 2H),
8.11 (s, 1H), 7.94 (s, 1H), 7.71-7.68 (m, 2H), 4.82 (s, 2H), 3.96
(s, 3H), 3.95-3.84 (m, 2H), 4.90 (s, 3H), 3.91-3.74 (m, 2H),
2.23-2.21 (m, 3H). LCMS M/Z (M+H) 364.
Example 44
(S)-1-(3-(6-(1-methyl-1H-pyrazol-4-yl)-3,4-dihydronaphthalen-1-yl)-1-(tetr-
ahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
##STR00327##
[0614] Step 1:
6-(1-methyl-1H-pyrazol-4-yl)-3,4-dihydronaphthalen-1(2H)-one
##STR00328##
[0616] To a solution of 6-bromo-3,4-dihydronaphthalen-1(2H)-one
(0.9 g, 4.0 mmol) in 1,4-dioxane (8 mL) and water (2 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) (146
mg, 0.20 mmol), K.sub.2CO.sub.3 (1.38 mg, 10.0 mmol) and
1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole
(915 mg, 4.4 mmol). The mixture was heated to 120.degree. C. for 16
h under a nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
The crude residue was purified by silica gel chromatography
(petroleum ether/EtOAc=5:1) to give the title compound (800 mg,
88%) as yellow oil. LCMS M/Z (M+H) 227.
Step 2:
4-methyl-N'-(6-(1-methyl-1H-pyrazol-4-yl)-3,4-dihydronaphthalen-1(-
2H)-ylidene)benzenesulfonohydrazide
##STR00329##
[0618] To a solution of
6-(1-methyl-1H-pyrazol-4-yl)-3,4-dihydronaphthalen-1(2H)-one (0.8
g, 3.54 mmol) in MeOH (10 mL) was added
4-methylbenzenesulfonohydrazide (725 mg, 3.89 mmol). The mixture
was heated to 70.degree. C. for 3 h under a nitrogen atmosphere.
After cooling the reaction to room temperature, the mixture was
filtered to give the title compound (1.2 g, 86%) as a white
solid.
Step 3:
(S)-1-(3-(6-(1-methyl-1H-pyrazol-4-yl)-3,4-dihydronaphthalen-1-yl)-
-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)e-
thanone
##STR00330##
[0620] To a solution of
(S)-1-(3-bromo-1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate E, 670 mg, 2.13 mmol) in
1,4-dioxane (5 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (65 mg,
0.09 mmol), t-BuOLi (312 mg, 3.9 mmol) and
4-methyl-N-(6-(1-methyl-1H-pyrazol-4-yl)-3,4-dihydronaphthalen-1(2H)-ylid-
ene)benzenesulfonohydrazide (0.7 g, 1.77 mmol). The mixture was
heated to 100.degree. C. for 3 h under a nitrogen atmosphere. After
cooling the reaction to room temperature, the mixture was filtered
and concentrated in vacuo. The crude residue was purified by silica
gel chromatography (DCM/MeOH=20:1) to give the title compound (680
mg, 86%) as a brown solid. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.70-7.68 (m, 1H), 7.55-7.53 (m, 1H), 7.23-7.10 (m, 3H),
6.13-6.10 (m, 1H), 4.77-4.74 (m, 1H), 4.38-4.20 (m, 2H), 4.11-4.06
(m, 2H), 4.00-3.66 (m, 4H), 3.87 (s, 3H), 2.82-2.71 (m, 4H),
2.40-2.29 (m, 4H), 2.11-1.99 (m, 3H). LCMS M/Z (M+H) 444.
Example 45
N-methyl-3-(3-(2-methylthiazol-5-yl)isoquinolin-8-yl)-1-(1-oxidotetrahydro-
-2H-thiopyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxami-
de
##STR00331##
[0621] Step 1: tetrahydro-2H-thiopyran-4-yl Methanesulfonate
##STR00332##
[0623] To a solution of tetrahydro-2H-thiopyran-4-ol (10 g, 84.6
mmol) and triethylamine (35.4 mL, 253.8 mmol) in DCM (150 mL) at
0.degree. C. was added methanesulfonyl chloride (10.7 mL, 138.8
mmol) dropwise under a nitrogen atmosphere. The mixture was stirred
at 25.degree. C. for 16 h. Water (100 mL) was added and extracted
with DCM (100 mL.times.2). The combined organic layers were dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo
to give the title compound (17 g, crude) as yellow oil that
required no further purification. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 4.73-4.69 (m, 1H), 3.19 (s, 3H), 2.76-2.63
(m, 4H), 2.17-2.16 (m, 2H), 1.87-1.84 (m, 2H).
Step 2: tert-butyl
3-bromo-1-(tetrahydro-2H-thiopyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]py-
ridine-5(4H)-carboxylate
##STR00333##
[0625] To a solution of tert-butyl
3-bromo-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(Intermediate A, 10 g, 33.1 mmol) in DMF (50 mL) was added
Cs.sub.2CO.sub.3 (27 g, 82.7 mmol) and tetrahydro-2H-thiopyran-4-yl
methanesulfonate (8.4 g, 43.0 mmol). The mixture was heated to
80.degree. C. for 16 h under a nitrogen atmosphere. After cooling
the reaction to room temperature, the mixture was filtered. The
mixture was diluted with EtOAc (100 mL) and washed with brine (100
mL.times.2). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (petroleum
ether:tert-butyl methyl ether:THF=from 10:1:1 to 3:1:1) to give the
title compound (5.9 g, 44%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 4.17 (s, 2H), 4.09-4.04 (m, 1H), 3.62-3.59
(m, 2H), 2.83-2.77 (m, 2H), 2.71-2.68 (m, 4H), 2.13-2.10 (m, 2H),
2.03-1.93 (m, 2H), 1.44 (s, 9H).
Step 3: tert-butyl
3-bromo-1-(1-oxidotetrahydro-2H-thiopyran-4-yl)-6,7-dihydro-1H-pyrazolo[4-
,3-c]pyridine-5(4H)-carboxylate
##STR00334##
[0627] To a solution of tert-butyl
3-bromo-1-(tetrahydro-2H-thiopyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]py-
ridine-5(4H)-carboxylate (500 mg, 1.24 mmol) in THE (4 mL) and
water (1 mL) at 0.degree. C. was added Oxone (382 mg, 0.62 mmol)
portionwise. The mixture was stirred at 25.degree. C. for 1 h. The
reaction was quenched by sat. aq. Na.sub.2SO.sub.3 (5 mL) and
extracted with DCM (5 mL.times.3). The combined organic layers were
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo to give the title compound (511 mg, crude) as a white solid
that required no further purification. LCMS M/Z (M+H) 418.
Step 4: 5-(8-chloroisoquinolin-3-yl)-2-methylthiazole
##STR00335##
[0629] To a solution of 8-chloroisoquinolin-3-yl
trifluoromethanesulfonate (3.0 g, 9.6 mmol) in 1,4-dioxane (10 mL)
and water (2 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) (0.7
g, 0.96 mmol), Na.sub.2CO.sub.3 (3.1 g, 28.9 mmol) and
2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole
(2.2 g, 9.6 mmol). The mixture was heated to 90.degree. C. for 12 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, DCM (80 mL) was added and washed with water (50
mL.times.2), brine (30 mL). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography (petroleum
ether/EtOAc=2:1) to give the title compound (0.8 g, 32%) as a
yellow solid. LCMS M/Z (M+H) 261.
Step 5:
2-methyl-5-(8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquin-
olin-3-yl)thiazole
##STR00336##
[0631] To a solution of
5-(8-chloroisoquinolin-3-yl)-2-methylthiazole (300 mg, 1.1 mmol) in
1,4-dioxane (4 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (55 mg, 0.1
mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl-
)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (90 mg, 0.1 mmol),
KOAc (282 mg, 2.9 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (876
mg, 3.5 mmol). The mixture was heated to 90.degree. C. for 1 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 353.
Step 6: tert-butyl
3-(3-(2-methylthiazol-5-yl)isoquinolin-8-yl)-1-(1-oxidotetrahydro-2H-thio-
pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00337##
[0633] To the above step cooled solution was added tert-butyl
3-bromo-1-(1-oxidotetrahydro-2H-thiopyran-4-yl)-6,7-dihydro-1H-pyrazolo[4-
,3-c]pyridine-5(4H)-carboxylate (100 mg, 0.2 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (19 mg, 0.02 mmol),
K.sub.3PO.sub.4 (152 mg, 0.7 mmol), 1,4-dioxane (4 mL) and water (2
mL). The reaction mixture was heated to 90.degree. C. for 4 h under
a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. DCM (60 mL) was
added and washed with water (50 mL.times.2), brine (50 mL). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by silica
gel chromatography (DCM/MeOH=50:1) to give the title compound (55
mg, 41%) as a brown solid. LCMS M/Z (M+H) 564.
Step 7:
4-(3-(3-(2-methylthiazol-5-yl)isoquinolin-8-yl)-4,5,6,7-tetrahydro-
-1H-pyrazolo[4,3-c]pyridin-1-yl)tetrahydro-2H-thiopyran 1-oxide
##STR00338##
[0635] To a solution of tert-butyl
3-(3-(2-methylthiazol-5-yl)isoquinolin-8-yl)-1-(1-oxidotetrahydro-2H-thio-
pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(55 mg, 0.1 mmol) in DCM (2 mL) was added trifluoroacetic acid (1.0
mL, 12 mmol). The mixture was stirred at room temperature for 2 h
and concentrated in vacuo to give the title compound (40 mg, crude)
as a yellow solid that required no further purification. LCMS M/Z
(M+H) 464.
Step 8:
N-methyl-3-(3-(2-methylthiazol-5-yl)isoquinolin-8-yl)-1-(1-oxidote-
trahydro-2H-thiopyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-c-
arboxamide
##STR00339##
[0637] To a solution of
4-(3-(3-(2-methylthiazol-5-yl)isoquinolin-8-yl)-4,5,6,7-tetrahydro-1H-pyr-
azolo[4,3-c]pyridin-1-yl)tetrahydro-2H-thiopyran 1-oxide (40 mg,
0.09 mmol) in DCM (3 mL) was added triethylamine (0.06 mL, 0.43
mmol) and N-methyl-1H-imidazole-1-carboxamide (54 mg, 0.4 mmol).
The reaction was stirred at room temperature for 5 h and
concentrated in vacuo. DCM (30 mL) was added and washed with water
(10 mL.times.3), brine (10 mL). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by reverse phase chromatography
(acetonitrile 20-50%/0.05% NH.sub.4OH in water) to give the title
compound (0.7 mg, 2%) as a yellow solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.74 (s, 1H), 8.42 (s, 1H), 8.38 (s, 1H),
7.98 (d, J=8.0 Hz, 1H), 7.90-7.83 (m, 1H), 7.62 (d, J=7.2 Hz, 1H),
6.65-6.56 (m, 1H), 4.65-4.54 (m, 1H), 4.42 (s, 2H), 3.76-3.67 (m,
2H), 2.95-2.89 (m, 2H), 2.86-2.67 (m, 2H), 2.71 (s, 3H), 2.51-2.12
(m, 9H). LCMS M/Z (M+H) 521.
Example 46
1-(1-acetylpiperidin-4-yl)-N-methyl-3-(3-(2-methylthiazol-5-yl)isoquinolin-
-8-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxamide
##STR00340##
[0638] Step 1: 1-acetylpiperidin-4-yl Methanesulfonate
##STR00341##
[0640] To a solution of 1-(4-hydroxy-1-piperidyl)ethanone (200 mg,
1.4 mmol) in DCM (5 mL) at 0.degree. C. was added triethylamine
(212 mg, 2.1 mmol) and methanesulfonyl chloride (480 mg, 4.19
mmol). The mixture was stirred at 25.degree. C. for 2 h. Water (50
mL) was added and the mixture was extracted with DCM (50
mL.times.3). The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (300 mg, crude) as yellow oil that required no
further purification. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
4.98-4.92 (m, 1H), 3.83-3.81 (m, 1H), 3.67-3.65 (m, 1H), 3.58-3.56
(m, 1H), 3.43-3.41 (m, 1H), 3.06 (s, 3H), 2.01 (s, 3H), 2.00-1.88
(m, 4H).
Step 2: tert-butyl
1-(1-acetylpiperidin-4-yl)-3-bromo-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-
-5(4H)-carboxylate
##STR00342##
[0642] To a solution of tert-butyl
3-bromo-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
(Intermediate A, 5.0 g, 16.6 mmol) in DMF (40 mL) was added
Cs.sub.2CO.sub.3 (19.5 g, 59.6 mmol) and 1-acetylpiperidin-4-yl
methanesulfonate (5.5 g, 24.8 mmol). The mixture was heated to
90.degree. C. for 12 h under a nitrogen atmosphere. After cooling
to room temperature, the mixture was filtered and concentrated in
vacuo. DCM (100 mL) was added and washed with brine (80
mL.times.2). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (petroleum
ether:tert-butyl methyl ether:THF=from 10:1:1 to 2:1:1) to give the
title compound (2.0 g, 28%) as clear oil. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 4.50-4.41 (m, 1H), 4.38-4.29 (m, 1H), 4.16
(s, 2H), 3.94-3.85 (m, 1H), 3.64-3.57 (m, 2H), 3.21-3.09 (m, 1H),
2.75-2.58 (m, 3H), 2.03 (s, 3H), 1.91-1.80 (m, 3H), 1.73-1.61 (m,
1H), 1.41 (s, 9H).
Step 3: tert-butyl
1-(1-acetylpiperidin-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl-
)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00343##
[0644] To a solution of tert-butyl
1-(1-acetylpiperidin-4-yl)-3-bromo-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-
-5(4H)-carboxylate (500 mg, 1.17 mmol) in 1,4-dioxane (10 mL) was
added 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (56
mg, 0.12 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl-
)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (92 mg, 0.12 mmol),
KOAc (345 mg, 3.51 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (740
mg, 2.93 mmol). The mixture was heated to 80.degree. C. for 2 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 475.
Step 4: tert-butyl
1-(1-acetylpiperidin-4-yl)-3-(3-(2-methylthiazol-5-yl)isoquinolin-8-yl)-6-
,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00344##
[0646] To the above step cooled solution was added
5-(8-chloroisoquinolin-3-yl)-2-methylthiazole (302 mg, 1.16 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (45 mg, 0.06 mmol),
K.sub.3PO.sub.4 (738 mg, 3.48 mmol), 1,4-dioxane (5 mL) and water
(3 mL). The reaction mixture was heated to 90.degree. C. for 12 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. DCM (20 mL) was
added and washed with water (15 mL.times.2), brine (20 mL). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by silica
gel chromatography (DCM/MeOH=20:1) to give the title compound (200
mg, 30%) as a yellow solid. LCMS M/Z (M+H) 573.
Step 5:
1-(4-(3-(3-(2-methylthiazol-5-yl)isoquinolin-8-yl)-4,5,6,7-tetrahy-
dro-1H-pyrazolo[4,3-c]pyridin-1-yl)piperidin-1-yl)ethanone
##STR00345##
[0648] To a solution of tert-butyl
1-(1-acetylpiperidin-4-yl)-3-(3-(2-methylthiazol-5-yl)isoquinolin-8-yl)-6-
,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate (200 mg,
0.35 mmol) in DCM (3 mL) was added trifluoroacetic acid (0.27 mL,
3.6 mmol). The mixture was stirred at room temperature for 2 h and
concentrated in vacuo to give the title compound (100 mg, crude) as
yellow oil that required no further purification. LCMS M/Z (M+H)
473.
Step 6:
1-(1-acetylpiperidin-4-yl)-N-methyl-3-(3-(2-methylthiazol-5-yl)iso-
quinolin-8-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxamide
##STR00346##
[0650] To a solution of
1-(4-(3-(3-(2-methylthiazol-5-yl)isoquinolin-8-yl)-4,5,6,7-tetrahydro-1H--
pyrazolo[4,3-c]pyridin-1-yl)piperidin-1-yl)ethanone (100 mg, 0.21
mmol) in DCM (2 mL) was added triethylamine (0.1 mL, 0.70 mmol) and
N-methyl-1H-imidazole-1-carboxamide (58 mg, 0.47 mmol). The
reaction was stirred at room temperature for 12 h and concentrated
in vacuo. The crude residue was purified by reverse phase
chromatography (acetonitrile 30-60%/0.05% NH.sub.4OH in water) to
give the title compound (62 mg, 55%) as a white solid. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 9.58 (s, 1H), 8.16 (s, 1H), 7.93 (s,
1H), 7.81 (d, J=8.4 Hz, 1H), 7.73-7.68 (m, 1H), 7.51 (d, J=7.2 Hz,
1H), 4.83-4.78 (m, 1H), 4.65-4.59 (m, 1H), 4.34 (s, 2H), 4.32-4.22
(m, 1H), 4.07-4.00 (m, 1H), 3.94-3.82 (m, 2H), 3.30-3.21 (m, 1H),
2.88-2.83 (m, 2H), 2.78 (d, J=4.8 Hz, 1H), 2.76 (s, 3H), 2.42-2.30
(m, 1H), 2.21-2.00 (m, 6H). LCMS M/Z (M+H) 530.
Example 47
3-isopropyl-N-methyl-1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-4,5--
dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxamide
##STR00347##
[0651] Step 1: ethyl 2-(benzyl(3-cyanopropyl)amino)acetate
##STR00348##
[0653] To a solution of ethyl 2-(benzylamino)acetate (50 g, 258.7
mmol) and K.sub.2CO.sub.3 (71.5 g, 517.5 mmol) in MeCN (500 mL) was
added 4-bromobutyronitrile (28.7 mL, 284.6 mmol) dropwise. The
mixture was heated to 80.degree. C. for 12 h under a nitrogen
atmosphere. After cooling to room temperature, the mixture was
filtered and concentrated in vacuo. The crude residue was purified
by silica gel chromatography (petroleum ether/EtOAc=5:1) to give
the title compound (56 g, 83%) as light yellow oil. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.37-7.28 (m, 5H), 4.18 (q, J=7.2 Hz,
2H), 3.80 (s, 2H), 3.32 (s, 2H), 2.80 (t, J=6.4 Hz, 2H), 2.47 (t,
J=7.2 Hz, 2H), 1.87-1.77 (m, 2H), 1.29 (t, J=7.2 Hz, 3H).
Step 2: ethyl
2-((tert-butoxycarbonyl)(3-cyanopropyl)amino)acetate
##STR00349##
[0655] To a solution of ethyl 2-(benzyl(3-cyanopropyl)amino)acetate
(22 g, 84.5 mmol) and Boc.sub.2O (38.8 mL, 169 mmol) in EtOH (150
mL) was added 10% Pd/C (2.3 g). The mixture was stirred at room
temperature for 6 h under a hydrogen atmosphere (40 Psi). The
mixture was filtered and the filtrate was concentrated in vacuo.
The crude residue was purified by silica gel chromatography
(petroleum ether/EtOAc=3:1) to give the title compound (17 g, 74%)
as colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
4.25-4.14 (m, 2H), 3.94-3.82 (m, 2H), 3.43-3.39 (m, 2H), 2.49-2.41
(m, 2H), 1.96-1.84 (m, 2H), 1.51-1.37 (m, 9H), 1.30-1.27 (m,
2H).
Step 3: tert-butyl 4-cyano-3-oxopiperidine-1-carboxylate
##STR00350##
[0657] To a solution of t-BuOK (8.5 g, 75.5 mmol) in toluene (100
mL) at 0.degree. C. was added ethyl
2-((tert-butoxycarbonyl)(3-cyanopropyl)amino)acetate (17 g, 62.9
mmol). The mixture was stirred at room temperature for 30 min under
a nitrogen atmosphere. Sat. aq. NH.sub.4Cl (200 mL) was added and
washed with hexanes (200 mL). The aqueous phase was acidified with
HCl (2 N) to pH 6 and then extracted with EtOAc (200 mL.times.2).
The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound (13 g, 92%) as yellow oil that required no further
purification.
Step 4: tert-butyl
3-amino-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate
##STR00351##
[0659] A mixture of tert-butyl
4-cyano-3-oxopiperidine-1-carboxylate (13 g, 57.9 mmol) and
hydrazine mono-hydrate (85%, 6.6 mL, 116 mmol) in EtOH (100 mL) was
heated to 60.degree. C. for 3 h. The mixture was concentrated in
vacuo to give the crude product that was dissolved in EtOAc (150
mL) and washed with sat. aq. Na.sub.2CO.sub.3 (100 mL), water (100
mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude residue was purified
by silica gel chromatography (DCM/MeOH=20:1) to give the title
compound (10 g, 72%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 4.28 (s, 2H), 3.48 (d, J=5.6 Hz, 2H), 2.27
(d, J=5.6 Hz, 2H), 1.40 (s, 9H).
Step 5: tert-butyl
3-bromo-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate
##STR00352##
[0661] To a stirred mixture of tert-butyl
3-amino-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate
(10 g, 42 mmol), CuBr.sub.2 (10.3 g, 46.2 mmol) in MeCN (80 mL) at
0.degree. C. was added isopentyl nitrite (7.4 mL, 54.6 mmol)
dropwise and the reaction mixture stirred for 20 min. The
temperature was raised to 60.degree. C. and the reaction mixture
was stirred for an additional 5 h. After cooling the reaction to
room temperature, the reaction mixture was quenched with water (200
mL) and the mixture was extracted with EtOAc (200 mL.times.2). The
combined organic layers were dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude residue was purified
by silica gel chromatography (petroleum ether/EtOAc=1:1) to give
the title compound (6 g, 47%) as a white solid. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 4.45 (s, 2H), 3.55 (d, J=5.6 Hz, 2H),
2.35 (d, J=5.6 Hz, 2H), 1.41 (s, 9H).
Step 6: tert-butyl
3-(prop-1-en-2-yl)-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxyla-
te
##STR00353##
[0663] To a solution of tert-butyl
3-bromo-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate (2
g, 6.6 mmol) in 1,4-dioxane (25 mL) and water (5 mL) was added
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (484
mg, 0.7 mmol), Na.sub.2CO.sub.3 (2.1 g, 19.9 mmol) and
4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (5.6 g,
33.1 mmol). The mixture was heated to 100.degree. C. for 12 h under
a nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
EtOAc (150 mL) was added and washed with water (140 mL.times.2).
The organic layer was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude residue was purified
by silica gel chromatography (petroleum ether/EtOAc=3:1) to give
the title compound (1.3 g, 73%) as a yellow solid. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 12.72-12.35 (m, 1H), 5.23 (s, 1H), 5.08
(s, 1H), 4.41 (s, 2H), 3.60-3.49 (m, 2H), 2.67-2.55 (m, 2H), 2.05
(s, 3H), 1.41 (s, 9H).
Step 7: tert-butyl
3-isopropyl-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate
##STR00354##
[0665] a solution of tert-butyl
3-(prop-i-en-2-yl)-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxyla-
te (1.2 g, 4.6 mmol) in MeOH (8 mL) was added 10% Pd/C (485 mg).
The mixture was stirred at room temperature for 12 h under a
hydrogen atmosphere (15 Psi). The mixture was filtered and the
filtrate was concentrated in vacuo to give the title compound (860
mg, 71%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 12.24 (s, 1H), 4.37 (s, 2H), 3.52 (d, J=6.0 Hz, 2H),
2.98-2.85 (m, 1H), 2.48 (d, J=6.0 Hz, 2H), 1.41 (s, 9H), 1.18 (d,
J=7.2 Hz, 6H).
Step 8: tert-butyl
3-isopropyl-1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-4,5-dihydro--
1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate
##STR00355##
[0667] To a solution of tert-butyl
3-isopropyl-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate
(240 mg, 0.9 mmol) in toluene (8 mL) was added
tris(dibenzylideneacetone)-dipalladium (83 mg, 0.09 mmol),
2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (38 mg, 0.09
mmol), t-BuONa (261 mg, 2.7 mmol) and
8-chloro-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline (286 mg, 1.2
mmol). The mixture was heated to 110.degree. C. for 12 h under a
nitrogen atmosphere. After cooling the reaction to room
temperature, the mixture was filtered and concentrated in vacuo.
EtOAc (50 mL) was added and washed with water (40 mL) and brine (40
mL). The organic phase was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude residue was purified
by silica gel chromatography (petroleum ether/EtOAc=2:1) to give
the title compound (150 mg, 35%) as a yellow solid. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 9.02 (s, 1H), 8.05 (s, 1H), 8.01 (s,
1H), 7.89-7.76 (m, 2H), 7.73-7.59 (m, 1H), 7.44 (d, J=7.6 Hz, 1H),
4.36 (s, 2H), 3.99 (s, 3H), 3.76-3.67 (m, 2H), 3.12-3.05 (m, 1H),
2.76-2.67 (m, 2H), 1.44 (s, 9H), 1.37 (d, J=6.4 Hz, 6H).
Step 9:
8-(3-isopropyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-1-yl)--
3-(1-methyl-1H-pyrazol-4-yl)isoquinoline
##STR00356##
[0669] To a solution of tert-butyl
3-isopropyl-1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-4,5-dihydro--
1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate (150 mg, 0.32 mmol) in
DCM (0.5 mL) was added trifluoroacetic acid (0.46 mL, 6.35 mmol).
The mixture was stirred at room temperature for 12 h and
concentrated in vacuo to give the title compound (120 mg, crude) as
brown oil that required no further purification. LCMS M/Z (M+H)
373.
Step 10:
3-isopropyl-N-methyl-1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-
-yl)-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxamide
##STR00357##
[0671] To a solution of
8-(3-isopropyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-1-yl)-3-(1-me-
thyl-1H-pyrazol-4-yl)isoquinoline (120 mg, 0.32 mmol) in DCM (2 mL)
was added triethylamine (0.09 mL, 0.64 mmol) and
N-methyl-1H-imidazole-1-carboxamide (81 mg, 0.64 mmol). The
reaction was stirred at room temperature for 12 h and concentrated
in vacuo. DCM (50 mL) was added and washed with water (40 mL),
brine (40 mL). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by Prep-TLC (DCM/MeOH=10:1) to give the title
compound (62 mg, 45%) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.99 (s, 1H), 8.03 (s, 1H), 8.00 (s, 1H), 7.83
(d, J=8.4 Hz, 1H), 7.80 (s, 1H), 7.70-7.66 (m, 1H), 7.43 (d, J=7.6
Hz, 1H), 4.62-4.46 (m, 1H), 4.35 (s, 2H), 3.97 (s, 3H), 3.65 (d,
J=5.6 Hz, 2H), 3.12-3.02 (m, 1H), 2.77 (d, J=4.4 Hz, 3H), 2.74 (d,
J=5.6 Hz, 2H), 1.35 (d, J=6.8 Hz, 6H). LCMS M/Z (M+H) 430.
Example 48
N-methyl-1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-3-morpholino-4,5-
-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxamide
##STR00358##
[0672] Step 1:
3-(1-methyl-1H-pyrazol-4-yl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)isoquinoline
##STR00359##
[0674] To a solution of
8-chloro-3-(1-methyl-1H-pyrazol-4-yl)isoquinoline (10.0 g, 41.04
mmol) in 1,4-dioxane (220 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (1.96 g,
4.1 mmol) and
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphe-
nyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (3.23 g, 4.1 mmol),
KOAc (10.1 g, 102.59 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (31.26
g, 123.11 mmol). The mixture was heated to 90.degree. C. for 1 h
under a nitrogen atmosphere. After cooling to room temperature,
EtOAc (400 mL) was added and washed with water (200 mL.times.2).
The organic layer was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude residue was purified
by silica gel chromatography (petroleum ether/EtOAc=1:1) to give
the title compound (11.0 g, 22% purity) as a brown solid that
required no further purification. LCMS M/Z (M+H) 336.
Step 2: (3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)boronic
acid
##STR00360##
[0676] To a solution of
3-(1-methyl-1H-pyrazol-4-yl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)isoquinoline (10 g, crude) in acetone (50 mL) and water (50 mL)
was added NaIO.sub.4 (16.0 g, 74.6 mmol) and NH.sub.4OAc (5.7 g,
74.6 mmol). The mixture was stirred at room temperature for 48 h.
The reaction was filtered and concentrated in vacuo. EtOAc (100 mL)
was added and washed with water (70 mL), brine (70 mL). The organic
layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (DCM/MeOH=10:1) to give the title compound (440 mg,
4% over 2 steps) as a yellow solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 9.64 (s, 1H), 8.57-8.51 (m, 2H), 8.31 (s,
1H), 8.06 (s, 1H), 8.01 (s, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.78 (d,
J=5.6 Hz, 1H), 7.73-7.63 (m, 1H), 3.91 (s, 3H).
Step 3: tert-butyl
3-bromo-1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-4,5-dihydro-1H-p-
yrazolo[3,4-c]pyridine-6(7H)-carboxylate
##STR00361##
[0678] To a solution of tert-butyl
3-bromo-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate
(400 mg, 1.3 mmol) in THE (10 mL) was added
(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)boronic acid (469 mg,
1.6 mmol), triethylamine (0.92 mL, 6.6 mmol), pyridine (1.43 mL,
13.2 mmol) and copper(II) acetate (721 mg, 4.0 mmol). The mixture
was heated to 60.degree. C. for 12 h under an oxygent atmosphere.
After cooling to room temperature, the mixture was filtered and
concentrated in vacuo. EtOAc (100 mL) was added and washed with
water (80 mL.times.2), brine (80 mL). The organic layer was dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The crude residue was purified by silica gel chromatography
(petroleum ether/EtOAc=1:1) to give the title compound (100 mg,
15%) as a yellow solid. LCMS M/Z (M+H) 511.
Step 4: tert-butyl
1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-3-morpholino-4,5-dihydro-
-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate
##STR00362##
[0680] To a solution of tert-butyl
3-bromo-1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-4,5-dihydro-1H-p-
yrazolo[3,4-c]pyridine-6(7H)-carboxylate (100 mg, 0.2 mmol) in
1,4-dioxane (4 mL) was
dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridy-
l)palladium(II), t-BuONa (57 mg, 0.6 mmol) and morpholine (0.035
mL, 0.4 mmol). The mixture was heated to 110.degree. C. for 16 h
under an argon atmosphere. After cooling to room temperature, DCM
(50 mL) was added and washed with water (40 mL.times.2), brine (40
mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The crude residue was purified
by silica gel chromatography (petroleum ether/EtOAc=1:1) to give
the title compound (42 mg, 41%) as a yellow solid. LCMS M/Z (M+H)
516.
Step 5:
4-(1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-4,5,6,7-tetrah-
ydro-1H-pyrazolo[3,4-c]pyridin-3-yl)morpholine
##STR00363##
[0682] To a solution of tert-butyl
1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-3-morpholino-4,5-dihydro-
-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxylate (42 mg, 0.08 mmol) in
DCM (1 mL) was added trifluoroacetic acid (0.06 mL, 0.8 mmol). The
mixture was stirred at room temperature for 3 h and concentrated in
vacuo to give the title compound (35 mg, crude) as brown oil that
required no further purification. LCMS M/Z (M+H) 416.
Step 6:
N-methyl-1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-3-morpho-
lino-4,5-dihydro-1H-pyrazolo[3,4-c]pyridine-6(7H)-carboxamide
##STR00364##
[0684] To a solution of
4-(1-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-4,5,6,7-tetrahydro-1H-
-pyrazolo[3,4-c]pyridin-3-yl)morpholine (35 mg, 0.08 mmol) in DCM
(3 mL) was added triethylamine (0.04 mL, 0.25 mmol) and
N-methyl-1H-imidazole-1-carboxamide (16 mg, 0.13 mmol). The
reaction was stirred at room temperature for 12 h and concentrated
in vacuo. EtOAc (50 mL) was added and washed with water (40
mL.times.2). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by reverse phase chromatography (acetonitrile
17-47%/0.05% HCl in water) to give the title compound (6 mg, 15%)
as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.38
(s, 1H), 8.49 (s, 1H), 8.40 (s, 1H), 8.22 (s, 1H), 8.01 (d, J=8.4
Hz, 1H), 7.98-7.90 (m, 1H), 7.62 (d, J=6.8 Hz, 1H), 6.61 (s, 1H),
4.43 (s, 2H), 3.94 (s, 3H), 3.76-3.70 (m, 4H), 3.65-3.55 (m, 2H),
3.23-3.13 (m, 4H), 2.65-2.55 (m, 2H), 2.52 (s, 3H). LCMS M/Z (M+H)
473.
Example 49
2-cyano-N-methyl-3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetra-
hydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrrolo[3,2-c]pyridine-5(4H)-carboxami-
de
##STR00365##
[0685] Step 1: 2-((tetrahydro-2H-pyran-4-yl)amino)acetonitrile
##STR00366##
[0687] To a solution of tetrahydro-2H-pyran-4-amine hydrochloride
(45 g, 327 mmol) and K.sub.2CO.sub.3 (135.6 g, 981 mmol) in MeCN
(300 mL) was added 2-chloroacetonitrile (24.9 mL, 392 mmol)
dropwise. The mixture was heated to 80.degree. C. for 16 h under a
nitrogen atmosphere. After cooling to room temperature, the mixture
was filtered and concentrated in vacuo. The crude residue was
purified by silica gel chromatography (petroleum ether/EtOAc=1:1)
to give the title compound (26 g, 57%) as brown oil. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 3.95-3.88 (m, 2H), 3.59 (s, 2H),
3.42-3.35 (m, 2H), 2.95-2.82 (m, 1H), 1.82-1.68 (m, 2H), 1.42-1.29
(m, 2H).
Step 2: tert-butyl
3-cyano-4-(((trifluoromethyl)sulfonyl)oxy)-5,6-dihydropyridine-1(2H)-carb-
oxylate
##STR00367##
[0689] To a solution of tert-butyl
3-cyano-4-oxopiperidine-1-carboxylate (21 g, 93.6 mmol) in DCM (200
mL) at 0.degree. C. was added triethylamine (17 mL, 121 mmol),
N,N-dimethylpyridin-4-amine (2.3 g, 18.7 mmol) and
trifluoromethanesulfonic anhydride (20.4 mL, 121 mmol). The
reaction was stirred at room temperature for 6 h. DCM (100 mL) was
added and washed with water (200 mL), brine (200 mL). The organic
layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (petroleum ether/EtOAc=3:1) to give the title
compound (20 g, 60%) as a yellow solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 4.26 (s, 2H), 3.62-3.56 (m, 2H), 2.73-2.62
(m, 2H), 1.42 (s, 9H).
Step 3: tert-butyl
3-amino-2-cyano-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrrolo[3,2-c-
]pyridine-5(4H)-carboxylate
##STR00368##
[0691] To a solution of tert-butyl
3-cyano-4-(((trifluoromethyl)sulfonyl)oxy)-5,6-dihydropyridine-1(2H)-carb-
oxylate (5 g, 14 mmol) in t-BuOH (70 mL) was added
2-((tetrahydro-2H-pyran-4-yl)amino)acetonitrile (2.4 g, 16.8 mmol)
and Cs.sub.2CO.sub.3 (13.7 g, 42 mmol). The mixture was heated to
90.degree. C. for 16 h under a nitrogen atmosphere. After cooling
to room temperature, the mixture was filtered and concentrated in
vacuo. EtOAc (250 mL) was added and washed with water (200
mL.times.2), brine (200 mL). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography (petroleum
ether/EtOAc=1:1) to give the title compound (270 mg, 6%) as a
yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.23 (s,
2H), 4.15-4.09 (m, 2H), 4.06-4.00 (m, 1H), 3.75-3.65 (m, 2H),
3.55-3.41 (m, 4H), 2.70-2.58 (m, 2H), 2.41-2.30 (m, 2H), 1.84-1.74
(m, 2H), 1.48 (s, 9H). LCMS M/Z (M+H) 347.
Step 4: tert-butyl
3-bromo-2-cyano-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrrolo[3,2-c-
]pyridine-5(4H)-carboxylate
##STR00369##
[0693] To a stirred mixture of tert-butyl
3-amino-2-cyano-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrrolo[3,2-c-
]pyridine-5(4H)-carboxylate (510 mg, 1.47 mmol), CuBr.sub.2 (428
mg, 1.91 mmol) in MeCN (30 mL) at 0.degree. C. was added isopentyl
nitrite (0.26 mL, 1.91 mmol) dropwise and the reaction mixture
stirred for 20 min. The temperature was raised to 60.degree. C. and
the reaction mixture was stirred for an additional 5 h. After
cooling the reaction to room temperature, the reaction mixture was
quenched with water (60 mL) and the mixture was extracted with
EtOAc (60 mL.times.2). The combined organic layers were washed with
brine (40 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (petroleum ether/EtOAc=3:2) to give the title
compound (170 mg, 28%) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 4.28 (s, 2H), 4.24-4.18 (m, 1H), 4.16-4.12 (m,
2H), 3.80-3.68 (m, 2H), 3.51-3.45 (m, 2H), 2.78-2.68 (m, 2H),
2.51-2.33 (m, 2H), 1.87-1.80 (m, 2H), 1.50 (s, 9H).
Step 5: tert-butyl
2-cyano-3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-2H-
-pyran-4-yl)-6,7-dihydro-1H-pyrrolo[3,2-c]pyridine-5(4H)-carboxylate
##STR00370##
[0695] To a solution of tert-butyl
3-bromo-2-cyano-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrrolo[3,2-c-
]pyridine-5(4H)-carboxylate (170 mg, 0.41 mmol) in THE (15 mL) and
water (3 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (20 mg,
0.041 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (33 mg, 0.041 mmol),
Na.sub.2CO.sub.3 (134 mg, 1.24 mmol) and
3-(1-methyl-1H-pyrazol-4-yl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)isoquinoline (181 mg, 0.54 mmol). The mixture was heated to
60.degree. C. for 12 h under a nitrogen atmosphere. After cooling
to room temperature, the mixture was filtered and concentrated in
vacuo. EtOAc (55 mL) was added and washed with water (40
mL.times.2). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (petroleum
ether/EtOAc=3:2) to give the title compound (192 mg, 86%) as a
yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.07 (s,
1H), 8.06 (s, 1H), 8.00 (s, 1H), 7.86-7.78 (m, 2H), 7.71-7.66 (m,
1H), 7.46 (d, J=6.8 Hz, 1H), 4.46-4.32 (m, 1H), 4.29-4.08 (m, 4H),
4.05-3.65 (m, 5H), 3.58-3.52 (m, 2H), 2.93-2.80 (m, 2H), 2.58-2.49
(m, 2H), 2.01-1.94 (m, 2H), 1.62 (s, 9H). LCMS M/Z (M+H) 539.
Step 6:
3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-2H--
pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine-2-carbonitrile
##STR00371##
[0697] To a solution of tert-butyl
2-cyano-3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-2H-
-pyran-4-yl)-6,7-dihydro-1H-pyrrolo[3,2-c]pyridine-5(4H)-carboxylate
(192 mg, 0.36 mmol) in DCM (0.5 mL) was added trifluoroacetic acid
(0.52 mL, 7.12 mmol). The mixture was stirred at room temperature
for 3 h and concentrated in vacuo to give the title compound (145
mg, crude) as brown oil that required no further purification. LCMS
M/Z (M+H) 439.
Step 7:
2-cyano-N-methyl-3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)--
1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrrolo[3,2-c]pyridine-5(4H)-c-
arboxamide
##STR00372##
[0699] To a solution of
3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-2H-pyran-4-
-yl)-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine-2-carbonitrile
(145 mg, 0.33 mmol) in DCM (3 mL) was added triethylamine (0.1 mL,
0.66 mmol) and N-methyl-1H-imidazole-1-carboxamide (83 mg, 0.66
mmol). The reaction was stirred at room temperature for 12 h and
concentrated in vacuo. DCM (30 mL) was added and washed with water
(30 mL.times.2). The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by Prep-TLC (DCM/MeOH=10:1) to give the title
compound (73 mg, 55%) as a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 9.05 (s, 1H), 8.04 (s, 1H), 7.99 (s, 1H),
7.86-7.80 (m, 2H), 7.73-7.68 (m, 1H), 7.46 (d, J=6.4 Hz, 1H),
4.46-4.32 (m, 2H), 4.25-4.15 (m, 2H), 4.12-3.95 (m, 6H), 3.75-3.64
(m, 1H), 3.55 (t, J=12.0 Hz, 2H), 2.96-2.84 (m, 2H), 2.73 (d, J=3.6
Hz, 3H), 2.62-2.45 (m, 2H), 2.03-1.93 (m, 2H). LCMS M/Z (M+H)
496.
Example 50
8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,-
3-c]pyridin-3-yl)imidazo[1,2-a]pyridine-2-carboxylic Acid
##STR00373##
[0701] To a solution of methyl
8-(5-acetyl-1-tetrahydropyran-4-yl-6,7-dihydro-4H-pyrazolo[4,3-c]pyridin--
3-yl)imidazo[1,2-a]pyridine-2-carboxylate (50 mg, 0.15 mmol),
obtained in a similar fashion as procedure S, in THE (0.35 mL) was
added lithium chloride (3.3 mg, 0.078 mmol) and sodium hydroxide
3.7 mol/L in water (77 .mu.L). The mixture was stirred at
55.degree. C. for 3 h. The reaction mixture was then cooled to room
temperature, concentrated HCl was added (20 .mu.L) and the solution
was concentrated in vacuo. The mixture obtained was purified by
reverse phase chromatography (acetonitrile 2-20%/0.1% ammonium
hydroxide in water) to give the title compound (5.0 mg, 17%) as a
white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6, 22/23 H) .delta.
8.58 (dt, J=6.9, 1.5 Hz, 1H), 8.47 (s, 1H), 7.53-7.40 (m, 1H), 7.03
(td, J=6.9, 3.8 Hz, 1H), 4.89 (s, 1H), 4.60 (s, 1H), 4.39 (tq,
J=11.9, 4.2 Hz, 1H), 3.98 (dt, J=10.1, 4.6 Hz, 2H), 3.79 (t, J=6.0
Hz, 2H), 3.55-3.43 (m, 2H), 2.95-2.75 (m, 2H), 2.19-2.01 (m, 5H),
1.87 (dt, J=13.5, 3.3 Hz, 2H). LCMS M/Z (M+H) 410.
Example 51
1-(3-(2-(pyridin-3-yl)-1H-benzo[d]imidazol-4-yl)-1-(tetrahydro-2H-pyran-4--
yl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one
##STR00374##
[0702] Step 1: 4-chloro-2-(pyridin-3-yl)-1H-benzo[d]imidazole
##STR00375##
[0704] To a solution of pyridine-3-carbonyl chloride hydrochloride
(157 mg, 0.884 mmol) and trimethylamine (224 mg, 2.21 mmol) in DCM
(5.3 mL) was added 3-chlorobenzene-1,2-diamine (105 mg, 0.736
mmol). The mixture was stirred at room temperature for 16 h. The
reaction mixture was then cooled to room temperature and saturated
aqueous NaHCO.sub.3 solution (20 mL) and brine solution (5 mL) were
added and the two layers were separated. The aqueous layer was
washed with DCM (2.times.10 mL). The combined organic layers were
dried over anhydrous MgSO.sub.4, filtered and concentrated in
vacuo. The residue obtained was dissolved in acetic acid (1.8 mL)
and concentrated HCl was added (60 .mu.L, 0.74 mmol). The mixture
was stirred at 100.degree. C. for 2 h. The reaction mixture was
then cooled to room temperature and concentrated in vacuo. The
crude residue was partitioned between DCM (20 mL) and saturated
aqueous NaHCO.sub.3 solution (20 mL). The two phases were separated
and the aqueous layer was extracted with DCM (2.times.15 mL). The
combined organic layers were washed with brine, dried over
anhydrous MgSO.sub.4, filtered and concentrated in vacuo. The
mixture obtained was purified by silica gel chromatography
(acetone/heptane=1:19 to 2:3) to give the title compound (97.4 mg,
58%) as a beige solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6, 7/8 H)
.delta. 8.75 (dd, J=4.8, 1.7 Hz, 1H), 7.53-7.45 (m, 1H), 7.39-7.27
(m, 2H), 9.30 (dd, J=2.3, 1.0 Hz, 1H), 8.42 (dt, J=8.0, 2.0 Hz,
1H), 7.74 (d, J=7.8 Hz, 1H). LCMS M/Z (M+H) 230.
Step 2:
1-(3-(2-(pyridin-3-yl)-1H-benzo[d]imidazol-4-yl)-1-(tetrahydro-2H--
pyran-4-yl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one
##STR00376##
[0706] To a solution of
1-(3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyr-
idin-5(4H)-yl)ethanone (Intermediate H, 50 mg, 0.15 mmol) and
bis(pinacolato)diboron (77 mg, 0.30 mmol) in dioxane (0.76 mL) was
added KOAc (45 mg, 0.46 mmol),
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-
-1,1'-biphenyl)]palladium(II) methanesulfonate (3.7 mg, 0.0046
mmol) and 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl
(4.4 mg, 0.0091 mmol). The mixture was stirred at 80.degree. C. for
16 h under a nitrogen atmosphere. The reaction mixture was cooled
to room temperature and 4-chloro-2-(3-pyridyl)-1H-benzimidazole (30
mg, 0.13 mmol), K.sub.3PO.sub.4.H.sub.2O (40 mg, 0.19 mmol), water
(0.3 mL) and
(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-
-1,1'-biphenyl)]palladium(II) methanesulfonate (3.7 mg, 0.0046
mmol) were added. The reaction mixture was stirred at 90.degree. C.
for 4 h under a nitrogen atmosphere. The reaction mixture was then
cooled to room temperature and concentrated in vacuo. The crude
residue was dissolved in dichloromethane (5 mL), dried over
anhydrous MgSO.sub.4, filtered through celite and concentrated in
vacuo. The mixture obtained was purified by reverse phase
chromatography (acetonitrile 5-50%/0.1% ammonium hydroxide in
water) to give the title compound (30.4 mg, 45%) as a white solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 13.49-12.78 (m, 1H),
9.52-9.31 (m, 1H), 8.68 (dt, J=3.9, 1.6 Hz, 1H), 8.60 (s, 1H), 7.60
(dt, J=8.1, 4.3 Hz, 2H), 7.45 (s, 1H), 7.34-7.29 (m, 1H), 4.85 (s,
2H), 4.41 (s, 1H), 4.04-3.97 (m, 2H), 3.88-3.77 (m, 2H), 3.51 (t,
J=11.8 Hz, 2H), 3.39-3.33 (m, 2H), 2.99-2.79 (m, 2H), 2.25-2.07 (m,
2H), 2.01 (s, 1H), 1.91 (d, J=12.7 Hz, 2H). LCMS M/Z (M+H) 443.
Example 52
N-methyl-3-(2-(1-methyl-1H-pyrazol-4-yl)-2,3,4,5-tetrahydro-1H-benzo[c]aze-
pin-6-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyrid-
ine-5(4H)-carboxamide
##STR00377##
[0707] Step 1:
6-bromo-2,3,4,5-tetrahydro-1H-benzo[c]azepin-1-one
##STR00378##
[0709] To a solution of 5-bromo-1-tetralone (3.0 g, 13.33 mmol) in
conc. HCl (30 mL) at 0.degree. C. was added sodium azide (1.79 g,
27.53 mmol) portionwise. The reaction mixture was heated to
50.degree. C. for 16 h under a nitrogen atmosphere. After cooling
to room temperature, the mixture was basified with K.sub.2CO.sub.3
(1 M) to pH 10 and then extracted with EtOAc (100 mL.times.2). The
combined organic layers were washed with sat. aq. NaHCO.sub.3 (50
mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (petroleum ether/EtOAc=3:2) to give the title
compound (1.2 g, 37%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.22-8.13 (m, 1H), 7.75-7.71 (m, 1H), 7.48
(d, J=6.8 Hz, 1H), 7.29-7.25 (m, 1H), 2.93 (t, J=6.8 Hz, 2H),
2.90-2.84 (m, 2H), 1.90-1.82 (m, 2H).
Step 2:
6-bromo-2-(1-methyl-1H-pyrazol-4-yl)-2,3,4,5-tetrahydro-1H-benzo[c-
]azepin-1-one
##STR00379##
[0711] To a solution of
6-bromo-2,3,4,5-tetrahydro-1H-benzo[c]azepin-1-one (1.5 g, 6.25
mmol) in 1,4-dioxane (50 mL) was added copper(I) iodide (119 mg,
0.62 mmol), (1R,2R)-cyclohexane-1,2-diamine (142 mg, 1.25 mmol),
K.sub.3PO.sub.4 (3.9 g, 18.74 mmol) and 4-iodo-1-methyl-1H-pyrazole
(1.95 g, 9.37 mmol). The reaction mixture was heated to 120.degree.
C. for 48 h under a nitrogen atmosphere. After cooling to room
temperature, the mixture was filtered and concentrated in vacuo.
Water (100 mL) was added and extracted with EtOAc (50 mL.times.3).
The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by silica gel chromatography (DCM/MeOH=20:1)
to give the title compound (900 mg, 45%) as a yellow solid. LCMS
M/Z (M+H) 320.
Step 3:
6-bromo-2-(1-methyl-1H-pyrazol-4-yl)-2,3,4,5-tetrahydro-1H-benzo[c-
]azepine
##STR00380##
[0713] To a solution of
6-bromo-2-(1-methyl-1H-pyrazol-4-yl)-2,3,4,5-tetrahydro-1H-benzo[c]azepin-
-1-one (900 mg, 2.81 mmol) in THE (50 mL) was added borane dimethyl
sulfide complex (1.41 mL, 14.1 mmol, 10 M). The reaction mixture
was heated to 70.degree. C. for 16 h under a nitrogen atmosphere.
After cooling to room temperature, the reaction was quenched with
MeOH (20 mL) and heated to 70.degree. C. for an additional 1 h.
After cooling to room temperature, the mixture was filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (DCM/MeOH=10:1) to give the title compound (700 mg,
81%) as gray oil. LCMS M/Z (M+H) 306.
Step 4: tert-butyl
1-(tetrahydro-2H-pyran-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxylate
##STR00381##
[0715] To a solution of tert-butyl
3-bromo-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridi-
ne-5(4H)-carboxylate (Intermediate G, 400 mg, 1.04 mmol) in
1,4-dioxane (8 mL) was added
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (30 mg,
0.06 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (25 mg, 0.03 mmol), KOAc (300
mg, 3.12 mmol) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (526
mg, 2.08 mmol). The mixture was heated to 80.degree. C. for 2 h
under a nitrogen atmosphere. After cooling the reaction to room
temperature, the reaction mixture was used to the next step
directly without further purification. LCMS M/Z (M+H) 434.
Step 5: tert-butyl
3-(2-(1-methyl-1H-pyrazol-4-yl)-2,3,4,5-tetrahydro-1H-benzo[c]azepin-6-yl-
)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H-
)-carboxylate
##STR00382##
[0717] To the above step cooled solution was added
6-bromo-2-(1-methyl-1H-pyrazol-4-yl)-2,3,4,5-tetrahydro-1H-benzo[c]azepin-
e (141 mg, 0.46 mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-am-
ino-1,1'-biphenyl-2-yl) palladium(II) (25 mg, 0.03 mmol),
K.sub.3PO.sub.4 (244 mg, 1.15 mmol), 1,4-dioxane (2 mL) and water
(2 mL). The reaction mixture was heated to 90.degree. C. for 1 h
under a nitrogen atmosphere. After cooling to room temperature, the
mixture was filtered and concentrated in vacuo. Water (50 mL) was
added and extracted with EtOAc (30 mL.times.3). The combined
organic layers were washed with brine (30 mL.times.3), dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
crude residue was purified by silica gel chromatography
(DCM/MeOH=20:1) to give the title compound (150 mg, 27%) as a
yellow solid. LCMS M/Z (M+H) 533.
Step 6:
2-(1-methyl-1H-pyrazol-4-yl)-6-(1-(tetrahydro-2H-pyran-4-yl)-4,5,6-
,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-2,3,4,5-tetrahydro-1H-benzo[-
c]azepine
##STR00383##
[0719] To a solution of tert-butyl
3-(2-(1-methyl-1H-pyrazol-4-yl)-2,3,4,5-tetrahydro-1H-benzo[c]azepin-6-yl-
)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H-
)-carboxylate (150 mg, 0.28 mmol) in DCM (10 mL) at 0.degree. C.
was added trifluoroacetic acid (0.42 mL, 5.63 mmol). The mixture
was stirred at 0.degree. C. for 2 h and concentrated in vacuo to
give the title compound (120 mg, crude) as brown oil that required
no further purification. LCMS M/Z (M+H) 433.
Step 7:
N-methyl-3-(2-(1-methyl-1H-pyrazol-4-yl)-2,3,4,5-tetrahydro-1H-ben-
zo[c]azepin-6-yl)-1-(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo[4,3-
-c]pyridine-5(4H)-carboxamide
##STR00384##
[0721] To a solution of
2-(1-methyl-1H-pyrazol-4-yl)-6-(1-(tetrahydro-2H-pyran-4-yl)-4,5,6,7-tetr-
ahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)-2,3,4,5-tetrahydro-1H-benzo[c]azepi-
ne (120 mg, 0.28 mmol) in DCM (15 mL) was added triethylamine (0.12
mL, 0.83 mmol) and N-methyl-1H-imidazole-1-carboxamide (69 mg, 0.55
mmol). The reaction was stirred at room temperature for 16 h. DCM
(40 mL) was added and washed with brine (30 mL.times.3). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The crude residue was purified by
reverse phase chromatography (acetonitrile 8-38%/0.2% formic acid
in water) to give the title compound (38 mg, 28%) as a white solid.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.26-7.21 (m, 1H),
7.17-7.07 (m, 3H), 6.79 (s, 1H), 4.42 (s, 2H), 4.25-4.08 (m, 5H),
3.83 (t, J=5.2 Hz, 2H), 3.76 (s, 3H), 3.58-3.48 (m, 4H), 3.02-2.95
(m, 2H), 2.85-2.75 (m, 5H), 2.39-2.25 (m, 2H), 1.92-1.84 (m, 2H),
1.82-1.74 (m, 2H). LCMS M/Z (M+H) 490.
Example 53
1-(3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro-2H-pyran-
-4-yl)-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridin-5-yl)ethan-1-one
##STR00385##
[0722] Step 1:
5-benzyl-1-(triisopropylsilyl)-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridi-
ne
##STR00386##
[0724] To a solution of
5-benzyl-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine (1.50 g, 7.06
mmol) in THE (18 mL) at 0.degree. C. was added sodium hydride 60%
in mineral oil (424 mg, 10.6 mmol). The solution was stirred at
0.degree. C. for 30 min, followed by the addition of
triisopropylsilyl chloride (1.82 mL, 8.48 mmol). The mixture was
stirred at 0.degree. C. for 15 min and then heated at 60.degree. C.
for 2 h. The reaction mixture was then cooled to room temperature
and concentrated in vacuo. Water (50 mL) and iPrOAc (50 mL) were
added and the two layers were separated. The aqueous layer was
extracted with iPrOAc (40 mL). The combined organic layers were
dried over anhydrous MgSO.sub.4, filtered and concentrated in
vacuo. The residue obtained was purified by silica gel
chromatography (iPrOAc/heptane=1:9) to give the title compound
(2.60 g, quant.) as a pale yellow oil. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.43-7.20 (m, 5H), 6.67 (d, J=2.9 Hz, 1H), 6.00
(d, J=2.8 Hz, 1H), 3.69 (s, 2H), 3.54 (s, 2H), 2.73 (s, 4H),
1.60-1.54 (m, 2H), 1.51-1.39 (m, 3H), 1.10 (d, J=7.5 Hz, 18H). LCMS
M/Z (M+H) 369.
Step 2:
1-(triisopropylsilyl)-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine
##STR00387##
[0726] To a 100 mL round bottom flask was added
5-benzyl-1-(triisopropylsilyl)-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridi-
ne (1.00 g, 2.71 mmol) and palladium 10% on carbon (433 mg, 0.407
mmol) in EtOH (9.0 mL) and AcOH (2.7 mL). The solution was purged
three times with hydrogen gas and stirred at 30.degree. C. for 4 h
under 1 atm of hydrogen. The hydrogen was removed in vacuo, the
reaction mixture was filtered through celite and the solvent was
concentrated in vacuo to afford the title compound (870 mg, quant.)
as an orange oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 6.73
(d, J=2.9 Hz, 1H), 6.06 (d, J=2.8 Hz, 1H), 4.15 (s, 2H), 3.39 (t,
J=6.0 Hz, 2H), 2.96 (t, J=5.8 Hz, 2H), 1.51-1.39 (m, 3H), 1.10 (d,
J=7.4 Hz, 18H). LCMS M/Z (M+H) 279.
Step 3:
1-(1-(triisopropylsilyl)-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyrid-
in-5-yl)ethan-1-one
##STR00388##
[0728] To a solution of
1-(triisopropylsilyl)-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine
(1.50 g, 7.06 mmol) in DCM (6.2 mL) at 0.degree. C. was added
triethylamine (947 mg, 9.36 mmol) followed by acetic anhydride (382
mg, 3.74 mmol). The solution was stirred at room temperature for 2
h. To the reaction mixture was then added saturated aqueous
NaHCO.sub.3 solution (40 mL) and DCM (10 mL) and the two layers
were separated. The aqueous layer was washed with DCM (2.times.30
mL). The combined organic layers were washed with brine, dried over
anhydrous MgSO.sub.4, filtered and concentrated in vacuo. The
residue obtained was purified by silica gel chromatography
(iPrOAc/heptane=3:7) to give the title compound (837 mg, 84%) as a
white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 6.76-6.68
(m, 1H), 6.07 (d, J=2.8 Hz, 1H), 4.63-4.46 (m, 2H), 3.90-3.63 (m,
2H), 2.82-2.68 (m, 2H), 2.19-2.11 (m, 3H), 1.56-1.39 (m, 3H),
1.13-1.07 (m, 18H). LCMS M/Z (M+H) 321.
Step 4:
1-(3-bromo-1-(triisopropylsilyl)-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-
-c]pyridin-5-yl)ethan-1-one
##STR00389##
[0730] To a solution of
1-(1-(triisopropylsilyl)-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridin-5-yl-
)ethan-1-one (400 mg, 1.25 mmol) in THE (4.2 mL) at -78.degree. C.
under a nitrogen atmosphere was added N-bromosuccinimide (244 mg,
1.37 mmol). The reaction mixture was stirred at -78.degree. C. for
2h30. The reaction mixture was then added to stirring MeOH (30 mL)
at -78.degree. C. and the the mixture was concentrated in vacuo.
Water (40 mL) and iPrOAc (40 mL) and the two layers were separated.
The aqueous layer was washed with iPrOAc (40 mL). The combined
organic layers were washed with brine, dried over anhydrous
MgSO.sub.4, filtered and concentrated in vacuo. The residue
obtained was purified by silica gel chromatography
(iPrOAc/heptane=3:7 to 7:3) to give the title compound (239 mg,
48%) as a pale yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 6.74-6.66 (m, 1H), 4.55-4.32 (m, 2H), 3.88-3.61 (m, 2H),
2.78-2.65 (m, 2H), 2.25-2.15 (m, 3H), 1.53-1.36 (m, 3H), 1.13-1.07
(m, 18H). LCMS M/Z (M+H) 399.
Step 5:
1-(3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(triisopropy-
lsilyl)-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridin-5-yl)ethan-1-one
##STR00390##
[0732] To a solution of
1-(3-bromo-1-(triisopropylsilyl)-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyri-
din-5-yl)ethan-1-one (225 mg, 0.563 mmol) and
3-(1-methyl-1H-pyrazol-4-yl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)isoquinoline (227 mg, 0.676 mmol) in 1,4-dioxane (1.9 mL) and
water (0.47 mL) was added K.sub.3PO.sub.4.H.sub.2O (334 mg, 1.41
mmol),
chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-
-amino-1,1'-biphenyl)]palladium(II) (23 mg, 0.028 mmol) and
2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (13 mg,
0.028 mmol). The mixture was stirred at 140.degree. C. for 12 min
in a microwave under a nitrogen atmosphere. The crude mixture was
diluted in DCM (30 mL), filtered through celite and concentrated in
vacuo. The residue obtained was purified by silica gel
chromatography (MeOH/DCM=1:9 to 3:7) to give the title compound
(149 mg, 50%) as a brown-orange solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 9.31 (s, 1H), 8.07-7.84 (m, 2H), 7.86-7.56 (m,
3H), 7.39 (dt, J=6.9, 1.3 Hz, 1H), 6.86 (d, J=2.3 Hz, 1H),
4.52-4.44 (m, 1H), 4.33 (s, 1H), 3.98 (s, 3H), 3.94 (t, J=5.7 Hz,
1H), 3.76 (t, J=5.7 Hz, 1H), 2.77-2.95 (m, 2H), 2.03-1.98 (m, 3H),
1.58-1.46 (m, 3H), 1.17 (dd, J=7.6, 2.6 Hz, 18H). LCMS M/Z (M+H)
528.
Step 6:
1-(3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1,4,6,7-tetrah-
ydro-5H-pyrrolo[3,2-c]pyridin-5-yl)ethan-1-one
##STR00391##
[0734] To a solution of
1-(3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(triisopropylsilyl)-
-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridin-5-yl)ethan-1-one (145
mg, 0.275 mmol) in THE (0.92 mL) was added tetrabutylammonium
fluoride 1 M in THE (0.330 mL, 0.330 mmol). The reaction mixture
was stirred at room temperature for 3 h. Water (2 mL) was added and
the mixture was concentrated in vacuo. The crude residue obtained
was dissolved in DMSO (2 mL), filtered and purified by reverse
phase chromatography (acetonitrile 30-70%/0.1% formic acid in
water) to give the title compound (77 mg, 75%) as a yellow oil.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.46 (d, J=13.6 Hz, 1H),
8.08-8.01 (m, 2H), 7.83 (d, J=5.3 Hz, 1H), 7.78-7.62 (m, 2H), 7.41
(t, J=10.1, 6.7 Hz, 1H), 6.89 (s, 1H), 4.54 (s, 1H), 4.37 (s, 1H),
3.99 (d, J=2.1 Hz, 4H), 3.81 (t, J=5.6 Hz, 1H), 2.91-2.77 (m, 2H),
2.17 (s, 1H), 2.05-1.98 (m, 3H). LCMS M/Z (M+H) 372.
Step 7: tetrahydro-2H-pyran-4-yl Trifluoromethanesulfonate
##STR00392##
[0736] To a solution of tetrahydropyran-4-ol (270 mg, 2.64 mmol) in
DCM (2.9 mL) was added pyridine (230 mg, 2.91 mmol) followed by
trifluoromethanesulfonic anhydride (810 mg, 2.87 mmol) at 0.degree.
C. The reaction mixture was stirred at 0.degree. C. for 20 min. The
reaction mixture was filtered and the solid was washed with DCM (2
mL). The filtrate was washed with water, 1.0 N HCl, water and
brine. The organic layer was dried over anhydrous MgSO.sub.4,
filtered and concentrated in vacuo to afford the title compound
(462 mg, 76%) as a black liquid which was used directly in the next
step. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 1.99 (m, 2H), 2.11
(m, 2H), 3.58 (m, 2H), 3.96 (m, 2H), 5.17 (m, 1H).
Step 8:
1-(3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1-(tetrahydro--
2H-pyran-4-yl)-1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridin-5-yl)ethan-1-on-
e
##STR00393##
[0738] To a solution of
1-(3-(3-(1-methyl-1H-pyrazol-4-yl)isoquinolin-8-yl)-1,4,6,7-tetrahydro-5H-
-pyrrolo[3,2-c]pyridin-5-yl)ethan-1-one (75 mg, 0.202 mmol) in DMF
(1.0 mL) was added sodium hydride 60% in mineral oil (9.7 mg, 0.24
mmol) at 0.degree. C. followed by tetrahydro-2H-pyran-4-yl
trifluoromethanesulfonate (200 mg, 0.854 mmol). The reaction
mixture was slowly warmed up and stirred at room temperature for 2
h. Then, the reaction mixture was stirred at 150.degree. C. for 10
min. The reaction mixture was cooled down to room temperature,
diluted with DCM (10 mL), filtered through celite and concentrated
in vacuo. The crude residue obtained was dissolved in DMSO (2 mL),
filtered and purified by reverse phase chromatography (acetonitrile
20-60%/0.1% ammonium hydroxide in water) to give the title compound
(1.5 mg, 2%) as a yellow oil. LCMS M/Z (M+H) 456.
The Following were Prepared in a Similar Fashion to the Above
Compounds:
TABLE-US-00001 Example Compound Name and Structure NMR; m/z
Procedure Example 54 1-(3-(1H-indazol-4-yl)-1-methyl- .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. A 6,7-dihydro-1H-pyrazolo[4,3-c] 8.53
(s, 1H), 7.55-7.48 (m, 2H), pyridin-5(4H)-yl)ethanone 7.22-7.19 (m,
1H), 4.79-4.75 ##STR00394## (m, 2H), 3.99-3.68 (m, 5H), 2.93- 2.81
(m, 2H), 2.24-2.16 (m, 3H); m/z 296. Example 55
1-(3-(1H-indol-4-yl)-1-methyl-6,7- .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. A dihydro-1H-pyrazolo[4,3-c]pyridin- 7.44-7.42
(m, 1H), 7.30 5(4H)-yl)ethanone (d, J = 6.0 Hz, 1H), 7.20-7.16
##STR00395## (m, 1H), 7.09 (d, J = 7.2 Hz, 1H), 4.60-4.66 (m, 2H),
3.99- 3.88 (m, 2H), 3.86 (s, 3H), 2.95-2.82 (m, 2H), 2.21- 2.08 (m,
3H); m/z 295. Example 56 1-(1-(cyclopropylmethyl)-3-(1H- .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. A indol-3-yl)-6,7-dihydro-1H-
8.08-8.03 (m, 1H), 7.43-7.40
pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (m, 2H), 7.17-7.09 (m,
2H), ##STR00396## 4.68-4.64 (m, 2H), 4.01-3.99 (m, 2H),3.95-3.85
(m,2H), 2.93-3.82 (m, 2H), 2.23-2.17 (m, 3H), 1.35-1.32 (m, 1H),
0.62- 0.59 (m, 2H), 0.47-0.43 (m, 2H); m/z 335. Example 57
1-(3-(1H-indol-3-yl)-1-(tetrahydro- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. A 2H-pyran-4-yl)-6,7-dihydro-1H- 11.23 (s,
1H), 8.31-8.20 (m, 1H), pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
7.56-7.35 (m,2H), 7.17-7.05 ##STR00397## (m, 2H), 4.59 (s, 2H),
4.39-4.23 (m, 1H), 4.08-3.95 (m, 2H), 3.80-3.74 (m, 2H), 3.50 (t, J
= 12.0 Hz, 2H), 2.91-2.71 (m, 2H), 2.22-2.09 (m, 5H), 1.90-1.78 (m,
2H); m/z 365. Example 58 1-(3-(1-isopropyl-1H-indol-3-yl)-1-
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. B
methyl-6,7-dihydro-1H- 8.24-8 19 (m, 1H), 7.54-7.47 (m,
pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone 2H), 7.20-7.16 (m,
1H),7.10-7.06 ##STR00398## (m, 1H), 4.84-4.77 (m, 1H), 4,60 (s,
2H), 3.80-3.73 (m, 5H), 2.82-2.68 (m, 2H), 2.12 (s, 3H), 1.53-1.51
(m, 6H); m/z 337. Example 59 1-(3-(1-methyl-1H-indol-3-yl)-1-
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. B
(tetrahydro-2H-pyran-4-yl)-6,7-dihydro- 8.33-8.23 (m, 1H),
7.58-7.38 1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (m, 2H),
7.23-7.17 (m, 1H), ##STR00399## 7.15-7.07 (m, 1H), 4.59 (s, 2H),
4.40-4.27 (m, 1H), 4.07-3.95 (m, 2H), 3.85 (s, 3H), 3.83-3.71 (m,
2H), 3.50 (t, J = 12.0 Hz, 2H), 2.90-2.71 (m, 2H), 2.23-2.08 (m,
5H), 1.93- 1.79 (m, 2H); m/z 379. Example 60
1-(1-methyl-3-(5-(1-methyl-1H- .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. C pyrazol-4-yl)-1H-indol-3-yl)-6,7- 8.22 (s, 1H), 7.91 (s,
1H), 7.81 dihydro-1H-pyrazolo[4,3-c]pyridin- (s, 1H), 7.43-7.38 (m,
3H), 5(4H)-yl)ethanone 4.69-4.65 (m,2H), 3.99-3.85 ##STR00400## (m,
2H), 3.93 (s, 3H), 3.84 (s, 3H), 2.90-2.78 (m, 2H), 2.24- 2.16 (m,
3H); m/z 375. Example 61 methyl 4-(5-acetyl-1-methyl-4,5,6,7-
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. C
tetrahydro-1H-pyrazolo[4,3-c] 11.60 (s, 1H), 8.04 (s, 1H),
pyridin-3-yl)-1H-indole-6-carboxylate 7.65-7.63 (m, 2H), 7.06-7.02
##STR00401## (m, 1H), 4.64-4.61 (m, 2H), 3.88 (s, 3H), 3.81-3.77
(m, 5H), 2.86-2.73 (m, 2H), 2.11-2.02 (m, 3H); m/z 353. Example 62
1-(3-(7-chloro-1H-indol-4-yl)-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. C methyl-6,7-dihydro-1H- 11.50 (s, 1H), 7.42
(s, 1H), 7.25-7.21 pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (m,
1H),7.08-7.00 (m, 2H), 4.61 ##STR00402## (s, 2H), 3.80-3.75 (m,
5H), 2.86-2.71 (m, 2H), 2.11- 2.04 (m, 3H); m/z 329. Example 63
1-(3-(6-methoxy-1H-indol-4-yl)-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. C methyl-6,7-dihydro-1H- 10.94 (s, 1H), 7.19
(s, 1H), pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone 6.88-6.84 (m,
1H), 6.64-6.61 ##STR00403## (m, 1H),4.60 (s, 2H), 3.80-3.75 (m,
8H), 2.84-2.70 (m, 2H), 2.10- 2.04 (m, 3H); m/z 325. Example 64
1-(1-methyl-3-(7-methyl-1H-indol- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. C 4-yl)-6,7-dihydro-1H-pyrazolo[4,3-c] 11.10
(s, 1H), 7.34-7.32 (m, 1H), pyridin-5(4H)-yl)ethanone 6.97-6.89 (m,
3H), 4.58 (s, 2H), ##STR00404## 3.81 -3.74 (m, 5H), 2.84-2.69 (m,
2H), 2.10- 2.02 (m, 3H); m/z 309. Example 65
1-(1-methyl-3-(3-(1-methyl-1H- .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. C pyrazol-4-yl)phenyl)-6,7-dihydro- 7.81-7.79 (m, 2H), 7.67
(s, 1H), 1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone 7.45-7.27 (m,
3H), 4.83-4.66 ##STR00405## (m, 2H), 3.99 (s, 3H), 3.95-3.75 (m,
2H), 3.82 (s, 3H), 2.80-2.72 (m, 2H), 2.21-2.15 (m, 3H); m/z 336.
Example 66 1-(1-(cyclopropylmethyl)-3-(7- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. C methyl-1H-indol-4-yl)-6,7-dihydro- 11.10
(s, 1H), 7.33 (s, 1H), 7.01
1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (s, 1H), 6.95-6.91 (m,
2H), 4.60 ##STR00406## (s, 2H), 3.96 (d, J = 6.8 Hz), 3.82-3.74 (m,
2H), 2.87-2.74 (m, 2H), 2.11-2.04 (m, 3H), 1.29-1.25 (m, 1H),
0.55-0.52 (m, 2H), 0.42-0.40 (m, 2H); m/z 349. Example 67
1-(1-(cyclopropylmethyl)-3-(1H- .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. C indol-4-yl)-6,7-dihydro-1H- 11.15 (s, 1H), 7.39-7.36
pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (m, 2H), 7.18-7.14 (m,
1H), ##STR00407## 7.06-7.00 (m, 2H), 4.64 (s, 2H), 3.98 (d, J = 7.2
Hz), 3.82-3.76 (m, 2H), 2.89-2.76 (m, 2H), 2.12-2.07 (m, 3H),
1.29-1.25 (m, 1H), 0.55 -0.52 (m, 2H), 0.45-0.41 (m, 2H); m/z 335.
Example 68 1-(3-(6-(1-methyl-1H-pyrazol-4-yl)- .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. C 1H-indol-3-yl)-1-(oxetan-3-yl)-6,7-
11.25 (s, 1H), 8.29-8.23 (m, 1H),
dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone 8.07 (s, IH),
7.81 (s, 1H), 7.52 ##STR00408## (s, 1H), 7.41 (s, 1H), 7.32-7.30
(m, 1H), 5.52-5.48 (m, 1H), 5.08- 5.04 (m, 2H), 4.91-4.87 (m, 2H),
4.56 (s, 2H), 3.85 (s, 3H), 3.75- 3.69 (m, 2H), 2.77-2.64 (m, 2H),
2.09-1.94 (m, 3H); m/z 417. Example 69
1-(3-(6-(1-methyl-1H-pyrazol-4-yl)-1H-indol-3-yl)- .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. C
1-(tetrahydrofuran-3-yl)-6,7-dihydro-1H- 11.29(s, 1H), 8.17-8.12
(m, 1H), pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone 8.06 (s, 1H),
7.81 (s, 1H), 7.50 ##STR00409## (s, 1H), 7.48-7.36 (m, 1H),
7.29-7.26 (m, 1H), 5.02-4 91 (m, 1H), 4.56 (s, 2H), 4.12-4.03 (m,
2H), 3.92-3.87 (m, 2H), 3.85 (s, 3H), 3.79-3.68 (m,2H), 2.89-2.68
(m, 2H), 2.41-2.23 (m, 2H), 2.10 (s, 3H); m/z 431. Example 70
1-(3-(2-methyl-6-(1-methyl-1H- .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. C pyrazol-4-yl)-1H-indol-3-yl)-1-((S)- 1.11 (s, 1H), 8.07
(s, 1H), 7.81 tetrahydrofuran-3-yl)-6,7-dihydro- (s, 1H), 7.43 (s,
1H), 7.42-7.37 1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (m, 1H),
7.20-7.17 (m, 1H), 4.98-4.94 ##STR00410## (m, 1H), 4.38-4.35 (m,
2H), 4.07-4.03 (m, 2H), 3.92-3.71 (m, 4H), 3.88 (s, 3H), 2.89-2.77
(m, 2H), 2.50-2.32 (m, 5H), 2.08-1.97 (m, 3H); m/z 445. Example 71
1-(3-(2-methyl-6-(1-methyl-1H- .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. C pyrazol-4-yl)-1H-indol-3-yl)-1- 11.11 (s, 1H), 8.06 (s,
1H), 7.81 (tetrahydrofuran-3-yl)-6,7-dihydro- (s, 1H), 7.44 (s,
1H), 7.40-7.37 1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (m,
1H),7.20-7.17 (m, 1H), 4.98-4.94 ##STR00411## (m, 1H), 4.37-4.35
(m, 2H), 4.07-4.02 (m, 2H), 3.92-3.71 (m, 4H), 3.88 (s, 3H),
2.88-2.77 (m, 2H), 2.50-2.32 (m, 5H), 2.08-1.97 (m, 3H); m/z 445.
Example 72 1-(3-(2-methyl-1H-indol-3-yl)-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. C (tetrahydro-2H-pyran-4-yl)-6,7-dihydro-
11.10 (s, 1H), 7.48-7.36 (m, 1H),
1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone 7.29 (d, J = 8.0 Hz,
1H), 7.05-6.99 ##STR00412## (m, 1H), 6.98-6.87 (m, 1H), 4.37-4.24
(m, 3H), 3.98-3.95 (m, 2H), 3.85-3.71 (m, 2H), 3.54-3.44 (m, 2H),
2.93-2.71 (m, 2H), 2.40-2.33 (m, 3H), 2.15-1.93 (m, 5H), 1.87-1.83
(m, 2H); m/z 379. Example 73 1-(3-(1H-indol-7-yl)-1-(tetrahydro-
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. C
2H-pyran-4-yl)-6,7-dihydro-1H- 10.74 (s, 1H), 7.54 (d, J = 8.0 Hz,
pyrazolo[4,3-c]pyridin-5(4H-yl)ethanone 1H), 7.45 (s, 1H),
7.24-7.17 (m, 1H), ##STR00413## 7.15-7.07 (m, 1H), 6.50 (s, 1H),
4.75 (s, 2H), 4.48-4.40 (m, 1H), 4.04-4.00 (m, 2H), 3.84-3.77 (m,
2H), 3.53 (t, J = 12.0 Hz, 2H), 2.97-2.75 (m, 2H), 2.40-2.28 (m,
2H), 2.17-2.09 (m, 3H), 1.87-1.83 (m, 2H); m/z 365. Example 74
1-(3-(5-chloroisoquinolin-8-yl)-1- .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. D (tetrahydro-2H-pyran-4-yl)-6,7-dihydro-
9.71-9.67 (m, 1H), 8.71-8.64 (m, 1H),
1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone 8.13-8.06 (m, 1H),
7.86-7.79 (m, 1H), ##STR00414## 7.55-7.47 (m, 1H), 4.63- 4.40 (m,
2H), 4.33-4.23 (m, 1H), 4.19-4.12 (m, 2H), 4.04-3.81 (m, 2H),
3.59-3.52 (m, 2H), 2.96-2.82 (m, 2H), 2.49-2.35 (m, 2H), 2.23-2.04
(m, 3H), 2.00-1.92 (m, 2H); m/z 411. Example 75
1-(3-(5-methylisoquinolin-8-yl)-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. D (tetrahydro-2H-pyran-4-yl)-6,7- 9.77 (s,
1H), 8.62-8.52 (m, 1H), dihydro-1H-pyrazolo[4,3-c]pyridin-
7.97-7.88 (m, 1H), 7.72-7.64 (m, 1H), 5(4H)-yl)ethanone 7.60-7.49
(m, 1H), 4 49 (s, 2H), ##STR00415## 4.48-4 38 (m, 1H), 4.05-3.95
(m, 2H), 3 .88-3.74 (m, 2H), 3.51 (t, J = 11.2 Hz, 2H), 3.00-2.78
(m, 2H), 2.69 (s, 3H), 2.20-2.03 (m, 5H), 2.00-1.87 (m, 2H); m/z
391. Example 76 1-(3-(5-methoxyisoquinolin-8-yl)-1- .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. E (tetrahydro-2H-pyran-4-yl)-6,7-
9.64-9.55 (m, 1H), 8.61-8.52 (m, 1H),
dihydro-1H-pyrazolo[4,3-c]pyridin- 8.11-8.04 (m, 1H), 7.54-7.45 (m,
1H), 5(4H)-yl)ethanone 7.09-7.02 (m, 1H), 4.63-4.40 (m,
##STR00416## 2H), 4.31- 4.21 (m, 1H), 4.20-4.13 (m, 2H), 4 06 (s,
3H), 3.98-3.80 (m, 2H), 3.57 (t, J = 12.0 Hz, 2H), 2.95-2.82 (m,
2H), 2.50-2.37 (m, 2H), 2.24-2.03 (m, 3H), 2.00-1.93 (m, 2H); m/z
407. Example 77 1-(3-(4-methoxyisoquinolin-8-yl)-1- .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. E (tetrahydro-2H-pyran-4-yl)-6,7-
9.41 (s, 1H), 8.22 (s, 1H), 8.17 dihydro-1H-pyrazolo[4,3-c]pyridin-
(d, J = 8 4 Hz, 1H), 7.87-7.80 5(4H)-yl)ethanone (m, 1H), 7.73
-7.61 (m, 1H), 4.50 ##STR00417## (s, 2H), 4.49-4 39 (m, 1H),4.08
(s, 3H), 4.04-3.95 (m, 2H), 3.86-3.76 (m, 2H), 3.51 (t, J = 12.0
Hz, 2H), 3.02-2.79 (m, 2H), 2.15-2.02 (m, 5H), 2.01-1.89 (m, 2H);
m/z 407. Example 78 1-(1-(tetrahydro-2H-pyran-4-yl)-3- .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. F
(4-vinylisoquinolin-8-yl)-6,7-dihydro- 9.69 (s, 1H), 8.69 (s, 1H),
8.21 1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (d, J = 8.4 Hz,
1H), 7.93-7.84 ##STR00418## (m, 1H), 7.74 -7.62 (m, 1H), 7.55- 7.48
(m, 1H), 5.98 (d, J = 17.6 Hz, 1H), 5.60 (d, J = 12.4 Hz, 1H), 4.50
(s, 2H), 4.49-4.39 (m, 1H), 4.05-3.95 (m, 2H), 3.86-3.77 (m, 2H),
3.51 (t, J = 12.0 Hz, 2H), 2.98-2.80 (m, 2H), 2.20-2.04 (m, 5H),
2.02-1.92 (m, 2H); m/z 403. Example 79
1-(3-(5-ethylisoquinolin-8-yl)-1- .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. G (tetrahydro-2H-pyran-4-yl)-6,7-dihydro- 9.66-9.55 (m,
1H), 8.61-8.52 (m, 1H), 1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
7.88-7.84 (m, 1H), 7.63-7.56 (m, 1H), ##STR00419## 7.55-7.48 (m,
1H), 4.62-4.40 (m, 2H), 4.32-4.23 (m, 1H),4.21-4.07 (m, 2H),
4.04-3.82 (m, 2H), 3.56 (t, J = 12.0 Hz, 2H), 3.09 (q, J = 7.2 Hz,
2H), 2.96-2.82 (m, 2H), 2.49-2.36 (m, 2H), 2.23-2.04 (m, 3H),
2.00-1.92 (m, 2H), 1.41 (t, J =7.2 Hz, 3H); m/z 405. Example 80
1-(3-(4-ethylisoquinolin-8-yl)-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. G (tetrahydro-2H-pyran-4-yl)-6,7-dihydro-
9.60 (s, 1H), 8.41 (s, 1H), 8.13
1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (d, J = 8 8 Hz, 1H),
7.92-7.83 ##STR00420## (m, 1H), 7.72-7.57 (m, 1H), 4.50 (s, 2H),
4.49-4.39 (m, 1H), 4.06-3.92 (m, 2H), 3.89-3.75 (m, 2H), 3.52 (t, J
= 12.0 Hz, 2H), 3.09 (q, J = 7.6 Hz, 2H), 3.00-2.81 (m, 2H),
2.19-2.05 (m, 5H), 2.04-1.89 (m, 2H), 1.33 (t, J = 7.6 Hz, 3H); m/z
405. Example 81 1-(3-(5-(hydroxymethyl)isoquinolin- .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. H
8-yl)-1-(tetrahydro-2H-pyran-4-yl)- 9.78 (s, 1H), 8.55 (d, J = 6.0
Hz, 1H), 6,7-dihydro-1H-pyrazolo[4,3-c] 7.97 (d, J = 6.4 Hz, 1H),
7.85 pyridin-5(4H)-yl)ethanone (d, J = 7.6 Hz, 1H), 7.67-7.56
##STR00421## (m, 1H), 5.50-5.41 (m, 1H), 4.99 (d, J = 5.6 Hz, 2H),
4 50
(s, 2H), 4.49-4.39 (m, 1H), 4.05-3.95 (m, 2H), 3.87-3.76 (m, 2H),
3.51 (t, J = 12.0 Hz, 2H), 2.98-2.82 (m, 2H), 2.14-2.02 (m, 5H),
2.00-1.88 (m, 2H); m/z 407. Example 82
1-(3-(4-(hydroxymethyl)isoquinolin- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. H 8-yl)-1-(tetrahydro-2H-pyran-4-yl)- 9.67
(s, 1H), 8.51 (s, 1H), 8.15 6,7-dihydro-1H-pyrazolo[4,3-c] (d, J =
8 4 Hz, 1H), 7.91-7.83 pyridin-5(4H)-yl)ethanone (m, 1H), 7.72-7.60
(m, 1H), 5.42 ##STR00422## (s, 1H),4.98 (s, 2H), 4.49 (s, 2H),
4.48-4.41 (m, 1H), 4.06-3.92 (m, 2H), 3.87-3.77 (m, 2H), 3.51 (t, J
= 12.0 Hz, 2H), 3.00-2.81 (m, 2H), 2.18-2.02 (m, 5H), 2.01=1.88 (m,
2H); m/z 407. Example 83 1-(3-(6-methyl-3-(1-methyl-1H-pyrazol-
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. I
4-ypisoquinolin-8-yl)-1-(tetrahydro- 9.54-9.48 (m, 1H), 8.10-7.96
2H-pyran-4-yl)-6,7-dihydro-1H-pyrazolo (m, 2H), 7.75-7.72 (m, 1H),
[4,3-c]pyridin-5(4H)-yl)ethanone 7.59-7.54 (m, 1H), 7.34-7.32
##STR00423## (m, 1H), 4.63-4.44 (m, 2H), 4.30-4.22 (m, 1H),
4.20-4.12 (m, 2H), 4.07-3.79 (m, 2H), 4.00 (s, 3H), 3.60-3.54 (m,
2H), 2.94-2.83 (m, 2H), 2.65-2.52 (m, 3H), 2.51-2.36 (m, 2H),
2.22-2.04 (m, 3H), 1.97-1.94 (m, 2H); m/z 471. Example 84
1-(3-(7-methyl-3-(1-methyl-1H- .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. I pyrazol-4-yl)isoquinolin-8-yl)-1- 8.86-8.84 (m, 1H), 8.03
(s, 1H), (tetrahydro-2H-pyran-4-yl)-6,7-dihydro- 7.98-7.96 (m, 1H),
7.78-7.72 1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (m, 2H),
7.61-7.57 (m, 1H), 4.31-4.25 ##STR00424## (m, 1H), 4.16-4.07
(m,4H), 3.98 (s, 3H) 3.96-3.66 (m, 2H), 3.57 (t, J = 12.0 Hz, 2H),
2.94-2.86 (m, 2H), 2.41-2.35 (m, 2H), 3.37 (s, 3H), 2.19-2.00 (m,
3H), 1.97-1.93 (m, 2H); m/z 471. Example 85
1-(3-(7-fluoro-3-(1-methyl-1H- .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. I pyrazol-4-ypisoquinolin-8-yl)-1- 9.39- 9.36 (m, 1H), 8.34
(s, 1H), (tetrahydro-2H-pyran-4-yl)-6,7-dihydro- 8.14 (s, 1H), 8.07
(s, 1H), 8.05-8.00 1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (m,
1H), 7.80-7.75 (m, 1H), ##STR00425## 4.49-4.46 (m, 1H), 4.37 (s,
2H), 4.05-3.97 (m, 2H), 3.91 (s, 3H), 3.85-3.79 (m, 2H), 3.52 (t, J
= 12.0 Hz, 2H), 3.02-2.85 (m, 2H), 2.16-1.93 (m, 7H); m/z 475.
Example 86 5-(8-(5-acetyl-1-(tetrahydro-2H- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. I pyran-4-yl)-4,5,6,7-tetrahydro-1H-
9.58-9.55 (m, 1H), 9.43 (s, 1H), pyrazolo[4,3-c]pyridin-3-yl)-7-
8.88-8.83 (m, 1H), 8.77 (s, 1H),
fluoroisoquinolin-3-yl)-N-methylpicolinamide 8.76-8 74 (m, 1H),
8.25-8.21 (m, 1H), ##STR00426## 8.20-8.17 (m, 1H), 7.94-7.89 (m,
1H), 4.53-4.50 (m, 1H), 4.39 (s, 2H), 4.05-3.97 (m, 2H), 3.90-3.78
(m, 2H), 3.53 (t, J = 12.4 Hz, 2H), 3.02-2.85 (m, 2H), 2.86 (d, J =
5.2 Hz, 3H), 2.16-1.93 (m, 7H); m/z 529. Example 87
5-(8-(5-acetyl-1-(tetrahydro-2H- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. I pyran-4-yl)-4,5,6,7-tetrahydro-1H- 10.00
(s, 1H), 9.45 (s, 1H), 8 87 pyrazolo[4,3-c]pyridin-3- (s, 1H),
8.79-8.76 (m, 1H), 8.71 yl)isoquinolin-3-yl)-N-methylpicolinamide
(s, 1H), 8.17 (d, J = 8.0 Hz, 1H), ##STR00427## 8.08 (d, J = 8.0
Hz, 1H), 7.94-7.89 (m, 1H), 7.75 -7.66 (m, 1H), 4.58 (s, 2H),
4.53-4.42 (m, 1H), 4.07-3.97 (m, 2H), 3.89- 3.79 (m, 2H), 3.56-3.50
(m, 2H), 3.00-2.80 (m, 2H), 2.86 (s, 3H), 2.22-1.90 (m, 7H); m/z
511. Example 88 1-(3-(3-(1-methyl-1H-pyrazol-4- .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. I yl)isoquinolin-8-yl)-1-(tetrahydro-
9.81 (s, 1H), 8.36 (s, 1H), 8.12-8.07
2H-pyran-4-yl)-6,7-dihydro-1H- (m, 2H), 7.90-7.86 (m, 1H),
7.81-7.76 pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (m, 1H),
7.59-7.50 (m, 1H), 4.55 ##STR00428## (s, 2H), 4.48-4.43 (m, 1H),
4.03-4.00 (m, 2H), 3.91 (s, 3H), 3.85-3.80 (m, 2H), 3.55-3.49 (m,
2H), 2.96-2.82 (m, 2H), 2.20-2.02 (m, 5H), 1.99-1.91 (m, 2H); m/z
457. Example 89 (S)-1-(3-(3-(1-methyl-1H-pyrazol-4- .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. I yl)isoquinolin-8-yl)-1- 9.76 (s,
1H), 8.36 (s, 1H), 8.13-8.05 (tetrahydrofuran-3-yl)-6,7-dihydro-
(m, 1H), 7.90-7.87 (m, 1H), 7.81-7.76
1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone (m, 1H), 7.58-7.50 (m,
1H), 5.10-5.06 ##STR00429## (m, 1H), 4.54 (s, 2H), 4.11-4.05 (m,
2H), 3.98-3.78 (m, 4H), 3.95 (s, 3H), 2.98-2.78 (m, 2H), 2.42-2.37
(m, 2H), 2.10-2.01 (m, 3H); m/z 443. Example 90
1-(3-(3-(1,5-dimethyl-1H-pyrazol-4- .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. I yl)isoquinolin-8-yl)-1-(tetrahydro- 9.72-9.65
(m, 1H), 7.92 (s, 1H), 2H-pyran-4-yl)-6,7-dihydro-1H- 7.85-7.64 (m,
3H), 7.54-7.46 (m, 1H), pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
4.65- 4.47 (m, 2H), 4.37-4.23 ##STR00430## (m, 1H), 4.21-4.15 (m,
2H), 4.07-3.82 (m, 2H), 3.89 (s, 3H), 3.63-3.54 (m, 2H), 2.96-2.84
(m, 2H), 2.67 (s, 3H), 2.54-2.39 (m, 2H), 2.21-2.08 (m, 3H),
2.01-1.93 (m, 2H); m/z 471. Example 91
3-(7-fluoro-3-(1-methyl-1H-pyrazol- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. J 4-yl)isoquinolin-8-yl)-N-methyl-1- 9.35 (s,
1H), 8 33 (s, 1H), 8.14 (tetrahydro-2H-pyran-4-yl)-6,7- (s, 1H),
8.07 (s, 1H), 8.06-8.00
dihydro-1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxamide (m, 1H),
7.80-7.75 (m, 1H), ##STR00431## 6.55-6.47 (m, 1H), 4.50-4.43 (m,
1H), 4.22(s, 2H), 4.02-3.98 (m, 2H), 3.91 (s, 3H), 3.73-3.67 (m,
2H), 3.55-3.48 (m, 2H), 2.87-2.79 (m, 2H), 2.55-2.53 (m, 3H),
2.18-2.08 (m, 2H), 1.97-1.91 (m, 2H); m/z 490. Example 92
N-methyl-3-(3-(6-(methylcarbamoyl)pyridin-3- .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. J yl)isoquinolin-8-yl)-1-(tetrahydro- 9.84 (s,
1H), 9.29 (s, 1H), 8.54-8.51 2H-pyran-4-yl)-6,7-dihydro-1H- (m,
1H), 8.27(d, J = 8.4 Hz, 1H),
pyrazolo[4,3-c]pyridine-5(4H)-carboxamide 8.15 (s, 1H), 8 12-8.05
(m, 1H), ##STR00432## 7.90 (d, J = 8.4 Hz, 1H), 7.80-7.75 (m, 1H),
7.61 (d, J = 7.2 Hz, 1H), 4.61-4.50 (m, 1H), 4 40 (s, 2H), 4.34-
4.27 (m, 1H), 4.19-4.15 (m, 2H), 3.90-3.87 (m, 2H), 3.61-3.55 (m,
2H), 3.08 (d, J = 5.2 Hz, 3H), 2.90-2.87 (m, 2H), 2.80 (d, J = 4.4
Hz, 3H), 2.50-2.40 (m, 2H), 2.04-1.93 (m, 2H); m/z 526. Example 93
N-methyl-3-(3-(1-methyl-1H- .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. J pyrazol-4-yl)isoquinolin-8-yl)-1- 9.87 (s, 1H), 8.38 (s,
1H), 8.16-8.13 (tetrahydro-2H-pyran-4-yl)-6,7-dihydro- (m, 1H),
8.11 (s, 1H), 7.90 (d, J =
1H-pyrazolo[4,3-c]pyridine-5(4H)-carboxamide (d, J = 8.0 Hz, 1H),
6.65-6.53 (m, ##STR00433## 1H), 4.52-4.42 (m, 3H), 4.05-3.98 (m,
2H), 3.92 (s, 3H), 3.73-3.67 (m, 2H), 3.55-3.48 (m, 2H), 2.87- 2.79
(m, 2H), 2.55-2.53 (m, 3H), 2.18-2.08 (m, 2H), 1.97-1.91 (m, 2H);
m/z 472. Example 94 N-methyl-3-(3-(2-methylthiazol-5- .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. J yl)isoquinolin-8-yl)-1-(tetrahydro-
9.65 (s, 1H), 8.19 (s, 1H), 7.96 2H-pyran-4-yl)-6,7-dihydro-1H- (s,
1H), 7.83 (d, J = 8 4 Hz, 1H),
pyrazolo[4,3-c]pyridine-5(4H)-carboxamide 7.75-7.70 (m, 1H), 7.54
(d, J = 7.2 ##STR00434## Hz, 1H), 4.52-4.44 (m, 1H), 4.34 (s, 2H),
4.30-4.26 (m, 1H), 4.21-4.13 (m, 2H), 3.89-3.86 (m, 2H), 3.60-3.64
(m, 2H), 2.88-2.86 (m, 2H), 2.80-2.77 (m, 6H), 2.48-2.38 (m, 2H),
1.97-1.94 (m, 2H); m/z 489. Example 95
N-methyl-3-(3-phenoxyphenyl)-1- .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. J (tetrahydro-2H-pyran-4-yl)-6,7- 7.57-7.23 (m, 5H),
7.22-6.83 (m, 3H), dihydro-1H-pyrazolo[4,3-c]pyridine- 6.75-6.57
(m, 1H), 4.48 (s, 2H), 4.41- 5(4H)-carboxamide 4.24 (m, 1H),
4.07-3.83 (m, 2H), ##STR00435## 3.75-3.50 (m, 4H), 2.85-2.65 (m,
2H), 2.59 (s, 3H), 2.17-1.92 (m, 2H), 1.90-1.71 (m, 2H); m/z 433.
Example 96 4-(8-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)- .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. K
4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3- 9.89-9.80 (m, 1H),
8.40 (s, 1H),
yl)isoquinolin-3-yl)-1-methyl-1H-pyrazole-5-carbonitrile 8.34 (s,
1H), 7.99 (d, J = 8.0 Hz, 1H), ##STR00436## 7.93-7.82 (m, 1H),
7.71-7.59 (m, 1H), 4.55 (s, 2H), 4.54-4.41 (m, 1H), 4.09 (s, 3H),
4.06-3.98 (m, 2H), 3.88-3.77 (m, 2H), 3.52 (t, J = 12.0 Hz, 2H),
3.01-2.80 (m, 2H), 2.19-2.02 (m, 5H), 2.01-1.88 (m, 2H); m/z 482.
Example 97 1-(3-(3-(3-(difluoromethyl)-1-methyl- .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. K 1H-pyrazol-4-yl)isoquinolin-8-yl)-1-
9.83-9.76 (m, 1H), 8.50 (s, 1H), 8.11
(tetrahydro-2H-pyran-4-yl)-6,7-dihydro-1H- (s, 1H), 7.93 (d, J =
8.0 Hz, 1H), pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone 7.86-7.79 (m,
1H), 7.65-7.54 (m, 1H), ##STR00437## 7.58 (t, J = 54.0 Hz, 1H),
4.54 (s, 2H), 4.52-4 41 (m, 1H), 4.07-3.93 (m, 2H), 3.97 (s, 3H),
3.88-3.77 (m, 2H), 3.52 (t, J = 12.0 Hz, 2H), 3.01-2.81 (m, 2H),
2.20-2.01 (m, 5H), 2.00-1.88 (m, 2H); m/z 507. Example 98
1-(3-(3-(5-(difluoromethyl)-1- .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. K methyl-1H-pyrazol-4-yl)isoquinolin- 9.81 (s, 1H), 8.30
(s, 1H), 8.22 8-yl)-1-(tetrahydro-2H-pyran-4-yl)- (s, 1H), 8.13 (t,
J = 54.0 Hz, 1H),
6,7-dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone 8.04-7.93
(m, 1H), 7.90-7.82 (m, 1H), ##STR00438## 7.69-7.56 (m, 1H), 4.54
(s, 2H), 4.53-4.41 (m, 1H), 4 06 (s, 3H), 4.05-4.00 (m, 2H),
3.89-3.73 (m, 2H), 3.52 (t, J = 12.0 Hz, 2H), 3.02-2.81 (m, 2H),
2.23-2.01 (m, 5H), 2.00-1.88 (m, 2H); m/z 507. Example 99
5-(5-acetyl-1-(tetrahydro-2H-pyran- .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. L 4-yl)-4,5,6,7-tetrahydro-1H- 7.35-7.27 (m,
1H), 7.25-7.12 (m, 2H), pyrazolo[4,3-c]pyridin-3-yl)-1,2- 6.71-6.59
(m, 1H), 4.60-4.38 (m, 4H), dihydroisoquinolin-3(4H)-one 4.26-4.07
(m, 3H), 3.96-3.75 (m, 2H), ##STR00439## 3.74-3.67 (m, 2H),
3.60-3.48 (m, 2H), 2.86-2.76 (m, 2H), 2.40-2.29 (m, 2H), 2.20-2.10
(m, 3H), 1.99-1.82 (m, 2H); m/z 395. Example 100
5-(5-acetyl-1-(tetrahydro-2H-pyran-4-yl)- .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. L 4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-
8.83-8.72 (m, 1H), 7.92-7.78 (m, 1H), 3-yl)isoquinolin-3(2H)-one
7.57 (d, J = 6.4 Hz, 1H), 7.45-7.34 ##STR00440## (m, 2H), 4.62-4.40
(m, 2H), 4.31- 4.23 (m, 1H), 4.21-4.12(m, 2H), 4.04-3.82 (m, 2H),
3.58 (t, J = 12.0 Hz, 2H), 2.95- 2.84 (m, 2H), 2.49-2.38 (m, 2H),
2.24-2.05 (m, 3H), 2.01-1.91 (m,2H); m/z 393. Example 101
5-(5-acetyl-1-(tetrahydro-2H-pyran- .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. L 4-yl)-4,5,6,7-tetrahydro-1H- 7.99 (d,J = 8.4
Hz, 1H), 7.58 (s, 1H), pyrazolo[4,3-c]pyridin-3-yl)-2-(1- 7.40-7.17
(m, 3 H), 4.90-4.87 methyl-1H-pyrazol-4-yl)-1,2- (m, 2H), 4.57-4.42
(m, 2H), dihydroisoquinolin-3(4H)-one 4.27-4.08 (m, 3H), 4.00-3.75
##STR00441## (m, 4H), 3.91 (s, 3H), 3.57 (t, J = 12.0 Hz, 2H),
2.90-2.76 (m, 2H), 2.43-2.25 (m, 2H), 2.20-2.11 (m, 3H), 1.95-1.85
(m, 2H); m/z 475. Example 102 5-(5-acetyl-1-(tetrahydro-2H-pyran-
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. L
4-yl)-4,5,6,7-tetrahydro-1H- 7.44-7.42 (m, 1H), 7.35-7.31 (m, 2H),
pyrazolo[4,3-c]pyridin-3-yl)-2-(1,5- 7.23-7.17 (m, 1H), 4.78-4.76
(m, 2H), dimethyl-1H-pyrazol-4-yl)-1,2- 4.57-4.45 (m, 2H),
4.25-4.13 (m, 3H), dihydroisoquinolin-3(4H)-one 3.98-3.79 (m, 7H),
3.55 (t, J = 12.0 ##STR00442## Hz, 2H), 2.87-2.80 (m, 2H),
2.43-2.28 (m, 2H), 2.21-2.10 (m, 6H), 1.97-1.76 (m,2H); m/z 489.
Example 103 5-(5-acetyl-1-(tetrahydro-2H-pyran- .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. L 4-yl)-4,5,6,7-tetrahydro-1H- 7.92 (d,
J = 2.8 Hz, 1H), 7.37-7.24 pyrazolo[4,3-c]pyridin-3-yl)-2-(6- (m,
4H), 6.58-6.52 (m, 1H), 6.48-6.41 (methylamino)pyridin-3-yl)-1,2-
(m, 1H), 4 83 (s, 2H), 4.46 (s, 2H), dihydroisoquinolin-3(4H)-one
4.42-4.33 (m, 1H), 4.03-3.94 (m, 2H), ##STR00443## 3.85 (s, 2H),
3.83-3.73 (m, 4H), 3.55-3.40 (m, 2H), 2.92-2.77 (m, 2H), 2.76 (d, J
= 5.2 Hz, 3H), 2.14-1.99 (m, 5H), 1.92-1.82 (m, 2H); m/z 501.
Example 104 5-(5-acetyl-1-(tetrahydro-2H-pyran- .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. L 4-yl)-4,5,6,7-tetrahydro-1H- 8.04 (d,
J = 2.4 Hz, 1H) ,7.49-7.45 pyrazolo[4,3-c]pyridin-3-yl)-2-(6- (m,
1H), 7.39-7.24 (m, 3H), 6.65 (dimethylamino)pyridin-3-yl)-1,2- (d,
J = 9.2 Hz, 1H), 4.86 (s, 2H), 4.47 dihydroisoquinolin-3(4H)-one
(s, 2H), 4.43-7.31 (m, 1H), 4.06-3.95 ##STR00444## (m, 2H),
3.89-3.84 (m, 2H), 3.83-3.72 (m, 2H), 3.46 (d, J = 11.6 Hz, 2H),
3.02 (s, 6H), 2.94-2.75 (m, 2H), 2.12-2.01 (m, 5H), 1.93-1.82 (m,
2H); m/z 515. Example 105 5-(5-(5-acetyl-1-(tetrahydro-2H- .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. L
pyran-4-yl)-4,5,6,7-tetrahydro-1H- 8.64 (s, 1H), 8.26 (d, J = 8.8
Hz, 2H), pyrazolo[4,3-c]pyridin-3-yl)-3-oxo-3,4- 8.00-7.91 (m, 1H),
7.84 (d, J =
8.4 dihydroisoquinolin-2(1H)-yl)-N-methylpicolinamide Hz, 1H),
7.39-7.33 (m, 2H), 4.97-4.93 ##STR00445## (m, 2H), 4.60-4.48 (m,
2H), 4.25-4.10 (m, 3H), 4.02-3.82 (m, 4H), 3.56 (t, J = 12.0 Hz,
2H), 3.05 (d, J = 5.2 Hz, 3H), 2.89-2.80 (m, 2H), 2.45-2.30 (m,
2H), 2.22-2.14 (m, 3H), 1.97-1.87 (m, 2H); m/z 529. Example 106
1-(1-methyl-3-(5-(1-methyl-1H- .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. M pyrazol-4-yl)-1H-indol-1-yl)-6,7- 8.09 (s, 1H), 7.84 (s,
1H), 7.80-7.71 dihydro-1H-pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone
(m, 2H), 7.55-7.41 (m, 2H), 6.66 ##STR00446## (s, 1H), 4.50-4 47
(m, 2H), 3.87 (s, 3H), 3.82-3.72 (m, 5H), 2.88-2.73 (m, 2H),
2.11-2.05 (m, 3H); m/z 375. Example 107
1-(1-methyl-3-(5-(1-methyl-1H- .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. M pyrazol-4-yl)-1H-benzo[d]imidazol- 8.46-8.40 (m, 1H),
8.17 (s, 1H), 7.96 1-yl)-6,7-dihydro-1H-pyrazolo[4,3-c] (s, 1H),
7.92 (s, 1H), 7.81-7.69 pyridin-5(4H)-yl)ethanone (m, 1H), 7.56 (d,
J = 8.4 Hz, 1H), ##STR00447## 4.57-4.51 (m, 2H), 3.88 (s, 3H),
3.83-3.72 (m, 5H), 2.90-2.74 (m, 2H), 2.11-2.06 (m, 3H); m/z 376.
Example 108 1-(1-methyl-3-(6-(1-methyl-1H- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. M pyrazol-4-yl)-1H-benzo[d]imidazol-
8.42-8.36 (m, 1H), 8.18 (s, 1H),
1-yl)-6,7-dihydro-1H-pyrazolo[4,3-c] 7.91-7.73 (m, 3H), 7.53 (d, J
= 8.4 pyridin-5(4H)-yl)ethanone Hz, 1H), 4.54-4.49 (m, 2H), 3.87
##STR00448## (s, 3H), 3.85-3.75 (m, 5H), 2.92-2.76 (m, 2H),
2.13-2.04 (m, 3H); m/z 376. Example 109
1-(3-(1H-indol-1-yl)-1-(tetrahydro- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. M 2H-pyran-4-yl)-6,7-dihydro-1H- 7.88-7.76
(m, 1H), 7.64 (d, J = 7.6 Hz,
pyrazolo[4,3-c]pyridin-5(4H)-yl)ethanone 1H), 7.60-7.47 (m, 1H),
7.26-7.18 (m, ##STR00449## 1H), 7.17-7.09 (m, 1H), 6.69 (s, 1H),
4.54-4.46 (m, 2H), 4.45- 4.34 (m, 1H),4.08- 3.94 (m, 2H), 3.80-3.76
(m, 2H), 3.49 (t, J = 12.0 Hz, 2H), 2.98-2.77 (m, 2H), 2.14- 2.02
(m, 5H), 1.96-1.82 (m, 2H); m/z 365. Example 110
1-(3-(3-methyl-1H-indol-1-yl)-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. M (tetrahydro-2H-pyran-4-yl)-6,7- 7.85-7.75
(m, 1H), 7.57 (d, J = 8.0 Hz, dihydro-1H-pyrazolo[4,3-c]pyridin-
1H), 7.38-7.26 (m, 1H), 7.25-7.18 5(4H)-yl)ethanone (m, 1H),
7.17-7.09 (m, 1H), 4.57-4.45 ##STR00450## (m, 2H), 4.44-4.30 (m,
1H), 4.02-3.93 (m, 2H), 3.86-3.72 (m, 2H), 3.49 (t, J = 12.0 Hz,
2H), 2.95-2.73 (m, 2H), 2.33 (s, 3H), 2.16-1.99 (m, 5H), 1.93-1.81
(m, 2H); m/z 379. Example 111 1-[3-[3-(3R)-methoxypyrrolidin-1-yl)-
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. N
8-isoquinolyl]-1-tetrahydropyran- 9.46 (t, J = 0.9 Hz, 1H), 7.61
(dt, J = 4-yl-6,7-dihydro-4H-pyrazolo 8.5, 1.0 Hz, 1H), 7.57-7.47
(m, 1H), [4,3-c]pyridin-5-yl]ethanone 7.22-7.04 (m, 1H), 6.69-6.57
(m, 1H), (single unknown stereoisomer) 4.51 (s, 2H), 4.43 (d, J =
4.2 Hz, 1H), ##STR00451## 4.16-4.08 (m, 1H), 4.08-3.97 (m, 2H),
3.87-3.75 (m, 2H), 3.61-3.40 (m, 6H), 3.29 (s, 3H), 2.98- 2.77 (m,
2H), 2.18-1.97 (m, 7H), 1.93 (d, J = 9.3 Hz, 2H); m/z 476.3.
Example 112 1-[3-[3-(3S)-methoxypyrrolidin-1-yl)- .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. N 8-isoquinolyl]-1-tetrahydropyran- 9.46
(t, J = 0.9 Hz, 1H), 7.61 (dt, J = 4-yl-6,7-dihydro-4H- 8.5, 1.0
Hz, 1H), 7.57-7.47 (m, 1H), pyrazolo[4,3-c]pyridin-5-yl]ethanone
7.22-7.04 (m, 1H), 6.69-6.57 (m, 1H), (single unknown stereoisomer)
4.51 (s, 2H), 4.43 (d, J = 4.2 Hz, 1H), ##STR00452## 4.16-4.08 (m,
1H), 4.08-3.97 (m, 2H), 3.87-3.75 (m, 2H), 3.61-3.40 (m, 6H), 3.29
(s, 3H), 2.98- 2.77 (m, 2H), 2.18-1.97 (m, 7H), 1.93 (d, J = 9.3
Hz, 2H); m/z 476.4. Example 113
1-[3-[3-(2R)-methylpyrrolidin-1-yl)- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. N 8-isoquinolyl]-1-tetrahydropyran-4- 9.42
(t, J = 0.8 Hz, 1H), 7.60 (dt, J =
yl-6,7-dihydro-4H-pyrazolo[4,3-c] 8.4, 0.9 Hz, 1H), 7.51 (ddd, J =
8.9, pyridin-5-yl]ethanone 6.9, 2.1 Hz, 1H), 7.20-7.03 (m, 1H),
(single unknown stereoisomer) 6.60 (s, 1H), 4.49 (d, J = 1.3 Hz,
2H), ##STR00453## 4.43 (td, J = 11.3, 9.6, 5.7 Hz, 1H), 4.30-4.20
(m, 1H), 4.06-3.97 (m, 2H), 3.82 (dt, J = 17.8, 6.1 Hz, 2H),
3.59-3.47 (m, 3H), 3.37- 3.32 (m, 1H), 2.88 (dt, J = 48.5, 5.8 Hz,
2H), 2.14-1.90 (m, 10H), 1.70 (d, J = 5.1 Hz, 1H), 1.20 (d, J = 6.2
Hz, 3H); m/z 460.2. Example 114
1-[3-[3-(2S)-methylpyrrolidin-1-yl)- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. N 8-isoquinolyl]-1-tetrahydropyran-4- 9.42
(t, J = 0.8 Hz, 1H), 7.60 (dt, J =
yl-6,7-dihydro-4H-pyrazolo[4,3-c] 8.4, 0.9 Hz, 1H), 7.51 (ddd, J =
8.9, pyridin-5-yl]ethanone 6.9, 2.1 Hz, 1H), 7.20-7.03 (m, 1H),
(single unknown stereoisomer) 6.60 (s, 1H), 4.49 (d, J = 1.3 Hz,
2H), ##STR00454## 4.43 (td, J = 11.3, 9.6, 5.7 Hz, 1H), 4.30-4.20
(m, 1H), 4.06-3.97 (m, 2H), 3.82 (dt, J = 17.8, 6.1 Hz, 2H),
3.59-3.47 (m, 3H), 3.37-3.32 (m, 1H), 2.88 (dt, J = 48.5, 5.8 Hz,
2H), 2.14-1.90 (m, 10H), 1.70 (d, J = 5.1 Hz, 1H), 1.20 (d, J = 6.2
Hz, 3H); m/z 460.2. Example 115
1-[3-[3-(3-fluoropyrrolidin-1-yl)-8- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. N isoquinolyl]-1-tetrahydropyran-4-yl- 9.48
(t, J = 0.9 Hz, 1H), 8.53 (s, 1H), 6,7-dihydro-4H-pyrazolo[4,3-c]
7.68-7.58 (m, 1H), 7.55 (ddd, J = pyridin-5-yl]ethanone 9.0, 7.0,
2.3 Hz, 1H), 7.23-7.10 (m, (mixture of enantiomers) 1H), 6.70 (s,
1H), 4.51 (s, 2H), 4.43 ##STR00455## (d, J = 4.0 Hz, 1H), 4.04-3.98
(m, 2H),3.83-3.68 (m, 4H) ,3.56-3.46 (m, 4H), 2.97-2.81 (m,
2H),2.31- 2.08 (m, 7H), 1.96-1.90 (m, 2H); m/z 464.2. Example 116
1-[3-[3-[(3S)-3-(dimethylamino)pyrrolidin-1-yl]-8- .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. N isoquinolyl]-1-tetrahydropyran-4-yl-
9.46 (t, J = 0.8 Hz, 1H), 7.60 (dt, J =
6,7-dihydro-4H-pyrazolo[4,3-c] 8.5, 1.0 Hz, 1H), 7.53 (ddd, J =
8.8, pyridin-5-yl]ethanone 6.9, 2.1 Hz, 1H), 7.21-7.04 (m, 1H),
(single known stereoisomer) 6.64 (d, J = 0.9 Hz, 1H), 4.50
##STR00456## (s, 2H), 4.47-4.39(m1, 1H), 4.07-3.94 (m, 2H),
3.85-3.72 (m, 3H), 3.66 (t, J = 9.2 Hz, 1H), 3.57-3.46 (m, 2H),
3.41 (td, J = 10.1, 6.8 Hz, 1H), 3.19 (dd, J = 10.0, 8.2 Hz, 1H),
2.94 (t, J = 5.8 Hz, 1H), 2.81 (t, J = 8.2 Hz, 2H), 2.23 (s, 6H),
2.21-1.99 (m, 6H), 1.92 (dd, J = 11.9, 6.4 Hz, 2H), 1.89-1.81 (m,
1H); m/z 489.3. Example 117
1-[3-[3-[3-(difluoromethyl)pyrrolidin-1-yl]-8- Not Determined; m/z
496.2. N isoquinolyl]-1-tetrahydropyran-4-yl-
6,7-dihydro-4H-pyrazolo[4,3-c] pyridin-5-yl]ethanone (mixture of
enantiomers) ##STR00457## Example 118
1-[3-[3-[(3S)-3-methylpyrrolidin-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. N yl]-8-isoquinolyl]-1- 9.43 (d, J = 0.8 Hz,
1H), 7.59 (dt, J = tetrahydropyran-4-yl-6,7-dihydro- 8.6, 1.0 Hz,
1H), 7.56-7.47 (m, 1H), 4H-pyrazolo[4,3-c]pyridin-5- 7.20-7.04 (m,
1H), 6.59 (d, J = 0.9 yl]ethanone Hz, 1H), 4.50 (d, J = 2.2 Hz,
2H), (single known stereoisomer) 4.47-4.38 (m, 1H), 4.01 (dt, J =
9.5, ##STR00458## 4.5 Hz, 2H), 3.81 (dt, J = 15.2, 5.8 Hz, 2H),
3.69 (dd, J = 10.1, 7.2 Hz, 1H), 3.59 (dd, J = 3.7, 2.3 Hz, 1H),
3.55-3.49 (m, 2H), 3.47-3.41 (m, 1H), 3.01 (dd, J = 10.1, 7.5 Hz,
1H), 2.98-2.78 (m, 2H), 2.43-2.33 (m, 1H), 2.21-2.01 (m, 6H), 1.93
(d, J = 12.6 Hz, 2H), 1.63 (dq, J = 12.0, 8.2 Hz, 1H), 1.11 (d, J =
6.6 Hz, 3H); m/z 460.3. Example 119
1-[3-[3-[(3S)-3-methylpyrrolidin-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. N yl]-8-isoquinolyl]-1- 9.49-9.37 (m, 1H),
7.59 (dt, J = 8.6, tetrahydropyran-4-yl-6,7-dihydro- 1.0 Hz, 1H),
7.51 (ddd, J = 8.8, 6.9, 4H-pyrazolo[4,3-c]pyridin-5-yl]ethanone
2.0 Hz, 1H), 7.17-7.05 (m, 1H), 6.59 (single known enantiomer) (d,
J = 0.9 Hz, 1H), 4.50 (d, J = 2.1 ##STR00459## Hz, 2H), 4.43 (q, J
= 5.6, 4.2 Hz, 1H), 4.04-3.97 (m, 2H), 3.85-3.76 (m, 2H), 3.72-3.65
(m, 1H), 3.58 (ddd, J = 4.5, 3.8, 1.3 Hz, 1H), 3.55-3.48 (m, 2H),
3.47-3.41 (m, 1H), 3.06-2.97 (m, 1H), 2.97-2.77 (m, 2H), 2.45-2.33
(m, 1H), 2.16- 2.00 (m, 6H), 1.93 (d, J = 12.2 Hz, 2H), 1.69-1.57
(m, 1H), 1.11 (d, J = 6.6 Hz, 3H); m/z 460.3. Example 120
1-[8-(5-acetyl-1-tetrahydropyran-4- .sup.1H NMR (400 MHz,
DMSO-d.sub.6, 34/36 H) N yl-6,7-dihydro-4H-pyrazolo[4,3-c] .delta.
9.51-9.41 (m, 1H), 7.61 (dt, J = 8.5,
pyridin-3-yl)-3-isoquinolyl]-N,N- 1.0 Hz, 1H), 7.53 (ddd, J = 8.8,
6.9, 2.1 dimethyl-pyrrolidine-3-carboxamide Hz, 1H), 7.22-7.07 (m,
1H), 6.64 (mixture of enantiomers) (d, J = 0.9 Hz, 1H), 4.51 (d, J
= 1.7 ##STR00460## Hz, 2H), 4.48-4.37 (m, 1H), 4.08-3.96 (m, 2H),
3.88-3.71 (m, 3H), 3.65-3.45 (m, 6H), 3.10 (s, 3H), 2.98-2.78 (m,
5H), 2.10 (s, 5H), 1.97-1.88 (m, 2H); m/z 517.3. Example 121
1-[3-[3-(3,3-dimethylpyrrolidin-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. N yl)-8-isoquinolyl]-1- 9.43 (t, J = 0.9 Hz,
1H), 7.59 (dt, J = tetrahydropyran-4-yl-6,7-dihydro- 8.5, 1.0 Hz,
1H), 7.55-7.48 (m, 1H), 4H-pyrazolo[4,3-c]pyridin-5-yl]ethanone
7.17-7.06 (m, 1H), 6.62-6.55 (m, 1H), ##STR00461## 4.50 (s, 2H),
4.46-4.39 (m, 1H), 4.01 (d, J = 11.5 Hz, 2H), 3.86-3.78 (m, 2H),
3.57-3.51 (m, 4H), 2.94 (t, J = 5.7 Hz, 2H), 2.14-2.06 (m, 5H),
1.96-1.91 (m, 2H), 1.13 (s, 6H), 1.06-1.01 (m, 2H), 0.90 (dd, J =
4.9, 2.0 Hz, 2H); m/z 474.3. Example 122
1-[3-[3-[(3R,4S)-3,4-difluoropyrrolidin-1-yl]-8- .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. N
isoquinolyl]-1-tetrahydropyran-4-yl-6,7- 9.51 (t, J = 0.8 Hz, 1H),
7.66 (dt, J = dihydro-4H-pyrazolo[4,3-c]pyridin-5-yl]ethanone 8.5,
1.0 Hz, 1H), 7.58 (ddd, J = 8.3, 6.9, (single meso isomer) 2.5 Hz,
1H), 7.27-7.11 (m, 1H), 6.76 ##STR00462## (d, J = 1.0 Hz, 1H),
5.61-5.48 (m, 1H), 5.48-5.34 (m, 1H), 4.51 (s, 2H), 4.44 (dt, J =
8.3, 4.2 Hz, 1H), 4.07-3.97 (m, 2H), 3.97-3.87 (m, 2H), 3.85-3.78
(m, 2H), 3.73-3.63 (m, 2H), 3.57- 3.46 (m, 2H), 2.98-2.77 (m, 2H),
2.16-2.00 (m, 5H), 1.93 (d, J = 11.5 Hz, 2H); m/z 482.2. Example
123 1-[3-[3-(2,2-dimethylpyrrolidin-1-yl)-8-isoquinolyl]- Not
Determined; m/z 474.3. N 1-tetrahydropyran-4-yl-6,7-dihydro-
4H-pyrazolo[4,3-c]pyridin-5-yl]ethanone ##STR00463## Example 124
1-[3-[3-(2,5-dihydropyrrol-1-yl)-8- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. N isoquinolyl]-1-tetrahydropyran-4-yl- 9.49
(t, J = 0.8 Hz, 1H), 7.69-7.59 6,7-dihydro-4H-pyrazolo[4,3-c] (m,
1H), 7.55 (ddd, J = 8.9, 7.0, 2.3 pyridin-5-yl]ethanone Hz, 1H),
7.24-6.96 (m, 1H), 6.65 ##STR00464## (d, J = 0.9 Hz, 1H), 6.08 (s,
2H), 4.51 (d, J = 1.7 Hz, 2H), 4.48-4.38 (m, 1H), 4.28 (s, 4H),
4.07-3.97 (m, 2H), 3.87-3.77 (m, 2H), 3.57-3.50 (m, 2H), 2.98-2.77
(m, 2H), 2.16-2.00 (m, 5H), 1.93 (d, J = 9.4 Hz, 2H); m/z 444.2.
Example 125 1-[3[3-(3,3-difluoropyrrolidin-1-yl)-8- Not Determined;
m/z 482.2. N isoquinolyl]-1-tetrahydropyran-4-yl-6,7-
dihydro-4H-pyrazolo[4,3-c]pyridin-5-yl]ethanone ##STR00465##
Example 126 1-[8-(5-acetyl-1-tetrahydropyran-4-yl-6,7- Not
Determined; m/z 471.2. N dihydro-4H-pyrazolo[4,3-c]pyridin-3-yl)-3-
isoquinolyl]pyrrolidine-3-carbonitrile (mixture of enantiomers)
##STR00466## Example 127
1-[3[3-(4-methylpiperazin-1-yl)-8-isoquinolyl]-1- .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. N tetrahydropyran-4-yl-6,7-dihydro-4H-
9.50 (d, J = 1.0 Hz, 1H), 7.70-7.63
pyrazolo[4,3-c]pyridin-5-yl]ethanone (m, 1H), 7.63-7.51 (m, 1H),
7.31-7.17 ##STR00467## (m, 1H), 7.02 (d, J = 0.9 Hz, 1H), 4.52 (d,
J = 2.7 Hz, 2H), 4.44 (dd, J = 11.3, 4.1 Hz, 1H), 4.07-3.95 (m,
2H), 3.88- 3.75 (m, 2H), 3.53 (ddd, J = 8.9, 7.4, 3.4 Hz, 6H),
3.01- 2.77 (m, 2H), 2.46 (t, J = 5.0 Hz, 4H), 2.24 (s, 3H),2.15-
2.00 (m, 5H), 1.93 (d, J = 11.4 Hz, 2H); m/z 475.3. Example 128
1-[3[3-(1-piperidyl)-8-isoquinolyl]- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. N 1-tetrahydropyran-4-yl-6,7-dihydro- 9.47
(d, J = 0.8 Hz, 1H), 7.76-7.48
4H-pyrazolo[4,3-c]pyridin-5-yl]ethanone (m, 2H), 7.19 (ddd, J =
26.4, 7.1, ##STR00468## 1.2 Hz, 1H), 6.99 (d, J = 1.0 Hz, 1H), 4.51
(d, J = 2.5 Hz, 2H), 4.43 (td, J = 11.2, 5.4 Hz, 1H), 4.09-3.95 (m,
2H), 3.81 (dt, J = 14.5, 5.8 Hz, 2H), 3.66- 3.45 (m, 6H), 3.01-2.77
(m, 2H), 2.22-1.98 (m, 5H), 1.98- 1.85 (m, 2H), 1.61 (d, J = 7.6
Hz, 6H); m/z 460.3. Example 129 1-[3-(3-pyrrolidin-1-yl-8- .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. N
isoquinolyl)-1-tetrahydropyran-4-yl-6,7- 9.44 (t, J = 0.8 Hz, 1H),
7.60 (dt, J =
dihydro-4H-pyrazolo[4,3-c]pyridin-5-yl]ethanone 8.5, 1.0 Hz, 1H),
7.52 (ddd, J = 8.8, ##STR00469## 6.9, 2.1 Hz, 1H), 7.12 (ddd, J =
26.7, 6.9, 1.2 Hz, 1H), 6.67-6.47 (m, 1H), 4.50 (s, 2H), 4.47- 4.35
(m, 1H), 4.10-3.92 (m, 2H), 3.81 (dt, J = 15.2, 5.8 Hz, 2H),
3.57-3.41 (m, 6H), 2.88 (dt, J = 48.6, 5.9 Hz, 2H), 2.19-2.04 (m,
4H), 2.02- 1.96 (m, 5H), 1.97-1.89 (m, 2H); m/z 446.2. Example 130
1-[3-(3-morpholino-8-isoquinolyl)- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. N 1-tetrahydropyran-4-yl-6,7-dihydro- 9.53
(d, J = 0.9 Hz, 1H), 7.73-7.65
4H-pyrazolo[4,3-c]pyridin-5-yl]ethanone (m, 1H), 7.65-7.56 (m, 1H),
7.39-7.19 ##STR00470## (m, 1H), 7.04 (d, J = 0.9 Hz, 1H), 4.52 (d,
J = 2.2 Hz, 2H), 4.49-4.36 (m, 1H), 4.09-3.93 (m, 2H), 3.88-3.72
(m, 6H), 3.52 (q, J = 6.8, 5.2 Hz, 6H), 2.99-2.77 (m, 2H), 2.16-
1.99 (m, 5H), 1.93 (d, J = 11.0 Hz, 2H); m/z 462.2. Example 131
1-(3-(quinolin-5-yl)-1-(tetrahydro- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. O 2H-pyran-4-yl)-1,4,6,7-tetrahydro- 8.93
(dd, J = 4.1, 1.7 Hz, 1H), 8.87
5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one (ddt, J = 9.0, 2.5, 1.2
Hz, 1H), ##STR00471## 8.04 (dt, J = 8.5, 1.0 Hz, 1H), 7.83 (ddd, J
= 8.4, 7.1, 3.2 Hz, 1H), 7.61-7.53 (m, 2H), 4.49 (s, 2H), 4.44
(ddd, J = 11.3, 7.4, 4.0 Hz, 1H), 4.05- 3.96 (m, 2H), 3.82 (dt, J =
13.9, 5.8 Hz, 2H), 3.59-3.46 (m, 2H), 2.95 (t, J = 5.9 Hz, 1H),
2.89-2.77 (m, 1H), 2.20- 2.06 (m, 4H), 2.00 (s, 1H), 1.97-1.88 (m,
2H); m/z 377. Example 132 1-(3-(2-methyl-2H-indazol-4-yl)-1-
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. O
(tetrahydro-2H-pyran-4-yl)-1,4,6,7- 8.67 (d, J = 2.8 Hz, 1H), 7.54
tetrahydro-5H-pyrazolo[4,3-c] (d, 7= 8.6 Hz, 1H), 7.30
pyridin-5-yl)ethan-1-one (ddd, J = 8.5, 6.9,3 4 Hz, ##STR00472##
1H), 7.12-7.01 (m, 1H), 4.71 (s, 2H), 4.39 (tt, J = 11.5, 4.2 Hz,
1H), 4.23 (s, 3H), 4.03 (dd, J = 11.4, 4.3 Hz, 2H), 3.85-3.74 (m,
2H), 3.53 (td, J = 11.9, 1.8 Hz, 2H), 2.94- 2.74 (m, 2H), 2.29-2.15
(m, 2H), 2.15-2.08 (m, 3H), 1.89 (dd, J = 12.2, 4.9 Hz, 2H); m/z
380. Example 133 1-(3-(naphthalen-1-yl)-1-(tetrahydro- .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. O
2H-pyran-4-yl)-1,4,6,7-tetrahydro- 8.36-8.28 (m, 1H), 8.02-7.91 (m,
2H), 5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one 7.62-7.43 (m, 4H),
4.41 (s, 3H), ##STR00473## 4.01 (dd, J = 10.6, 4.8 Hz, 2H),
3.87-3.75 (m, 2H), 3.57-3.46 (m, 2H), 2.99-2.79 (m, 2H), 2.20-2.03
(m, 4H), 2.00-1.88 (m, 3H); m/z 376. Example 134
1-(3-(1H-indol-6-yl)-1-(tetrahydro- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. O 2H-pyran-4-yl)-1,4,6,7-tetrahydro- 11.09
(d, J = 10.7 Hz, 1H), 5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one
7.69-7.60 (m, 1H), 7.57 ##STR00474## (d, J = 8.2 Hz, 1H), 7.41-
7.28 (m, 2H), 6.42 (ddd, J = 2.9, 1.9, 0.9 Hz, 1H), 4.69 (d, J =
8.4 Hz, 2H), 4.33 (ddt, J = 11.3, 7.3, 4.1 Hz, 1H), 4.05- 3.94 (m,
2H), 3.78 (dt, J = 19.0, 5.9 Hz, 2H), 3.57-3.43 (m, 2H), 2.91-2.71
(m, 2H), 2.18-2.05 (m, 5H), 1.85 (d, J = 12.9 Hz, 2H); m/z 365.
Example 135 1-(3-(benzo[b]thiophen-3-yl)-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. O (tetrahydro-2H-pyran-4-yl)-1,4,6,7- 8.77-
8.63 (m, 1H), 8.17-7.99 (m, 1H), tetrahydro-5H-pyrazolo[4,3-c]
7.88-7.63 (m, 1H), 7.50-7.34 (m, pyridin-5-yl)ethan-1-one 2H), 4.66
(d, J = 4.4 Hz, 2H), ##STR00475## 4.41 (td, J = 11.3, 5.6 Hz, 1H),
4.07-3.97 (m, 2H), 3.80 (dt, J = 13.7, 5.8 Hz, 2H), 3.58- 3.46 (m,
2H), 2.91- 2.70 (m, 2H), 2.22-2.07 (m, 5H), 1.91 (d, J = 13.3 Hz,
2H); m/z 382. Example 136 1-(3-(quinolin-4-yl)-1-(tetrahydro-
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. O
2H-pyran-4-yl)-1,4,6,7-tetrahydro-5H- 8.95 (dd, J = 4.5, 2.4 Hz,
1H), 8.61 pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one (dt, J = 8.7, 2.0
Hz, 1H), 8.11-8.03 ##STR00476## (m, 1H), 7.79 (ddd, J = 8.4, 6.8,
1.4 Hz, 1H), 7.65 (dddd, J = 8.3, 6.8, 2.4, 1.3 Hz, 1H), 7.61-7.38
(m, 1H), 4.56 (d, J = 5.2 Hz, 2H), 4.47 (ddt, J = 11.3, 7.3, 4.1
Hz, 1H), 4.10-3.95 (m, 2H), 3.82 (dt, J = 11.9, 5.8 Hz, 2H),
3.58-3.47 (m, 2H), 3.01-2.79 (m, 2H), 2.20- 2.05 (m, 4H), 2.01 (s,
1H), 1.95 (td, J = 6.1, 3.7 Hz, 2H); m/z 377. Example 137
1-(3-(5-fluoro-1H-indol-4-yl)-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. O (tetrahydro-2H-pyran-4-yl)-1,4,6,7- 11.21
(d, J = 2.9 Hz, 1H), 7.45-7.34 tetrahydro-5H-pyrazolo[4,3-c] (m,
2H), 7.01 (ddd, J = 11.2, 8.8, 3.1 pyridin-5-yl)ethan-1-one Hz,
1H), 6.71-6.58 (m, 1H), 4.39 ##STR00477## (d, J = 2.1 Hz, 3H),
4.06- 3.96 (m, 2H), 3.86-3.74 (m, 2H), 3.57-3.45 (m, 2H), 2.97-
2.75 (m, 2H), 2.17-2.04 (m, 4H), 1.99 (s, 1H), 1.96- 1.86 (m, 2H);
m/z 383. Example 138 5-(5-acetyl-1-(tetrahydro-2H-pyran- .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. O 4-yl)-4,5,6,7-tetrahydro-1H-
11.30 (d, J = 5.6 Hz, 1H), 8.24 pyrazolo[4,3-c]pyridin-3- (dd, J =
8.0, 1.3 Hz, 1H), 7.74-7.60 yl)isoquinolin-1(2H)-one (m, 1H), 7.54
(td, J = 7.7, 1.8 Hz, 1H), ##STR00478## 7.17 (dt, J = 7.3, 5.1Hz,
1H), 6.93 (t, J = 8.1 Hz, 1H), 4.47- 4.34 (m, 3H), 3.99 (dd, J =
10.4, 4.7 Hz, 2H), 3.79 (dt, J = 12.0, 5.7 Hz, 2H), 3.50 (td, J =
12.0, 2.3 Hz, 2H), 2.96-2.76 (m, 2H), 2.12-2.05 (m, 3H), 2.00 (s,
1H), 1.94-1.85 (m, 3H); m/z 393. Example 139
1-(1-(tetrahydro-2H-pyran-4-yl)-3- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. O (5,6,7,8-tetrahydronaphthalen-1-yl 7.12 (d,
J = 7.6 Hz, 1H), 7.06 1,4,6,7-tetrahydro-5H-pyrazol[4,3-c] (d, J =
8.0 Hz, 1H), 7.02-6.96 (m, 1H), pyridin-5-yl)ethan-1-one 4.34 (d, J
= 3.3 Hz, 2H), 3.96 ##STR00479## (d, J = 12.3 Hz, 2H), 3.76 (dt, J
= 16.8, 5.8 Hz, 2H), 3.47 (tt, J = 12.0, 2.2 Hz, 2H), 2.87 (dd, J =
10.0, 4.3 Hz, 1H), 2.81-2.60 (m, 5H), 2.08 (s, 2H), 2.06-1.98 (m,
3H), 1.84 (dd, J = 12.8, 5.2 Hz, 2H), 1.78-1.61 (m, 5H); m/z 380.
Example 140 1-(3-(1H-indazol-4-yl)-1- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. O (tetrahydro-2H-pyran-4-yl)-1,4,6,7- 13.09
(s, 1H), 8.54 (d, J = 6.2 Hz, 1H), tetrahydro-5H-pyrazol[4,3-c]
7.49 (d, J = 8.3 Hz, 1H), 7.47-7.37 pyridin-5-yl)ethan-1-one (m,
1H), 7.14 (ddd, J = 18.2, 7.1, 0.8 ##STR00480## Hz, 1H), 4.71 (s,
2H), 4.41 (ddd, J = 11.3, 8.9, 4.3 Hz, 1H), 4.08-3.98 (m, 2H),
3.85-3.74 (m, 2H), 3.59-3.47 (m, 2H), 2.96-2.76 (m, 2H), 2.25-2.03
(m, 5H), 1.92 (d, J = 13.0 Hz, 2H); m/z 366. Example 141
1-(3-(1H-indol-4-yl)-1-(tetrahydro- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. O 2H-pyran-4-yl)-1,4,6,7-tetrahydro- 10.74
(s, 1H), 7.54 (d, J = 7.7 Hz, 1H),
5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one 7.45 (t, J =2.8 Hz, 1H),
7.26-7.15 ##STR00481## (m, 1H), 7.11 (td, J = 7.6, 1.4Hz, 1H), 6.50
(dd, J = 3,1, 2.1 Hz,1H), 4.75 (d, J = 3.2 Hz, 2H), 4.41 (ddt, J =
11.6, 7.6, 4.0 Hz, 1H), 4.10-3.97 (m, 2H), 3.81 (dt, J = 15.5, 5.8
Hz, 2H), 3.60-3.44 (m, 2H), 2.97-2.74 (m, 2H), 2.37 (tq, J = 12.2,
6.1,5.5 Hz, 2H), 2.13 (d, J = 5.2 Hz, 3H), 1.87(t, J = 8.3 Hz, 2H);
m/z 365. Example 142 1-(1-(tetrahydro-2H-pyran-4-yl)-3- .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. O
(5-(trifluoromethyl)-1H-pyrrolo[2,3-b] 8.84 (dd, J = 12.2, 2.3 Hz,
1H), 8.63 pyridin-3-yl)-1,4,6,7-tetrahydro-5H- (d, J = 2.1 Hz, 1H),
7.95-7.75 (m, 1H), pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one 4.64 (s,
2H), 4.37 (ddq, J = 11.2, 7.8, ##STR00482## 4.0 Hz, 1H), 4.03 (dd,
J = 10.6, 4.3 Hz, 2H), 3,78 (dt, J = 15.3, 5.7 Hz, 2H), 3.51 (t, J
= 11,7 Hz, 2H), 2.92-2.71 (m, 2H), 2.20-2 04 (m, 6H), 1.94-1.84 (m,
2H); m/z 434. Example 143 1-(3-(2-aminoquinazolin-5-yl)-1- .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. P
(tetrahydro-2H-pyran-4-yl)-1,4,6,7- 9.59 (d, J = 4.5 Hz, 1H), 7.73
tetrahydro-5H-pyrazolo[4,3-c] (ddd, J = 8.5, 7.2, 3.6 Hz, 1H), 7.41
pyridin-5-yl)ethan-1-one (d, J = 8 4 Hz, 1H), 7.28-7.10 (m, 1H),
##STR00483## 6.82 (s, 2H), 4.55 (s, 2H), 4.43 (tq, J = 11.4, 4.2
Hz, 1H), 4.01 (dd, J = 10.7, 4.7 Hz, 2H), 3.81 (dt, J = 12.1, 5.8
Hz, 2H), 3.51 (dt, J = 11.7, 2.1 Hz, 2H), 2.94-2.74 (m, 2H),
2.18-2.00 (m, 5H), 1.98-1.87(m, 2H); m/z 393. Example 144
1-(3-(isoquinolin-8-yl)-1- .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. P (tetrahydro-2H-pyran-4-yl)-1,4,6,7- 9.81 (d, J = 1.0 Hz,
1H), 8.54 tetrahydro-5H-pyrazolo[4,3-c] (d, J = 5.6 Hz, 1H), 7.99
(d, J = 8.2 pyridin-5-yl)ethan-1-one Hz, 1H), 7.92-7.80 (m, 2H),
7.73-7.59 ##STR00484## (m, 1H), 4.57-4.40 (m, 3H), 4.05-3.96 (m,
2H), 3.83 (dt, J = 12.0, 5 8 Hz, 2H), 3.58-3.46 (m, 2H), 3.02-2.80
(m, 2H), 2.20-2.05 (m, 4H), 2.01 (s, 1H), 1.96 (t, J = 8.3 Hz, 2H);
m/z 377. Example 145 4-(5-acetyl-1-(tetrahydro-2H-pyran- .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. Q 4-yl)-4,5,6,7-tetrahydro-1H-
7.81-7.61 (m, 5H), 7.10-7.01 (m, 2H),
pyrazolo[4,3-c]pyridin-3-yl)-2-(4- 5.19-5.13 (m, 2H), 4.75 (s, 2H),
methoxyphenyl)isoindolin-1-one 4.42 (td, J = 11.1, 5.5 Hz, 1H),
##STR00485## 4.02 (d, J = 11.7 Hz, 2H), 3.85-3.74 (m, 2H), 3.79 (s,
3H), 3.57-3.47 (m, 2H), 2.95-2.75 (m, 2H), 2.18- 2.05 (m, 4H), 1.91
(d, J = 13.0 Hz, 3H); m/z 487. Example 146
4-(5-acetyl-1-(tetrahydro-2H-pyran- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. Q 4-yl)-4,5,6,7-tetrahydro-1H- 8.14 (d, J =
1.7 Hz, 1H), 7.76-7.60 pyrazolo[4,3-c]pyridin-3-yl)-2-(1- (m, 4H),
5.06-5.00 (m, 2H), 4.74 methyl-1H-pyrazol-4-yl)isoindolin-1-one (s,
2H), 4 43 (td, J = 11.3, 5.4 Hz, ##STR00486## 1H), 4.09-4.01 (m,
2H), 3.89 (s, 3H), 3.85-3.74 (m, 2H), 3.53 (dd, J = 12.8, 10.6 Hz,
2H), 2.96-2.75 (m, 2H), 2.19- 2.09 (m, 5H), 1.92 (d, J = 12.9 Hz,
2H); m/z 461. Example 147
1-(3-(6-(1-methyl-1H-pyrazol-4-yl)-1H-indol-1-yl)- Not Determined;
m/z 445. R 1-(tetrahydro-2H-pyran-4-yl)-1,4,6,7-tetrahydro-
5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one ##STR00487## Example 148
1-(3-(2-(4-(pyrrolidin-1- Not Determined; m/z 511. S
yl)phenyl)imidazo[1,2-a]pyridin-8-
yl)-1-(tetrahydro-2H-pyran-4-yl)-
1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c] pyridin-5-yl)ethan-1-one
##STR00488## Example149 1-(1-(tetrahydro-2H-pyran-4-yl)-3- .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. S
(2-(3-(trifluoromethyl)phenyl)imidazo[1,2-a] 8.69 (d, J = 1.3 Hz,
1H), 8.57 (td, J = pyridin-8-yl)-1,4,6,7-tetrahydro- 6.9, 1.3 Hz,
1H), 8.45- 8.26 (m, 2H), 5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one
7.69 (ddd, J = 8.8, 4 9, 2.5 Hz, ##STR00489## 2H), 7.48 (ddd, J =
14 9, 7.1, 1.2 Hz, 1H), 7.01 (q, J = 7.1 Hz, 1H), 4.83 (d, J = 14.2
Hz, 2H), 4.40 (ddt, J = 15.4, 11.0, 5.4 Hz, 1H), 4.05-3.96 (m, 2H),
3.81 (q, J = 5.9 Hz, 2H), 3.51 (dd, J = 12.8, 10.8 Hz, 2H),
2.94-2.74 (m, 2H), 2.20- 2.04 (m, 4H), 1.98 (s, 1H), 1.95-1.85 (m,
2H); m/z 510. Example 150
1-(3-(2-(2-fluoro-4-methoxyphenyl)imidazo[1,2-a] .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. S
pyridin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-1,4,6,7- 8.61 (dd, J =
6.6, 2.4 Hz, 1H), 8.40-8.11
tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one (m, 2H),
7.46(dd, J = 10.3, 6.9 Hz, 1H), ##STR00490## 7.02-6.85 (m, 3H),
4.84 (s, 2H), 4.40 (tq, J = 10.4, 4.7 Hz, 1H), 4.00 (dd, J = 11.1,
4.2 Hz, 2H), 3.87-3.77 (m, 5H), 3.50 (t, J = 11.7 Hz, 2H),
2.94-2.75 (m, 2H), 2.19- 2.03 (m, 4H), 1.97 (s, 1H), 1.89 (dd, J =
11.8, 3.3 Hz, 2H); m/z 490. Example 151
1-(3-(2-(3-methoxyphenyl)imidazo[1,2-a] .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. S pyridin-8-yl)-1-(tetrahydro-2H- 8.58-8.52
(m, 1H), 8.51 (s, 1H), pyran-4-yl)-1,4,6,7-tetrahydro-5H- 7.66-7.50
(m, 2H), 7.51-7.40 (m, 1H), pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one
7.36 (q, J = 8.1 Hz, 1H), 6.97 (q, J = 6.7 ##STR00491## Hz, 1H),
6.93-6.85 (m, 1H), 4.89-4.76 (m, 2H), 4.40 (dq, J = 11.3, 6.1 Hz,
1H), 4.03-3.96 (m, 2H), 3.87- 3.76 (m, 5H), 3.50 (t, J = 11.9 Hz,
2H), 2.98-2.78 (m, 2H), 2.18-2.04 (m, 4H), 1.98 (s, 1H), 1.89 (d, J
= 13.0 Hz, 2H); m/z 472. Example 152
1-(3-(2-(3-fluoro-4-methoxyphenyl)imidazo[1,2-a] .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. S
pyridin-8-yl)-1-(tetrahydro-2H-pyran-4-yl)-1,4,6,7- 8.53 (ddd, J =
6.7, 4.4, 1.3 Hz, 1H),
tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one 8.44 (d, J =
1.9 Hz, 1H), 7.92-7.82 ##STR00492## (m, 1H), 7.82-7.75 (m, 1H),
7.45 (ddd, J = 13.0, 7.1, 1.2 Hz, 1H), 7.24 (dt, J = 14.5, 9.0 Hz,
1H), 6.97 (q, J = 6.8 Hz, 1H), 4.84 (d, J = 5.1 Hz, 2H), 4.39 (dt,
J = 11.2, 6.0 Hz, 1H), 4.00 (d, J = 11.4 Hz, 2H), 3.88 (s, 3H),
3.82 (dt, J = 11.6, 5.8 Hz, 2H), 3.55-3.43 (m, 2H), 2.97-2.75 (m,
2H), 2.17-2.05 (m, 4H), 1.99 (s, 1H), 1.89 (d, J = 12.6 Hz, 2H);
m/z 490. Example 153 1-(3-(2-(1H-indol-3-yl)imidazo[1,2-a] .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. S
pyridin-8-yl)-1-(tetrahydro-2H- 11.42- 11.29 (m, 1H), 8.54 (td, J =
pyran-4-yl)-1,4,6,7-tetrahydro-5H- 6.6, 6.2, 1.3 Hz, 1H), 8.42 (s,
1H), pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one 8.20-7.81 (m, 2H),
7.49-7.37 (m,
2H), ##STR00493## 7.21-7.08 (m, 2H), 6.93 (q, J = 7.1 Hz, 1H),
4.98-4.85 (m, 2H), 4.40 (tt, J = 10.4, 4.7 Hz, 1H), 4.05-3.96 (m,
2H), 3.82 (dt, J = 12.1, 5.7 Hz, 2H), 3.51 (td, J = 11.9, 2.0 Hz,
2H), 3.01-2.79 (m, 2H), 2.21- 2.05 (m, 4H), 1.98-1.86 (m, 3H); m/z
481. Example 154 1-(3-(2-(3-hydroxyphenyl)imidazo[1,2-a] .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. S
pyridin-8-yl)-1-(tetrahydro-2H- 9.44 (s, 1H), 8.54 (ddd, J = 6.7,
2.8, 1.2 pyran-4-yl)-1,4,6,7-tetrahydro-5H- Hz, 1H), 8.40 (d, J =
3.1 Hz, 1H), 7.49- pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one 7.35 (m,
3H), 7.22 (td, J = 7.8, 3.0 Hz, ##STR00494## 1H), 7.01-6.91 (m,
1H), 6.76-6.68 (m, 1H), 4.86 (s, 1H), 4.76 (s, 1H), 4.44-4.34 (m,
1H), 4.00 (d, J = 11.3 Hz, 2H), 3.83 (q, J = 5.6 Hz, 2H), 3.50 (dd,
J = 12.6, 10.8 Hz, 2H), 2.94 (d, J = 5.8 Hz, 1H), 2.83 (d, J = 5.9
Hz, 1H), 2.20-2.04 (m, 4H), 1.99 (s, 1H), 1.89 (d, J = 12.7 Hz,
2H); m/z 458. Example 155 1-(3-(2-(4-methoxyphenyl)imidazo[1,2-a]
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. S
pyridin-8-yl)-1-(tetrahydro-2H- 8.53 (ddd, J = 6.7, 4.0, 1.3 Hz,
1H), pyran-4-yl)-1,4,6,7-tetrahydro-5H- 8.37 (d, J = 1.8 Hz, 1H),
8.03-7.96 pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one (m, 1H),
7.95-7.88 (m, 1H), 7.43 ##STR00495## (ddd, J = 13.4, 7.1, 1.3 Hz,
1H), 7.08-6.98 (m, 2H), 6.95 (q, J = 6.8 Hz, 1H), 4.85 (d, J = 13.8
Hz, 2H), 4,39 (tt, J = 10.5, 5.1 Hz, 1H), 4.00 (d, J = 11.7 Hz,
2H), 3.87-3.77 (m, 5H), 3.50 (t, J = 11.7 Hz, 2H), 2.98-2.77 (m,
2H), 2.19- 2.04 (m, 4H), 1.99 (s, 1H), 1.94-1.85 (m, 2H); m/z 472.
Example 156 1-(1-(tetrahydro-2H-pyran-4-yl)-3-(2-(4- .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. S
(trifluoromethoxy)phenyl)imidazo[1,2-a] 8.61-8.55 (m, 1H), 8.54 (s,
1H), pyridin-8-yl)-1,4,6,7-tetrahydro- 8.24-8.15 (m, 1H), 8.15-8.07
(m, 1H), 5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one 7.52-7.38 (m,
3H), 6.99 (q, J = 6.6 ##STR00496## Hz, 1H), 4.84 (s, 2H), 4.40 (tt,
J = 10.7, 4,8 Hz, 1H), 4.00 (dd, J = 11.3, 4.4 Hz, 2H), 3.82 (dt, J
= 11.8, 5.8 Hz, 2H), 3.50 (td, J = 12.1, 2.1 Hz, 2H), 2.98- 2.75
(m, 2H), 2.19-2.03 (m, 4H), 1.97 (s, 1H), 1.89 (d, J = 13.6 Hz,
2H); m/z 526. Example 157 1-(3-(2-(4-methoxyphenyl)imidazo[1,2-a]
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. S
pyridin-5-yl)-1-(tetrahydro-2H- 9.01 (d, J = 4.7 Hz, 1H), 7.93-7.85
pyran-4-yl)-1,4,6,7-tetrahydro-5H- (m, 2H), 7.62 (d, J = 8.9 Hz,
1H), 7.38 pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one (dt, J = 9.0, 6.9
Hz, 1H), 7.09-6.93 ##STR00497## (m, 3H), 4.67 (s, 2H), 4.53 (tt, J
= 11.2, 4.0 Hz, 1H), 4.06 (dd, J = 10.8, 4.7 Hz, 2H), 3.88-3.77 (m,
5H), 3.55 (tt, J = 11.6, 2.1 Hz, 2H), 2.97- 2.78 (m, 2H), 2.24-2.04
(m, 5H), 2.05-1.94 (m, 2H); m/z 472. Example 158
1-(3-(2-(pyridin-3- .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. T
yl)benzo[d]oxazol-7-yl)-1- 9.51-9.37 (m, 1H), 8.82 (dt, J = 4.9,
1.6 (tetrahydro-2H-pyran-4-yl)-1,4,6,7- Hz, 1H), 8.72-8.54 (m, 1H),
7.84 (dd, tetrahydro-5H-pyrazolo[4,3-c] J = 7.9, 1.1 Hz, 1H),
7.73-7.63 (m, 2H), pyridin-5-yl)ethan-1-one 7.52 (t, J = 7.8 Hz,
1H), 4.86-4.72 ##STR00498## (m, 2H), 4.44 (ddt, J = 11.3, 8.3, 4.2
Hz, 1H), 4.07-3.98 (m, 2H), 3.92-3.79 (m, 2H), 3.52 (td, J = 11.9,
2.1 Hz, 2H), 3.03- 2.79 (m, 2H), 2.22-2.04 (m, 4H), 2.01 (s, 1H),
1.97-1.88 (m, 2H); m/z 444. Example 159 1-(3-(2-(4- .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. T methoxyphenyl)benzo[d]oxazol-7-
8.32-8.11 (m, 2H), 7.74 (dd, J = 8.0, 1.1
yl)-1-(tetrahydro-2H-pyran-4-yl)- Hz, 1H), 7.62 (dd, J = 7.6, 1.2
Hz, 1H), 1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c] 7.45 (t, J = 7.8 Hz,
1H), 7.23-7.12 (m, pyridin-5-yl)ethan-1-one 2H), 4.80-4.70 (m, 2H),
4.49-4.37 (m, ##STR00499## 1H), 4.07-3.98 (m, 2H), 3.91- 3.79 (m,
5H), 3.52 (td, J = 1.9, 2.1 Hz, 2H), 2.95-2.75 (m, 2H), 2.22-2.06
(m, 4H), 2.01 (s, 1H), 1.92 (dd, J = 13.1, 3.8 Hz, 2H); m/z 473.
Example 160 3-(5-acetyl-1-tetrahydrofuran-3-yl- .sup.1H NMR (400
MHz, DMSO-d.sub.6, 24/26 H) U 6,7-dihydro-4H-pyrazolo[4,3-c]
.delta. 10.50 (d, J = 3.6 Hz, 1H), 8.29-8.10
pyridin-3-yl)-N-(1-methylpyrazol-4-yl)benzamide (m, 1H), 8.04 (d, J
= 0.8 Hz, 1H), ##STR00500## 7.95-7.84 (m, 1H), 7.84- 7.71 (m, 1H),
7.68-7.51 (m, 2H), 5.00 (dd, J = 6 7, 4.7 Hz, 1H), 4,70 (s, 2H),
4.16-3.99 (m, 2H), 3.98-3.67 (m, 6H), 3.00-2.84 (m, 1H), 2.44- 2.26
(m, 2H), 2.18-1.97 (m, 3H); m/z 435.2. Example 161
3-(5-acetyl-1-tetrahydrofuran-3-yl- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. U 6,7-dihydro-4H-pyrazolo[4,3-c] 10.76 (s,
1H), 8.25-8.09 (m, 1H), 8.01 pyridin-3-yl)-N-(4- (dd, J = 8.8, 1.7
Hz, 2H), 7.96-7.76 cyanophenyl)benzamide (m, 4H), 7.63 (td, J =
7.7, 3.3 Hz, 1H), ##STR00501## 5.15-4.86 (m, 1H), 4.70 (s, 2H),
4.24-3.99 (m, 2H), 3.99 - 3.82 (m, 4H), 3.77 (dt, J = 11.3, 4.2 Hz,
4H), 2.11 (d, J = 9.7 Hz, 3H); m/z 456.2. Example 162
3-(5-acetyl-1-tetrahydrofuran-3-yl- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. U 6,7-dihydro-4H-pyrazolo[4,3-c] 10.68 (d, J
= 4.4 Hz, 1H), 8.82 (dd, J = pyridin-3-yl)-N-(6-quinolyl)benzamide.
4.2, 1.7 Hz, 1H), 8.66-8.49 (m, 1H), 8.40- ##STR00502## 8.29 (m,
1H), 8.29-8.14 (m, 1H), 8.12-7.91 (m, 3H), 7.84 (dd, J = 7.9, 1.5
Hz, 1H), 7.64 (td, J = 7.7, 2.7 Hz, 1H), 7.51 (dd, J = 8.3, 4,2 Hz,
1H), 5.01 (qd, J = 6.7, 4.6 Hz, 1H), 4.73 (d, J = 3.0 Hz, 2H),4.19-
4.00 (m, 2H), 4.00-3.84 (m, 3H), 3.78 (d, J =5.9 Hz, 3H), 2.36 (qd,
J = 7.1, 2.9 Hz, 2H), 2.21-1.98 (m, 3H); m/z 482.2. Example 163
3-(5-acetyl-1-tetrahydrofuran-3-yl- .sup.1H NMR (400 MHz,
DMSO-d.sub.6, 26/28 H) U 6,7-dihydro-4H-pyrazolo[4,3-c] .delta.
10.53 (d, J = 3.6 Hz, 1H), 8.29
pyridin-3-yl)-N-(4-imidazol-1-ylphenyl)benzamide (dt, J = 1.9, 1.0
Hz, 1H), 8.19 (dt, J = ##STR00503## 11.5, 1.8 Hz, 1H), 8.05-7.91
(m, 2H), 7.91-7.78 (m, 1H), 7.71 (dd, J = 8.6, 0.9 Hz, 1H), 7.63(t,
J = 7.7 Hz, 1H), 7.40 (ddd, J = 8.7, 2.9, 1.7 Hz, 1H), 5.08-4.90
(m, 1H), 4.72 (t, J = 2.3 Hz, 2H), 4.23- 3.97 (m, 5H), 3.98-3.82
(m, 2H), 3.78 (d, J = 5.9 Hz, 1H), 2.42-2.29 (m, 3H), 2.12 (d, J =
7.5 Hz, 3H); m/z 485.2. Example 164
3-(5-acetyl-1-tetrahydrofuran-3-yl- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. U 6,7-dihydro-4H-pyrazolo[4,3-c] 10.51 (d, J
= 3.6 Hz, 1H), 8.31-8.10
pyridin-3-yl)-N-(4-imidazol-1-ylphenyl)benzamide (m, 2H), 8.06-7.89
(m, 3H), 7.86-7.77 ##STR00504## (m, 1H), 7.72 (t, J = 1.3 Hz, 2H),
7.71-7.56 (m, 3H), 5.09- 4.91 (m, 1H), 4.71 (s, 2H), 4.08 (tt, J =
9.0, 7 0 Hz, 2H), 3.97-3.82 (m, 2H), 3.77 (q, J = 6.4, 5.5 Hz, 2H),
2.43- 2.25 (m, 4H), 2.11 (d, J = 7.9 Hz, 3H); m/z 497.2. Example
165 3-(5-acetyl-1-tetrahydrofuran-3-yl- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. U 6,7-dihydro-4H-pyrazolo[4,3-c] 10.39(d, J =
3.6 Hz, 1H), 8.64 (s, 1H), pyridin-3-yl)-N[4-(1,2,4-triazol- 8.14
(dt, J = 9.2, 1.8 Hz, 1H), 7.98 1-ylmethyl)phenyl]benzamide (s,
1H), 7.95-7.84 (m, 2H), 7.77 (dd, ##STR00505## J = 8.6, 1.9 Hz,
2H), 7.60 (td, J = 7.8, 2.5 Hz, 1H), 7.36- 7.25 (m, 2H), 5.38 (s,
2H), 4.99 (qd, J = 6.6, 4.6 Hz, 1H), 4.70 (d, J = 3.7 Hz, 2H),
4.16- 3.97 (m, 2H), 3.97-3.82 (m, 4H), 3.77 (q, J = 7.0, 6.0 Hz,
2H), 2.34 (qd, J = 6.8, 2.7 Hz, 2H), 2.16-2.02 (m, 3H); m/z 512.2.
Example 166 3(5-acetyl-1-tetrahydrofuran-3-yl- Not Determined; m/z
511.2. U 6,7-dihydro-4H-pyrazolo[4,3-c]
pyridin-3-yl)-N-(1-methyl-5- phenyl-imidazol-2-yl)benzamide
##STR00506## Example 167 3-(5-acetyl-1-tetrahydrofuran-3-yl-
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. U
6,7-dihydro-4H-pyrazolo[4,3-c] 12.77 (s, 1H), 8.57 (s, 1H),
8.36-8.07 pyridin-3-yl)-N(1-methylbenzimidazol-2-yl)benzamide (m,
1H), 8.00-7.68 (m, 1H), 7.68-7.39 ##STR00507## (m, 3H), 7.25 (dqd,
J = 8.8, 7.4, 1.3 Hz, 2H), 5.18-4.89 (m, 1H), 4.76 (d, J = 14.1 Hz,
2H), 4.27-3.98 (m, 2H), 3.99- 3.84 (m, 2H), 2.97-2.83 (m, 1H), 2.77
(s, 3H), 2.42- 2.28 (m, 5H), 2.13 (d, J = 11.2 Hz, 3H); m/z 485.2
Example 168 3-(5-acetyl-1-tetrahydrofuran-3-yl- .sup.1H NMR (400
MHz, DMSO-d.sub.6, 26/27 H) U 6,7-dihydro-4H-pyrazolo[4,3-c]
.delta. 10.82 (s, 1H), 9.16 (dd, J = 2.5, 0.8
pyridin-3-yl)-N-(3-quinolyl)benzamide Hz, 1H), 8.87 (dt, J =2.5,
1.3 Hz, 1H), ##STR00508## 8.42-8.10 (m, 1H), 8.16- 7.97 (m, 3H),
7.93-7.79 (m, 1H), 7.76-7.53 (m, 2H), 5.01 (qd, J = 6.7, 4.6 Hz,
1H), 4.73 (d, J = 3.7 Hz, 2H), 4.20- 3.98 (m, 2H), 3.98-3.84 (m,
2H), 2.89 (t, J = 5.9 Hz, 1H), 2.78 (d, J = 6.2 Hz, 1H), 2.44- 2.27
(m, 4H), 2.12 (d, J = 6.3 Hz, 3H); m/z 482.2. Example169
3-(5-acetyl-1-tetrahydrofuran-3-yl-6,7-dihydro-4H- Not Determined;
m/z 446.2. U
pyrazolo[4,3-c]pyridin-3-yl)-N-(2-methyl-4-pyridyl)benzamide
##STR00509## Example 170 3-(5-acetyl-1-tetrahydrofuran-3-yl-
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. U
6,7-dihydro-4H-pyrazolo[4,3-c] 10.76 (s, 1H), 9.18 (d, J = 1.4 Hz,
2H), pyridin-3-yl)-N-pyrimidin-5-yl-benzamide 8.93 (s, 1H), 8.21
(dt, J = 12.9, 1.7 Hz, ##STR00510## 1H), 8.07-7.78 (m, 2H), 7.64
(td, J = 7.8, 3.4 Hz, 1H), 5,00 (qd, J = 6.7, 4.7 Hz, 1H), 4.21-
3.98 (m, 2H), 3.98-3.66 (m, 6H), 2.35 (qd, J = 6.8, 2,3 Hz, 4H),
2.11 (d, J = 8.9 Hz, 3H); m/z 433.2. Example 171
3-(5-acetyl-1-tetrahydrofuran-3-yl- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. U 6,7-dihydro-4H-pyrazolo[4,3-c] 10.56 (s,
1H), 8.94 (dd, J = 2.5, 0.9 Hz,
pyridin-3-yl)-N-(3-pyridyl)benzamide 1H), 8.32 (dd, J = 4.6, 1.5
Hz, 1H), ##STR00511## 8.27-8.12 (m, 2H), 8.02-7.88 (m, 1H),
7.88-7.77 (m, 1H), 7.62 (td, J = 7.7, 2.9 Hz, 1H), 7.41 (ddt, J =
8.4, 4.8, 0.8 Hz, 1H), 5.00 (qd, J = 6.6, 4.6 Hz, 1H), 4.71 (d, J =
3.6 Hz, 2H), 4.19- 3.99 (m, 2H), 3.99-3.81 (m, 3H), 3.77 (q, J =
6.9, 5.7 Hz, 2H), 2.98-2.85 (m, 1H), 2.35 (qd, J = 6.7, 2.4 Hz,
2H), 2.18-1.98 (m, 3H); m/z 432.2. Example 172
3-(5-acetyl-1-tetrahydrofuran-3-yl- .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. U 6,7-dihydro-4H-pyrazolo[4,3-c] 10.38 (d, J
= 4.7 Hz, 1H), 8.30-8.22
pyridin-3-yl)-N-(1-methylindazol-5-yl)benzamide (m, 1H), 8.18 (dt,
J = 10.6, 1.7 Hz, ##STR00512## 1H), 8.03 (dd, J = 1.9, 0.9 Hz, 1H),
7.92 (dq, J =7.8, 1.6 Hz, 1H), 7.87-7.76 (m, 1H), 7.69 (d, J = 1.9
Hz, 1H), 7.66-7.54 (m, 2H), 5.00 (qd, J = 6.6, 4.7 Hz, 1H), 4.72
(d, J = 3.8 Hz, 2H), 4.17-3.97 (m, 5H), 3.99- 3.82 (m, 2H), 3.77
(q, J =7.0, 5.7 Hz, 2H), 2.89 (t, J = 5.8 Hz, 1H), 2.77 (s, 1H),
2.43- 2.25 (m, 2H), 2.11 (d, J =7.6 Hz, 3H); m/z 485.2. Example 173
5-(5-acetyl-1-(tetrahydro-2H-pyran- .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. V 4-yl)-4,5,6,7-tetrahydro-1H- 7.34-7.20 (m,
3H), 4.99-4.87 (m, 1H), pyrazolo[4,3-c]pyridin-3-yl)-2- 4.55- 4.27
(m, 4H), 4.24-4.04 (m, 3H),
isopropyl-1,2-dihydroisoquinolin-3(4H)-one 4.00-3.75 (m, 2H),
3.74-3.65 (m, ##STR00513## 2H), 3,54 (t, J = 12.0 Hz, 2H),
2.89-2.72 (m, 2H), 2.42- 2.26 (m, 2H), 2.20-2.11 (m, 3H), 1.95-1.83
(m, 2H), 1.19 (t, J = 6 8 Hz, 6H); m/z 437. Example 174
5-(5-acetyl-1-(tetrahydro-2H-pyran- .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. V 4-yl)-4,5,6,7-tetrahydro-1H- 7.34-7.18 (m,
3H), 4.54 (s, 2H), 4.53- pyrazolo[4,3-c]pyridin-3-yl)-2- 4.45 (m,
1H), 4.43-4.34 (m, 2H), 4.25-
cyclohexyl-1,2-dihydroisoquinolin-3(4H)-one 4.07 (m, 3H), 3.99-3.76
(m, 2H), 3.75- ##STR00514## 3.66 (m, 2H), 3.54 (t, J = 12.0 Hz,
2H), 2.90-2.73 (m, 2H), 2.42-2.27 (m, 2H), 2.20- 2.11 (m, 3H),
1.94-1.77 (m, 4H), 1.74-1.58 (m, 4H), 1.56- 1.36 (m, 4H); m/z 477.
Example 175 5-(5-acetyl-1-(tetrahydro-2H-pyran- .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. V 4-yl)-4,5,6,7-tetrahydro-1H-
7.30-7.15(m, 3H), 5.09-4.95 pyrazolo[4,3-c]pyridin-3-yl)-2- (m,
1H), 4.49-4.32 (m, 1H), cyclobutyl-1,2-dihydroisoquinolin-3(4H)-one
4.16-4.02 (m, 3H), 3.91- ##STR00515## 3.69 (m, 2H), 3.65-3.61 (m,
2H), 3.49 (t, J = 12.0 Hz, 2H), 2.80-2.68 (m, 2H), 2.35- 2.20 (m,
2H), 2.18-2.03(m, 7H), 1.88-1.79 (m, 2H), 1.75- 1.59 (m, 2H); m/z
449.
Example 176
IC.sub.50 Measurements for Inhibitors Using CBP TR-FRET Binding
Assay
[0739] His/Flag epitope tagged CBP was cloned, expressed, and
purified to homogeneity. CBP binding and inhibition was assessed by
monitoring the engagement of a biotinylated small molecule compound
with the target using the TR-FRET assay technology (Perkin-Elmer).
Specifically, in a 384 well ProxiPlate CBP (4 nM final) was
combined with biotin-ligand (60 nM final) in 50 mM HEPES (pH 7.5),
50 mM NaCl, 1 mM TCEP, 0.0100 (w/v) BSA, and 0.0080% (w/v) Brij-35
either in the presence of DMSO (final 0.20% DMSO) or compound
dilution series in DMSO. After 10 minutes incubation at room
temperature, a mixture Eu-W 1024 Anti-6.times.His antibody (Perkin
Elmer ADO 110) and SureLight.TM. All ophycocyanin-Streptavidin
(APC-SA, Perkin Elmer CR130-100) were added to a final
concentrations of 0.2 nMolar antibody and 50 nMolar APC-SA,
respectively. After twenty minutes of equilibration, the plates
were read on an Envision instrument and IC.sub.50s calculated using
a four parameter non-linear curve fit.
MYC_RPL19 QuantiGene Assay in MV-4-11 Cells
[0740] QuantiGene 2.0 Reagent system, Affymetrix: HUMAN MYCN; V-myc
myelocytomatosis viral related oncogene, neuroblastoma derived
(avian); NM_005378 SA-15008. 10,000 MV-4-11 cells (GNE in-house)
were plated in 75 ul complete media: RPMI-1640 (GNE in-house), 10%
FBS (Life Technologies, cat. no. 10082), 1% Pen-strep (GNE
in-house), in 96 well clear flat bottom plates (Costar, cat. no.
3595). 25 ul compound was added for 4 hours at 37 deg C. in a 1:3
serial dilution 10-point dose response, with a final DMSO
concentration=0.2%. The cells were then lysed according to the
assay kit's protocol and frozen at -80 deg C. The following day, an
appropriate volume of Working Probe Set was prepared by combining
the following reagents in the order listed: Nuclease-free water,
Lysis Mixture, Blocking Reagent, and 2.0 Probe Set (MYC or RPL19).
20 ul of the working probe set was added into each assay well on
the capture plate, and then 80 ul of the lysates were transferred
into the assay plates. The capture plate was placed in a 55 deg C.
incubator for overnight hybridization (16-20 hours). The following
day, wash buffer was prepared according to manufacturer's
recommendations. The capture plates were washed with 300 ul per
well of 1.times. wash buffer three times. Then 100 ul Pre-Amplifier
was added to the plate for a 60 minute incubation at 55 deg C.
After the incubation, the capture plate was washed with 300 ul per
well of 1.times. wash buffer three times, and 100 ul Amplifier was
added to the plate for a 60 minute incubation at 55 deg C. The
capture plate was again washed with 300 ul per well of 1.times.
wash buffer three times, and 100 ul Label Probe was added to the
plate for a 60 minute incubation at 50 deg C. Then the capture
plate was washed with 300 ul per well of 1.times. wash buffer three
times, and 100 ul 2.0 Substrate was added to each well of the
plate. The plates were incubated at RT for 5 minutes in the dark
and read on the Envision using the luminescence protocol, with an
integration time set at 0.2 seconds.
[0741] Data for representatitive compounds from the assays
described above is provided in the following table.
TABLE-US-00002 CBP HTRF Myc Example Compound IC.sub.50 (.mu.M)
IC.sub.50 (.mu.M) 1 ##STR00516## 0.0935 2 ##STR00517## 0.0972 3
##STR00518## 0.0174 4 ##STR00519## 0.0581 5 ##STR00520## 0.0687 6
##STR00521## 0.0493 7 ##STR00522## 0.0240 8 ##STR00523## 0.0687 9
##STR00524## 0.00113 0.0321 10 ##STR00525## 0.00115 0.0153 11
##STR00526## 0.00238 0.114 12 ##STR00527## 0.123 13 ##STR00528##
0.0598 14 ##STR00529## 0.00342 0.203 15 ##STR00530## 0.00232 0.0540
16 ##STR00531## 0.0899 17 ##STR00532## 0.0553 18 ##STR00533##
0.0676 19 ##STR00534## 0.00674 1.09 20 ##STR00535## 0.0813 21
##STR00536## 0.0263 6.16 22 ##STR00537## 0.0248 23 ##STR00538##
0.0167 24 ##STR00539## 0.00205 0.0263 25 ##STR00540## 0.00107
0.00596 26 ##STR00541## 0.00643 27 ##STR00542## 0.0167 28
##STR00543## 0.0416 29 ##STR00544## 0.0595 30 ##STR00545## 0.00502
0.198 31 ##STR00546## 0.00171 0.0646 32 ##STR00547## 0.00564 0.0728
33 ##STR00548## 0.00237 0.0988 34 ##STR00549## 0.00145 0.0592 35
##STR00550## 0.0182 36 ##STR00551## 0.00139 0.0465 37 ##STR00552##
0.00242 0.164 38 ##STR00553## 0.0165 1.43 39 ##STR00554## 0.00577
0.617 40 ##STR00555## 0.0176 0.727 41 ##STR00556## 0.0265 42
##STR00557## 0.0104 43 ##STR00558## 0.0547 44 ##STR00559## 0.00629
45 ##STR00560## 0.00154 0.481 46 ##STR00561## 0.00097 0.00903 47
##STR00562## 0.00191 0.0619 48 ##STR00563## 0.00961 0.568 49
##STR00564## 0.00142 0.0559 50 ##STR00565## 0.124 51 ##STR00566##
0.121 52 ##STR00567## 53 ##STR00568## 0.00238 0.0520 54
##STR00569## 0.124 55 ##STR00570## 0.0999 56 ##STR00571## 0.0308 57
##STR00572## 0.0431 58 ##STR00573## 0.109 59 ##STR00574## 0.0525 60
##STR00575## 0.0080 61 ##STR00576## 0.103 62 ##STR00577## 0.0430 63
##STR00578## 0.0999 64 ##STR00579## 0.0378 65 ##STR00580## 0.0849
66 ##STR00581## 0.0205 67 ##STR00582## 0.0300 68 ##STR00583##
0.0159 69 ##STR00584## 0.0155 70 ##STR00585## 0.00514 71
##STR00586## 0.00657 72 ##STR00587## 0.125 73 ##STR00588## 0.141 74
##STR00589## 0.0902 75 ##STR00590## 0.0134 0.523 76 ##STR00591##
0.0272 77 ##STR00592## 0.0233 78 ##STR00593## 0.0294 79
##STR00594## 0.0308 80 ##STR00595## 0.0378 81 ##STR00596## 0.0648
82 ##STR00597## 0.0805 83 ##STR00598## 0.00270 0.0662 84
##STR00599## 0.0222 8.87 85 ##STR00600## 0.00212 0.052 86
##STR00601## 0.00233 0.0305 87 ##STR00602## 0.000997 0.0180 88
##STR00603## 0.00116 0.0319 89 ##STR00604## 0.00192 90 ##STR00605##
0.00110 0.0168 91 ##STR00606## 0.00117 0.0283 92 ##STR00607##
0.000760 0.00735 93 ##STR00608## 0.000910 94 ##STR00609## 0.00100
0.00722 95 ##STR00610## 0.0357 96 ##STR00611## 0.00294 0.0755 97
##STR00612## 0.00334 0.221 98 ##STR00613## 0.00278 0.109 99
##STR00614## 0.0814 100 ##STR00615## 0.0788 101 ##STR00616##
0.00641 0.330 102 ##STR00617## 0.0568 103 ##STR00618## 0.0410 104
##STR00619## 0.0345 105 ##STR00620## 0.0338 106 ##STR00621## 0.0111
107 ##STR00622## 0.0954 108 ##STR00623## 0.0576 109 ##STR00624##
0.0912 110 ##STR00625## 0.0677 111 ##STR00626## 0.00362 0.152 112
##STR00627## 0.00349 0.249 113 ##STR00628## 0.00299 0.863 114
##STR00629## 0.00349 1.15 115 ##STR00630## 0.00898 0.942 116
##STR00631## 0.0245 117 ##STR00632## 0.0128 1.51 118 ##STR00633##
0.00281 0.850 119 ##STR00634## 0.00282 1.18 120 ##STR00635##
0.00369 0.119 121 ##STR00636## 0.00582 1.12 122 ##STR00637##
0.00408 0.563
123 ##STR00638## 0.0132 2.49 124 ##STR00639## 0.00215 0.491 125
##STR00640## 0.00417 0.925 126 ##STR00641## 0.00771 0.374 127
##STR00642## 0.0145 128 ##STR00643## 0.00244 0.280 129 ##STR00644##
0.00185 0.605 130 ##STR00645## 0.00725 0.285 131 ##STR00646## 0.094
132 ##STR00647## 0.133 133 ##STR00648## 0.0168 0.623 134
##STR00649## 0.123 135 ##STR00650## 0.0354 1.01 136 ##STR00651##
0.133 137 ##STR00652## 0.054 138 ##STR00653## 0.132 139
##STR00654## 0.131 140 ##STR00655## 0.0445 2.71 141 ##STR00656##
0.115 142 ##STR00657## 0.0423 0.504 143 ##STR00658## 0.0581 144
##STR00659## 0.0295 0.761 145 ##STR00660## 0.013 4.42 146
##STR00661## 0.0298 147 ##STR00662## 0.00609 0.263 148 ##STR00663##
0.0797 149 ##STR00664## 0.123 150 ##STR00665## 0.0667 151
##STR00666## 0.128 152 ##STR00667## 0.0461 >10 153 ##STR00668##
0.0957 154 ##STR00669## 0.101 155 ##STR00670## 0.0691 156
##STR00671## 0.107 157 ##STR00672## 0.00446 1.04 158 ##STR00673##
0.0193 5.32 159 ##STR00674## 0.00411 1.58 160 ##STR00675## 0.0737
161 ##STR00676## 0.082 162 ##STR00677## 0.0329 163 ##STR00678##
0.0497 164 ##STR00679## 0.0831 165 ##STR00680## 0.0442 166
##STR00681## 0.0365 167 ##STR00682## 0.011 168 ##STR00683## 0.0326
169 ##STR00684## 0.0592 170 ##STR00685## 0.123 171 ##STR00686##
0.104 172 ##STR00687## 0.0392 173 ##STR00688## 0.0305 174
##STR00689## 0.0119 175 ##STR00690## 0.0104 0.122
Exemplification of CBP/EP300 Inhibitors for the Treatment of
Fibrotic Disease
[0742] Cell culture: Collagen 1-coated 384-well plates (BD
Biosciences cat #356667) were seeded with Normal Human Lung
Fibroblasts (Lonza cat #CC-2512) at 2000 cells per well in 50 .mu.l
DMEM (Genentech) containing 0.5% fetal bovine serum (Sigma cat
#F2442). After 16 hours, the indicated compounds were added to
cells at final concentrations ranging from 10 .mu.M to 0.005 nM in
an 8-fold dilution series. After one hour, TGF beta (Genentech) was
added to cells to a final concentration of 10 ng/ml. All treatments
were performed in duplicate.
[0743] Animal study: Bleomycin was administered to mice via
subcutaneous implantation of an osmotic pump (Alzet cat #1007D).
After bleomycin administration, mice were treated with compounds by
oral gavage. Mice received either MCT vehicle (0.5% w/v
methylcellulose, 0.2% w/v polysorbate 80), G0272 in MCT at 5 mg/kg
twice daily, G0272 in MCT at 15 mg/kg twice daily, G5049 in MCT at
5 mg/kg twice daily, G5049 in MCT at 15 mg/kg twice daily, G3486 in
MCT at 15 mg/kg twice daily, or G3486 in MCT at 45 mg/kg twice
daily. To label newly synthesized collagen, mice were injected
intraperitoneally with 35 ml/kg heavy water (Sigma Aldrich, cat
#151882) in two doses and heavy water was provided in drinking
water. At study termination, blood samples were collected by
retro-orbital bleed under isoflurane anesthesia and mice were
euthanized. Upper right lung lobes were placed in glass vials and
snap frozen in liquid nitrogen for mass spectrometry. The lower
right lung lobe was placed in RNAlater for expression analysis, and
frozen at -20.degree. C.
[0744] Lung hydroxyproline determination: Lungs were thawed, dried
overnight at 80.degree. C., then hydrolyzed at 110.degree. C.
overnight in 6N HCl. The remainder of this paragraph was performed
by KineMed, Emeryville Calif.). A 100 .mu.l aliquot of tissue
hydrolysate received a spike containing 1 .mu.g
.sup.2H.sub.3-labeled hydroxyproline (D3-OH1P;
trans-4-Hydroxy-L-proline-2,5,5-d3; CDN), and then dried under
vacuum and re-suspended in a solution of 50% acetonitrile, 50 mM
K.sub.2HPO.sub.4 and pentafluorobenzyl bromide before incubation.
Derivatives were extracted into ethyl acetate, and the top layer
was removed and dried by vacuum centrifugation. In order to
acetylate the hydroxyl moiety of hydroxyproline, samples were
incubated with a solution of acetonitrile,
N-Methyl-N-[tert-butyldimethyl-silyl]trifluoroacetamide and
methylimidizole. This material was extracted in petroleum ether and
dried with Na.sub.2SO.sub.4. The derivatized hydroxyproline was
analyzed by GC/MS, performed in the negative chemical ionization
mode. Selected ion monitoring was performed on ions with
mass-to-charge ratios (m/z) 445, 446, 447, and 448 which include
all of the carbon-hydrogen bonds from hydroxyproline. Incorporation
of .sup.2H into hydroxyproline was calculated as the molar fraction
of molecules with one excess mass unit above the natural abundance
fraction (EMi). Fractional collagen synthesis (f) was calculated as
the ratio of the EMi value in protein-bound hydroxyproline to the
maximal value possible at the body water enrichment present. This
method has previously been described (Gardner, J. L., et al.,
Measurement of liver collagen synthesis by heavy water labeling:
effects of profibrotic toxicants and antifibrotic interventions. Am
J Physiol Gastrointest Liver Physiol, 2007. 292(6): p. G1695-705).
Additionally, hydroxyproline content in each tissue sample was
determined by comparing the abundance in the m3 448 m/z channel
representing the D3-OHP internal standard in each sample with that
the m0 445 m/z ion. A set of standards with known OHP/D3-OHP
concentration ratios was analyzed alongside the samples.
.sup.2H.sub.2O enrichment in plasma was determined using a
previously described method (Previs S F, Hazey J W, Diraison F,
Beylot M, David F, Brunengraber H (1996) Assay of the deuterium
enrichment of water via acetylene. J Mass Spectrom 31:639-642.).
Briefly body water is evaporated from plasma by overnight
incubation at 80.degree. C. Samples are then mixed in 10M NaOH and
acetone followed by a second overnight incubation. This material
was extracted in hexane and dried with Na.sub.2SO.sub.4 prior to
GCMS analysis.
[0745] RNA isolation: For cultured cells, after 24 hours of
treatment with TGF beta and CBP/p300 inhibitor, mRNA was isolated
with the Turbocapture 384 mRNA kit (Qiagen cat #72271) according to
the manufacturers' instructions and eluted with 30 .mu.l elution
buffer. For lungs, tissues were thawed, removed from RNAlater,
homogenized in GentleMACS M tubes (Miltenyi Biotec cat
#130-093-236) and RNA extracted with the RNeasy 96 kit (Qiagen cat
#74182) according to the manufacturers instructions.
Expression analysis: First-strand cDNA was synthesized using 14
.mu.l mRNA for cultured cells and 150 ng RNA for lung. The High
Capacity cDNA Reverse Transcription Kit (Life Technologies cat
#4368814) was used according to the manufacturers protocol.
Specific target amplification was performed using 1.25 .mu.l cDNA,
Taqman assays (Life Technologies cat #4331182) at a final
concentration of 0.2.times., and Taqman Preamp Master Mix (Life
Technologies cat #4488593) and subsequently diluted according to
the protocol for Fluidigm qPCR (Fluidigm Corp). Samples and assays
were mixed with loading buffers and loaded onto 192.24 IFCs
(Fluidigm cat #100-6266) according to the manufacturers
instructions. Reactions were mixed using the IFC controller RX
(Fluidigm) then amplified and measured using the Biomark system
(Fluidigm). For cultured cells, relative expression of each target
gene was determined using the .DELTA.Ct method, normalizing to the
Ct for HPRT1 using Excel software (Microsoft). To generate heat
maps, TGF beta-mediated expression increase for each gene in the
presence of CBP/p300 inhibitor was divided by the increase in the
absence of CBP/p300 inhibitor using Excel (i.e. (2.sup.-.DELTA.Ct,
SMI+TGFb-2.sup.-.DELTA.Ct, SMI, no TGFb)/(2.sup.-.DELTA.Ct,
TGFb-2.sup.-.DELTA.Ct, no TGFb)) Line graphs of 2.sup.-.DELTA.Ct
values were generated using Prism software (Graphpad). For lung,
relative expression of each target gene was determined using the
.DELTA..DELTA.Ct method, normalizing to the Ct for GAPDH and the
vehicle control group. Heat maps were generated with Excel software
(Microsoft).
[0746] While a number of embodiments have been described, these
examples may be altered to provide other embodiments that utilize
the compounds and methods described herein. Therefore, the scope of
this invention is to be defined by the appended claims rather than
by the specific embodiments that have been represented by way of
example.
* * * * *