U.S. patent application number 17/687259 was filed with the patent office on 2022-09-29 for inhibitors of cyclin dependent kinase 7 (cdk7).
The applicant listed for this patent is Syros Pharmaceuticals, Inc.. Invention is credited to Claudio Edmundo Chuaqui, Stephane Ciblat, Anzhelika Kabro, Kate-Lyn Lund, Jason J. Marineau, Henri Piras, Kenneth Matthew Whitmore.
Application Number | 20220305014 17/687259 |
Document ID | / |
Family ID | 1000006379521 |
Filed Date | 2022-09-29 |
United States Patent
Application |
20220305014 |
Kind Code |
A1 |
Marineau; Jason J. ; et
al. |
September 29, 2022 |
INHIBITORS OF CYCLIN DEPENDENT KINASE 7 (CDK7)
Abstract
The present invention provides, inter alia, compounds having the
structures of formulas described herein; pharmaceutically
acceptable salts, solvates, hydrates, tautomers, and isotopic forms
thereof; and compositions (e.g., pharmaceutical compositions and
kits) containing one or more of the foregoing. Also provided are
methods of administering and uses involving the compounds and/or
pharmaceutical compositions for treating or preventing disease. The
disease can be a proliferative disease, such as a cancer (e.g., a
blood cancer (e.g., a leukemia or lymphoma), a brain cancer, a
breast cancer, melanoma, multiple myeloma, or an ovarian cancer) a
benign neoplasm, pathologic angiogenesis, or a fibrotic disease.
While no aspect of the invention is limited by the biological
events that may transpire, administering a compound or other
composition described herein may selectively inhibit the aberrant
expression or activity of cyclin-dependent kinase 7 (CDK7) and,
thereby, induce cellular apoptosis and/or inhibit the transcription
of disease-related genes in the patient (or in a biological
sample).
Inventors: |
Marineau; Jason J.;
(Franklin, MA) ; Chuaqui; Claudio Edmundo;
(Arlington, MA) ; Ciblat; Stephane; (Montreal,
CA) ; Kabro; Anzhelika; (Lachine, CA) ; Piras;
Henri; (Montreal, CA) ; Whitmore; Kenneth
Matthew; (Montreal, CA) ; Lund; Kate-Lyn;
(Oro-Medonte, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Syros Pharmaceuticals, Inc. |
Cambridge |
MA |
US |
|
|
Family ID: |
1000006379521 |
Appl. No.: |
17/687259 |
Filed: |
March 4, 2022 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16962808 |
Jul 16, 2020 |
11311542 |
|
|
PCT/US2019/013845 |
Jan 16, 2019 |
|
|
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17687259 |
|
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|
62617884 |
Jan 16, 2018 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 45/06 20130101;
C07D 401/14 20130101; C07D 471/04 20130101; C07D 413/14 20130101;
A61K 31/506 20130101; C07D 417/14 20130101 |
International
Class: |
A61K 31/506 20060101
A61K031/506; A61K 45/06 20060101 A61K045/06; C07D 401/14 20060101
C07D401/14; C07D 413/14 20060101 C07D413/14; C07D 417/14 20060101
C07D417/14; C07D 471/04 20060101 C07D471/04 |
Claims
1. A pharmaceutical composition, wherein the composition is
formulated for oral administration and comprises a pharmaceutically
acceptable excipient and a compound of Formula (I): ##STR00305## or
a pharmaceutically acceptable salt, stereoisomer, or isotopic form
thereof, wherein: ring A is ##STR00306## ring B is ##STR00307##
wherein each of R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are,
independently, hydrogen or --CH.sub.3; X and Y are, independently,
N or CH; R.sup.1 is hydrogen or fluoro; and the compound is other
than: ##STR00308##
2. The pharmaceutical composition of claim 1, wherein ring A is
##STR00309##
3.-4. (canceled)
5. The pharmaceutical composition of claim 1, wherein ring B is
##STR00310##
6.-7. (canceled)
8. The pharmaceutical composition of claim 1, wherein ring A is
##STR00311## and ring B is ##STR00312##
9. The pharmaceutical composition of claim 1, wherein X is CH and Y
is CH.
10. The pharmaceutical composition of claim 1, wherein X is N.
11. A pharmaceutical composition, wherein the composition is
formulated for oral administration and comprises a pharmaceutically
acceptable excipient and a compound of Formula II: ##STR00313## or
a pharmaceutically acceptable salt, stereoisomer, or isotopic form
thereof, wherein: ring A is ##STR00314## and R.sup.1 is hydrogen or
fluoro.
12. The pharmaceutical composition of claim 1, wherein the compound
of Formula (I) is selected from the group consisting of:
##STR00315## ##STR00316## ##STR00317## or a pharmaceutically
acceptable salt, stereoisomer, or isotopic form thereof.
13. A pharmaceutical composition, wherein the composition is
formulated for oral administration and comprises a pharmaceutically
acceptable excipient and a compound selected from the group
consisting of: ##STR00318## ##STR00319## ##STR00320## ##STR00321##
or a pharmaceutically acceptable salt, stereoisomer, or isotopic
form thereof.
14. (canceled)
15. A method of treating a patient suffering from a disease
associated with aberrant activity of CDK7, wherein the disease is a
proliferative disease, an infectious disease, or a disease caused
by or associated with expanded repeats of simple nucleotide tracts,
and the method comprises a step of administering to the patient a
therapeutically effective amount of the pharmaceutical composition
of claim 1.
16.-17. (canceled)
18. The method of claim 15, wherein the proliferative disease is a
cancer or benign neoplasm.
19. The method of claim 18, wherein the cancer is a blood cancer, a
bone cancer, a brain cancer, a breast cancer, a colorectal cancer,
a lung cancer, a melanoma, a neuroblastoma, an ovarian cancer, or a
pancreatic cancer.
20. The method of claim 19, wherein the blood cancer is chronic
lymphocytic leukemia (CLL), chronic myelomonocytic leukemia (CMML),
acute lymphoblastic leukemia (ALL), T-cell acute lymphoblastic
leukemia (T-ALL), chronic myelogenous leukemia (CML), lymphoma, or
multiple myeloma.
21.-25. (canceled)
26. The method of claim 19, wherein the blood cancer is acute
myelogenous leukemia (AML).
27. The method of claim 19, wherein the bone cancer is osteosarcoma
or Ewing's sarcoma.
28. The method of claim 19, wherein the breast cancer is
triple-negative breast cancer (TNBC).
29. The method of claim 19, wherein the cancer is a brain
cancer.
30. The method of claim 19, wherein the lung cancer is small cell
lung cancer (SCLC).
31. The method of claim 19, wherein the cancer is a colorectal
cancer, a melanoma, a neuroblastoma, an ovarian cancer, or a
pancreatic cancer.
32. The method of claim 15, wherein the method further comprises a
step of administering, to the patient, a therapeutically effective
amount of an anti-proliferative agent, an anti-cancer agent, an
immunosuppressant agent, or a pain-relieving agent.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of the filing date of
U.S. Provisional Application No. 62/617,884, filed Jan. 16, 2018,
the entire content of which is hereby incorporated by reference
herein.
BACKGROUND OF THE INVENTION
[0002] Kinases that are members of the cyclin-dependent kinase
(CDK) family play critical roles in cellular proliferation. Among
the mammalian CDKs, CDK7 has uniquely consolidated kinase
activities that help regulate both the cell cycle and gene
transcription. In the cytosol, CDK7 exists within a heterotrimeric
complex and is thought to function as a CDK1/2-activating kinase
(CAK); phosphorylation of conserved residues in CDK1/2 by CDK7 is
required for full catalytic activity and cell cycle progression. In
the nucleus, CDK7 forms the kinase core of the RNA polymerase
(RNAP) II general transcription factor complex and phosphorylates
its C-terminal domain (CTD), a requisite step in initiating gene
transcription. By both activating CDK1/2 and phosphorylating the
CTD of RNAP II, CDK7 supports critical facets of cellular
proliferation, cell cycling, and gene transcription.
[0003] Although some progress has been made, it is difficult to
develop selective inhibitors of CDK7 because its sequence and
structure are similar to the sequences and structures of other
CDKs. Thus, there is still an unmet need for selective CKD7
inhibitors.
SUMMARY OF THE INVENTION
[0004] Described herein are selective CDK7 inhibitors (compounds)
of Formula (I) and pharmaceutically acceptable salts, solvates
(e.g., hydrates), tautomers, and isotopic forms thereof. These
compounds preferably demonstrate greater specificity for CDK7 than
for one or more of CDK2, CDK9, and CDK12 (e.g., at least 10-, 100-,
or 1,000-fold greater specificity) when assessed in an enzymatic
assay that measures the IC.sub.50 of the compound. Also described
are compositions containing a compound described herein (e.g., a
pharmaceutical composition or kit) and methods of using the
compounds (or salts, solvates (e.g., hydrates), tautomers, and
isotopic forms thereof), pharmaceutical compositions, or kits to
treat or prevent a disease associated with aberrant CDK7 expression
(e.g., overexpression or misexpression) or activity (e.g.,
overactivity). The disease can be a proliferative disease (e.g., a
cancer such as a blood cancer (e.g., leukemia) breast cancer,
melanoma, multiple myeloma, ovarian cancer (or any other cancer
described further below), a benign neoplasm, or a condition
characterized by pathologic angiogenesis, or a fibrotic disease).
The fibrotic disease can be NASH (non-alcoholic steatohepatitis) or
NAFLD (non-alcoholic fatty liver disease), which can progress to
cirrhosis of the liver and eventual liver failure; any other
disease or chronic damage to the liver that results in fibrosis
(e.g., alcoholism or hepatitis); scleroderma, which can progress to
systemic scleroderma (also known as systemic sclerosis (SSc)); any
other disease characterized by cutaneous fibrosis or resulting in
pulmonary fibrosis (e.g., cystic fibrosis, multiple sclerosis,
rheumatoid arthritis, systemic lupus erythematosus (SLE), or
idiopathic pulmonary fibrosis); kidney fibrosis (e.g., as occurs in
connection with chronic kidney disease such as Alport Syndrome,
glomerulonephritis, polycystic kidney disease, and reflux
nephropathy); and cardiac (e.g., endomyocardial) fibrosis, which is
a common phenomenon in heart or cardiovascular diseases such as
ischemic heart disease, atherosclerosis, arteriosclerosis, and
inherited cardiomyopathies, as well as in Behcet's disease,
diabetes, and ageing. The disease can also be an infectious disease
(e.g., a viral infection caused by an influenza virus, human
immunodeficiency virus (HIV), herpes virus, or human papilloma
virus (HPV)) or a disease caused by or associated with expanded
repeats of simple nucleotide tracts including, but not limited to,
Huntington's Disease (HD), myotonic dystrophy (e.g., DM1 and DM2),
and some forms of amyotrophic lateral sclerosis (ALS). Diseases
caused by or associated with an expanded repeat are also known in
the art as trinucleotide repeat disorders, trinucleotide repeat
expansion disorders, triplet repeat expansion disorders, and codon
reiteration disorders. In these genetic diseases, trinucleotide
repeats in certain genes or introns exceed the stable threshold
observed in healthy patients.
[0005] In one aspect, the present invention provides compounds of
formula (I)
##STR00001##
and pharmaceutically acceptable salts, solvates (e.g., hydrates),
tautomers, and isotopically labeled derivatives thereof, wherein
Ring A, Ring B, X, Y, R.sup.1 and subvariables thereof are as
defined herein. For ease of reading, we may not refer to both a
compound of the invention and a pharmaceutically acceptable salt
thereof when describing each and every composition, method, and use
within the scope of the invention. It is to be understood that
where a compound of the invention can be used, a pharmaceutically
acceptable salt thereof may also be useful, and making that
determination is well within the ability of one of ordinary skill
in the art.
[0006] While pharmaceutical compositions within the scope of the
invention are described further below, we note here that they can
contain a compound of Formula (I) (or a pharmaceutically acceptable
salt, solvate (e.g., hydrate), tautomer, or isotopic form thereof)
and a pharmaceutically acceptable excipient. The active ingredient
(e.g., a compound of Formula (I)), regardless of its precise
chemical form (e.g., isomeric or isotopic forms), can be present in
a therapeutically or prophylactically effective amount, and the
pharmaceutical compositions described herein can be packaged in
unit dosages, fractions thereof or multiples thereof.
[0007] In addition to administering a composition described herein
to a patient who has been diagnosed as having a disease described
herein, the compositions of the invention can be used ex vivo to
alter CDK7 expression or activity in a biological sample (e.g., a
cultured cell line or a blood or tissue sample obtained from a
patient). In any circumstance in which a compound or composition
described herein is administered to a patient, the patient may have
been diagnosed with a disease that is associated with aberrant
expression or activity of CDK7, and any of the methods of treatment
or uses described herein can include a step of determining whether
CDK7 expression or activity is aberrant in a biological sample
obtained from the patient. This information may also be obtained
indirectly. Thus, the methods of treatment and uses described
herein can include a step of administering/using a compound or
composition described herein, where, prior to the
administration/use, a biological sample obtained from the patient
has been determined to exhibit aberrant (e.g., elevated) CDK7
expression or activity.
[0008] The kits can include a container with a compound of Formula
(I) or a pharmaceutically acceptable salt, solvate (e.g., hydrate),
tautomer, or isotopic form thereof, or a pharmaceutical composition
thereof. In certain embodiments, the kits described herein further
include instructions (in written or other form) for administering a
compound described herein (e.g., a compound of Formula (I) or a
pharmaceutically acceptable salt, solvate (e.g., hydrate),
tautomer, or isotopic form thereof, or the pharmaceutical
composition thereof. Paraphernalia (e.g., a syringe, needles,
tubing, gloves, bandages, tape, local anestheics, etc. . . . ) may
also be included.
[0009] The following definitions apply to the compositions and
methods described herein unless the context clearly indicates
otherwise. It will be evident to one of ordinary skill in the art
that the definitions apply to grammatical variants of these terms,
some of which are particularly mentioned below (e.g.,
"administration" and "administering"). The chemical elements are
identified in accordance with the Periodic Table of the Elements,
CAS version, Handbook of Chemistry and Physics, 75.sup.th Ed.
Additionally, general principles of organic chemistry that may
facilitate the production of the compounds described herein can be
found in "Organic Chemistry," Thomas Sorrell, University Science
Books, Sausalito, 1999; "March's Advanced Organic Chemistry,"
5.sup.th Ed., Ed. Smith and March, John Wiley & Sons, New York,
2001; Larock, "Comprehensive Organic Transformations," VCH
Publishers, Inc., New York, 1989; and Carruthers, "Some Modern
Methods of Organic Synthesis," 3.sup.rd Edition, Cambridge
University Press, Cambridge, 1987.
[0010] The term "about," when used in reference to a value,
signifies any value or range of values that is plus-or-minus 10% of
the stated value (e.g., within plus-or-minus 1%, 2%, 3%, 4%, 5%,
6%, 7%, 8%, 9% or 10% of the stated value). For example, a dose of
about 10 mg means any dose as low as 10% less than 10 mg (9 mg),
any dose as high as 10% more than 10 mg (11 mg), and any dose or
dosage range therebetween (e.g., 9-11 mg; 9.1-10.9 mg; 9.2-10.8 mg;
and so on). In case of any doubt, the stated value is included;
about 10 mg includes 10 mg. Where a stated value cannot be exceeded
(e.g., 100%), "about" means a value or range of values that is up
to and including 10% less than the stated value (e.g., a purity of
about 100% means 90%-100% pure (e.g., 95%-100% pure, 96%-100% pure,
97%-100% pure etc. . . . )).
[0011] The term "administration" and variants thereof, such as
"administering," refer to the application of a compound described
herein, a form thereof (e.g., a pharmaceutically acceptable salt or
solvate) or a composition containing any such compounds or forms
(e.g., a pharmaceutical composition) to a patient (e.g., a human
patient) or system (e.g., a cell- or tissue-containing system
maintained ex vivo (e.g., any cell, tissue, or organ culture, which
may be maintained by conventional or new techniques). One of
ordinary skill in the art will know a variety of routes that may,
in appropriate circumstances, be utilized for administration to a
patient or system. For example, the route of administration to a
patient may be oral (i.e., by swallowing a pharmaceutical
composition) or parenteral (a term encompassing any route of
administration that is not oral; e.g., intra-arterial or
intravenous, intra-articular, intracranial, intralesional,
intramuscular, intraperitoneal, or intrathecal). Preferably, the
compositions are administered orally, subcutaneously,
intralesionally, intraperitoneally or intravenously. Any
pharmaceutical composition described herein can be sterile and/or
in an orally available or injectable form made using techniques and
excipients known in the art (including those described further
herein).
[0012] Further, the route of administration to a patient can be
bronchial (e.g., by bronchial instillation), by mouth (i.e., oral),
dermal (which may be or comprise topical application to the dermis
or intradermal, interdermal, or transdermal administration),
intragastric or enteral (i.e., directly to the stomach or
intestine, respectively), intramedullary, intramuscular,
intranasal, intraperitoneal, intrathecal, intravenous (or
intra-arterial), intraventricular, by application to or injection
into a specific organ (e.g., intrahepatic), mucosal (e.g., buccal,
rectal, sublingual, or vaginal), subcutaneous, tracheal (e.g., by
intratracheal instillation), or ocular (e.g., topical,
subconjunctival, or intravitreal). Administration can involve
continuous dosing (e.g., by oral administration or perfusion) for a
selected time (e.g., over or every 1-3 hours; 3-6 hours; over a
period of 12 hours; over a period of 24 hours; etc. . . . ),
intermittent dosing (e.g., a plurality of doses separated in time),
and/or periodic dosing (e.g., doses separated by a common period of
time (e.g., every so many hours, daily, weekly, twice per week,
etc.)).
[0013] The term "angiogenesis" refers to the formation and growth
of new blood vessels. Normal angiogenesis occurs in healthy
patients during development and in the context of wound healing.
However, patients suffering from many different diseases, including
cancer, diabetes (particularly the progression to blindness
associated therewith), age-related macular degeneration, rheumatoid
arthritis, and psoriasis, experience excessive and detrimental
angiogenesis. Angiogenesis is detrimental when it produces blood
vessels that support diseased cells (e.g., tumor cells), destroy
normal tissues (e.g., tissue within the eye), or facilitates tumor
metastases. We may refer to such unwanted angiogenesis as
"pathologic angiogenesis."
[0014] Two events or entities are "associated" with one another if
one or more features of the first (e.g., its presence, level,
activity, and/or form) are correlated with a feature of the second.
For example, a first entity (e.g., a CDK7), gene expression
profile, genetic signature (i.e., a single or combined group of
genes in a cell with a uniquely characteristic pattern of gene
expression), metabolite, or event (e.g., loss of cell cycle control
in CDK7-positive cells)) is associated with a particular disease,
if its presence, level, activity, and/or form correlates with the
incidence of, severity of, and/or susceptibility to the disease
(e.g., a cancer disclosed herein). Associations are typically
assessed across a relevant population. Two or more entities are
physically "associated" with one another if they interact, directly
or indirectly, so that they are and/or remain in physical proximity
with one another in a given circumstance (e.g., within a cell
maintained under physiological conditions (e.g., within cell
culture) or within a pharmaceutical composition). Entities that are
physically associated with one another can be covalently linked to
one another or non-covalently associated by, for example, hydrogen
bonds, van der Waals forces, hydrophobic interactions, magnetism,
or combinations thereof.
[0015] The terms "binding" and variants thereof (such as "bound"
and "bind(s)") refer to a covalent or non-covalent association of
two or more entities (e.g., a compound and an agent within a
pharmaceutical composition or a compound and its target within a
cell). "Direct" binding or direct association occurs when the two
entities physically contact one another (e.g., through a chemical
bond) whereas indirect binding or indirect association occurs when
one of the entities physically contacts one or more intermediate
entities that bring the entities into physical proximity with one
another (e.g., within a complex). Binding can be assessed in a
variety of contexts (e.g., in full or partial isolation or in more
complex, naturally occurring or model systems (e.g., in a tissue,
organ, or cell in vivo or maintained in a tissue culture
environment)).
[0016] The term "biologically active" describes an agent (e.g., a
compound described herein) that produces an observable biological
effect or result in a biological system or model thereof (e.g., in
a human, other animal, or a system maintained in vitro). The
"biological activity" can result from binding between the agent and
a target (e.g., a CDK7), and it may result in modulation (e.g.,
induction, enhancement, or inhibition) of a biological pathway or
event (e.g., a cellular activity (e.g., immunostimulation) or
proliferation). The presence of biological activity and,
optionally, its extent, can be assessed using known and/or standard
methods to detect an immediate or downstream product or event
associated with the biological activity, and any of the methods
described herein can include a step of assessing such activity.
[0017] The term "biological sample" refers to a sample obtained or
derived from a biological source of interest (e.g., a tissue or
organism (e.g., an animal or human patient) or cell culture). The
biological sample can contain a biological cell, tissue or fluid or
any combination thereof. For example, a biological sample can be or
can include ascites; blood; blood cells; bodily fluid(s), any of
which may include or exclude cells; bone marrow; cerebrospinal
fluid (CSF); feces; flexural fluid; free floating nucleic acids;
gynecological fluids; immune infiltrates; lymph; peritoneal fluid;
plasma; saliva; sputum; surgically-obtained specimens; tissue
scraped or swabbed from the skin or a mucus membrane (e.g., in the
nose, mouth, or vagina); tissue or fine needle biopsy samples;
urine; washings or lavages such as a ductal lavage or
broncheoalveolar lavage; or other body fluids, tissues, secretions,
and/or excretions. A biological sample may include cancer cells or
immune cells, such as NK cells and/or macrophages, which are found
in many tissues and organs, including the spleen and lymph nodes.
Cells (e.g., NK cells, macrophages, and cancer cells) within the
sample may have been obtained from an individual for whom a
treatment is intended. Samples used in the form in which they were
obtained may be referred to as "primary" samples, and samples that
have been further manipulated (e.g., by adding or removing one or
more components to/from the sample) may be referred to as
"secondary" or "processed" samples. Such processed samples can
contain or be enriched for a particular cell type (e.g., a
CDK7-expressing cell such as a macrophage or tumor cell), cellular
component (e.g., a membrane fraction), or cellular material (e.g.,
one or more cellular proteins, including one or more of a CDK7,
DNA, or RNA (e.g., mRNA), which may have been subjected to
amplification).
[0018] The term "cancer" refers to a disease in which cells exhibit
an aberrant growth phenotype characterized by loss of control of
cell proliferation to an extent that will be detrimental to a
patient having the disease; such cells can be referred to as a
"cancer cell," a "tumor cell," or a "malignant cell." A cancer can
be classified by the type of tissue in which it originated
(histological type) and/or by the primary site in the body in which
the cancer first developed. Based on histological type, cancers are
generally grouped into six major categories: carcinomas; sarcomas;
myelomas; leukemias; lymphomas; and mixed types. A cancer treated
as described herein can be of any one of these types and may
comprise cells that are precancerous (e.g., benign), malignant,
pre-metastatic, metastatic, and/or non-metastatic. A patient who
has a malignancy or malignant lesion has a cancer. The present
disclosure specifically identifies certain cancers to which its
teachings may be particularly relevant, and one or more of these
cancers may be characterized by a solid tumor or by a hematologic
tumor, which may also be known as a blood cancer (e.g., of a type
described herein).
[0019] The term "comparable" refers to two or more items (e.g.,
agents, entities, situations, sets of conditions, etc.) that are
not identical to one another but are sufficiently similar to permit
comparison therebetween so that one of ordinary skill in the art
will appreciate that conclusions may reasonably be drawn based on
differences or similarities observed. Comparable sets of
conditions, circumstances, individuals, or populations are
characterized by a plurality of substantially identical features
and one or a small number of varied features. One of ordinary skill
in the art will understand, in context, what degree of identity is
required in any given circumstance for two or more items to be
considered comparable. For example, two items are comparable to one
another when they have in common a sufficient number and type of
substantially identical features to warrant a reasonable conclusion
that any differences in results obtained or phenomena observed with
the items are caused by or are indicative of the variation in those
features that are varied. A comparable item can serve as a
"control." For example, a "control patient/population" can be an
untreated patient/population who is afflicted with the same disease
as a patient/population being treated.
[0020] The term "combination therapy" refers to those situations in
which a patient is exposed to two or more therapeutic regimens
(e.g., two or more therapeutic agents) to treat a single disease
(e.g., a cancer). The two or more regimens may be administered
simultaneously or sequentially (e.g., all "doses" of a first
regimen are administered prior to administration of any dose(s) of
a second regimen by the same or a different route of
administration). For clarity, combination therapy does not require
that individual agents be administered together in a single
composition (or even necessarily at the same time), although in
some embodiments, two or more agents (e.g., compounds described
herein) may be administered together in a single composition or
even as a combination compound (e.g., associated in a single
chemical complex or covalent entity). In any of the compositions or
methods described herein, a compound of Formula I can be a "first"
agent.
[0021] The term "compound" means a chemical compound (e.g., a
compound represented by a structural Formula depicted herein, a
sub-genus thereof, or a species thereof). Any given compound can be
biologically, therapeutically, or prophylactically active (e.g.,
when contained in a pharmaceutical composition in an effective
amount) and can be provided and/or utilized (e.g., used in a
biological assay, administered to a patient, incorporated into a
medicament, or otherwise used as described herein) in any of a
variety of forms.
[0022] A "disease" is a pathologic state regardless of whether the
disease is commonly referred to as a condition, disorder, syndrome,
or the like (e.g., a myeloproliferative disorder is a disease).
[0023] The terms "dosage form," "formulation," and "preparation"
are used to refer to compositions containing a compound or other
biologically and/or therapeutically active agent that is suitable
for administration to a patient. The term "unit dosage form" refers
to a physically discrete unit of a compound or other biologically
and/or therapeutically active agent (e.g., a therapeutic or
diagnostic agent) formulated for administration to a patient.
Typically, each such unit contains a predetermined quantity of the
active agent, which may be the amount prescribed for a single dose
(i.e., an amount expected to correlate with a desired outcome when
administered as part of a therapeutic regimen) or a fraction
thereof. One of ordinary skill in the art will appreciate that the
total amount of a therapeutic composition or agent administered to
a particular patient is determined by one or more attending
physicians and may involve administration of multiple unit dosage
forms.
[0024] The term "dosing regimen" refers to the unit dosage form(s)
administered to, or prescribed for, a patient, and typically
includes more than one dose separated by periods of time (e.g., as
described elsewhere herein). The dosage form(s) administered within
a dosing regimen can be of the same unit dose amount or of
different amounts. For example, a dosing regimen can comprises a
first dose in a first dose amount, followed by one or more
additional doses in a second dose amount that is the same as or
different from the first dose amount.
[0025] An "effective amount" of a compound of Formula (I) refers to
an amount sufficient to elicit the desired biological response
(e.g., treating or preventing the disease). As will be appreciated
by one of ordinary skill in the art, the effective amount of a
compound described herein may vary depending on such factors as the
desired biological endpoint, the pharmacokinetics of the compound,
the disease being treated, the mode of administration, and the age
and health of the patient. An effective amount encompasses
therapeutic and prophylactic treatment. For example, in treating
cancer, an effective amount of an inventive compound may reduce the
tumor burden or stop the growth or spread of a tumor.
[0026] The term "excipient" refers to an adjuvant, carrier,
diluent, or other vehicle with which a compound or composition
described herein is administered or otherwise used. The excipient
can be a sterile or sterilizable liquid, such as a water (e.g.,
water for injection), an aqueous solution (e.g., an isotonic salt
solution (e.g., 0.9% NaCl), Ringer's solution, or a solution
comprising 1,3-butanediol), or a natural or synthetic oil (e.g., a
petroleum-based or mineral oil, an animal oil, or a vegetable oil
(e.g., a peanut, soybean, sesame, or canola oil)). The oil may also
be a nonvolatile oil of any animal or plant origin (i.e., a "fixed"
oil). Sterile, fixed oils are conventionally employed as solvents
and suspending media.
[0027] The excipient can be a solid; a liquid that includes one or
more solid components (e.g., a salt, for example, a "normal
saline"); a mixture of solids; or a mixture of liquids. The
excipient may have characteristics that make it useful as a binding
agent, buffering agent, diluent, disintegrating agent, dispersing
agent, emulsifier, granulating agent, lubricating agent,
preservative, or surface active agent. Excipients include, but are
not limited to, alumina, aluminum stearate, a buffer (e.g., a
phosphate salt (e.g., disodium hydrogen phosphate or potassium
hydrogen phosphate) a sodium salt (e.g., sodium chloride), a zinc
salt, or other salts or electrolytes), a cellulose-based substance
(e.g., sodium carboxymethylcellulose), colloidal silica, glycine,
an ion exchanger, lecithin, polyethylene glycol, potassium sorbate,
protamine sulfate, a serum protein (e.g., human serum albumin),
sorbic acid, a partial glyceride mixture of saturated vegetable
fatty acids, magnesium trisilicate, polyvinyl pyrrolidone, a
polyethylene-polyoxypropylene-block polymer, a polyacrylate, water,
wax, and wool fat.
[0028] The term "hydrate" refers to a compound that is bound to
water. The amount of water contained in the hydrate can be
expressed as a ratio of the number of water molecules to the number
of compound molecules. Thus, a hydrate of a compound may be
represented by a general formula such as Rx H.sub.2O, where R is
the compound and x is a number greater than 0. For example, where x
is 1, the hydrate is a monohydrate; where x is 0.5, the hydrate is
a hemihydrate; where x is 2, the hydrate is a dihydrate; and where
x is 6, the hydrate is a hexahydrate. A hydrate is a type of
solvate.
[0029] "Improve(s)," "increase(s)" or "reduce(s)/decrease(s)" are
terms used to characterize the manner in which a value has changed
relative to a reference value. For example, a measurement obtained
from a patient (or a biological sample obtained therefrom) prior to
treatment can be increased or reduced/decreased relative to that
measurement obtained during or after treatment in the same patient,
a control patient, on average in a patient population, or
biological sample(s) obtained therefrom. The value may be improved
in either event, depending on whether an increase or decrease is
associated with a positive therapeutic outcome.
[0030] The term "inhibitor" refers to an agent, including a
compound described herein, whose presence (e.g., at a certain level
or in a certain form) correlates with a decrease in the expression
or activity of another agent (i.e., the inhibited agent or target)
or a decrease in the occurrence of an event (e.g., tumor
progression or metastasis). In some embodiments, an inhibitor
exerts its influence on a target by binding to the target, directly
or indirectly. In other embodiments, an inhibitor exerts its
influence by binding and/or otherwise altering a regulator of the
target, so that the expression and/or activity of the target is
reduced. Inhibition can be assessed in silico, in vitro (e.g., in a
cell, tissue, or organ culture or system), or in vivo (e.g., in a
patient or animal model).
[0031] An "isotopic form" of a compound described herein (e.g., a
compound of Formula (I)) is a form in which one or more elements of
the compound have been replaced with an isotopic variant of that
element. Where a compound contains an isotopic substitution, it can
be, e.g., .sup.2H or .sup.3H for H; .sup.11C, .sup.13C or .sup.14C
for .sup.12C; .sup.13N or .sup.15N for .sup.14N; .sup.17O or
.sup.18O for .sup.16O; .sup.36Cl for .sup.35C; .sup.18F for
.sup.19F; .sup.131I for .sup.127I; etc. Such compounds have use,
for example, as analytical tools, as probes in biological assays,
and/or as therapeutic or prophylactic agents for use in accordance
with the present invention. In particular, an isotopic substitution
of deuterium (2H) for hydrogen is known to potentially slow down
metabolism, shift metabolism to other sites on the compound, slow
down racemization and/or have other effects on the pharmacokinetics
of the compound that may be therapeutically beneficial. In
particular, deuterated forms of a compound described herein (or
other forms thereof, such as salts or solvates) are embodiments of
the present invention; such forms include deuterium in place of one
or more of the hydrogen atoms in the compound but no other isotopic
variants (e.g., no isotopes of .sup.12C, .sup.14N, or
.sup.18O).
[0032] The terms "neoplasm" and "tumor" are used herein
interchangeably and refer to an abnormal mass of tissue in which
the growth of the mass surpasses and is not coordinated with the
growth of a normal tissue. A neoplasm or tumor may be "benign" or
"malignant" depending on the following characteristics: the degree
of cellular differentiation (including morphology and
functionality), rate of growth, local invasion, and metastasis. A
"benign neoplasm" is generally well differentiated, has a slower
growth rate than a malignant neoplasm, and remains localized to the
site of origin (i.e., does not have the capacity to infiltrate,
invade, or metastasize to distant sites). Benign neoplasms include,
but are not limited to, acrochordons, adenomas, chondromas,
intraepithelial neoplasms, lentigos, lipoma, sebaceous
hyperplasias, seborrheic keratoses, and senile angiomas. The benign
neoplasm can also be tuberous sclerosis, or tuberous sclerosis
complex (TSC) or epiloia (derived from "epilepsy, low intelligence,
adenoma sebaceum"). Benign neoplasms can later give rise to
malignant neoplasms (believed to occur as a result of genetic
changes in a subpopulation of the tumor's neoplastic cells), and
such neoplasms are referred to as "pre-malignant neoplasms." An
exemplary pre-malignant neoplasm is a teratoma. In contrast, a
"malignant neoplasm" is generally poorly differentiated (anaplasia)
and grows rapidly with progressive infiltration, invasion, and
destruction of surrounding tissue. Malignant neoplasms also
generally have the capacity to metastasize to distant sites.
[0033] A "patient" is any organism to which a compound described
herein (or any variant thereof, as also described herein (e.g., a
salt or solvate) is administered in accordance with the present
invention e.g., for experimental, diagnostic, prophylactic, and/or
therapeutic purposes. Typical patients include animals (e.g.,
mammals such as mice, rats, rabbits, non-human primates, and
humans; birds; insects; worms; etc.). A patient can be suffering
from a disease described herein (e.g., a proliferative disease,
such as cancer or a benign neoplasm).
[0034] A "pharmaceutical composition" is a composition in which an
active agent (e.g., an active pharmaceutical ingredient (e.g., a
compound described herein)) is formulated together with one or more
pharmaceutically acceptable excipients. The active agent can be
present in a unit dose amount appropriate for administration in a
therapeutic regimen that shows a statistically significant
probability of achieving a predetermined therapeutic effect when
administered to patients of a relevant population. The
pharmaceutical composition may be specially formulated for
administration in solid or liquid form, including forms made for
oral or parenteral (e.g., intravenous) administration. For
parenteral administration, the composition can be formulated, for
example, as a sterile solution or suspension for subcutaneous,
intramuscular, intravenous, intra-arterial, intraperitoneal, or
epidural injection. Pharmaceutical compositions comprising an
active agent (e.g., a compound described herein) can also be
formulated as sustained-release formulations or as a cream,
ointment, controlled-release patch, or spray for topical
application. Creams, ointments, foams, gels, and pastes can also be
applied to mucus membranes lining the nose, mouth, vagina, and
rectum. Any of the compounds described herein and any
pharmaceutical composition containing such a compound may also be
referred to as a "medicament."
[0035] The term "pharmaceutically acceptable," when applied to an
excipient used to formulate a composition disclosed herein (e.g., a
pharmaceutical composition), means an excipient that is compatible
with the other ingredients of the composition and not prohibitively
deleterious to a patient (e.g., it is sufficiently non-toxic in the
amount required and/or administered (e.g., in a unit dosage form).
When applied to a salt (e.g., a salt of a compound described
herein), "pharmaceutically acceptable" refers to the salt form of a
compound that is, within the scope of sound medical judgment,
suitable for use in contact with the tissues of humans and lower
animals without prohibitive toxicity, irritation, allergic response
and the like, and that is commensurate with a reasonable
benefit/risk ratio. Pharmaceutically acceptable salts are well
known in the art (see, e.g., Berge et al., J. Pharmaceutical
Sciences, 66:1-19, 1977). Pharmaceutically acceptable salts of the
compounds described herein include those derived from suitable
inorganic and organic acids and bases. Examples of pharmaceutically
acceptable, nontoxic acid addition salts are salts of an amino
group formed with inorganic acids such as hydrochloric acid,
hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric
acid or with organic acids such as acetic acid, oxalic acid, maleic
acid, tartaric acid, citric acid, succinic acid, or malonic acid or
by using other methods known in the art such as ion exchange. Other
pharmaceutically acceptable salts include adipate, alginate,
ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate,
borate, butyrate, camphorate, camphorsulfonate, citrate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, formate, fumarate, glucoheptonate,
glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate,
hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate,
laurate, lauryl sulfate, malate, maleate, malonate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,
oleate, oxalate, palmitate, pamoate, pectinate, persulfate,
3-phenylpropionate, phosphate, picrate, pivalate, propionate,
stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like.
Salts derived from appropriate bases include alkali metal, alkaline
earth metal, ammonium and N.sup.+(C.sub.1-4 alkyl).sub.4.sup.-
salts. Representative alkali or alkaline earth metal salts include
sodium, lithium, potassium, calcium, magnesium, and the like.
Further pharmaceutically acceptable salts include, when
appropriate, nontoxic ammonium, quaternary ammonium, and amine
cations formed using counterions such as lower alkyl sulfonate,
aryl sulfonate, carboxylate, halide, hydroxide, nitrate, phosphate,
and sulfate.
[0036] When applied to a composition (e.g., a pharmaceutical
composition), the term "pharmaceutically acceptable" indicates that
the composition is suitable for administration to a patient by
virtue of being non-contaminated (e.g., sterile) and non-toxic
(i.e., generally safe; not expected to be poisonous).
[0037] A "prophylactically effective amount" of a compound
described herein is an amount sufficient to delay the onset of a
disease for a period of time that is substantially longer than
expected (e.g., as evidenced by the absence of the signs and
symptoms associated with the disease). A prophylactically effective
amount can be an amount that improves overall prophylaxis or
enhances the prophylactic efficacy of another prophylactic agent (a
"second" prophylactic agent (e.g., a "second" compound)).
[0038] A "proliferative disease" is a disease that occurs due to
abnormal growth or extension by the multiplication of cells
(Walker, Cambridge Dictionary of Biology; Cambridge University
Press: Cambridge, UK, 1990). A proliferative disease can be
associated with the pathological proliferation of normally
quiescent or normally dividing cells; the pathological migration of
cells from their normal location (e.g., metastasis of neoplastic
cells); the pathological expression of proteolytic enzymes such as
the matrix metalloproteinases (e.g., collagenases, gelatinases, and
elastases); and/or pathologic angiogenesis, as occurs in
proliferative retinopathies and tumor metastases. Exemplary
proliferative diseases include cancers, which may also be referred
to as "malignant neoplasms," benign neoplasms, and pathologic
angiogenesis.
[0039] A "sign or symptom is reduced" when one or more objective
signs or subjective symptoms of a disease are reduced in magnitude
(e.g., intensity, severity, etc.) and/or frequency. A delay in the
onset of a particular sign or symptom is one form of reducing the
frequency of that sign or symptom. Reducing a sign or symptom can
be achieved by, e.g., a "therapeutically active" compound
optionally administered in an effective amount, as described
herein.
[0040] The term "solvate" refers to a compound bound to a solvent,
usually by a solvolysis reaction. This physical association may
include hydrogen bonding. Solvents that can be used in the reaction
include water, methanol, ethanol, acetic acid, DMSO (dimethyl
sulfoxide), THF (tetrahydrofuran), diethyl ether, and the like. A
compound of Formula I or (Ia) can be prepared, e.g., in crystalline
form, and then solvated. The solvate can be pharmaceutically
acceptable and can be either a stoichiometric or non-stoichiometric
solvate. In certain instances, the solvate will be capable of
isolation, for example, when one or more solvent molecules are
incorporated in the crystal lattice of a crystalline solid.
"Solvate" encompasses both solution-phase and isolable solvates,
and representative solvates include hydrates, ethanolates, and
methanolates.
[0041] The term "specific," as used herein with reference to an
agent (e.g., a compound) having an activity (e.g., inhibition of a
target), means that the agent discriminates between potential
target entities or states. For example, an agent binds
"specifically" to its intended target (or otherwise specifically
inhibits its target) if it preferentially binds or otherwise
inhibits the expression or activity of that target in the presence
of one or more alternative targets. Although the invention is not
so limited, a specific and direct interaction can depend upon
recognition of a particular structural feature of the target (e.g.,
an epitope, a cleft, or a binding site). Specificity need not be
absolute; the degree of specificity need only be enough to result
in an effective treatment without unacceptable side effects. The
specificity of an agent can be evaluated by comparing the effect of
the agent on an intended target or state relative to its effect on
a distinct target or state. The effects on the intended and
distinct targets can each be determined or the effect on the
intended target can be determined and compared to a reference
standard developed at an earlier time (e.g., a reference specific
binding agent or a reference non-specific binding agent). In some
embodiments, the agent does not detectably bind the competing
alternative target under conditions in which it detectably (and,
preferably, significantly) binds its intended target and/or does
not detectably inhibit the expression or activity of the competing
target under conditions in which it detectably (and, preferably,
significantly) inhibits the expression or activity of its intended
target. A compound of the invention may exhibit, with respect to
its target(s), a higher on-rate, lower off-rate, increased
affinity, decreased dissociation, and/or increased stability
compared with the competing alternative target, and any of these
parameters can be assessed in methods of the invention.
[0042] As used herein with regard to a compound, the term "stable,"
means the compound is not rendered inactive or substantially
inactive when it is subjected to conditions that allow for its
production, detection, recovery, purification, or use as described
herein.
[0043] The invention encompasses "stereoisomeric forms" of a
compound described herein (e.g., an optical and/or structural
isomer). The stereoisomers of any referenced or depicted structure
can be enantiomers and diastereomers (e.g., cis trans isomers and
conformational isomers). These include the R and S configurations
for each asymmetric center, Z and E double bond isomers, and Z and
E conformational isomers. Compositions containing a single type of
stereochemical isomer as well as enantiomeric, diastereomeric, and
geometric (or conformational) mixtures of the present compounds are
within the scope of the invention. Where a particular enantiomer is
preferred, it can be provided substantially free of the
corresponding enantiomer, and may also be referred to as "optically
enriched," meaning that the compound is made up of a significantly
greater proportion of one enantiomer. For example, a plurality of
the compound molecules can be made up of at least about 90% by
weight of the preferred enantiomer (e.g., at least about 95%, 98%,
or 99% by weight). Preferred enantiomers can be isolated from
racemic mixtures by methods known in the art, including chiral high
pressure liquid chromatography (HPLC) and the formation and
crystallization of chiral salts. They can also be prepared by
asymmetric syntheses. If needed, one could consult, for example,
Jacques et al., Enantiomers, Racemates and Resolutions (Wiley
Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725
(1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill,
NY, 1962); and Wilen, Tables of Resolving Agents and Optical
Resolutions p. 268 (E.L. Eliel, Ed., Univ. of Notre Dame Press,
Notre Dame, Ind. 1972).
[0044] The term "substantially" refers to the qualitative condition
of exhibiting a characteristic or property of interest to a total
or near-total extent or degree. One of ordinary skill in the art
will understand that biological and chemical phenomena rarely, if
ever, go to completion and/or proceed to completeness or achieve or
avoid an absolute result. The term "substantially" may therefore be
used to capture the potential lack of completeness inherent in many
biological and chemical phenomena. For example, a chemical reaction
may be characterized as substantially complete even though the
yield is well below 100%. Certain features may also be deemed
"substantially identical" when they are about the same and/or
exhibit about the same activity. For example, two nearly identical
compounds that produce about the same effect on an event (e.g.,
cellular proliferation) may be described as substantially similar.
With regard to the purity of a compound or composition,
"substantially pure" is defined below.
[0045] The term "substantially pure," when used to refer to a
compound described herein, means that a preparation of the compound
is more than about 85% (w/w) compound (e.g., more than about 90%,
95%, 97%, 98%, 99%, or 99.9%).
[0046] An individual who is "susceptible to" a disease (e.g., a
proliferative disease, such as cancer) has a greater than average
risk for developing the disease. Such an individual may not yet
display any symptoms of the disease and may not have not been
diagnosed with the disease. Such an individual may have been
exposed to conditions associated with development of the disease
(e.g., exposure to a carcinogen). Susceptibility can be assessed by
one of ordinary skill in the art and can be determined relative to
a population-based risk.
[0047] The term "tautomer" refers to a structural or constitutional
isomer of a compound; tautomers are compounds having the same
molecular formula but different atomic organization and bonding
patterns. Tautomeric compounds readily interconvert and vary from
one another in the displacement of hydrogen atoms and electrons.
Thus, two tautomeric compounds may be in equilibrium through the
movement of .pi. electrons and an atom (usually H). For example,
enols and ketones are tautomers because they are rapidly
interconverted by treatment with either acid or base. Tautomeric
compounds may exhibit improved chemical reactivity and biological
activity relative to the comparable non-tautomeric compound.
[0048] A "therapeutic regimen" refers to a dosing regimen that,
when administered across a relevant population, is correlated with
a desired therapeutic outcome.
[0049] The term "treatment," and linguistic variants thereof, such
as "treat(s)" and "treating," refer to any use of a compound or
pharmaceutical composition described herein to partially or
substantially completely alleviate, ameliorate, relieve, inhibit,
reduce the severity of, and/or reduce the incidence of one or more
signs or symptoms of a particular disease (e.g., a proliferative
disease such as cancer). The patient being treated (or who has been
identified as a candidate for treatment) may exhibit only early
signs or symptoms of the disease or may exhibit one or more
established or advanced signs or symptoms of the disease.
"Treatment" is distinguished from "prophylaxis," which relates to
delaying the onset of one or more signs or symptoms of a disease.
In that case, the patient may not exhibit signs and/or symptoms of
the disease and/or may be known to have one or more susceptibility
factors that are statistically correlated with increased risk of
development of the relevant disease. However, once a patient
exhibits signs or symptoms of a disease and has been treated,
treatment may be continued to delay or prevent recurrence.
[0050] A "therapeutically effective amount" of a compound described
herein is an amount sufficient to treat a disease in a population
of patients. A therapeutically effective amount is an amount that
provides a therapeutic benefit in the treatment of a disease (e.g.,
by treating one or more signs or symptoms associated with the
disease). A therapeutically effective amount of a compound
described herein can also be an amount that enhances the
therapeutic efficacy of another therapeutic agent (a "second"
therapeutic agent (e.g., a "second" compound)).
[0051] The terms "neoplasm" and "tumor" are used herein
interchangeably and refer to an abnormal mass of tissue wherein the
growth of the mass surpasses and is not coordinated with the growth
of a normal tissue. A neoplasm or tumor may be "benign" or
"malignant," depending on the following characteristics: degree of
cellular differentiation (including morphology and functionality),
rate of growth, local invasion, and metastasis. A "benign neoplasm"
is generally well differentiated, has characteristically slower
growth than a malignant neoplasm, and remains localized to the site
of origin. In addition, a benign neoplasm does not have the
capacity to infiltrate, invade, or metastasize to distant sites.
Exemplary benign neoplasms include, but are not limited to, lipoma,
chondroma, adenomas, acrochordon, senile angiomas, seborrheic
keratoses, lentigos, and sebaceous hyperplasias. In some cases,
certain "benign" tumors may later give rise to malignant neoplasms,
which may result from additional genetic changes in a subpopulation
of the tumor's neoplastic cells, and these tumors are referred to
as "pre-malignant neoplasms." An exemplary pre-malignant neoplasm
is a teratoma. In contrast, a "malignant neoplasm" is generally
poorly differentiated (anaplasia) and has characteristically rapid
growth accompanied by progressive infiltration, invasion, and
destruction of the surrounding tissue. Furthermore, a malignant
neoplasm generally has the capacity to metastasize to distant
sites.
[0052] The details of one or more embodiments of the invention are
set forth herein. Other features, objects, and advantages of the
invention will be apparent from the Detailed Description, the
Figures, the Examples, and the claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0053] FIG. 1 is a Table disclosing exemplary compounds of the
invention.
[0054] FIG. 2 is a Table disclosing further exemplary compounds of
the invention.
[0055] FIG. 3 is a Table providing additional details of the
synthesis of certain compounds of the invention, as well as their
NMR and MS values.
DETAILED DESCRIPTION
[0056] In one aspect of the present invention, provided are
compounds of formula (I):
##STR00002##
or a pharmaceutically acceptable salt, solvate (e.g., hydrate),
tautomer, stereoisomer, or isotopic form thereof, wherein:
[0057] ring A is:
##STR00003##
[0058] ring B is:
##STR00004##
wherein each of R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are,
independently, hydrogen or --CH.sub.3;
[0059] X and Y are, independently, N or CH;
[0060] R.sup.1 is hydrogen or fluoro; and
wherein the compound is other than:
##STR00005##
[0061] In some embodiments, ring A is:
##STR00006##
In more specific embodiments, ring A is
##STR00007##
In even more specific embodiments, ring A is
##STR00008##
[0062] In some embodiments, ring A is
##STR00009##
[0063] In some embodiments, ring B is:
##STR00010##
In some embodiments, ring B is:
##STR00011##
In some embodiments, ring B is:
##STR00012##
[0064] In various embodiments of Formula I: R.sup.1 is hydrogen;
R.sup.1 is fluoro; X is CH and Y is CH; X is N and Y is CH; X is N
and Y is N; R.sup.2 and R.sup.3 is hydrogen; each of R.sup.4 and
R.sup.5 are --CH.sub.3; R.sup.4 is hydrogen and R.sup.5 is
--CH.sub.3.
[0065] Also described herein are compounds of Formula II:
##STR00013##
and pharmaceutically acceptable salts, solvates (e.g., hydrates),
tautomers, stereoisomers, and isotopic forms thereof, wherein:
[0066] ring A is:
##STR00014##
and
[0067] R.sup.1 is selected from hydrogen and fluoro.
[0068] In some embodiments of a compound of Formula II, ring A
is:
##STR00015##
In more specific embodiments, ring A is:
##STR00016##
In even more specific embodiments, ring A is:
##STR00017##
In some embodiments of Formula II, ring A is
##STR00018##
[0069] In various embodiments of Formula II: R.sup.1 is hydrogen;
R.sup.1 is fluoro.
[0070] The compound can be selected from any of the compounds in
the Table of FIG. 1 or the Table of FIG. 2 or can be a
pharmaceutically acceptable salt, solvate (e.g., hydrate),
tautomer, stereoisomer, or isotopic form thereof.
[0071] Any compound described herein can be prepared using methods
described herein and/or known in the art. Techniques useful in
synthesizing these compounds are accessible to one of ordinary
skill in the art, and the discussion below illustrates certain of
the diverse methods available for use in assembling them. The
discussion is not intended to limit the scope of useful reactions
or reaction sequences. Any pharmaceutical composition described
herein can be prepared by methods known in the art of pharmacology.
In general, such preparatory methods include the steps of bringing
the compound (e.g., a compound of formula (I); the "active
ingredient") into association with an excipient and/or one or more
other accessory ingredients or "second" agents, and then, if
necessary and/or desirable, shaping and/or packaging the product
into a desired single- or multi-dose unit and/or packaging the
composition within a kit.
[0072] Compounds and/or other compositions provided herein (e.g.,
pharmaceutical compositions) have a variety of uses, including in
research and/or clinical settings (e.g., in methods of providing a
diagnosis or prognosis and in prophylactic or therapeutic treatment
methods).
[0073] In some embodiments, a provided compound and/or composition
is considered to be specific for a given kinase or set of kinases
when it shows at least or about 2-fold, 3-fold, 4-fold, 5-fold,
10-fold, 15-fold, 20-fold, 50-fold, 100-fold or more activity for
the specific kinase(s) than for one or more appropriate comparator
kinase(s) (e.g., for CDK7 relative to one or more of CDK2, CDK9,
and/or CDK12). One of ordinary skill in the art will recognize the
evaluating specificity in terms of "fold difference" is only one
applicable measure. For example, specificity can be expressed as a
"percent difference." For example, a provided compound and/or
composition is considered to be specific for CDK7 when it shows at
least 101%, 105%, 110%, 120%, 130%, 140%, 150%, 200%, 300%, 400%,
500% or more activity for CDK7 than for one or more of CDK2, CDK9,
and/or CDK12.
[0074] Compounds of the present disclosure can be prepared and
administered in a wide variety of oral and parenteral dosage forms.
Thus, the compounds of the present disclosure can be formulated for
administration by injection (e.g. intravenously, intramuscularly,
intracutaneously, subcutaneously, or intraperitoneally). The
compounds can also be formulated for administration by inhalation
(e.g., intranasally) or by insufflation. In other embodiments, the
compounds described herein can be formulated for topical or
transdermal administration (i.e., they can be in a dosage form
suitable for administration by the various routes just
described).
[0075] For preparing pharmaceutical compositions including a
compound described herein, pharmaceutically acceptable carriers can
be added in either solid or liquid form or a combination thereof.
Solid dosage form preparations within the scope of the present
invention include powders, tablets, pills, capsules, cachets,
suppositories, and dispersible granules. A solid carrier can be a
substance that may also act as a diluent, flavoring agent, binder,
preservative, tablet disintegrating agent, or an encapsulating
material. In powders, the carrier is a finely divided solid in a
mixture with the finely divided active component (e.g., a compound
described herein, e.g., a compound conforming to the structure of
Formula II). In tablets, the active component is mixed with the
carrier having the necessary binding properties in suitable
proportions and compacted in the shape and size desired.
Pharmaceutical compositions, including those formulated as powders
and tablets, can contain from about 5% to about 70% of the active
compound. Suitable carriers are magnesium carbonate, magnesium
stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin,
tragacanth, methylcellulose, sodium carboxymethylcellulose, a low
melting wax, cocoa butter, and the like. Tablets, powders,
capsules, pills, cachets, and lozenges can be used as solid dosage
forms suitable for oral administration. For preparing
suppositories, a low melting wax, such as a mixture of fatty acid
glycerides or cocoa butter, is melted and the active component is
dispersed therein.
[0076] Liquid form preparations include solutions, suspensions, and
emulsions, for example, water or water/propylene glycol solutions.
For parenteral injection, liquid preparations can be formulated in
solution in aqueous polyethylene glycol solution. When parenteral
application is needed or desired, particularly suitable admixtures
for the compounds of the invention are injectable, sterile
solutions, preferably oily or aqueous solutions, as well as
suspensions, emulsions, or implants, including suppositories. In
some embodiments, suitable carriers for parenteral administration
will be selected for human administration.
[0077] In particular, carriers for parenteral administration
include aqueous solutions of dextrose, saline, pure water, ethanol,
glycerol, glycerol formal, polyethylene glycol, propylene glycol,
peanut oil, sesame oil, polyoxyethylene-block polymers,
pyrrolidine, N-methyl pyrrolidione, and the like. Ampoules are
convenient unit dosage forms. The compounds of the present
disclosure can also be incorporated into liposomes or administered
via transdermal pumps or patches. Pharmaceutical admixtures
suitable for use in the present disclosure include those described,
for example, in Pharmaceutical Sciences (17th Ed., Mack Pub. Co.,
Easton, Pa.) and WO 96/05309, the teachings of both of which are
hereby incorporated by reference.
[0078] Aqueous solutions suitable for oral use can be prepared by
dissolving the active component in water and adding suitable
colorants, flavors, stabilizers, and thickening agents as desired.
Aqueous suspensions suitable for oral use can be made by dispersing
the finely divided active component in water with viscous material,
such as natural or synthetic gums, resins, methylcellulose, sodium
carboxymethylcellulose, and other well-known suspending agents.
[0079] Also included are solid form preparations that are intended
to be converted, shortly before use, to liquid form preparations
for oral administration. Such liquid forms include solutions,
suspensions, and emulsions. These preparations may contain, in
addition to the active component, colorants, flavors, stabilizers,
buffers, artificial and natural sweeteners, dispersants,
thickeners, solubilizing agents, and the like.
[0080] The pharmaceutical compositions are preferably in unit
dosage form. The unit dosage form can be a packaged preparation,
the package containing discrete quantities of preparation, such as
packeted tablets, capsules, and powders in vials or ampoules. Unit
dosage forms can also be capsules, tablets, cachets, lozenges, or
the appropriate number of any of these in packaged form.
[0081] The quantity of active component in a unit dose preparation
may be varied or adjusted from 0.1 mg to 10000 mg, more typically
1.0 mg to 1000 mg, most typically 10 mg to 500 mg, according to the
particular application and the potency of the active component. The
composition can, if desired, also contain other compatible
therapeutic agents.
[0082] Some compounds may have limited solubility in water and
therefore may require a surfactant or other appropriate co-solvent
in the composition (e.g., polysorbate 20, 60, and 80; Pluronic.RTM.
F-68, F-84, and P-103; cyclodextrin; and polyoxyl 35 castor oil).
Such co-solvents are typically employed at a level between about
0.01% and about 2% by weight.
[0083] Viscosity greater than that of simple aqueous solutions may
be desirable to decrease variability in dispensing the
formulations, to decrease physical separation of components of a
suspension or emulsion of formulation, and/or otherwise to improve
the formulation. Such viscosity building agents include, for
example, polyvinyl alcohol, polyvinyl pyrrolidone, methyl
cellulose, hydroxy propyl methylcellulose, hydroxyethyl cellulose,
carboxymethyl cellulose, hydroxy propyl cellulose, chondroitin
sulfate and salts thereof, hyaluronic acid and salts thereof, and
combinations of the foregoing. Such agents are typically employed
at a level between about 0.01% and about 2% by weight.
[0084] Compositions of the present invention may additionally
include components to provide sustained release and/or comfort
(e.g., high molecular weight, anionic mucomimetic polymers, gelling
polysaccharides, and finely-divided drug carrier substrates). These
components are discussed in detail in U.S. Pat. Nos. 4,911,920;
5,403,841; 5,212,162; and 4,861,760. The entire contents of these
patents are incorporated herein by reference in their entirety for
all purposes.
[0085] Pharmaceutical compositions provided by the present
disclosure include compositions wherein the active ingredient is
contained in a therapeutically effective amount, i.e., in an amount
effective to achieve its intended purpose. The actual amount
effective for a particular application will depend, inter alia, on
the condition being treated. For example, when administered in
methods to a subject with cancer, such compositions will contain an
amount of active ingredient effective to achieve the desired
result.
[0086] The dosage and frequency (single or multiple doses) of
compound administered can vary depending on a variety of factors,
including route of administration; the size, age, sex, health, body
weight, body mass index, and diet of the recipient; nature and
extent of symptoms of the disease being treated; any concurrent
treatment; and complications from any disease or treatment regimen.
Other therapeutic regimens or agents can be used in conjunction
with the methods and compounds of the present disclosure.
[0087] For any compound, any variant form thereof (e.g., any salt
or solvate), or any pharmaceutical composition described herein,
the therapeutically effective amount can be initially determined
from, or informed by data generated in, cell culture assays and/or
animal models of disease. For example, a dose for humans can be
formulated to achieve a concentration that has been found to be
effective in animals. The dosage in humans can be adjusted by, for
example, monitoring kinase (CDK7) inhibition, the signs an symptoms
of the disease being treated, and side effects and subsequently
adjusting the dosage upwards or downwards.
[0088] Dosages may be varied depending upon the requirements of the
patient and the compound being employed. The dose administered to a
patient should be sufficient to effect a beneficial therapeutic
response in the patient over time. The size of the dose also will
also be determined by the existence, nature, and extent of any
adverse side effects. Generally, treatment is initiated with
smaller dosages, which are less than the optimum dose of the
compound. Thereafter, the dosage is increased by small increments
until the optimum effect under a desired circumstance is reached.
In some embodiments, the concentration of compound is about 0.001%
to 10% w/v (e.g., about 0.1% to about 5% w/v). In some embodiments,
the concentration range is 0.1% to 5% w/v. Concentrations, dosage
amounts, and intervals can be adjusted in each individual patient
to provide levels of the administered compound effective for the
particular disease being treated. This will provide a therapeutic
regimen commensurate with the severity of the patient's
disease.
[0089] Methods of Treatment and Uses: The disease to be treated or
prevented using the compounds of Formula (I), pharmaceutically
acceptable salts, solvates (e.g., hydrates), tautomers,
stereoisomers, isotopically labeled derivatives thereof, or
compositions containing any of the foregoing will typically be
associated with aberrant expression or activity of CDK7. For
example, a gene encoding CDK7 can be overexpressed or misexpressed
(i.e., expressed at abnormally high levels in a tissue where it is
normally expressed; expressed or overexpressed in a tissue where it
is not normally expressed; or expressed at a time during which it
is not normally expressed). The activity of the encoded protein can
also be increased, and aberrant expression or activity can be
relative to the expression or activity in a comparable patient,
population of patients, biological sample, or plurality of
biological samples that do not manifest signs or symptoms of the
disease in question. Both expression and activity may be abberant,
or only activity may be abberant.
[0090] Methods of testing patients and biological samples for
levels of CDK7 expression or activity are within the scope of the
invention and may be performed prior to, during, or after treatment
as described herein. CDK7 levels can be tested and compared between
healthy patients and patients having a disease; between treated and
untreated patients; between biological samples from healthy tissues
and diseased tissues (whether treated or untreated); and between
biological samples that represent models of a disease disclosed
herein. Although no aspect of the invention is limited by the
cellular mechanisms that result from contacting a cell described
herein (whether in a patient or a biological sample ex vivo) with a
compound or other composition described herein, this contact may
cause cytotoxicity via induction of apoptosis, and assessing
cytotoxicity can be a part of the methods of testing patients and
biological samples prior to, during, or after treatment.
[0091] As noted, the proliferative disease to be treated or
prevented using the compounds described herein or pharmaceutically
acceptable salts, solvates (e.g., hydrates), tautomers,
stereoisomers, isotopically labeled derivatives, and compositions
thereof include the cancers described herein, particularly when a
given cancer is known to be or found to be associated with aberrant
CDK7 expression or activity. The proliferative disease can be a
cancer associated with dependence on BCL-2 anti-apoptotic proteins
(e.g., MCL-1 and/or XIAP); with overexpression of MYC; with
overexpression of CDK18; with overexpression of CDK19; with
overexpression of FGFR1; with overexpression of CDK6; with
overexpression of CCND2; or with overexpression of CDKN2A. In
certain embodiments, the proliferative disease is a cancer
associated with the absence of, or suppression of, a wild-type RB1
gene.
[0092] In certain embodiments, the proliferative disease is a blood
cancer, which may also be referred to as a hematopoietic or
hematological cancer or malignancy. More specifically and in
various embodiments, the blood cancer can be a leukemia such as
acute lymphocytic leukemia (ALL; e.g., B cell ALL or T cell ALL),
acute myelocytic leukemia (AML; e.g., B cell AML or T cell AML),
chronic myelocytic leukemia (CML; e.g., B cell CML or T cell CML),
chronic lymphocytic leukemia (CLL; e.g., B cell CLL (e.g., hairy
cell leukemia) or T cell CLL), chronic neutrophilic leukemia (CNL),
or chronic myelomonocytic leukemia (CMML). The blood cancer can
also be a lymphoma such as Hodgkin lymphoma (HL; e.g., B cell HL or
T cell HL), non-Hodgkin lymphoma (NHL; e.g., B cell NHL or T cell
NHL), follicular lymphoma, chronic lymphocytic leukemia/small
lymphocytic lymphoma (CLL/SLL), mantle cell lymphoma (MCL), a
marginal zone B cell lymphoma (e.g., splenic marginal zone B cell
lymphoma), primary mediastinal B cell lymphoma (e.g., splenic
marginal zone B cell lymphoma), primary mediastinal B cell
lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (i.e.,
Waldenstrom's macroglobulinemia), immunoblastic large cell
lymphoma, precursor B lymphoblastic lymphoma, or primary central
nervous system (CNS) lymphoma. The B cell NHL can be diffuse large
cell lymphoma (DLCL; e.g., diffuse large B cell lymphoma), and the
T cell NHL can be precursor T lymphoblastic lymphoma or a
peripheral T cell lymphoma (PTCL). In turn, the PTCL can be a
cutaneous T cell lymphoma (CTCL) such as mycosis fungoides or
Sezary syndrome, angioimmunoblastic T cell lymphoma, extranodal
natural killer T cell lymphoma, enteropathy type T cell lymphoma,
subcutaneous anniculitis-like T cell lymphoma, or anaplastic large
cell lymphoma. While the invention is not limited to treating or
preventing blood cancers having any particular cause or
presentation, stem cells within the bone marrow may proliferate,
thereby becoming a dominant cell type within the bone marrow and a
target for a compound described herein. Leukemic cells can
accumulate in the blood and infiltrate organs such as the lymph
nodes, spleen, liver, and kidney. In some embodiments, a compound
of the present disclosure is useful in the treatment or prevention
of a leukemia or lymphoma.
[0093] In other embodiments, the proliferative disease is
characterized by a solid tumor considered to be either of its
primary location or metastatic. Exemplary cancers include, but are
not limited to, acoustic neuroma; adenocarcinoma; adrenal gland
cancer; anal cancer; angiosarcoma (e.g., lymphangiosarcoma,
lymphangioendotheliosarcoma, hemangiosarcoma); appendix cancer;
benign monoclonal gammopathy (also known as monoclonal gammopathy
of unknown significance (MGUS); biliary cancer (e.g.,
cholangiocarcinoma); bladder cancer; breast cancer (e.g.,
adenocarcinoma of the breast, papillary carcinoma of the breast,
mammary cancer, or medullary carcinoma of the breast, any of which
may be present in subjects having a particular profile, such as an
HR+ (ER+ or PR+), HR- (having neither estrogen nor progesterone
receptors), a triple negative breast cancer (TNBC; ER-/PR-/HER2-),
or a triple-positive breast cancer); brain cancer (e.g.,
meningioma, glioblastoma, glioma (e.g., astrocytoma,
oligodendroglioma), medulloblastoma, or neuroblastoma); bronchus
cancer; carcinoid tumor carcinoid tumor, which may be benign;
cervical cancer (e.g., cervical adenocarcinoma); choriocarcinoma;
chordoma; craniopharyngioma; colorectal cancer (e.g., colon cancer,
rectal cancer, or colorectal adenocarcinoma); connective tissue
cancer; epithelial carcinoma; ependymoma; endotheliosarcoma (e.g.,
Kaposi's sarcoma or multiple idiopathic hemorrhagic sarcoma);
endometrial cancer (e.g., uterine cancer or uterine sarcoma);
esophageal cancer (e.g., adenocarcinoma of the esophagus or
Barrett's adenocarcinoma; Ewing's sarcoma (or other pediatric
sarcoma, such as embryonal rhabdomyosarcoma or alveolar
rhabdomyosarcoma); eye cancer (e.g., intraocular melanoma or
retinoblastoma); familial hypereosinophilia; gallbladder cancer;
gastric cancer (e.g., stomach adenocarcinoma); gastrointestinal
stromal tumor (GIST); germ cell cancer; head and neck cancer (e.g.,
head and neck squamous cell carcinoma, oral cancer (e.g., oral
squamous cell carcinoma)), throat cancer (e.g., laryngeal cancer,
pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer));
hemangioblastoma; hypopharynx cancer; inflammatory myofibroblastic
tumors; immunocytic amyloidosis; kidney cancer (e.g.,
nephroblastoma a.k.a. Wilms' tumor, renal cell carcinoma); liver
cancer (e.g., hepatocellular cancer (HCC), malignant hepatoma);
lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer
(SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the
lung); leiomyosarcoma (LMS); mastocytosis (e.g., systemic
mastocytosis); muscle cancer; myelodysplastic syndrome (MDS);
mesothelioma; myeloproliferative disorder (MPD) (e.g., polycythemia
vera (PV), essential thrombocytosis (ET), agnogenic myeloid
metaplasia (AMM) a.k.a. myelofibrosis (MF), chronic idiopathic
myelofibrosis, chronic myelocytic leukemia (CML), chronic
neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES));
neuroblastoma; neurofibroma (e.g., neurofibromatosis (NF) type 1 or
type 2, schwannomatosis); neuroendocrine cancer (e.g.,
gastroenteropancreatic neuroendocrine tumor (GEP-NET), carcinoid
tumor); osteosarcoma (e.g., bone cancer); ovarian cancer (e.g.,
cystadenocarcinoma, ovarian embryonal carcinoma, ovarian
adenocarcinoma); papillary adenocarcinoma; pancreatic cancer (e.g.,
pancreatic adenocarcinoma, intraductal papillary mucinous neoplasm
(IPMN), Islet cell tumors); penile cancer (e.g., Paget's disease of
the penis and scrotum); pinealoma; primitive neuroectodermal tumor
(PNT); plasma cell neoplasia; paraneoplastic syndromes;
intraepithelial neoplasms; prostate cancer (e.g., prostate
adenocarcinoma); rectal cancer; rhabdomyosarcoma; salivary gland
cancer; skin cancer (e.g., squamous cell carcinoma (SCC),
keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)); small
bowel cancer or small intestine cancer (e.g., appendix cancer);
soft tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH),
liposarcoma, malignant peripheral nerve sheath tumor (MPNST),
chondrosarcoma, fibrosarcoma, myxosarcoma); sebaceous gland
carcinoma; testicular cancer (e.g., seminoma, testicular embryonal
carcinoma); thyroid cancer (e.g., papillary carcinoma of the
thyroid, papillary thyroid carcinoma (PTC), medullary thyroid
cancer); urethral cancer; vaginal cancer; and vulvar cancer (e.g.,
Paget's disease of the vulva).
[0094] The proliferative disease can be associated with pathologic
angiogenesis, and the methods of the invention (and uses of the
compounds and other compositions described herein) encompass
inhibiting pathologic angiogenesis in the context of cancer
treatment.
[0095] As noted, the patient for which administration is
contemplated includes, but is not limited to, a human (i.e., a
male, female or transgendered person of any age group, e.g., a
pediatric patient (e.g., infant, child, adolescent) or adult
patient (e.g., young adult, middle-aged adult, or senior adult))
and/or other non-human animals, for example, mammals (e.g.,
primates (e.g., cynomolgus monkeys, rhesus monkeys); commercially
relevant mammals such as cattle, pigs, horses, sheep, goats, cats,
and/or dogs) and birds (e.g., commercially relevant birds such as
chickens, ducks, geese, and/or turkeys). Thus, the patient may be a
domesticated animal. The non-human animal may be a transgenic
animal (e.g., an animal valuable in research, such as a transgenic
mouse or transgenic pig).
[0096] A cell used or treated as described herein may be a healthy
cell or an unhealthy or abnormal cell (e.g., a cancer cell, tumor
cell, or other cell that is proliferating uncontrollably). A
compound of the invention may be administered to any such cell in
vitro or in vivo. In various embodiments, the cell is: a cancer
cell; a tumor cell; a proliferating cell; a blood cell (e.g., a
lymphocyte); an endothelial cell; or an immune cell. In various
embodiments, the cancer cell is: a leukemia cell; a CLL cell; a
CMML cell; an AML cell; a breast cancer or ovarian cancer cell; a
melanoma cell; a multiple myeloma cell; or a cell of any other
cancer or disease disclosed herein.
[0097] In certain embodiments, the methods described herein
comprise the additional step of administering one or more
additional pharmaceutical agents in combination with the compound
of formula (I) a pharmaceutically acceptable salt thereof, or
compositions comprising such compound or pharmaceutically
acceptable salt thereof. Such additional pharmaceutical agents
include, but are not limited to, anti-proliferative agents,
anti-cancer agents, anti-diabetic agents, anti-inflammatory agents,
immunosuppressant agents, and a pain-relieving agent. The
additional pharmaceutical agent(s) may synergistically augment
inhibition of CDK7 induced by the inventive compounds or
compositions of this invention in the biological sample or subject.
Thus, the combination of the inventive compounds or compositions
and the additional pharmaceutical agent(s) may be useful in
treating proliferative diseases resistant to a treatment using the
additional pharmaceutical agent(s) without the inventive compounds
or compositions.
[0098] As indicated, the present invention provides the compounds
described herein (e.g., a compound of Formula (I)) and
pharmaceutically acceptable salts, solvates (e.g., hydrates),
tautomers, isotopic forms, and compositions thereof (e.g.,
pharmaceutical compositions), for use in the treatment of a
proliferative disease or any other disease described herein in a
patient. While the compounds, variants thereof, compositions
containing them, and methods of use are not limited according to
the underlying mechanism of action, we note that use may result in
an inhibition of cell growth by, for example, promoting apoptosis
or altering the expression or activity of disease-related
genes.
EXAMPLES
[0099] The compounds provided herein can be prepared from readily
available starting materials using modifications to the specific
synthesis protocols set forth below that would be well known to
those of skill in the art. It will be appreciated that where
typical or preferred process conditions (i.e., reaction
temperatures, times, mole ratios of reactants, solvents, pressures,
etc.) are given, other process conditions can also be used unless
otherwise stated. Optimum reaction conditions may vary with the
particular reactants or solvents used, but such conditions can be
determined by those skilled in the art by routine optimization
procedures.
[0100] Additionally, as will be apparent to those skilled in the
art, conventional protecting groups may be necessary to prevent
certain functional groups from undergoing undesired reactions. The
choice of a suitable protecting group for a particular functional
group as well as suitable conditions for protection and
deprotection are well known in the art. For example, numerous
protecting groups, and their introduction and removal, are
described in Greene et al., Protecting Groups in Organic Synthesis,
Second Edition, Wiley, New York, 1991, and references cited
therein.
Example 1. Synthesis of
(S)-4-(6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl)-N-(6,6-dimethylpiperi-
din-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (Compound 104)
Step 1:
6-bromo-3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole
##STR00019##
[0102] To a solution of 2,4-dichloro-5-(trifluoromethyl)pyrimidine
(4.87 g, 22.44 mmol) in dichloroethane (29 mL) was added aluminum
trichloride (3.26 g, 24.49 mmol). The resulting suspension was
stirred at 70.degree. C. for 15 min to provide a clear yellow
solution. Reaction mixture was then slightly cooled down (to
60.degree. C.) followed by addition of 6-bromoindole (4.0 g, 20.40
mmol). The resulting orange solution was stirred at 80.degree. C.
for 2 h. The reaction mixture was cooled down to RT (room
temperature), and ice-cold water was then added followed by
stirring for 30 min. The resulting slurry was further diluted with
EtOAc (ethyl acetate) and water, then filtered. Aqueous layer was
extracted with EtOAc (twice), the organics were combined, dried
over Na.sub.2SO.sub.4, and concentrated under vacuum. The crude red
solid was dissolved in minimal amount of MeOH (methanol) and, after
stirring for 30 min at RT, was then filtered to provide the desired
regioisomer as a beige solid (2.20 g, 5.85 mmol, 29% yield). The
product was used in the next step without further purification.
Step 2:
6-bromo-3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-1-(phenylsu-
lfonyl)-1H-indole
##STR00020##
[0104] To a solution of
6-bromo-3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole
(1.19 g, 3.16 mmol) in dry THF (tetrahydrofuran; 15.8 mL) was added
sodium tert-butoxide, 2 M in THF (1.90 mL, 3.79 mmol), and the
resulting mixture was stirred for 30 min at 0.degree. C.
Benzenesulfonyl chloride (0.48 mL, 3.79 mmol) was then added.
Reaction mixture was stirred for 15 min at 0.degree. C. and then at
RT for overnight. The mixture was diluted with EtOAc and washed
with water. The organic phase was separated, dried over
Na.sub.2SO.sub.4, and concentrated under vacuum to provide the
crude title compound as a beige solid (1.40 g, 2.71 mmol, 86%
yield). The product was used in the next step without further
purification.
Step 3: (S)-6,6-dimethylpiperidin-3-amine
##STR00021##
[0106] (S)-tert-Butyl (6-oxopiperidin-3-yl)carbamate (1.00 g, 4.67
mmol) (Tetrahedron Letters 36:8204, 1995) was dissolved in THF (47
mL), and the solution was cooled to -10.degree. C. Zirconium(IV)
chloride (2.61 g, 11.22 mmol) was added, and the mixture was
stirred for 30 min at this temperature. Methylmagnesium bromide 3 M
solution in ether (20.25 mL, 60.75 mmol) was added, and then
reaction mixture was allowed to slowly warm up to RT and then
stirred for overnight. The solution was quenched with 30% NaOH,
diluted with EtOAc, filtered, and then extracted 3 times with
EtOAc. The organics were combined, dried over Na.sub.2SO.sub.4,
filtered, and concentrated under vacuum to provide crude product as
a yellow oil that was used without further purification. The
obtained oil was dissolved in DCM (dichloromethane; 47 mL) and TFA
(trifluoroacetic acid; 3.58 mL, 46.73 mmol) was added. The reaction
mixture was stirred at RT for 16 h. The mixture was concentrated
under reduced pressure and co-evaporated for few times with DCM to
provide a brown oil of crude title compound, which was used in the
next step without further purification.
Step 4:
(S)-4-(6-bromo-1-(phenylsulfonyl)-1H-indol-3-yl)-N-(6,6-dimethylpi-
peridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
##STR00022##
[0108] To a solution of
6-bromo-3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-1-(phenylsulfonyl)-
-1H-indole (1.40 g, 2.71 mmol) and crude
(S)-6,6-dimethylpiperidin-3-amine from the previous step (4.67
mmol) in dry THF (21 mL) was added DIPEA (N,N-diisopropyl
ethylamine; 1.42 mL, 8.13 mmol). The reaction mixture was then
stirred at RT for 16 h. The reaction mixture was diluted with EtOAc
and washed with water. Organic phase was separated, dried over
Na.sub.2SO.sub.4, filtered, and concentrated under vacuum to
provide a yellow oil of crude product that was purified by reverse
phase chromatography (C18, MeCN in aq. (aqueous) 10 mM ammonium
bicarbonate, 30 to 100% gradient, where product was washed out from
the column at 100% MeCN with 10 mL of IPA (isopropanol)). The title
compound was obtained as a brown foam (330 mg, 0.54 mmol, 20%
yield).
Step 5:
(S)-4-(6-(3,5-dimethylisoxazol-4-yl)-1-(phenylsulfonyl)-1H-indol-3-
-yl)-N-(6,6-dimethylpiperidin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
##STR00023##
[0110] To a solution of
(S)-4-(6-bromo-1-(phenylsulfonyl)-1H-indol-3-yl)-N-(6,6-dimethylpiperidin-
-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (100 mg, 0.164 mmol)
and (3,5-dimethylisoxazol-4-yl)boronic acid (69 mg, 0.493 mmol) in
previously degassed 2:1 mixture of dioxane/H.sub.2O (3 mL) was
added Cs.sub.2CO.sub.3 (161 mg, 0.493 mmol), and the mixture was
degassed for another 15 min. Then, Pd(PPh.sub.3).sub.4 (19 mg,
0.016 mmol) was added, and the reaction mixture was stirred at
100.degree. C. for 1.5 h. The mixture was allowed to cool down to
RT, diluted with EtOAc and washed with water and brine. The organic
phase was separated, dried over Na.sub.2SO.sub.4, filtered, and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography (EtOAc in hexanes, 0 to 100%
gradient followed by 100% of IPA) to give the title compound as a
yellowish oil (56 mg, 0.09 mmol, 55% yield).
Step 6:
(S)-4-(6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl)-N-(6,6-dimethy-
lpiperidin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
##STR00024##
[0112] To a solution of
(S)-4-(6-(3,5-dimethylisoxazol-4-yl)-1-(phenylsulfonyl)-1H-indol-3-yl)-N--
(6,6-dimethylpiperidin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
(56 mg, 0.09 mmol) in dioxane (1 mL) was added 5M aq. NaOH (0.18
mL, 0.90 mmol). The reaction mixture was stirred at 70.degree. C.
for 3 h. The mixture was then concentrated, and the residue was
purified by reverse phase chromatography (C18, MeCN in aq. 10 mM
ammonium bicarbonate, 0 to 100% gradient) to provide the title
compound as an off-white solid (26 mg, 0.052 mmol, 58% yield).
Example 2. Synthesis of
(S)--N-(6,6-dimethylpiperidin-3-yl)-4-(6-(thiazol-2-yl)-1H-indol-3-yl)-5--
(trifluoromethyl)pyrimidin-2-amine (Compound 106)
Step 1:
(S)--N-(6,6-dimethylpiperidin-3-yl)-4-(1-(phenylsulfonyl)-6-(thiaz-
ol-2-yl)-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
##STR00025##
[0114] To a solution of
(S)-4-(6-bromo-1-(phenylsulfonyl)-1H-indol-3-yl)-N-(6,6-dimethylpiperidin-
-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (Example 1; 220 mg,
0.36 mmol) in toluene (3.6 mL) was added
2-(tributylstannyl)thiazole (169 mg, 0.45 mmol), and this
suspension was degassed for 20 min.
Bis(triphenylphosphine)palladium(II) dichloride (13 mg, 0.02 mmol)
was then added, and the reaction mixture was stirred at 100.degree.
C. for 2 h. After cooling down to RT, the mixture was filtered
through Celite, then diluted with water (20 mL) and EtOAc (30 mL)
and basified with sat. (saturated) aq. NaHCO.sub.3 (5 mL). Organic
phase was separated, washed with water (20 mL) and brine
(2.times.10 mL), dried over MgSO.sub.4, filtered, and concentrated
under reduced pressure. The residue was purified by silica gel
column chromatography (THF in hexanes, 0 to 100% gradient) to
obtain the title compound as a yellow oil (101 mg, 0.16 mmol, 45%
yield).
Step 2:
(S)--N-(6,6-dimethylpiperidin-3-yl)-4-(6-(thiazol-2-yl)-1H-indol-3-
-yl)-5-(trifluoromethyl)pyrimidin-2-amine
##STR00026##
[0116] To a solution of
(S)--N-(6,6-dimethylpiperidin-3-yl)-4-(1-(phenylsulfonyl)-6-(thiazol-2-yl-
)-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (101 mg, 0.16
mmol) in dioxane (1.6 mL) was added 5M aq. NaOH (0.32 mL, 1.60
mmol), and the reaction mixture was stirred at 70.degree. C. for 2
h. The mixture was then concentrated, and the residue was purified
by reverse phase chromatography (C18, MeCN in aq. 10 mM ammonium
formate pH 3.8, 0 to 100% gradient) to obtain the title compound as
a pale yellow solid (20.3 mg, 0.043 mmol, 27% yield).
Example 3. Synthesis of
(S)-4-(6-(oxazol-2-yl)-1H-indol-3-yl)-N-(piperidin-3-yl)-5-(trifluorometh-
yl)pyrimidin-2-amine (Compound 110) and
(S)-4-(6-(oxazol-5-yl)-1H-indol-3-yl)-N-(piperidin-3-yl)-5-(trifluorometh-
yl)pyrimidin-2-amine (Compound 111)
Step 1: (S)-tert-butyl
3-((4-(6-bromo-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(trifluoromethyl)pyrim-
idin-2-yl)amino)piperidine-1-carboxylate
##STR00027##
[0118] A mixture of
1-(benzenesulfonyl)-6-bromo-3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl-
]indole (Example 1, step 2; 1.00 g, 1.94 mmol, 1.00 eq),
(S)-tert-butyl (3S)-3-aminopiperidine-1-carboxylate (388.54 mg,
1.94 mmol, 1.00 eq) and DIEA (N,N-diisopropylethylamine; 752.18 mg,
5.82 mmol, 1.02 mL, 3.00 eq) in NMP (N-methyl-2-pyrrolidone; 10.00
mL) was stirred at 140.degree. C. for 1 hr. The mixture was diluted
with water (100 mL) and extracted with EtOAc (50 mL.times.2).
Combined organic layers were washed with brine (100 mL.times.2),
dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure to a yellow oil. The residue was purified by
SiO.sub.2 chromatography (PE/EtOAc, 10:1 to 5:1) to afford the
title compound (1.13 g, 86%) as a yellow solid.
Step 2: (S)-tert-butyl
3-((4-(6-(oxazol-2-yl)-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(trifluorometh-
yl)pyrimidin-2-yl)amino)piperidine-1-carboxylate and (S)-tert-butyl
3-((4-(6-(oxazol-5-yl)-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(trifluorometh-
yl)pyrimidin-2-yl)amino)piperidine-1-carboxylate
##STR00028##
[0120] To a microwave vial, (S)-tert-butyl
3-((4-(6-bromo-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(trifluoromethyl)pyrim-
idin-2-yl)amino)piperidine-1-carboxylate (150.0 mg, 0.220 mmol),
pivalic acid (9.0 mg, 0.088 mmol), K.sub.2CO.sub.3 (91.4 mg, 0.661
mmol), Pd(OAc).sub.2 (2.5 mg, 0.011 mmol; 5 mol %), and RuPhos
(2-dicyclohexylphosphino-2',6'-diisopropoxybiphenyl; 10.3 mg, 0.022
mmol; 10 mol %) were added. The microwave vial was sealed under
nitrogen. Dry, degassed toluene (1.0 mL) was then added followed by
oxazole (29 .mu.L, 0.441 mmol). The reaction mixture was stirred at
110.degree. C. in an oil bath for 16 h. After cooling down to RT,
the reaction mixture was concentrated, and the residue was purified
by silica gel column chromatography (EtOAc in hexanes, 0 to 100%
gradient) to give the title compounds: 23 mg (pale oil, 0.034 mmol,
16% yield) and 46 mg (pale oil, 0.069 mmol, 31% yield),
correspondingly.
Step 3:
(S)-4-(6-(oxazol-2-yl)-1H-indol-3-yl)-N-(piperidin-3-yl)-5-(triflu-
oromethyl)pyrimidin-2-amine
##STR00029##
[0122] To a solution of (S)-tert-butyl
3-((4-(6-(oxazol-2-yl)-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(trifluorometh-
yl)pyrimidin-2-yl)amino)piperidine-1-carboxylate (41 mg, 0.061
mmol) in dioxane (3 mL) was added 5M aq. NaOH (0.122 mL, 0.610
mmol), and the reaction mixture was stirred at 50.degree. C. for 3
h. The mixture was cooled down to RT, diluted with MeTHF
(2-methyltetrahydrofuran; 5 mL) and water (5 mL). Organic phase was
separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated
under reduced pressure. The obtained crude was then re-dissolved in
DCM (2 mL), and TFA was added (0.187 mL, 2.440 mmol). The reaction
mixture was stirred at RT for 1 h. The mixture was concentrated
under reduced pressure and azeotroped for few times with DCM. The
basified crude product was then purified by reverse phase
chromatography (C18, MeCN in aq. 10 mM ammonium formate pH 3.8, 0
to 100% gradient) to obtain the title compound as an off-white
solid (7.0 mg, 0.016 mmol, 27% yield).
Example 4. Synthesis of
(S)-4-(6-(3-methylisoxazol-4-yl)-1H-indol-3-yl)-N-(piperidin-3-yl)-5-(tri-
fluoromethyl)pyrimidin-2-amine (Compound 114)
Step 1: (S)-tert-butyl
3-((4-(6-(3-methylisoxazol-4-yl)-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(tri-
fluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carboxylate
##STR00030##
[0124] To a solution of (S)-tert-butyl
3-((4-(6-bromo-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(trifluoromethyl)pyrim-
idin-2-yl)amino)piperidine-1-carboxylate (Example 3, step 1; 150
mg, 0.22 mmol) in degassed 2:1 mixture of dioxane/water (1.5 mL)
were added NaHCO.sub.3 (37 mg, 0.44 mmol) and
3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole
(81 mg, 0.39 mmol), the resulted suspension was degassed for 15
min. Tetrakis(triphenylphosphine)pal-ladium(0) (25 mg, 0.02 mmol)
was added, and the reaction mixture was stirred at 100.degree. C.
for 1 h. After cooling down to RT, the mixture was diluted with
EtOAc (15 mL) and water (10 mL), the crude product was extracted
with EtOAc (2.times.10 mL). The combined organic phase was washed
with sat. aq. NaHCO.sub.3 (5 mL), water (2.times.5 mL), and brine
(2.times.10 mL), then dried over Na.sub.2SO.sub.4, filtered, and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography (EtOAc in hexanes, 0 to 100%
gradient) to obtain the title compound as a yellow oil (139 mg,
0.20 mmol, 93% yield).
Step 2: (S)-tert-butyl
3-((4-(6-(3-methylisoxazol-4-yl)-1H-indol-3-yl)-5-(trifluoromethyl)pyrimi-
din-2-yl)amino)piperidine-1-carboxylate
##STR00031##
[0126] To a solution of (S)-tert-butyl
3-((4-(6-(3-methylisoxazol-4-yl)-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(tri-
fluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carboxylate (139 mg,
0.20 mmol) in THF (2 mL) was added 1M TBAF (tetrabutylammonium
fluoride) solution in THF (2 mL, 2.04 mmol), and the reaction
mixture was stirred at 70.degree. C. for 30 min. The mixture was
then concentrated under vacuum, re-dissolved in MeTHF (5 mL) and
washed with sat. aq. NH.sub.4Cl (5.times.5 mL) and brine (5 mL).
Organic phase was separated, dried over MgSO.sub.4, filtered, and
concentrated under reduced pressure to provide a yellow oil of
crude title compound (111 mg, 0.20 mmol, quant. yield), which was
used in the next step without further purification.
Step 3:
(S)-4-(6-(3-methylisoxazol-4-yl)-1H-indol-3-yl)-N-(piperidin-3-yl)-
-5-(trifluoromethyl)pyrimidin-2-amine
##STR00032##
[0128] To a solution of (S)-tert-butyl
3-((4-(6-(3-methylisoxazol-4-yl)-1H-indol-3-yl)-5-(trifluoromethyl)pyrimi-
din-2-yl)amino)piperidine-1-carboxylate (111 mg, 0.20 mmol) in DCM
(6 mL) was added TFA (0.31 mL, 4.00 mmol), and the reaction mixture
was stirred at RT for 1 h. The mixture was then concentrated and
azeotroped for few times with DCM. The basified residue was
purified by reverse phase chromatography (C18, MeCN in aq. 10 mM
ammonium formate pH 3.8, 0 to 100% gradient) to obtain the title
compound as a white solid (49.6 mg, 0.11 mmol, 56% yield over 2
steps).
Example 5. Synthesis of
3,5-dimethyl-4-(3-(2-(methylsulfonyl)-5-(trifluoromethyl)-pyrimidin-4-yl)-
-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole for
Synthesis of Compound 116
Step 1: 3,5-dimethyl-4-(1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole
##STR00033##
[0130] To a solution of 6-bromo-1H-pyrrolo[2,3-b]pyridine (6.00 g,
30.5 mmol) in degassed 2:1 mixture of dioxane/water (150 mL) were
added (3,5-dimethylisoxazol-4-yl)boronic acid (4.72 g, 33.5 mmol)
and NaHCO.sub.3 (5.12 g, 60.9 mmol), and the resulted suspension
was degassed for 30 min. Pd(dppf)Cl.sub.2
([1,1'-Bis(diphenyl-phosphino)ferrocene]dichloropalladium(II)) DCM
complex (620 mg, 0.76 mmol) was then added, and the reaction
mixture was stirred at 100.degree. C. for 2 h. After cooling down
to RT, the mixture was extracted with EtOAc (3.times.30 mL), the
combined organic phase was then washed with sat. aq. NaHCO.sub.3
(2.times.20 mL) and brine (2.times.20 mL), dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure
to obtain the crude title compound as a tan solid (6.5 g, 30.5
mmol, quant. yield), which was used in the next step without
further purification.
Step 2:
4-(3-bromo-1H-pyrrolo[2,3-b]pyridin-6-yl)-3,5-dimethylisoxazole
##STR00034##
[0132] A suspension of
3,5-dimethyl-4-(1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole (6.5 g,
30.5 mmol) in DCM (150 mL) was cooled to 0.degree. C., and M amount
of the required NBS (N-bromosuccinimide; 2.71 g, 15.2 mmol) was
added portion wise followed by stirring for 30 min. Reaction was
followed by LCMS (liquid chromatography mass spectrometry). After
30 min (where the conversion of about 40% was observed), the
remaining % amount of NBS (2.71 g, 15.2 mmol) was added portion
wise, and the reaction mixture was stirred for another 20 minutes.
The reaction was then quenched by the addition of sat. aq.
Na.sub.2S.sub.2O.sub.3 and stirred vigorously for 16 h. After
overnight, the mixture was concentrated under reduced pressure and
re-dissolved in EtOAc (100 mL) and water (50 mL). Organic phase was
separated, washed with sat. aq. NaHCO.sub.3 (50 mL), water
(2.times.50 mL), and brine (2.times.50 mL), then dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure
to provide the crude title compound as a brown solid (8.54 g, 29.2
mmol, 96% yield), which was used in the next step without further
purification.
Step 3:
4-(3-bromo-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3,5-d-
imethylisoxazole
##STR00035##
[0134] A solution of
4-(3-bromo-1H-pyrrolo[2,3-b]pyridin-6-yl)-3,5-dimethylisoxazole
(8.54 g, 29.2 mmol) in dry THF (146 mL) at 0.degree. C. was treated
with NaOt-Bu (3.37 g, 35.1 mmol) and then was stirred for 30 min at
this temperature before adding benzenesulfonyl chloride (3.92 mL,
30.7 mmol). The reaction mixture was then stirred for 30 min at
0.degree. C. The mixture was concentrated to about 20 mL of THF
remaining and diluted with water (70 mL). After sonication, the
obtained suspension was filtered, the solid was rinsed with water
and then dried for to provide the crude title compound as a red
solid (12.64 g, 29.2 mmol, quant. yield), which was used in the
next step without further purification.
Step 4:
3,5-dimethyl-4-(1-(phenylsulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-di-
oxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole
##STR00036##
[0136] To a solution of
4-(3-bromo-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3,5-dimethyl-
isoxazole (8.69 g, 20.1 mmol) in degassed dioxane (67 mL) were
added bis(pinacolato)diboron (12.9 g, 50.7 mmol) and KOAc (9.91 g,
101.0 mmol), and the mixture was degassed for 30 min.
Pd(dppf)Cl.sub.2 DCM complex (1.63 g, 2.00 mmol) was added, and
then the reaction mixture in the sealed tube was stirred at
100.degree. C. for 1 h. The resulted black solution was cooled down
to RT, diluted with EtOAc (100 mL), filtered through Celite and
treated with sat. aq. NaHCO.sub.3 (75 mL). Crude product was
extracted with EtOAc (2.times.30 mL), combined organic phase was
then washed with water (2.times.50 mL) and brine (2.times.50 mL),
dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure to a black sticky solid of crude title compound
(9.61 g, 20.1 mmol, quant. yield), which was used in the next step
without further purification.
Step 5:
3,5-dimethyl-4-(3-(2-(methylthio)-5-(trifluoromethyl)pyrimidin-4-y-
l)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole
##STR00037##
[0138] To a solution of
3,5-dimethyl-4-(1-(phenylsulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboro-
lan-2-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole (9.61 g, 20.1
mmol) in degassed 2:1 mixture of dioxane/water (100 mL) was added
Cs.sub.2CO.sub.3 (13.06 g, 40.1 mmol), and obtained suspension was
degassed for 30 min.
4-Chloro-2-(methylthio)-5-(trifluoromethyl)pyrimidine
(US20130017194) (5.04 g, 22.1 mmol) and
tetrakis(triphenylphosphine)palladium(0) (2.32 g, 2.01 mmol) were
added, and the reaction mixture was stirred at 100.degree. C. After
2 h, another portion of
4-chloro-2-(methylthio)-5-(trifluoromethyl)pyrimidine (1.00 g, 4.38
mmol) was added, and the reaction mixture was heated for another 30
min at 100.degree. C. After cooling down to RT, the mixture was
diluted with EtOAc (100 mL) and water (75 mL), and pH was adjusted
to pH 8 with sat. aq. NaHCO.sub.3. Crude product was extracted with
EtOAc (3.times.75 mL), combined organic phase was then washed with
water (2.times.50 mL) and brine (75 mL), separated, dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced
pressure. The residue was purified by silica gel column
chromatography (EtOAc in hexanes, 0 to 100% gradient) to provide an
orange paste, which was then re-purified by reverse phase
chromatography (C18, MeCN in aq. 10 mM ammonium formate pH 3.8, 0
to 100% gradient) to give the title compound as a brown solid (2.05
g, 3.76 mmol, 19% yield).
Step 6:
3,5-dimethyl-4-(3-(2-(methylsulfonyl)-5-(trifluoromethyl)pyrimidin-
-4-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole
##STR00038##
[0140] To a solution of
3,5-dimethyl-4-(3-(2-(methylthio)-5-(trifluoromethyl)pyrimidin-4-yl)-1-(p-
henylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole (2.00 g,
3.67 mmol) in DME (dimethoxyethane; 37 mL) was added m-CPBA
(3-chloroperbenzoic acid; 77%) (1.65 g, 7.34 mmol), and the
reaction mixture was stirred at RT for 2 h. The suspension was then
filtered, and the solids were washed with DCM (20 mL) and EtOAc (20
mL). The collected organic filtrate was concentrated under reduced
pressure, and the residue was purified by silica gel column
chromatography (EtOAc in hexanes, 0 to 100% gradient). The title
compound was obtained as a pale orange solid (1.08 g, 1.87 mmol,
51% yield).
Example 6. Synthesis of
4-(6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-N-((3R,4R-
)-4-methoxypiperidin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 117)
Step 1: Racemic trans tert-butyl
3-azido-4-methoxypiperidine-1-carboxylate
##STR00039##
[0142] To a solution of racemic trans tert-butyl
3-azido-4-hydroxypiperidine-1-carboxylate (Angew. Chem., Int. Ed.
2016, 55, 11382) (504 mg, 2.08 mmol) in dry THF at 0.degree. C. in
an ice/water bath, was added sodium hydride (100 mg, 2.5 mmol)
portion wise. The suspension was stirred at this temperature for 15
min. Then, iodomethane (0.144 mL, 2.29 mmol) was added. The
resulting solution was allowed to reach RT and stirred for 16 h.
After full conversion of starting material, the reaction mixture
was quenched by addition of MeOH and then diluted with DCM. Water
was added, and organic layer was separated, washed with brine
(2.times.20 mL), dried over MgSO.sub.4, filtered, and concentrated
under reduced pressure. The yellowish residue was purified by
silica gel chromatography (EtOAc in hexanes, 0 to 30% gradient).
The title compound (511 mg, 1.99 mmol, 96% yield) was obtained as a
translucent oil.
Step 2: Racemic trans tert-butyl
3-amino-4-methoxypiperidine-1-carboxylate
##STR00040##
[0144] To a solution of racemic trans tert-butyl
3-azido-4-methoxypiperidine-1-carboxylate (511 mg, 1.99 mmol) in
EtOH (ethanol; 10 mL) was added Pd/C, 10 wt. % (400 mg). The
suspension was purged with hydrogen (three times). The resulting
mixture was then stirred at RT for 3 h under atmospheric pressure
of hydrogen. After full conversion of starting material, the
suspension was filtered through a pad of Celite. The filtrate was
concentrated under reduced pressure to provide the crude title
compound as a yellowish oil, which was used in the next step
without further purification.
Step 3: Racemic trans tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]-
pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-4-methoxypiperidine-
-1-carboxylate
##STR00041##
[0146] To a solution of
3,5-dimethyl-4-(3-(2-(methylsulfonyl)-5-(trifluoromethyl)pyrimidin-4-yl)--
1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole (Example
5; 105 mg, 0.182 mmol) and racemic trans tert-butyl
3-amino-4-methoxypiperidine-1-carboxylate (63 mg, 0.273 mmol) in
dry THF (0.9 mL) was added DIPEA (0.1 mL, 0.545 mmol). The reaction
mixture was then stirred for 16 h at RT. The mixture was diluted
with EtOAc (10 mL), washed with brine (2.times.10 mL). Organic
phase was separated, dried over Na.sub.2SO.sub.4, filtered, and
concentrated under reduced pressure. The obtained crude title
compound (130 mg, 0.179 mmol, 98% yield) was used in the next step
without further purification.
Step 4: Racemic trans tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(tr-
ifluoromethyl)pyrimidin-2-yl)amino)-4-methoxypiperidine-1-carboxylate
##STR00042##
[0148] To a solution of racemic trans tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]-
pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-4-methoxypiperidine-
-1-carboxylate (130 mg, 0.179 mmol) in dioxane (1.2 mL) was added
5M aq. NaOH (0.360 mL, 1.79 mmol). The reaction mixture was then
stirred at room temperature for 16 h. After overnight, the mixture
was concentrated under reduced pressure, and the residue was
purified by reverse phase chromatography (C18, MeCN in aq. 10 mM
ammonium formate pH 3.8, 0 to 100% gradient). The title compound
(65 mg, 0.111 mmol, 62% yield) was obtained as a pale yellow
solid.
Step 5: (3R,4R)-tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(tr-
ifluoromethyl)pyrimidin-2-yl)amino)-4-methoxypiperidine-1-carboxylate
(A)
##STR00043##
[0150] Racemic trans tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(tr-
ifluoromethyl)pyrimidin-2-yl)amino)-4-methoxypiperidine-1-carboxylate
(65 mg) was separated by chiral HPLC (high pressure liquid
chromatography) using a ChiralPak IA column and 8:92 mixture of
EtOH/Hexanes for elution. Two enantiomers were obtained: Peak 1 (A,
tentatively assigned as (3R,4R), 20 mg, pale yellow solid), and
Peak 2 (B, tentatively assigned as (3S,4S), 18 mg, pale yellow
solid).
Step 6:
4-(6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-N--
((3R,4R)-4-methoxypiperidin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
##STR00044##
[0152] To a solution of Compound A from the previous step assigned
as (3R,4R)-enantiomer (20 mg, 0.034 mmol) in DCM (1 mL) was added
TFA (0.22 mL, 2.87 mmol). After 1 h of stirring at RT, reaction
mixture was concentrated under reduced pressure, and the basified
residue was purified by reverse phase chromatography (C18, MeCN in
aq. 10 mM ammonium formate pH 3.8, 10 to 50% gradient) to afford
the title compound (9.4 mg, 0.019 mmol, 57% yield) as a white
solid.
Example 7. Synthesis of
4-(6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl)-N-((3R,4R)-4-methoxypiper-
idin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (Compound 127)
Step 1: (3R,4R)-tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-1-(phenylsulfonyl)-1H-indol-3-yl)-5--
(trifluoromethyl)pyrimidin-2-yl)amino)-4-methoxypiperidine-1-carboxylate
(A)
##STR00045##
[0154] Racemic trans tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-1-(phenylsulfonyl)-1H-indol-3-yl)-5--
(trifluoromethyl)pyrimidin-2-yl)amino)-4-methoxypiperidine-1-carboxylate
(Example 6; 197 mg, 0.271 mmol) was separated by chiral HPLC using
a ChiralPak IA column and 8:20:72 mixture of MeOH/IPA/hexanes for
elution. Two enantiomers were obtained: Peak 1 (A, tentatively
assigned as (3R,4R), 84 mg, pale yellow solid), and Peak 2 (B,
tentatively assigned as (3S,4S), 81 mg, pale yellow solid).
Step 2:
4-(6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl)-N-((3R,4R)-4-metho-
xypiperidin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
##STR00046##
[0156] To a solution of Compound A from the previous step assigned
as (3R,4R)-enantiomer (84 mg, 0.116 mmol) in dioxane (1 mL) was
added 5M aq. NaOH (1.2 mL, 5.8 mmol). The reaction mixture was then
stirred at 70.degree. C. until full conversion. After 4 h, mixture
was cooled down to RT and diluted in water. Aqueous layer was
extracted with MeTHF (3.times.25 mL). Combined organic phase was
washed with brine (50 mL), separated, dried over Na.sub.2SO.sub.4,
filtered, and concentrated under reduced pressure. The obtained
crude was re-dissolved in DCM (1.7 mL), and TFA (0.444 mL, 5.8
mmol) was added. After 1 h of stirring at RT, the reaction mixture
was concentrated under reduced pressure, and the basified residue
was purified by reverse phase chromatography (C18, MeCN in aq. 10
mM ammonium formate pH 3.8, 10 to 55% gradient) to afford the title
compound (39.2 mg, 0.081 mmol, 70% yield over 2 steps) as a white
solid.
Example 8. Synthesis of (S)-tert-butyl
3-((4-(6-(3-methylisothiazol-4-yl)-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(t-
rifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carboxylate useful
in the synthesis of Compound 128
Step 1: (S)-tert-butyl
3-((4-(1-(phenylsulfonyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-
-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carbo-
xylate
##STR00047##
[0158] To a solution of (S)-tert-butyl
3-((4-(6-bromo-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(trifluoromethyl)pyrim-
idin-2-yl)amino)piperidine-1-carboxylate (Example 3, step 1; 400
mg, 0.59 mmol) in degassed dioxane (4.2 mL) were added
bis(pinacolato)diboron (373 mg, 1.47 mmol) and KOAc (288 mg, 2.95
mmol), and the mixture was degassed for 15 min. Pd(dppf)Cl.sub.2
DCM complex (48 mg, 0.06 mmol) was added, and then the reaction
mixture was stirred at 95.degree. C. until full conversion
(followed by LCMS). After 2 h, the mixture was cooled down to RT,
diluted with EtOAc (50 mL), washed with sat. aq. NaHCO.sub.3 (50
mL), water (50 mL), and brine (50 mL). Organic phase was separated,
dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure to provide a brown semi-solid of crude title
compound, which was used in the next step without further
purification.
Step 2: (S)-tert-butyl
3-((4-(6-(3-methylisothiazol-4-yl)-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(t-
rifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carboxylate
[0159] To a solution of (S)-tert-butyl
3-((4-(1-(phenylsulfonyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-
-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carbo-
xylate (214 mg, 0.294 mmol) and 4-bromo-3-methylisothiazole (79 mg,
0.441 mmol) in previously degassed 2:1 mixture of dioxane/H.sub.2O
(6 mL) was added Na.sub.2CO.sub.3 (94 mg, 0.882 mmol), and the
mixture was degassed for 15 min. Pd(dppf)Cl.sub.2 DCM complex (24
mg, 0.029 mmol) was added, and the reaction mixture was stirred at
100.degree. C. for 2 h. The mixture was cooled down to RT, diluted
with EtOAc (30 mL), washed with sat. aq. NaHCO.sub.3 (30 mL), water
(30 mL), and brine (30 mL). The organic phase was separated, dried
over Na.sub.2SO.sub.4, filtered, and concentrated under reduced
pressure. The residue was purified by silica gel column
chromatography (EtOAc in 1:1 mixture of hexanes/DCM, 0 to 100%
gradient) to provide the title compound as a yellowish oil (152 mg,
74% yield).
Example 9. Synthesis of
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo-[2,3-b]-pyridin-3-yl]-N-[(3S)-
-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
101)
Step 1: 3,5-dimethyl-4-(1H-pyrrolo-[2,3-b]-pyridin-6-yl)
isoxazole
##STR00048##
[0161] A mixture of 6-bromo-1H-pyrrolo-[2,3-b]-pyridine (1 g, 5.08
mmol, 1 eq), (3,5-dimethylisoxazol-4-yl) boronic acid (1.43 g,
10.15 mmol, 2 eq), Pd(dppf)Cl.sub.2.CH.sub.2Cl.sub.2 (207.23 mg,
253.77 .mu.mol, 0.05 eq), NaHCO.sub.3 (1.28 g, 15.23 mmol, 592.17
.mu.L, 3 eq) in DME (10 mL) and H.sub.2O (3 mL) was degassed and
purged with N.sub.2 for 3 times, and then the mixture was stirred
at 100.degree. C. for 12 h under N.sub.2 atmosphere. The reaction
mixture was poured into H.sub.2O (20 mL) and extracted with EtOAc
(20 mL.times.3). The combined organic layers were washed with brine
(20 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by column chromatography (SiO.sub.2, PE/EtOAc=10/1 to
2/1) to afford the title compound (1.1 g, 90% purity) as a white
solid.
Step 2:
4-(3-bromo-1H-pyrrolo-[2,3-b]-pyridin-6-yl)-3,5-dimethyl-isoxazole
##STR00049##
[0163] To a solution of
3,5-dimethyl-4-(1H-pyrrolo-[2,3-b]-pyridin-6-yl) isoxazole (0.9 g,
4.22 mmol, 1 eq) in DMF (dimethylformamide; 10 mL) was added NBS
(676.10 mg, 3.80 mmol, 0.9 eq). The mixture was stirred at
25.degree. C. for 1 h. The reaction mixture was poured into
H.sub.2O (20 mL), and extracted with EtOAc (20 mL.times.3). The
combined organic layers were washed with brine (20 mL.times.2),
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. The residue was purified by
column chromatography (SiO.sub.2, PE/EtOAc=10/1 to 1/1) to afford
the title compound (600 mg, crude) as pink solid which was used in
the next step directly.
Step 3:
4-[1-(benzenesulfonyl)-3-bromo-pyrrolo-[2,3-b]-pyridin-6-yl]-3,5-d-
imethyl-isoxazole
##STR00050##
[0165] To a solution of
4-(3-bromo-1H-pyrrolo[2,3-b]-pyridin-6-yl)-3,5-dimethyl-isoxazole
(0.6 g, 2.05 mmol, 1 eq) in DMF (9 mL) and THF (1 mL) was added NaH
(98.58 mg, 2.46 mmol, 60% purity, 1.2 eq) and benzenesulfonyl
chloride (471.58 mg, 2.67 mmol, 341.73 .mu.L, 1.3 eq). The mixture
was stirred at 25.degree. C. for 1 h. The reaction mixture was
poured into H.sub.2O (20 mL) while white solid formed. The solid
was filtered and concentrated under reduced pressure to afford the
title compound (600 mg) as white solid, which used into the next
step without further purification.
Step 4: 4-[1-(benzenesulfonyl)
pyrrolo-[2,3-b]-pyridin-6-yl]-3,5-dimethyl-isoxazole;
4-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)
pyrrolo-[2,3-b]-pyridin-6-yl]-3,5-dimethyl-isoxazole
##STR00051##
[0167] A mixture of
4-[1-(benzenesulfonyl)-3-bromo-pyrrolo-[2,3-b]-pyridin-6-yl]-3,5-dimethyl-
-isoxazole (0.27 g, 624.58 .mu.mol, 1 eq), BPD (237.91 mg, 936.87
.mu.mol, 1.5 eq), Pd(dppf)Cl.sub.2 (45.70 mg, 62.46 .mu.mol, 0.1
eq), KOAc (122.60 mg, 1.25 mmol, 2 eq) in dioxane (3 mL) was
degassed and purged with N.sub.2 for 3 times, and then the mixture
was stirred at 100.degree. C. for 1 h under N.sub.2 atmosphere. The
reaction mixture was poured into H.sub.2O (20 mL) and extracted
with EtOAc (20 mL.times.3). The combined organic layers were washed
with brine (10 mL.times.2), dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to afford the title
compound (600 mg, crude, 2 Batches in parallel) as brown oil which
was used into the next step without further purification.
Step 5: Tert-butyl
(3S)-3-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)pyrrolo-[2,3-
-b]-pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-1-c-
arboxylate
##STR00052##
[0169] A mixture of 4-[1-(benzenesulfonyl)
pyrrolo-[2,3-b]-pyridin-6-yl]-3,5-dimethyl-isoxazole and
4-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)
pyrrolo-[2,3-b]-pyridin-6-yl]-3,5-dimethyl-isoxazole (600 mg, from
previous step), tert-butyl
(3S)-3-[[4-chloro-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-1--
carboxylate ((PCT Int. Appl., WO2014124230); 476.63 mg, 1.25 mmol),
Pd(dppf)Cl.sub.2 (91.59 mg, 125.17 .mu.mol), Na.sub.2CO.sub.3
(265.33 mg, 2.50 mmol) in dioxane (10 mL) and H.sub.2O (2 mL) was
degassed and purged with N.sub.2 for 3 times, and then the mixture
was stirred at 100.degree. C. for 12 h under N.sub.2 atmosphere.
The reaction mixture was poured into H.sub.2O (30 mL) and extracted
with EtOAc (30 mL.times.4). The combined organic layers were washed
with brine (100 mL.times.2), dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give a residue. The
residue was purified by column chromatography (SiO.sub.2,
PE/EtOAc=10/1 to 2/1) to afford the title compound (0.25 g, 286.65
.mu.mol, 22.90% yield, 80% purity) as yellow solid.
Step 6: Tert-butyl
(3S)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo-[2,3-b]-pyridin-3-yl-
]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-1-carboxylate
##STR00053##
[0171] To a solution of tert-butyl
(3S)-3-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)pyrrolo-[2,3-
-b]-pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-1-c-
arboxylate (0.2 g, 286.65 .mu.mol, 1 eq) in MeOH (1 mL) was added
NaOH (2 M, 0.8 mL, 5.58 eq). The mixture was stirred at 60.degree.
C. for 1 h. The reaction mixture was concentrated under reduced
pressure to afford the title compound (0.2 g, crude) as yellow
solid, which was used into the next step without further
purification.
Step 7:
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo-[2,3-b]-pyridin-3-yl]--
N-[(3S)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00054##
[0173] To a solution of tert-butyl
(3S)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo-[2,3-b]-pyridin-3-yl-
]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-1-carboxylate
(0.2 g, 358.70 .mu.mol, 1 eq) in EtOAc (1 mL) was added HCl/EtOAc
(4 M, 2 mL, 22.30 eq). The mixture was stirred at 25.degree. C. for
1 h then concentrated under reduced pressure to give a residue that
was purified by prep-HPLC (HCl condition) to afford the title
compound (31.6 mg, HCl salt, 100% purity) as a yellow solid.
Example 10. Synthesis of
2-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyr-
rolo-[2,3-b]pyridin-6-yl]thiazole for use in Synthesizing Compound
108
Step 1: 1-(benzenesulfonyl)-6-bromo-pyrrolo[2,3-b]pyridine
##STR00055##
[0175] To a solution of 6-bromo-1H-pyrrolo[2,3-b]pyridine (1 g,
5.08 mmol, 1 eq) in DMF (9 mL) and THF (1 mL) was added NaH (243.82
mg, 6.10 mmol, 60% purity, 1.2 eq) at 0.degree. C. for 0.5 h, then
benzenesulfonyl chloride (1.17 g, 6.60 mmol, 845.22 .mu.L, 1.3 eq)
was added to the solution at 0.degree. C. The mixture was stirred
at 20.degree. C. for 12 h. The reaction mixture was diluted with
water (100 mL) and extracted with EtOAc (30 mL.times.3). The
combined organic layers were washed with brine (30 mL.times.3),
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. The residue was purified by
column chromatography (SiO.sub.2, PE/EtOAc=5/1) to afford the title
compound (1.6 g, 4.27 mmol, 84.07% yield, 90% purity) as a white
solid.
Step 2:
2-[1-(benzenesulfonyl)pyrrolo[2,3-b]pyridin-6-yl]thiazole
##STR00056##
[0177] To a solution of
1-(benzenesulfonyl)-6-bromo-pyrrolo[2,3-b]pyridine (0.5 g, 1.48
mmol, 1 eq) in DMF (5 mL) was added tributyl (thiazol-2-yl)
stannane (610.32 mg, 1.63 mmol, 1.1 eq), Pd(PPh.sub.3).sub.4
(171.35 mg, 148.28 .mu.mol, 0.1 eq) and CuI (28.24 mg, 148.28
.mu.mol, 0.1 eq). The mixture was stirred at 130.degree. C. for 12
h under N.sub.2. The reaction mixture was diluted with saturated
aqueous of KF (50 mL) and extracted with EtOAc (10 mL.times.3). The
combined organic layers were washed with brine (10 mL.times.3),
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. The residue was purified by
column chromatography (SiO.sub.2, PE/EtOAc=3/1) to afford the title
compound (420 mg) as a white solid. (The reaction was combined with
another reaction in 50 mg scale for purification.)
Step 3:
2-[1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridin-6-yl]thiazole
##STR00057##
[0179] To a solution of
2-[1-(benzenesulfonyl)pyrrolo[2,3-b]pyridin-6-yl]thiazole (0.41 g,
1.20 mmol, 1 eq) in DMF (4 mL) was added NBS (213.74 mg, 1.20 mmol,
1 eq) at 0.degree. C. The mixture was stirred at 20.degree. C. for
12 h. The reaction mixture was diluted with water (50 mL) and
extracted with EtOAc (10 mL.times.3). The combined organic layers
were washed with brine (10 mL.times.3), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, PE/EtOAc=6/1 to EtOAc) to afford the
title compound (300 mg) as a pink red solid.
Step 4:
2-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
-yl)pyrrolo-[2,3-b]pyridin-6-yl]thiazole
##STR00058##
[0181] To a solution of
2-[1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridin-6-yl]thiazole
(0.8 g, 1.90 mmol, 1 eq) in dioxane (10 mL) was added BPD (725.01
mg, 2.86 mmol, 1.5 eq), KOAc (373.61 mg, 3.81 mmol, 2 eq) and
Pd(dppf)Cl.sub.2 (139.27 mg, 190.34 .mu.mol, 0.1 eq). The mixture
was stirred at 100.degree. C. for 2 h under N.sub.2. The reaction
mixture was diluted with water (100 mL) 10 and extracted with EtOAc
(15 mL.times.3). The combined organic layers were washed with brine
(15 mL.times.3), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to afford the title compound (1
g) as a brown oil.
Example 11. Synthesis of Tert-butyl
(5S)-5-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)
pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-3,3--
dimethyl-piperidine-1-carboxylate useful in the synthesis of
Compound 115)
Step 1: 4-chloro-2-methylsulfanyl-5-(trifluoromethyl)pyrimidine
##STR00059##
[0183] To a solution of 2,4-dichloro-5-(trifluoromethyl)pyrimidine
(10 g, 46.09 mmol, 1 eq) in THF (200 mL) was added dropwise
ZnCl.sub.2 (1 M, 59.91 mL, 1.3 eq) at 0.degree. C. After addition,
the mixture was stirred at this temperature for 2 h, and then NaSMe
(3.88 g, 55.31 mmol, 3.52 mL, 1.2 eq) was added dropwise at
0.degree. C. The resulting mixture was stirred at 15.degree. C. for
14 h. The mixture was quenched with aq. HCl (150 mL, 1M), and then
extracted with EtOAc (60 mL.times.3), dried over Na.sub.2SO.sub.4
and concentrated. The residue was purified by column chromatography
(SiO.sub.2, PE/DCM=1:0 to 50:1) to afford the title compound (10 g,
43.74 mmol, 94.91% yield) as a colorless oil.
Step 2:
4-[1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyri-
midin-4-yl]pyrrolo-[2,3-b]pyridin-6-yl]-3,5-dimethylisoxazole
##STR00060##
[0185] To a solution of
4-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyr-
rolo[2,3-b]pyridin-6-yl]-3,5-dimethyl-isoxazole (Example 9; 3 g,
3.76 mmol, 1 eq) and
4-chloro-2-methylsulfanyl-5-(trifluoromethyl)pyrimidine (1.43 g,
5.63 mmol, 1.5 eq) in dioxane (30 mL)/H.sub.2O (6 mL) was added
Na.sub.2CO.sub.3 (1.19 g, 11.27 mmol, 3 eq) and Pd(dppf)Cl.sub.2
(274.76 mg, 375.50 .mu.mol, 0.1 eq). The mixture was stirred at
100.degree. C. under N.sub.2 atmosphere for 1 h. The reaction
mixture was poured into water (150 mL). The aqueous phase was
extracted with ethyl acetate (50 mL.times.3). The combined organic
phase was washed with brine (50 mL.times.2), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue
was purified by silica gel chromatography (100-200 mesh silica gel,
PE/EtOAc=10/1,4/1) to afford the title compound (0.7 g, 769.86
.mu.mol, 20.50% yield, 60% purity) as a yellow solid.
Step 3:
4-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyri-
midin-4-yl]-pyrrolo[2,3-b]-pyridin-6-yl]-3,5-dimethylisoxazole
##STR00061##
[0187] To a solution of
4-[1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-
-yl]pyrrolo[2,3-b]pyridin-6-yl]-3,5-dimethyl-isoxazole (0.7 g, 1.28
mmol, 1 eq) in DCM (50 mL) was added m-CPBA (573.08 mg, 2.82 mmol,
2.2 eq). The mixture was stirred at 20.degree. C. for 12 h. The
residue was poured into a mixture of Sat.NaHCO.sub.3 (20 mL) and
Sat. Na.sub.2SO.sub.3 (20 mL), and the mixture was stirred for 5
min. The aqueous phase was extracted with DCM (30 mL.times.2). The
combined organic phase was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuum. The residue was purified by
silica gel chromatography (100-200 mesh silica gel,
PE/EtOAc=10/1,2/1, contained 10% DCM) to afford the title compound
(0.5 g, 822.43 .mu.mol, 64.10% yield, 95% purity) as a white
solid.
Step 4: Tert-butyl
(5S)-5-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)pyrrolo[2,3--
b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-3,3-dimethyl-pipe-
ridine-1-carboxylate
##STR00062##
[0189] To a solution of
4-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-
-yl]pyrrolo[2,3-b]pyridin-6-yl]-3,5-dimethyl-isoxazole (0.1 g,
173.14 .mu.mol, 1 eq), tert-butyl
(5S)-5-amino-3,3-dimethyl-piperidine-1-carboxylate (114.01 mg,
207.77 .mu.mol, 1.2 eq, FA) in THF (3 mL) was added DIEA (111.89
mg, 865.72 .mu.mol, 150.79 .mu.L, 5 eq). The mixture was stirred at
20.degree. C. under N.sub.2 atmosphere for 1 h. The reaction
mixture was poured into water (30 mL). The aqueous phase was
extracted with EtOAc (20 mL.times.3). The combined organic phase
was washed with brine (20 mL.times.2), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue
was purified by silica gel chromatography (100-200 mesh silica gel,
PE/EtOAc=10/1,2/1) to afford the title compound (0.12 g) as yellow
solid.
Example 12. Synthesis of
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrazolo-[3,4-b]-pyridin-3-yl]-N-[(3S-
)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
118)
Step 1: 3,5-dimethyl-4-(1H-pyrazolo-[3,4-b]-pyridin-6-yl)
isoxazole
##STR00063##
[0191] A mixture of 6-chloro-1H-pyrazolo-[3,4-b]-pyridine (1 g,
6.51 mmol, 1 eq), (3,5-dimethylisoxazol-4-yl) boronic acid (1.38 g,
9.77 mmol, 1.5 eq), Pd(dppf)Cl.sub.2.CH.sub.2Cl.sub.2 (531.77 mg,
651.17 .mu.mol, 0.1 eq), K.sub.2CO.sub.3 (5 M, 3.91 mL, 3 eq) in
dioxane (20 mL) was degassed and purged with N.sub.2 for 3 times,
and then the mixture was stirred at 100.degree. C. for 2 h under
N.sub.2 atmosphere. The reaction mixture was poured into water (100
mL), and then extracted with EtOAc (30 mL.times.3). The combined
organic layers were washed with brine (200 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, PE/EtOAc=10/1 to 1/1) to afford the
title compound (1 g) as a yellow solid.
Step 2:
4-(3-iodo-1H-pyrazolo-[3,4-b]-pyridin-6-yl)-3,5-dimethyl-isoxazole
##STR00064##
[0193] To a solution of
3,5-dimethyl-4-(1H-pyrazolo-[3,4-b]-pyridin-6-yl) isoxazole (0.9 g,
4.20 mmol, 1 eq) in DMF (10 mL) was added KOH (942.92 mg, 16.80
mmol, 4 eq) and molecular iodine (2.13 g, 8.40 mmol, 1.69 mL, 2
eq). The mixture was stirred at 140.degree. C. for 3 h. The
reaction mixture was quenched by addition 10% Na.sub.2SO.sub.3 (100
mL) at 15.degree. C., then extracted with EtOAc (50 mL.times.3),
dried over Na.sub.2SO.sub.4 and concentrated to afford the title
compound (1 g, crude) as a yellow solid.
Step 3:
4-(3-iodo-1-tetrahydropyran-2-yl-pyrazolo-[3,4-b]-pyridin-6-yl)-3,-
5-dimethyl-isoxazole
##STR00065##
[0195] To a solution of
4-(3-iodo-1H-pyrazolo-[3,4-b]-pyridin-6-yl)-3,5-dimethyl-isoxazole
(0.9 g, 2.65 mmol, 1 eq) in DCM (1 mL) was added
3,4-dihydro-2H-pyran (267.10 mg, 3.18 mmol, 290.32 .mu.L, 1.2 eq)
and PTSA (p-toluenesulfonic acid; 45.57 mg, 264.61 .mu.mol, 0.1
eq). The mixture was stirred at 25.degree. C. for 3 h. The mixture
was washed with Sat.NaHCO.sub.3 (100 mL.times.2) and brine (200
mL), dried over Na.sub.2SO.sub.4, filtered and the filtrate was
evaporated. The residue was purified by column chromatography
(SiO.sub.2, PE/EtOAc=10/1 to 1/1) to afford the title compound (0.8
g, 90% purity) as a white solid.
Step 4:
3,5-dimethyl-4-[3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-
-yl]-1-tetrahydropyran-2-yl-pyrazolo-[3,4-b]-pyridin-6-yl]-isoxazole
##STR00066##
[0197] A mixture of
4-(3-iodo-1-tetrahydropyran-2-yl-pyrazolo-[3,4-b]-pyridin-6-yl)-3,5-dimet-
hyl-isoxazole (0.25 g, 589.30 .mu.mol, 1 eq),
4-chloro-2-methylsulfanyl-5-(trifluoromethyl)pyrimidine (134.73 mg,
589.30 .mu.mol, 1 eq), Pd(PPh.sub.3).sub.4 (68.10 mg, 58.93
.mu.mol, 0.1 eq), trimethyl (trimethylstannyl) stannane (193.07 mg,
589.30 .mu.mol, 122.20 .mu.L, 1 eq) in Tol. (2 mL) was degassed and
purged with N.sub.2 for 3 times, and then the mixture was stirred
at 100.degree. C. for 16 h under N.sub.2 atmosphere. The reaction
mixture was poured into aq. KF (50 mL), and then extracted with
EtOAc (20 mL.times.3). The combined organic layers were washed with
brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by column chromatography (SiO.sub.2, PE/EtOAc=10/1 to
3/1) to afford the title compound (150 mg, 305.81 .mu.mol, 51.89%
yield) as a yellow solid.
Step 5:
3,5-dimethyl-4-[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-
-yl]-1-tetrahydropyran-2-yl-pyrazolo-[3,4-b]-pyridin-6-yl]-isoxazole
##STR00067##
[0199] To a solution of
3,5-dimethyl-4-[3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-yl]-1--
tetrahydropyran-2-yl-pyrazolo-[3,4-b]-pyridin-6-yl]-isoxazole (0.15
g, 305.81 .mu.mol, 1 eq) in DCM (1 mL) was added m-CPBA (155.22 mg,
764.53 .mu.mol, 85% purity, 2.5 eq). The mixture was stirred at
15.degree. C. for 16 h. The reaction mixture was quenched by
addition Sat.Na.sub.2SO.sub.3 (5 mL) and NaHCO.sub.3 (5 mL),
stirred at this temperature for 10 min and then extracted with DCM
(10 mL.times.2). The combined organic layers were washed with brine
(30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. The residue was
triturated with PE:EtOAc (5:1.5 mL). The solid was filtered and
dried to afford the title compound (100 mg, 191.39 .mu.mol, 62.58%
yield) as a yellow solid.
Step 6: Tert-butyl
(3S)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-tetrahydropyran-2-yl-pyrazolo-
-[3,4-b]-pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidin-
e-1-carboxylate
##STR00068##
[0201] A mixture of
3,5-dimethyl-4-[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl]-1--
tetrahydropyran-2-yl-pyrazolo-[3,4-b]-pyridin-6-yl]-isoxazole (0.1
g, 191.39 .mu.mol, 1 eq), tert-butyl
(3S)-3-aminopiperidine-1-carboxylate (57.50 mg, 287.08 .mu.mol, 1.5
eq), DIEA (74.20 mg, 574.16 .mu.mol, 100.01 .mu.L, 3 eq) in THF (1
mL) was as stirred at 15.degree. C. for 3 h. It was concentrated.
The residue was purified by column chromatography (SiO.sub.2,
PE/EtOAc=10/1 to 1/1) to afford the title compound (100 mg) as a
yellow gum.
Step 7:
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrazolo-[3,4-b]-pyridin-3-yl]-
-N-[(3S)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00069##
[0203] To a solution of tert-butyl
(3S)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-tetrahydropyran-2-yl-pyrazolo-
-[3,4-b]-pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidin-
e-1-carboxylate (0.06 g, 93.36 .mu.mol, 1 eq) in DCM (1 mL) was
added Et.sub.3SiH (32.57 mg, 280.08 .mu.mol, 44.73 .mu.L, 3 eq) and
TFA (212.90 mg, 1.87 mmol, 138.25 .mu.L, 20 eq). The mixture was
stirred at 20.degree. C. for 5 h. The reaction mixture was
concentrated under reduced pressure to give a residue. The residue
was purified by prep-HPLC (HCl condition) to afford the title
compound (20.4 mg, HCl salt, 100% purity) as a yellow solid.
Example 13. Synthesis of
(3R,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-y-
l]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-N,N-dimethyl-piperidine-3-carb-
oxamide (Compound 119)
Step 1:
O1-tert-butyl-O3-methyl(3R,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3,5-d-
imethylisoxazol-4-yl)-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrim-
idin-2-yl]amino]piperidine-1,3-dicarboxylate
##STR00070##
[0205] To a solution of
4-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-
-yl]-pyrrolo[2,3-b]pyridin-6-yl]-3,5-dimethyl-isoxazole (Example
11; 260 mg, 450.17 .mu.mol, 1 eq) in THF (5 mL) was added DIPEA
(290.91 mg, 2.25 mmol, 392.06 .mu.L, 5 eq) and O1-tert-butyl
O3-methyl (3R,5S)-5-aminopiperidine-1,3-dicarboxylate (174.43 mg,
675.26 .mu.mol, 1.5 eq). The mixture was stirred at 15.degree. C.
for 1 h. The reaction mixture was diluted with H.sub.2O (15 mL),
and then extracted with EtOAc (15 mL.times.3). The combined organic
layers were washed with brine (40 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
The residue was purified by column chromatography (SiO.sub.2,
PE/EtOAc=10:1 to 1:1) to afford the title compound (240 mg) as a
yellow solid.
Step 2:
(3R,5S)-1-tert-butoxycarbonyl-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-
-1H-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]pi-
peridine-3-carboxylic acid
##STR00071##
[0207] To a solution of
O1-tert-butyl-O3-methyl(3R,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethyl-
isoxazol-4-yl)-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2--
yl]amino]piperidine-1,3-dicarboxylate (220 mg, 291.10 .mu.mol, 1
eq) in MeOH (2 mL) was added NaOH (2 M, 2.00 mL, 13.74 eq). The
mixture was stirred at 15.degree. C. for 1 h. The reaction mixture
was concentrated under reduced pressure to remove MeOH. The residue
was diluted with H.sub.2O (15 mL) and acidified by 2N HCl to pH=2
and extracted with EtOAc (15 mL.times.3). The combined organic
layers were washed with brine (30 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to afford the
title compound (140 mg) as a yellow solid.
Step 3:
tert-butyl(3R,5S)-3-(dimethylcarbamoyl)-5-[[4-[6-(3,5-dimethylisox-
azol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-y-
l]amino]piperidine-1-carboxylate
##STR00072##
[0209] To a solution of
(3R,5S)-1-tert-butoxycarbonyl-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyr-
rolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-
e-3-carboxylic acid (60 mg, 99.74 .mu.mol, 1 eq) in THF (1 mL) was
added T.sub.3P (190.41 mg, 299.21 .mu.mol, 177.95 .mu.L, 50%
purity, 3 eq), DIPEA (64.45 mg, 498.69 .mu.mol, 86.86 .mu.L, 5 eq)
and N-methylmethanamine (2 M, 249.35 .mu.L, 5 eq). The mixture was
stirred at 15.degree. C. for 12 h. The reaction mixture was diluted
with H.sub.2O (15 mL), and then extracted with EtOAc (15
mL.times.3). The combined organic layers were washed with brine (40
mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to afford the title compound (50 mg) as yellow
solid.
Step 4:
(3R,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyri-
din-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-N,N-dimethyl-piperidine-
-3-carboxamide
##STR00073##
[0211] To a solution of
tert-butyl(3R,5S)-3-(dimethylcarbamoyl)-5-[[4-[6-(3,5-dimethylisoxazol-4--
yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino-
]piperidine-1-carboxylate (40 mg, 63.63 .mu.mol, 1 eq) was added
HCl/EtOAc (4 M, 2 mL). The mixture was stirred at 15.degree. C. for
1 h. The reaction mixture was concentrated under reduced pressure
to give a residue. The residue was purified by prep-HPLC to afford
the title compound (13.4 mg, FA) as white solid.
Example 14. Synthesis of Tert-butyl
(3S,5R)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-y-
l]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-5-(methylcarbamoyl)piperidine--
1-carboxylate useful in the synthesis of Compound 120
##STR00074##
[0213] To a solution of
(3R,5S)-1-tert-butoxycarbonyl-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyr-
rolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-
e-3-carboxylic acid (60 mg, 99.74 .mu.mol, 1 eq) in THF (1 mL) was
added T.sub.3P (190.41 mg, 299.21 .mu.mol, 177.95 .mu.L, 50%
purity, 3 eq), DIPEA (64.45 mg, 498.69 .mu.mol, 86.86 .mu.L, 5 eq)
and methanamine (2M, 249.35 .mu.L, 5 eq). The mixture was stirred
at 15.degree. C. for 12 h. The reaction mixture was diluted with
H.sub.2O (10 mL) and extracted with EtOAc (10 mL.times.3). The
combined organic layers were washed with brine (20 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, Petroleum ether/Ethyl acetate=1:1 to
1:20) to afford the title compound (50 mg) as white solid.
Example 15. Synthesis of
(3R,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl]-5-(trifluorom-
ethyl)pyrimidin-2-yl]-amino]-N, N-dimethyl-piperidine-3-carboxamide
(Compound 112)
Step 1:
2-[[6-bromo-3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-indol-1-
-yl]-methoxy]-ethyl-trimethyl-silane
##STR00075##
[0215] To a solution of
6-bromo-3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-1H-indole
(Example 19, step 1; 2 g, 5.31 mmol, 1 eq) in THF (20 mL) was added
NaH (254.94 mg, 6.37 mmol, 60% 25 purity, 1.2 eq) at -10.degree. C.
After addition, the mixture was stirred at this temperature for 30
min, and then 2-(chloromethoxy) ethyl-trimethyl-silane (1.33 g,
7.97 mmol, 1.41 mL, 1.5 eq) was added dropwise at -10.degree. C.
The resulting mixture was stirred at 0.degree. C. for 1 h. The
reaction mixture was diluted with water (150 mL) and extracted with
ethyl acetate (80 mL.times.2). The combined organic layers were
washed with brine (200 mL.times.2), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
The residue was purified by column chromatography (SiO.sub.2,
Petroleum ether/Ethyl acetate=50:1 to 20:1) to afford the title
compound (2.6 g) as a white solid.
Step 2:
O1-tert-butyl-O3-methyl-(3R,5S)-5-[[4-[6-bromo-1-(2-trimethylsilyl-
ethoxymethyl)
indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-1,3-dica-
rboxylate
##STR00076##
[0217] A mixture of
2-[[6-bromo-3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-indol-1-yl]-me-
thoxy]-ethyl-trimethyl-silane (1.46 g, 2.87 mmol, 1 eq),
O1-tert-butyl-O3-methyl-(3R,5S)-5-aminopiperidine-1,3-dicarboxylate
(0.89 g, 3.45 mmol, 1.2 eq) and DIEA (1.11 g, 8.61 mmol, 1.50 mL, 3
eq) in NMP (15 mL) was stirred at 140.degree. C. for 1 h. The
reaction mixture was diluted with water (100 mL) and extracted with
EtOAc (50 mL.times.2). The combined organic layers were washed with
brine (100 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by column chromatography (SiO2, PE/EtOAc=30:1 to 5:1)
to afford the title compound (1.68 g, 2.21 mmol, 77.09% yield, 96%
purity) as a white solid.
Step 3:
O1-tert-butyl-O3-methyl-(3R,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-y-
l)-1-(2-trimethylsilylethoxymethyl)indol-3-yl]-5-(trifluoromethyl)pyrimidi-
n-2-yl]-amino]-piperidine-1,3-dicarboxylate
##STR00077##
[0219] A mixture of
O1-tert-butyl-O3-methyl-(3R,5S)-5-[[4-[6-bromo-1-(2-trimethylsilylethoxym-
ethyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-1,3-
-dicarboxylate (0.7 g, 960.65 .mu.mol, 1 eq),
(3,5-dimethylisoxazol-4-yl)boronic acid (270.77 mg, 1.92 mmol, 2
eq), Pd(dppf)Cl.sub.2.CH.sub.2Cl.sub.2 (78.45 mg, 96.07 .mu.mol,
0.1 eq) and NaHCO.sub.3 (242.11 mg, 2.88 mmol, 112.09 .mu.L, 3 eq)
in DME (9 mL), H.sub.2O (3 mL) was degassed and purged with N.sub.2
for 3 times, and then the mixture was stirred at 100.degree. C. for
1 h under N.sub.2 atmosphere. The reaction mixture was diluted with
water (100 mL) and extracted with EtOAc (50 mL.times.2). The
combined organic layers were washed with brine (100 mL.times.2),
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. The residue was purified by
column chromatography (SiO.sub.2, PE/EtOAc=10:1 to 5:1) to afford
the title compound (600 mg) as a yellow solid.
Step 4:
(3R,5S)-1-tert-butoxycarbonyl-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-
-1-(2-trimethylsilylethoxymethyl)
indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-3-carbox-
ylic acid
##STR00078##
[0221] To a solution of
O1-tert-butyl-O3-methyl-(3R,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-(2-
-trimethylsilylethoxymethyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-
-amino]-piperidine-1,3-dicarboxylate (550 mg, 738.38 .mu.mol, 1 eq)
in MeOH (6 mL) was added NaOH (2 M, 3.69 mL, 10 eq). The mixture
was stirred at 20.degree. C. for 0.5 h. The reaction mixture was
adjusted pH to 4 with aqueous HCl (0.5 M, 80 mL) and extracted with
EtOAc (30 mL.times.3). The combined organic layers were washed with
brine (90 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue to afford the
title compound (600 mg, crude) was used into the next step without
further purification.
Step 5: Tert-butyl
(3R,5S)-3-(dimethylcarbamoyl)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-(2-t-
rimethylsilylethoxymethyl)
indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-ylamino]-piperidine-1-carboxyl-
ate
##STR00079##
[0223] To a solution of
(3R,5S)-1-tert-butoxycarbonyl-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-(2-t-
rimethylsilylethoxymethyl)
indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-3-carbox-
ylic acid (0.2 g, 273.65 .mu.mol, 1 eq) in THF (2 mL) was added
T.sub.3P (1.74 g, 2.74 mmol, 1.63 mL, 50% purity, 10 eq) and DIEA
(530.51 mg, 4.10 mmol, 714.97 .mu.L, 15 eq), N-methylmethanamine (2
M, 2.74 mL, 20 eq). The mixture was stirred at 20.degree. C. for 1
h. The reaction mixture was diluted with water (100 mL) and
extracted with EtOAc (50 mL.times.2). The combined organic layers
were washed with brine (100 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, PE/EtOAc=5:1 to 1:1) to afford the title
compound (200 mg) as a colorless oil.
Step 6:
(3R,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl]-5-(tri-
fluoromethyl)pyrimidin-2-yl]-amino]-N,
N-dimethyl-piperidine-3-carboxamide
##STR00080##
[0225] To a solution of tert-butyl
(3R,5S)-3-(dimethylcarbamoyl)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-(2-t-
rimethylsilylethoxymethyl)
indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-1-carbox-
ylate (0.18 g, 237.49 .mu.mol, 1 eq) in dioxane (2 mL) was added
H.sub.2SO.sub.4 (232.93 mg, 2.37 mmol, 126.59 .mu.L, 10 eq). The
mixture was stirred at 40.degree. C. for 2 h. The mixture was
adjusted pH to 8 with saturated aqueous K.sub.2CO.sub.3 and
extracted with EtOAc (20 mL.times.3). The combined organic layers
were washed with brine (30 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by prep-HPLC (HCl
condition) to afford the title compound (28.6 mg, HCl salt) as a
yellow solid.
Example 16. Synthesis of Tert-butyl
(3S,5R)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-(2-trimethylsilylethoxymet-
hyl)
indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-5-(methylcarbam-
oyl) piperidine-1-carboxylate useful in the synthesis of Compound
113
##STR00081##
[0227] To a solution of
(3R,5S)-1-tert-butoxycarbonyl-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-(2-t-
rimethylsilylethoxymethyl)
indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-3-carbox-
ylic acid (0.2 g, 273.65 .mu.mol, 1 eq) in THF (5 mL) was added
T.sub.3P (1.74 g, 2.74 mmol, 1.63 mL, 50% purity, 10 eq) and DIEA
(530.51 mg, 4.10 mmol, 714.97 .mu.L, 15 eq), methanamine (2 M, 2.74
mL, 20 eq). The mixture was stirred at 20.degree. C. for 1 h. The
reaction mixture was diluted with water (100 mL) and extracted with
ethyl acetate (50 mL.times.2). The combined organic layers were
washed with brine (100 mL.times.2), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
The residue was purified by column chromatography (SiO.sub.2,
Petroleum ether/Ethyl acetate=5:1 to 1:1) to afford the title
compound (200 mg) as a white solid.
Example 17. Synthesis of
Tert-butyl(5S)-3,3-dimethyl-5-[[4-[6-thiazol-2-yl-1-(2-trimethylsilyletho-
xymethyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1--
carboxylate useful in the Synthesis of Compound 109
Step 1:
N-[(3S)-1-benzyl-5,5-dimethyl-3-piperidyl]-4-[6-bromo-1-(2-trimeth-
ylsilylethoxy
methyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00082##
[0229] A mixture of
2-[[6-bromo-3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]indol-1-yl]meth-
oxy ethyl-trimethyl-silane (270 mg, 532.74 .mu.mol, 1 eq),
(3S)-1-benzyl-5,5-dimethyl-piperidin-3-amine (135.74 mg, 532.74
.mu.mol, 1 eq, HCl) and DIEA (344.26 mg, 2.66 mmol, 463.96 .mu.L, 5
eq) in NMP (2.7 mL) was heated to 75.degree. C. and stirred for 6
h. The mixture was poured into water 20 mL, and extracted with
EtOAc (20 mL.times.3). The combined organic layer was washed with
brine (50 mL), dried over Na.sub.2SO.sub.4, filtered. The filtrate
was concentrated under reduced pressure to give a residue. The
residue was purified by MPLC (SiO.sub.2, PE/EtOAc=10/1 to 1:1) to
afford the title compound (230 mg, 91.5% purity) as yellow oil.
Step
2N-[(3S)-1-benzyl-5,5-dimethyl-3-piperidyl]-4-[6-thiazol-2-yl-1-(2-tr-
imethylsilylethoxymethyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00083##
[0231] To a mixture of
N-[(3S)-1-benzyl-5,5-dimethyl-3-piperidyl]-4-[6-bromo-1-(2-trimethylsilyl-
ethoxymethyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine (200
mg, 290.40 .mu.mol, 1 eq) in DMF (0.5 mL) was added
tributyl(thiazol-2-yl)stannane (325.98 mg, 871.21 .mu.mol, 3 eq),
CuI (5.53 mg, 29.04 .mu.mol, 0.1 eq) and Pd(PPh.sub.3).sub.4 (33.56
mg, 29.04 .mu.mol, 0.1 eq). The mixture was heated to 130.degree.
C. and stirred for 12 h under N.sub.2. Saturated KF solution (20
mL) was added and the reaction mixture was stirred for about 1 hr.
Then the reaction mixture was extracted by EtOAc (20 mL.times.3).
The organic layer was combined, washed by brine (100 mL), dried
over Na.sub.2SO.sub.4, filtered and evaporated to afford the
residue. The residue was purified by MPLC (SiO.sub.2, PE/EtOAc=10/1
to 1:1) to afford the title compound (125 mg, 85.2% purity) as
yellow solid.
Step 3:
tert-butyl(5S)-3,3-dimethyl-5-[[4-[6-thiazol-2-yl-1-(2-trimethylsi-
lylethoxymethyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperi-
dine-1-carboxylate
##STR00084##
[0233] To a mixture of
N-[(3S)-1-benzyl-5,5-dimethyl-3-piperidyl]-4-[6-thiazol-2-yl-1-(2-trimeth-
ylsilylethoxymethyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
(95 mg, 137.10 .mu.mol, 1 eq) and Boc.sub.2O (di-t-butyl
dicarbonate; 149.61 mg, 685.51 .mu.mol, 157.49 .mu.L, 5 eq) in THF
(1 mL) was added Pd/C (15 mg, 10% purity) under N.sub.2. The
suspension was degassed under vacuum and purged with H.sub.2
several times. The mixture was stirred for 34 h at 25.degree. C.
under H.sub.2 (15 psi). The mixture was filtered, and the filtrate
was concentrated to afford the title compound (200 mg, crude, 73.4%
purity) as brown oil.
Example 18. Synthesis of
(3R,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl]-5-(trifluorom-
ethyl)pyrimidin-2-yl]amino]piperidin-3-ol (Compound 105)
Step 1: tert-butyl
N-[(3S,5R)-1-benzyl-5-hydroxy-3-piperidyl]carbamate
##STR00085##
[0235] To a solution of tert-butyl
N-[(3S,5R)-1-benzyl-5-[tert-butyl(dimethyl)silyl]oxy-3-piperidyl]carbamat-
e (8.6 g, 20.44 mmol, 1 eq) in THF (50 mL) was added TBAF (1 M,
22.49 mL, 1.1 eq). The reaction mixture was stirred at 30.degree.
C. for 12 h. The reaction mixture was evaporated to afford a
residue. The residue was purified by column chromatography
(SiO.sub.2, PE/EtOAc=3/1) to afford the title compound (5.4 g,
17.45 mmol, 85.34% yield, 99% purity) as white solid.
Step 2: (3R,5S)-5-amino-1-benzyl-piperidin-3-ol
##STR00086##
[0237] Tert-butyl
N-[(3S,5R)-1-benzyl-5-hydroxy-3-piperidyl]carbamate (5.4 g, 17.62
mmol, 1 eq) dissolved in HCl/EtOAc (25 mL) was stirred at
30.degree. C. for 1 h under N.sub.2 atmosphere. The reaction
mixture was evaporated to afford the product to afford the title
compound (4.2 g, 16.61 mmol, 94.25% yield, 96% purity, HCl salt) as
pink solid. It will be used directly in next step.
Step 3:
(3R,5S)-1-benzyl-5-[[4-(6-bromo-1H-indol-3-yl)-5-(trifluoromethyl)-
pyrimidin-2-yl]amino]piperidin-3-ol
##STR00087##
[0239] A flask was fitted with
6-bromo-3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-1H-indole
(Example 19; 500 mg, 1.33 mmol, 1 eq),
(3R,5S)-5-amino-1-benzyl-piperidin-3-ol (322.32 mg, 1.33 mmol, 1
eq, HCl) and DIPEA (858.05 mg, 6.64 mmol, 1.16 mL, 5 eq) in NMP (1
mL). The reaction mixture was heated to 70.degree. C. for 12 h. The
combined mixture was poured into H.sub.2O (50 mL). EtOAc (40
mL.times.3) was used to extract the product. The organic layer was
washed by brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and
evaporated to afford the crude product. The crude product was
purified by MPLC (SiO.sub.2, PE/EtOAc=3/1-1/1) to afford the title
compound (1 g, 32.6% purity) as light yellow oil.
Step 4:
(3R,5S)-1-benzyl-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-y-
l]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-3-ol
##STR00088##
[0241]
(3R,5S)-1-benzyl-5-[[4-(6-bromo-1H-indol-3-yl)-5-(trifluoromethyl)p-
yrimidin-2-yl]amino]piperidin-3-ol (700 mg, 1.28 mmol, 1 eq),
(3,5-dimethylisoxazol-4-yl)boronic acid (361.11 mg, 2.56 mmol, 2
eq), Pd(PPh.sub.3).sub.4 (296.09 mg, 256.23 .mu.mol, 0.2 eq) and
Na.sub.2CO.sub.3 (271.58 mg, 2.56 mmol, 2 eq) in DMF (10 mL) and
H.sub.2O (2 mL) was de-gassed and then heated to 130.degree. C. for
12 h under N.sub.2. The combined reaction mixture was poured into
H.sub.2O (40 mL). EtOAc (40 mL.times.3) was used to extract the
product. The organic layer was washed by brine (30 mL), dried over
Na.sub.2SO.sub.4, filtered and evaporated to afford the crude
product. The crude product was purified by MPLC (SiO.sub.2,
PE/EtOAc=1/1) to afford the title compound (460 mg, 44.2% purity)
as yellow solid. It contains some TPPO.
Step 5: tert-butyl
(3S,5R)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl]-5-(trifluorom-
ethyl)pyrimidin-2-yl]amino]-5-hydroxypiperidine-carboxylate
##STR00089##
[0243] To a solution of
(3R,5S)-1-benzyl-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl]-5-(t-
rifluoromethyl)pyrimidin-2-yl]amino]piperidin-3-ol (360 mg, 639.90
.mu.mol, 1 eq) and Boc.sub.2O (279.31 mg, 1.28 mmol, 294.01 .mu.L,
2 eq) in THF (10 mL) was added Pd/C (120 mg, 10% purity) under
N.sub.2. The suspension was degassed under vacuum and purged with
H.sub.2 several times. The mixture was stirred under H.sub.2 (15
psi) at 30.degree. C. for 8.5 h. The reaction mixture was filtered
through celite and the cake was washed by THF (15 mL). The filtrate
was evaporated to afford the title compound (400 mg, crude) as
yellow oil. It contains some TPPO and was used directly in next
step without any further purification.
Step 6:
(3R,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl]-5-(tri-
fluoromethyl)pyrimidin-2-yl]amino]piperidin-3-ol
##STR00090##
[0245] A mixture of tert-butyl
(3S,5R)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl]-5-(trifluorom-
ethyl)pyrimidin-2-yl]amino]-5-hydroxy-piperidine-1-carboxylate (400
mg, 698.60 .mu.mol, 1 eq) in HCl/EtOAc (15 mL) was stirred at
30.degree. C. for 1 h. The reaction mixture was evaporated to
afford a residue. Then H.sub.2O (10 mL) was added to the mixture
and PE/EtOAc (10/1, 15 mL.times.3) was used to extract undesired
TPPO. The aqueous was evaporated to afford the crude product. The
combined crude product was purified by acidic prep-HPLC (HCl
condition). The separated solution was lyophilized after removing
MeCN to afford the title compound (53.3 mg, 99.7% purity, HCl salt)
as yellow solid.
Example 19. Synthesis of
N-[(3S)-1-benzyl-5,5-dimethyl-3-piperidyl]-4-(6-bromo-1H-indol-3-yl)-5-(t-
rifluoromethyl)pyrimidin-2-amine useful in the synthesis of
Compound 102)
Step 1:
6-bromo-3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-1H-indole
##STR00091##
[0247] To a solution of 2,4-dichloro-5-(trifluoromethyl)pyrimidine
(22.58 g, 104.06 mmol, 1.2 eq) in DCE (1,2-dichloroethane; 200 mL)
was added AlCl.sub.3 (15.03 g, 112.73 mmol, 6.16 mL, 1.3 eq). The
mixture was stirred at 80.degree. C. for 0.5 h. Then
6-bromo-1H-indole (17 g, 86.72 mmol, 1 eq) was added to the
solution at 80.degree. C. The mixture was stirred at 80.degree. C.
for 11.5 h. The reaction mixture was diluted with water (1500 mL)
and extracted with EtOAc (500 mL.times.3). The combined organic
layers were washed with brine (200 mL.times.3), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, PE/EtOAc=10/1 to 3/1), then the residue
was washed with MeOH (200 mL), filtered to afford the title
compound (9.9 g, 26.29 mmol, 30.32% yield, 100% purity) as white
solid.
Step 2:
N-[(3S)-1-benzyl-5,5-dimethyl-3-piperidyl]-4-(6-bromo-1H-indol-3-y-
l)-5-(trifluoromethyl)pyrimidin-2-amine
##STR00092##
[0249] A mixture of (3S)-1-benzyl-5,5-dimethyl-piperidin-3-amine
(676.65 mg, 2.66 mmol, 1 eq, HCl),
6-bromo-3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-1H-indole (1
g, 2.66 mmol, 1 eq) and DIEA (1.72 g, 13.28 mmol, 2.31 mL, 5 eq) in
NMP (3 mL) was heated to 75.degree. C. and stirred for 3 h under
N.sub.2. The mixture was poured into water 20 mL, and extracted
with EtOAc (20 mL.times.3). The combined organic layer was washed
with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered. The
filtrate was concentrated under reduced pressure to give a residue.
The residue was purified by MPLC (SiO.sub.2, PE/EtOAc=7/1, 1500 mL)
to afford the title compound as yellow solid.
Example 20. Synthesis of
(3R,5S)-1-benzyl-5-[[4-[6-thiazol-2-yl-1-(2-trimethylsilylethoxymethyl)
indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidin-3-ol
Useful in the Synthesis of Compound 107)
##STR00093##
[0251] A mixture of
(3R,5S)-1-benzyl-5-[[4-[6-bromo-1-(2-trimethylsilylethoxymethyl)
indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidin-3-ol
(400 mg, 591.16 .mu.mol, 1 eq), tributyl (thiazol-2-yl) stannane
(663.58 mg, 1.77 mmol, 3 eq), CuI (11.26 mg, 59.12 .mu.mol, 0.1
eq), Pd(PPh.sub.3).sub.4 (68.31 mg, 59.12 .mu.mol, 0.1 eq) and in
DMF (5 mL) was degassed and purged with N.sub.2 for 3 times, and
then the mixture was stirred at 130.degree. C. for 16 h under
N.sub.2 atmosphere. The reaction mixture was quenched by addition
saturated CsF solution (20 mL), extracted with EtOAc (20
mL.times.3). The combined organic layers were washed with brine 20
mL, dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. The residue was purified by
column chromatography (SiO.sub.2, PE/EtOAc=5/1 to 4/1) to afford
the title compound (328 mg) as a yellow solid.
Example 21. Synthesis of
4-[6-(3-methyl-1,2,4-oxadiazol-5-yl)-1H-indol-3-yl]-N-[(3S)-3-piperidyl]--
5 (trifluoromethyl)pyrimidin-2-amine (Compound 121)
Step 1: Methyl 3-[2-chloro-5-(trifluoromethyl)
pyrimidin-4-yl]-1H-indole-6-carboxylate
##STR00094##
[0253] To a solution of 2, 4-dichloro-5-(trifluoromethyl)
pyrimidine (24.77 g, 114.16 mmol, 2.00 eq) in DCE (200.00 mL) was
added AlCl.sub.3 (15.98 g, 119.87 mmol, 6.55 mL, 2.10 eq). The
mixture was stirred at 90.degree. C. for 30 min, and then methyl
1H-indole-6-carboxylate (10.00 g, 57.08 mmol, 1.00 eq) was added at
90.degree. C. The resulting mixture was stirred at 90.degree. C.
for 15.5 h. The reaction mixture was filtered. The filtrate was
diluted with water (200 mL) and extracted with DCM (80 mL.times.3).
The combined organic layers were washed with brine (200
mL.times.2), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. The residue was washed
with MeOH (50 mL) and filtered to afford the title compound (4.00
g, 10.57 mmol, 18.52% yield, 94% purity) as a yellow solid.
Step 2: Methyl 3-[2-[[(3S)-1-tert-butoxycarbonyl-3-piperidyl]
amino]-5-(trifluoromethyl)pyrimidin-4-yl]-1H-indole-6-carboxylate
##STR00095##
[0255] A mixture of methyl
3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-1H-indole-6-carboxylate
(2.50 g, 7.03 mmol, 1.00 eq), tert-butyl
(3S)-3-aminopiperidine-1-carboxylate (1.83 g, 9.14 mmol, 1.30 eq),
DIEA (4.54 g, 35.15 mmol, 6.14 mL, 5.00 eq) in NMP (10.00 mL) was
stirred at 140.degree. C. for 1 h. The reaction mixture was poured
into water 200 mL, and then extracted with EtOAc (70 mL.times.3).
The combined organic layers were washed with brine (300 mL), dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to give a residue. The residue was purified by column
chromatography (SiO.sub.2, PE/EtOAc=5:1 to 1:1) to afford the title
compound (2.20 g, 3.39 mmol, 48.19% yield, 80% purity) as a yellow
solid.
Step 3: 3-[2-[[(3S)-1-tert-butoxycarbonyl-3-piperidyl]
amino]-5-(trifluoromethyl)pyrimidin-4-yl]-1H-indole-6-carboxylic
acid
##STR00096##
[0257] A mixture of methyl
3-[2-[[(3S)-1-tert-butoxycarbonyl-3-piperidyl]
amino]-5-(trifluoromethyl)pyrimidin-4-yl]-1H-indole-6-carboxylate
(2.20 g, 4.23 mmol, 1.00 eq), LiOH (5 M, 1.69 mL, 2.00 eq) in MeOH
(20.00 mL) was stirred at 25.degree. C. for 3 h. The reaction
mixture was concentrated, and then diluted with water 100 mL and
extracted with EtOAc (50 mL.times.3). The organic layers was
removed, and the aqueous phase was adjusted pH to 1 by HCl (1M),
extracted with EtOAc (50 mL.times.3). The combined organic layers
were washed with brine (300 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to afford the
title compound (1.80 g, 3.20 mmol, 75.57% yield, 90% purity) as a
yellow solid.
Step 4: Tert-butyl
(3S)-3-[[4-[6-[[(Z)--N-hydroxy-C-methyl-carbonimidoyl]-carbamoyl]-1H-indo-
l-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-1-carboxylate
##STR00097##
[0259] A mixture of
3-[2-[[(3S)-1-tert-butoxycarbonyl-3-piperidyl]-amino]-5-(trifluoromethyl)-
pyrimidin-4-yl]-1H-indole-6-carboxylic acid (200 mg, 395.66
.mu.mol, 1 eq), N-hydroxyacetamidine (58.62 mg, 791.31 .mu.mol, 2
eq), HOBt (1-hydroxybenzotriazole; 106.92 mg, 791.31 .mu.mol, 2
eq), EDCI (151.70 mg, 791.31 .mu.mol, 2 eq) and DIPEA (255.68 mg,
1.98 mmol, 344.58 .mu.L, 5 eq) in acetonitrile (ACN; 9 mL). The
mixture was stirred at 30.degree. C. for 7 h. The reaction mixture
was concentrated under reduced pressure to afford the title
compound (240 mg, crude) as white solid. The crude product was used
directly into the next step without purification.
Step 5: Tert-butyl (3S)-3-[[4-[6-(3-methyl-1,
2,4-oxadiazol-5-yl)-1H-indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-ami-
no]-piperidine-1-carboxylate
##STR00098##
[0261] A solution of tert-butyl
(3S)-3-[[4-[6-[[(Z)--N-hydroxy-C-methyl-carbonimidoyl]-carbamoyl]-1H-indo-
l-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-1-carboxylate
(240 mg, 427.38 .mu.mol, 1 eq) in Diglyme (10 mL) was stirred at
100.degree. C. for 12 h. The reaction mixture was concentrated
under reduced pressure to remove digylme. The residue was diluted
with H.sub.2O (50 mL) and extracted with EtOAc (20 mL.times.3). The
combined organic layers were washed with brine (60 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, PE/EtOAc=5/1 to 1/1) to afford the title
compound (100 mg) as yellow solid.
[0262] (The reaction was combined with another reaction in 70 mg
scale for purification and work up.)
Step 6:
4-[6-(3-methyl-1,2,4-oxadiazol-5-yl)-1H-indol-3-yl]-N-[(3S)-3-pipe-
ridyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00099##
[0264] A solution of tert-butyl
(3S)-3-[[4-[6-(3-methyl-1,2,4-oxadiazol-5-yl)-1H-indol-3-yl]-5-(trifluoro-
methyl)pyrimidin-2-yl]-amino]-piperidine-1-carboxylate (80 mg,
147.18 .mu.mol, 1 eq) in HCl/EtOAc (4 M, 8.00 mL, 217.42 eq) was
stirred at 15.degree. C. for 30 min. The reaction mixture was
concentrated under reduced pressure to give a residue. The residue
was purified by prep-HPLC (FA condition) to afford the title
compound (64 mg, FA, 100% purity) as white solid.
[0265] (The reaction was combined with another reaction in 20 mg
scale for purification and work-up.)
Example 22. Synthesis of
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-indazol-3-yl]-N-[(3S)-3-piperidyl]-5--
(trifluoromethyl)pyrimidin-2-amine (Compound 100)
Step 1: Tert-butyl
(3S)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-(2-trimethylsilylethoxy
methyl)
indazol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidin-
e-1-carboxylate
##STR00100##
[0267] A mixture of tert-butyl
(3S)-3-[[4-[6-bromo-1-(2-trimethylsilylethoxymethyl)indazol-3-yl]-5-(trif-
luoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate (0.3 g,
446.68 .mu.mol, 1 eq), (3,5-dimethylisoxazol-4-yl)boronic acid
(75.54 mg, 536.02 .mu.mol, 1.2 eq), Pd(PPh.sub.3).sub.4 (51.62 mg,
44.67 .mu.mol, 0.1 eq) and Na.sub.2CO.sub.3 (94.69 mg, 893.36
.mu.mol, 2 eq) in DMF (20 mL), H.sub.2O (4 mL) was degassed and
purged with N.sub.2 for 3 times, and then the mixture was stirred
at 130.degree. C. for 2 h under N.sub.2 atmosphere. The reaction
mixture was diluted with water (50 mL) and extracted with EtOAc (20
ml.times.3). The combined organic layers were washed with brine (50
mL.times.2), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. The residue was purified
by column chromatography (SiO.sub.2, PE/EtOAc=10:1 to 5:1) to
afford the title compound (260 mg) as a yellow oil.
[0268] (The reaction was combined with another reaction in 50 mg
scale for purification.)
Step 2:
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-indazol-3-yl]-N-[(3S)-3-piperi-
dyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00101##
[0270] To a solution of tert-butyl
(3S)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-(2-trimethylsilylethoxymethyl-
)
indazol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-piperidine-1-car-
boxylate (210 mg, 305.31 .mu.mol, 1 eq) in dioxane (4 mL) was added
H.sub.2SO.sub.4 (299.45 mg, 3.05 mmol, 162.74 .mu.L, 10 eq). The
mixture was stirred at 40.degree. C. for 2 h. The reaction mixture
was adjusted pH to 8 with saturated aqueous Na.sub.2CO.sub.3 and
extracted with EtOAc (10 mL.times.3). The combined organic layers
were washed with brine (30 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by prep-HPLC (FA
condition) to afford the title compound (49.4 mg, FA, purity
98.479%) as a white solid.
Example 23. Synthesis of Tert-butyl
(3S)-3-[[4-[6-thiazol-2-yl-1-(2-trimethylsilylethoxymethyl)
indazol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carbox-
ylate useful in the Synthesis of Compound 103
Step 1:
Tert-butyl(3S)-3-[[4-[6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2--
yl)-1-(2-trimethylsilylethoxymethyl)indazol-3-yl]-5-(trifluoromethyl)pyrim-
idin-2-yl]amino]piperidine-1-carboxylate
##STR00102##
[0272] A mixture of tert-butyl
(3S)-3-[[4-[6-bromo-1-(2-trimethylsilylethoxymethyl)indazol-3-yl]-5-(trif-
luoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate (400 mg,
595.57 .mu.mol, 1 eq),
4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-
-dioxaborolane (302.48 mg, 1.19 mmol, 2 eq), Pd(dppf)Cl.sub.2
(43.58 mg, 59.56 .mu.mol, 0.1 eq) and AcOK (175.35 mg, 1.79 mmol, 3
eq) in DME (4 mL) was degassed and purged with N.sub.2 for 3 times,
and then the mixture was stirred at 80.degree. C. for 2 h under
N.sub.2 atmosphere. The reaction mixture was diluted with water 40
mL and extracted with EtOAc (20 mL.times.3). The combined organic
layers were washed with brine (80 mL.times.1), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, PE/EtOAc=20:1 to 10:1) to afford the
title compound (400 mg) as a yellow oil.
Step 2: Tert-butyl
(3S)-3-[[4-[6-thiazol-2-yl-1-(2-trimethylsilylethoxymethyl)
indazol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carbox-
ylate
##STR00103##
[0274] A mixture of tert-butyl
(3S)-3-[[4-[6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2-trimethy-
lsilylethoxymethyl)indazol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]p-
iperidine-1-carboxylate (350 mg, 487.00 .mu.mol, 1 eq),
2-bromothiazole (103.84 mg, 633.10 .mu.mol, 57.06 .mu.L, 1.3 eq),
Pd(dppf)Cl.sub.2 (35.63 mg, 48.70 .mu.mol, 0.1 eq) and
Na.sub.2CO.sub.3 (51.62 mg, 487.00 .mu.mol, 1 eq) in dioxane (5
mL), H.sub.2O (1 mL) as degassed and purged with N.sub.2 for 3
times, and then the mixture was stirred at 80.degree. C. for 2 h
under N.sub.2 atmosphere. The reaction mixture was diluted with
water 20 mL and extracted with EtOAc (10 mL.times.3). The combined
organic layers were washed with brine (30 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, Petroleum ether/Ethyl acetate=10:1 to
2:1) to give a residue. The residue was purified again by prep-TLC
(thin layer chromatography; SiO.sub.2, PE:EtOAc=3:1) to afford the
title compound (160 mg) as a yellow solid.
Example 24. Synthesis of
4-[6-(1-methyltetrazol-5-yl)-1Hindol-3-yl]-N-[(3S)-3-piperidyl]-5-(triflu-
oromethyl)pyrimidin-2-amine (Compound 122)
Step 1: 5-bromo-1-methyl-tetrazole
##STR00104##
[0276] A suspension of 1-methyltetrazole-5-thiol (0.5 g, 4.30 mmol,
1 eq), ZnBr.sub.2 (1.94 g, 8.61 mmol, 430.88 .mu.L, 2 eq) in AcOH
(10 mL) was heated to 40.degree. C. for 0.5 h, then peroxyacetic
acid (13.10 g, 25.83 mmol, 11.39 mL, 6 eq) was added to the above
carefully and the resulting mixture was stirred at 80.degree. C.
for 11.5 h. The reaction mixture was cooled to 15.degree. C. and
poured into ice-water (1000 mL) and adjusted pH to 9 with
NaHCO.sub.3 (50 g). The aqueous phase was extracted with EtOAc (200
mL.times.3). The combined organic phase was washed with Sat.
Na.sub.2SO.sub.3 (500 mL.times.2). The combined organic phase was
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuum to afford the title compound (0.45 g, crude) as white solid
and used directly.
Step 2: Tert-butyl
(3S)-3-[[4-[6-(1-methyltetrazol-5-yl)-1H-indol-3-yl]-5-(trifluoromethyl)p-
yrimidin-2-yl]-amino]-piperidine-1-carboxylate
##STR00105##
[0278] To a solution of (S)-tert-butyl
3-((4-(1-(phenylsulfonyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-
-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carbo-
xylate (Example 8; 0.1 g, 137.44 .mu.mol, 1 eq),
5-bromo-1-methyl-tetrazole (33.60 mg, 206.16 .mu.mol, 1.5 eq) in
dioxane (2 mL)/H.sub.2O (0.4 mL) was added Pd(dppf)Cl.sub.2 (10.06
mg, 13.74 .mu.mol, 0.1 eq) and Na.sub.2CO.sub.3 (43.70 mg, 412.31
.mu.mol, 3 eq). The mixture was stirred at 100.degree. C. under
N.sub.2 atmosphere for 12 h. The reaction mixture was cooled to
15.degree. C. and poured into water (20 mL). The aqueous phase was
extracted with EtOAc (20 mL.times.3). The combined organic phase
was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuum. The residue was purified by silica gel
chromatography (100-200 mesh silica gel, PE/EtOAc=5/1, 1/1) to
afford the title compound (40 mg, 58.87 .mu.mol, 42.84% yield, 80%
purity) as yellow solid.
Step 3:
4-[6-(1-methyltetrazol-5-yl)-1Hindol-3-yl]-N-[(3S)-3-piperidyl]-5--
(trifluoromethyl)pyrimidin-2-amine
##STR00106##
[0280] A solution of tert-butyl
(3S)-3-[[4-[6-(1-methyltetrazol-5-yl)-1H-indol-3-yl]-5-(trifluoromethyl)p-
yrimidin-2-yl]-amino]-piperidine-1-carboxylate (40 mg, 73.59
.mu.mol, 1 eq) in HCl/EtOAc (10 mL) was stirred at 15.degree. C.
for 1 h. The reaction mixture was concentrated in vacuum. The
residue was purified by twice prep-HPLC (HCl condition) to afford
the title compound (9 mg, HCl salt 99% purity) as a yellow solid.
The reaction was combined with another reaction in 30 mg scale for
purification.
Example 25. Synthesis of
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo-[2,3-b]-pyridin-3-yl]-N-[(3S,-
6S)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 124) and
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo-[2,3-b]-pyridin-3-yl-
]-N-[(3R,6R)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 125)
Step 1: 4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)
pyrrolo-[2,3-b]-pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-ol
##STR00107##
[0282] To a solution of
4-[1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-
-yl]-pyrrolo[2,3-b]-pyridin-6-yl]-3,5-dimethyl-isoxazole (Example
11; 1.9 g, 2.44 mmol, 1 eq) in DCM (50 mL) was added m-CPBA (1.16
g, 5.36 mmol, 2.2 eq). The mixture was stirred at 20.degree. C. for
12 h. The residue was poured into a mixture of Sat.NaHCO.sub.3 (20
mL) and Sat. Na.sub.2SO.sub.3 (20 mL), and the mixture was stirred
for 5 min. The aqueous phase was extracted with DCM (30
mL.times.2). The combined organic phase was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue
was washed with a mixture of PE:EtOAc=5:1 (20 mL), filtered and the
filter cake was collected to afford the title compound (1 g, crude)
as yellow solid and used directly.
Step 2:
4-[1-(benzenesulfonyl)-3-[2-chloro-5-(trifluoromethyl)pyrimidin-4--
yl]-pyrrolo-[2,3-b]-pyridin-6-yl]-3,5-dimethylisoxazle
##STR00108##
[0284] A solution of
4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)
pyrrolo-[2,3-b]-pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-ol (1
g, 1.94 mmol, 1 eq) in POCl.sub.3 (15 mL) was stirred at 90.degree.
C. for 0.5 h. The solution was cooled to 15.degree. C. and
concentrated in vacuum. The residue was adjusted pH to 9 with sat.
NaHCO.sub.3 and extracted with ethyl acetate (50 mL.times.3). The
combined organic phase was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuum to afford the title compound
(0.6 g, crude) as a yellow solid and used directly.
Step 3: Benzyl
(2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)pyrrolo[2-
,3-b]-pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-pipe-
ridine-1-carboxylate and Benzyl
(2R,5R)-5-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)pyrrolo[2-
,3-b]-pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-pipe-
ridine-1-carboxylate
##STR00109##
[0286] To a solution of
4-[1-(benzenesulfonyl)-3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-pyr-
rolo-[2,3-b]pyridin-6-yl]-3,5-dimethyl-isoxazole (0.26 g, 486.97
.mu.mol, 1 eq) and benzyl 5-amino-2-methyl-piperidine-1-carboxylate
(241.85 mg, 973.95 .mu.mol, 2 eq) in NMP (0.4 mL) was added DIEA
(314.69 mg, 2.43 mmol, 424.11 .mu.L, 5 eq). The mixture was stirred
at 70.degree. C. for 2 h. The solution was cooled to 15.degree. C.,
diluted with water (50 mL) and extracted with EtOAc (20
mL.times.3). The combined organic phase was washed with brine (30
mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuum. The residue was purified by silica gel
chromatography (100-200 mesh silica gel, PE/EtOAc=5/1,2/1) to give
a racemate product (0.25 g) as a yellow solid. The product was
separated by SFC (column: Chiralpak AS-H 250*30 mm i.d. 5u Mobile
phase: A for CO2 and B for MeOH (0.1% NH.sub.3H.sub.2O) Gradient: B
%=40% Flow rate: 70 g/min Wavelength: 220 nm. Column temperature:
40.degree. C. System back pressure: 100 bar Cycle time: 5 min) to
afford the title compound 1 (Peak 1, RT=1.74 min, 120 mg, 160.91
.mu.mol, 33.04% yield) as a yellow solid. Compound 2 (Peak 2, 100
mg, 134.09 .mu.mol, 27.54% yield. It was used for example 4) as a
yellow solid.
Step 4: Benzyl
(2S,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo-[2,3-b]-pyridin-3-
-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carbox-
ylate
##STR00110##
[0288] To a solution of benzyl
(2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)pyrrolo[2-
,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piper-
idine-1-carboxylate (0.1 g, 134.09 .mu.mol, 1 eq) in MeOH (3 mL)
was added NaOH (2 M, 670.45 .mu.L, 10 eq). The mixture was stirred
at 40.degree. C. for 1 h. The solution was cooled to 15.degree. C.
and poured into water (10 mL). The aqueous phase was extracted with
EtOAc (10 mL.times.2). The combined organic phase was washed with
brine (10 mL.times.2), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuum to afford the title compound
(70 mg, crude) as a yellow solid and used directly.
Step 5:
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo-[2,3-b]-pyridin-3-yl]--
N-[(3S,6S)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00111##
[0290] To a solution of benzyl
(2S,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo-[2,3-b]-pyridin-3-
-yl]-5-(trifluoromethyl)pyrimidin-2-yl]-amino]-2-methyl-piperidine-1-carbo-
xylate (70 mg, 115.59 .mu.mol, 1 eq) in DCM (5 mL) was added
BBr.sub.3 (115.83 mg, 462.34 .mu.mol, 44.55 .mu.L, 4 eq) at
0.degree. C. The mixture was stirred at 15.degree. C. for 2 h. The
reaction mixture was concentrated in vacuum. The residue was washed
with PE (10 mL), filtered and the filter cake was collected. The
residue was purified by prep-HPLC (HCl condition) to afford the
title compound (26.8 mg, 100% purity, HCl salt) as a yellow
solid.
[0291]
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo-[2,3-b]-pyridin-3-yl]-N-
-[(3R,6R)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
is produced following steps 4 and 5, but starting with benzyl
(2R,5R)-5-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)pyrrolo[2-
,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piper-
idine-1-carboxylate.
Example 26. Synthesis of Tert-butyl
(3S)-3-[[4-[1-(benzenesulfonyl)-6-(3-methyltriazol-4-yl)indol-3-yl]-5-(tr-
ifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate Useful
in the Synthesis of Compound 126)
##STR00112##
[0293] A mixture of tert-butyl
(3S)-3-[[4-[1-(benzenesulfonyl)-6-bromo-indol-3-yl]-5-(trifluoromethyl)py-
rimidin-2-yl]amino]piperidine-1-carboxylate (300 mg, 440.83
.mu.mol, 1 eq), tributyl-(3-methyltriazol-4-yl)stannane (1.31 g,
1.76 mmol, 4 eq) and Pd(dppf)Cl.sub.2 (32.26 mg, 44.08 .mu.mol, 0.1
eq) in dioxane (0.5 mL) was stirred for 20 h at 100.degree. C.
under N.sub.2. The mixture was cooled to 15.degree. C. and poured
into water (10 mL) and extracted with EtOAc (20 mL.times.3). The
combined organic layer was washed with brine (20 mL.times.3), dried
over Na.sub.2SO.sub.4, filtered. The filtrate was concentrated
under reduced pressure to give a residue. The residue was purified
by MPLC (SiO.sub.2, PE:EtOAc=4/1 to 1/1) to afford the title
compound (280 mg, 83.6% purity) as a yellow solid. The reaction was
combined with another reaction in 20 mg scale for work up.
Example 27. Synthesis of
4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl]-N-[(3S)-3-piperidyl]--
5-(trifluoromethyl)pyrimidin-2-amine (Compound 129)
Step 1:
3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-1H-indole-6-carboni-
trile
##STR00113##
[0295] To a solution of 2, 4-dichloro-5-(trifluoromethyl)pyrimidine
(30.53 g, 140.69 mmol, 2 eq) in DCE (250 mL) was added AlCl.sub.3
(19.70 g, 147.72 mmol, 8.07 mL, 2.1 eq). The mixture was stirred at
90.degree. C. for 0.5 h. Then 1H-indole-6-carbonitrile (10 g, 70.34
mmol, 1 eq) was added and the resulting solution was stirred at
90.degree. C. for 2 h. The residue was dissolved in MeOH (100 mL)
and poured into ice-water (1000 mL) and stirred for 5 min. The
solids were formed and filtered to collect the cake. The cake was
washed with MeOH (500 mL), filtered and concentrated in vacuum. The
residue was purified by silica gel chromatography (PE:EtOAc=1:1) to
afford the title compound (10 g, 26.34 mmol, 37.45% yield, 85%
purity) as yellow solid.
Step 2: 3-[2-chloro-5-(trifluoromethyl)
pyrimidin-4-yl]-1-(2-trimethylsilylethoxymethyl)indole-6-carbonitrile
##STR00114##
[0297] To a solution of 3-[2-chloro-5-(trifluoromethyl)
pyrimidin-4-yl]-1H-indole-6-carbonitrile (1 g, 3.10 mmol, 1 eq) in
THF (20 mL) was added NaH (185.93 mg, 4.65 mmol, 60% purity, 1.5
eq) at 0.degree. C. The mixture was stirred at 0.degree. C. for 0.5
h. Then SEMCl (775.03 mg, 4.65 mmol, 822.75 .mu.L, 1.5 eq) was
added and the mixture was stirred at 0.degree. C. for 1 h. The
residue was poured into ice-water (50 mL). The aqueous phase was
extracted with EtOAc (50 mL.times.3). The combined organic phase
was washed with brine (50 mL.times.2), dried with anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue
was purified by silica gel chromatography PE:EtOAc=20:1-10:1-8:1 to
afford the title compound (0.7 g, 1.53 mmol, 49.37% yield, 99%
purity) as white solid.
Step 3: Tert-butyl
(3S)-3-[[4-[6-cyano-1-(2-trimethylsilylethoxymethyl)indol-3-yl]-5-(triflu-
oromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
##STR00115##
[0299] A mixture of
3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-1-(2-trimethylsilylethoxym-
ethyl)indole-6-carbonitrile (0.5 g, 1.10 mmol, 1 eq), tert-butyl
(3S)-3-aminopiperidine-1-carboxylate (287.42 mg, 1.44 mmol, 1.3 eq)
and DIEA (428.01 mg, 3.31 mmol, 576.83 .mu.L, 3 eq) in NMP (5 mL)
was stirred at 140.degree. C. for 1 h. The reaction mixture was
diluted with H.sub.2O (80 mL) and extracted with EtOAc (30
mL.times.2). The combined organic layers were washed with brine
(100 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated. The residue was purified by column chromatography
(SiO.sub.2, PE:EtOAc=10:1 to 2:1) to afford the title compound
(0.63 g, 817.19 umol, 74.03% yield, 80% purity) as a yellow
oil.
Step 4: Tert-butyl
(3S)-3-[[4-[6-[(Z)--N'-hydroxycarbamimidoyl]-1-(2-trimethylsilylethoxymet-
hyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carbo-
xylate
##STR00116##
[0301] To a solution of tert-butyl
(3S)-3-[[4-[6-cyano-1-(2-trimethylsilylethoxymethyl)indol-3-yl]-5-(triflu-
oromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate (0.5 g,
810.70 .mu.mol, 1 eq) in EtOH (5 mL) was added hydroxylamine
hydrochloride (563.36 mg, 8.11 mmol, 10 eq), Na.sub.2CO.sub.3
(687.41 mg, 6.49 mmol, 8 eq) and the mixture was stirred at
30.degree. C. for 12 h. The reaction mixture was diluted with
H.sub.2O (50 mL) and extracted with EtOAc (30 mL.times.3). The
combined organic layers were washed with brine (50 mL.times.2),
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to afford the title compound (0.56 g, crude) as a
yellow solid.
Step 5: Tert-butyl
(3S)-3-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1-(2-trimethylsilylethoxyme-
thyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carb-
oxylate
##STR00117##
[0303] To a solution of tert-butyl
(3S)-3-[[4-[6-[(Z)--N'-hydroxycarbamimidoyl]-1-(2-trimethylsilylethoxymet-
hyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amin]piperidine-1-carbox-
ylate (0.5 g, 769.49 .mu.mol, 1 eq) in EtOH (6 mL) and EtOAc (2 mL)
was added EtONa (104.73 mg, 1.54 mmol, 2 eq). The mixture was
stirred at 90.degree. C. for 30 min. The reaction mixture was
concentrated to give a residue. The residue was purified by column
chromatography (SiO.sub.2, Petroleum ether/Ethyl acetate=5:1 to
2:1) to afford the title compound (300 mg) as a yellow oil. (The
reaction was combined with another reaction in 80 mg scale for
purification.)
Step 6:
4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl]-N-[(3S)-3-pipe-
ridyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00118##
[0305] A mixture of tert-butyl
(3S)-3-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1-(2-trimethylsilylethoxyme-
thyl)indol-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carb-
oxylate (0.25 g, 371.03 .mu.mol, 1 eq) and H.sub.2SO.sub.4 (363.91
mg, 3.71 mmol, 197.78 .mu.L, 10 eq) in dioxane (5 mL) was stirred
at 40.degree. C. for 1 h. The mixture was adjusted pH to 8 with
saturated aqueous Na.sub.2CO.sub.3 and stirred at 20.degree. C. for
20 min. Then the aqueous was extracted with EtOAc (20 mL.times.3).
The combined organic layers were washed with brine (30 mL.times.2),
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. The residue was purified by
prep-HPLC (FA condition) to afford title compound (50.7 mg, FA) as
a white solid.
Example 28. Synthesis of Benzyl
(3S,5R)-3-amino-5-methoxy-piperidine-1-carboxylate Useful in the
Synthesis of Compound 131
Step 1: benzyl
(3S,5R)-3-(tert-butoxycarbonylamino)-5-methoxy-piperidine-1-carboxylate
##STR00119##
[0307] A benzyl
(3S,5R)-3-(tert-butoxycarbonylamino)-5-hydroxy-piperidine-1-carboxylate
(500 mg, 1.43 mmol, 1 eq) in THF (5 mL) was added NaH (74.19 mg,
1.85 mmol, 60% purity, 1.3 eq) at 0.degree. C. The mixture was
stirred at 0.degree. C. for 0.5 h. Then MeI (222.79 mg, 1.57 mmol,
97.71 .mu.L, 1.1 eq) was added. The mixture was stirred for another
1 h at 15.degree. C. It was poured into water (15 mL) and extracted
with EtOAc (25 mL.times.3). The combined organic layer was washed
with brine (25 mL.times.3), dried over Na.sub.2SO.sub.4, filtered.
The filtrate was concentrated under reduced pressure to give a
residue. The residue was purified by MPLC (SiO.sub.2, PE:EtOAc=4/1
to 1/1) to afford the title compound (450 mg, 987.83 .mu.mol,
69.23% yield, 80% purity) as brown solid.
Step 2: benzyl
(3S,5R)-3-amino-5-methoxy-piperidine-1-carboxylate
##STR00120##
[0309] A solution of benzyl (3S,5R)-3-(tert-butoxy
carbonylamino)-5-methoxy-piperidine-1-carboxylate (450 mg, 1.23
mmol, 1 eq) in HCl/EtOAc (50 mL) was stirred at 15.degree. C. for 1
h. The reaction mixture was concentrated in vacuum, the residue was
dissolved into MeOH (15 mL) and adjusted pH to 9 by Amberlyst 21 to
afford the title compound (350 mg, crude) as brown solid. It will
be used next step without further purification.
Example 29. Synthesis of
(3R,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-y-
l]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-3-ol (Compound
133)
Step 1:
(3R,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)py-
rrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-1-benzy-
l-piperidin-3-ol
##STR00121##
[0311] A mixture of
4-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-
-yl]pyrrolo[2,3-b]pyridin-6-yl]-3,5-dimethyl-isoxazole (280 mg,
484.80 umol, 1 eq), (3R,5S)-5-amino-1-benzyl-piperidin-3-ol (130.01
mg, 630.24 umol, 1.3 eq) and DIEA (313.29 mg, 2.42 mmol, 422.22 uL,
5 eq) in THF (3 mL) was stirred at 15.degree. C. for 1 h. It was
poured into water (10 mL) and extracted with EtOAc (20 mL.times.3).
The combined organic layer was washed with brine (20 mL.times.3),
dried over Na.sub.2SO.sub.4, filtered. The filtrate was
concentrated under reduced pressure. The residue was purified by
MPLC (SiO.sub.2, PE/EtOAc=3/1 to 1/1) to afford the title compound
(170 mg, 94% purity) as brown solid.
[0312] (Note: The reaction was combined with another reaction in 20
mg scale for work up)
Step 2: tert-butyl
(3S,5R)-3-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)pyrrolo[2-
,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-5-hydroxy-pipe-
ridine-1-carboxylate
##STR00122##
[0314] To a solution of
(3R,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)pyrrolo[2-
,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-1-benzyl-piper-
idin-3-ol (150 mg, 213.15 umol, 1 eq) and Boc.sub.2O (69.78 mg,
319.72 umol, 73.45 uL, 1.5 eq) in THF (2 mL) was added Pd/C (20 mg,
10% purity) under N.sub.2. The suspension was degassed under vacuum
and purged with H.sub.2 several times. It was stirred under H.sub.2
(15 psi) at 15.degree. C. for 1 h. Then the mixture was filtered
though a pad of celite. The filtrate was concentrated under reduced
pressure to give a residue. It was purified by MPLC (SiO.sub.2,
PE/EtOAc=4/1 to 1/1) to afford the title compound (100 mg) as
yellow solid. (Note: The reaction was combined with another
reaction in 20 mg scale for work up).
Step 3: tert-butyl
(3S,5R)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-y-
l]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-5-hydroxy-piperidine-1-carboxy-
late
##STR00123##
[0316] A mixture of tert-butyl
(3S,5R)-3-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethylisoxazol-4-yl)pyrrolo[2-
,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-5-hydroxy-pipe-
ridine-1-carboxylate (75 mg, 105.08 umol, 1 eq) and NaOH (2 M,
262.71 uL, 5 eq) in MeOH (0.2 mL) was stirred at 50.degree. C. for
1 h and then cooled to the room temperature. It was poured into the
water (10 mL), and extracted with EtOAc (20 mL.times.3). The
combined organic layer was washed with brine (20 mL.times.3), dried
over Na.sub.2SO.sub.4, filtered. The filtrate was concentrated
under reduced pressure to afford the title compound (70 mg, crude)
as brown solid which was used in next step without further
purification
[0317] (Note: The reaction was combined with another reaction in 10
mg scale for work up)
Step 4:
(3R,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyri-
din-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-3-ol
##STR00124##
[0319] A solution of tert-butyl
(3S,5R)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-y-
l]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-5-hydroxy-piperidine-1-carboxy-
late (50 mg, 87.17 umol, 1 eq) in HCl/EtOAc (5 mL) was stirred at
15.degree. C. for 1 h. It was concentrated. The crude product was
purified by prep-HPLC (HCl condition) to afford the title compound
(10 mg, 95.1% purity, HCl, 100% ee) as yellow solid.
[0320] (Note: The reaction was combined with another reaction in 20
mg scale for work up)
Example 30. Synthesis of
4-[6-(3-methyl-1,2,4-oxadiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3-
S)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
136)
Step 1: 1H-pyrrolo[2,3-b]pyridine-6-carboxylic acid
##STR00125##
[0322] To a solution of ethyl
1H-pyrrolo[2,3-b]pyridine-6-carboxylate (4.3 g, 22.61 mmol, 1 eq)
in dioxane (40 mL) was added NaOH (2 M, 50 mL, 4.42 eq). The
mixture was stirred at 80.degree. C. for 1 h. The solution was
concentrated under reduced pressure to remove dioxane and water.
The residue was diluted with H.sub.2O (50 mL) and adjusted pH to 2
with HCl (1M), with a lot of solid production, filtered to afford
the title compound (3.5 g, crude) as a yellow solid.
Step 2:
N-[(E)-N-hydroxy-C-methyl-carbonimidoyl]-1H-pyrrolo[2,3-b]pyridine-
-6-carboxamide
##STR00126##
[0324] To a solution of 1H-pyrrolo[2,3-b]pyridine-6-carboxylic acid
(1 g, 6.17 mmol, 1 eq) in MeCN (10 mL) was added HOBt (1.67 g,
12.33 mmol, 2 eq), EDCI (2.36 g, 12.33 mmol, 2 eq),
N-hydroxyacetamidine (913.77 mg, 12.33 mmol, 2 eq), and DIPEA (3.99
g, 30.84 mmol, 5.37 mL, 5 eq). The mixture was stirred at
30.degree. C. for 12 h. Then it was concentrated under reduced
pressure to afford the title compound (1.5 g, crude) as white solid
which was used directly without further purification.
Step 3:
3-methyl-5-(1H-pyrrolo[2,3-b]pyridin-6-yl)-1,2,4-oxadiazole
##STR00127##
[0326] A solution of
N-[(E)-N-hydroxy-C-methyl-carbonimidoyl]-1H-pyrrolo[2,3-b]pyridine-6-carb-
oxamide (1.5 g, 6.87 mmol, 1 eq) in Diglyme (20 mL) was stirred at
100.degree. C. for 12 h. The reaction mixture was poured into water
(200 mL) and the solid was formed. It was filtered to afford the
title compound (0.4 g, crude) as brown solid.
Step 4:
5-(3-bromo-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methyl-1,2,4-oxadiazol-
e
##STR00128##
[0328] To a solution of
3-methyl-5-(1H-pyrrolo[2,3-b]pyridin-6-yl)-1,2,4-oxadiazole (0.4 g,
2.00 mmol, 1 eq) in DMSO (dimethylsulfoxide; 4 mL) was added NBS
(391.18 mg, 2.20 mmol, 1.1 eq). The mixture was stirred at
15.degree. C. for 2 h. It was diluted with H.sub.2O (30 mL), with a
lot of solid formed. It was filtered to afford the title compound
(480 mg, crude) as brown solid.
Step 5:
5-[1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridin-6-yl]-3-methy-
l-1,2,4-oxadiazole
##STR00129##
[0330] To a solution of
5-(3-bromo-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methyl-1,2,4-oxadiazole
(675 mg, 2.42 mmol, 1 eq) in DMF (7 mL) was added NaH (125.76 mg,
3.14 mmol, 60% purity, 1.3 eq). The mixture was stirred at
0.degree. C. for 0.5 h. Then the benzenesulfonyl chloride (640.75
mg, 3.63 mmol, 464.31 uL, 1.5 eq) was added, the resulting mixture
was stirred at 15.degree. C. for 1.5 h. Then it was quenched by
addition water (40 mL), with a lot of solid formed, and then it was
filtered to afford the crude product. The crude product was
purified by column chromatography (SiO.sub.2, EtOAc) to afford the
title compound (480 mg, 801.43 umol, 33% yield, 70% purity) as an
orange solid.
Step 6: 5-[1-(benzenesulfonyl)-3-(4, 4,5,
5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrrolo[2,3-b]pyridin-6-yl]-3-methy-
l-1,2,4-oxadiazole
##STR00130##
[0332] A mixture of
5-[1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-1,2,4-
-oxadiazole (410 mg, 977.93 umol, 1 eq), BPD (372.50 mg, 1.47 mmol,
1.5 eq), KOAc (191.95 mg, 1.96 mmol, 2 eq), Pd(dppf)Cl.sub.2
(107.33 mg, 146.69 umol, 0.15 eq) in dioxane (4 mL) was degassed
and purged with N.sub.2 for 5 times, and then the mixture was
stirred at 100.degree. C. for 1 h under N.sub.2 atmosphere. The
solution was diluted with H.sub.2O (25 mL) and extracted with EtOAc
(15 mL.times.3). The combined organic layers were washed with brine
(45 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. The residue was purified
by column chromatography (SiO.sub.2, PE/EtOAc=10/1 to 5/1) to
afford the title compound (240 mg, 411.74 umol, 42% yield, 80%
purity) as a yellow solid.
Step 7: Tert-butyl
(3S)-3-[[4-[1-(benzenesulfonyl)-6-(3-methyl-1,2,4-oxadiazol-5-yl)pyrrolo[-
2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-c-
arboxylate
##STR00131##
[0334] A mixture of
5-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyr-
rolo[2,3-b]pyridin-6-yl]-3-methyl-1,2,4-oxadiazole (240 mg, 411.74
umol, 1 eq), tert-butyl
(3S)-3-[[4-chloro-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-ca-
rboxylate (172.46 mg, 452.91 umol, 1.1 eq), Na.sub.2CO.sub.3 (87.28
mg, 823.47 umol, 2 eq), Pd(dppf)Cl.sub.2 (45.19 mg, 61.76 umol,
0.15 eq) in dioxane (2.5 mL) and H.sub.2O (0.5 mL) was degassed and
purged with N.sub.2 for 3 times, and then the mixture was stirred
at 100.degree. C. for 1.5 h under N.sub.2 atmosphere. The solution
was concentrated under reduced pressure to remove dioxane. The
residue was diluted with H.sub.2O (30 mL) and extracted with EtOAc
(15 mL.times.2). The combined organic layers were washed with brine
(30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. The residue was purified
by column chromatography (SiO.sub.2, PE/EtOAc=3/1 to 1/1) to afford
the title compound (340 mg, 70% purity) as yellow solid.
Step 8: Tert-butyl
(3S)-3-[[4-[6-(3-methyl-1,2,4-oxadiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-3--
yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
##STR00132##
[0336] To a solution of tert-butyl
(3S)-3-[[4-[1-(benzenesulfonyl)-6-(3-methyl-1,2,4-oxadiazol-5-yl)pyrrolo[-
2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-c-
arboxylate (80 mg, 116.84 umol, 1 eq) in MeOH (2.5 mL) was added
NaOH (2 M, 292.10 uL, 5 eq). The mixture was stirred at 60.degree.
C. for 1 h. The solution was concentrated under reduced pressure to
remove MeOH. The residue was diluted with H.sub.2O (15 mL) and
adjusted pH to 7 by added HCl (1M), then extracted with EtOAc (15
mL.times.2). The combined organic layers were washed with brine (30
mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to afford the crude product (80 mg, crude) as
yellow solid.
Step 9:
4-[6-(3-methyl-1,2,4-oxadiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl-
]-N-[(3S)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00133##
[0338] To a solution of tert-butyl
(3S)-3-[[4-[6-(3-methyl-1,2,4-oxadiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-3--
yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
(60 mg, 110.19 umol, 1 eq) in EtOAc (0.5 mL) was added HCl/EtOAc (4
M, 1 mL, 36.30 eq). The mixture was stirred at 14.degree. C. for 1
h. The solution was concentrated under reduced pressure to give a
residue. The residue was purified by prep-HPLC (FA condition) to
afford the title compound (27.3 mg, FA, 99% purity) as white
solid.
Example 31. Synthesis of N-[(3S)-5,
5-dimethyl-3-piperidyl]-4-(6-thiazol-2-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)--
5-(trifluoromethyl)pyrimidin-2-amine (Compound 137)
Step 1:
2-[1-(benzenesulfonyl)pyrrolo[2,3-b]pyridin-6-yl]thiazole
##STR00134##
[0340] A mixture of
1-(benzenesulfonyl)-6-bromo-pyrrolo[2,3-b]pyridine (3.1 g, 9.19
mmol, 1 eq), tributyl(thiazol-2-yl)stannane (3.78 g, 10.11 mmol,
1.1 eq), CuI (175.09 mg, 919.36 umol, 0.1 eq) and
Pd(PPh.sub.3).sub.4 (1.06 g, 919.36 umol, 0.1 eq) in DMF (30 mL)
was degassed and purged with N.sub.2 for 3 times, and then the
mixture was stirred at 130.degree. C. for 12 h under N.sub.2
atmosphere. The mixture was poured into water (300 mL) and EtOAc
(200 mL), and filtered to remove the CuI. The organic layers were
separated, washed with brine (150 mL), dried over Na.sub.2SO.sub.4
and concentrated to give a residue. It was suspended in MeOH (20
mL), and stirred 10 min at room temperature and filtered. The
filter cake was collected and dried to dryness to afford the title
compound (2 g, 5.27 mmol, 57% yield, 90% purity) as a brown
solid.
Step 2:
2-[1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridin-6-yl]thiazole
##STR00135##
[0342] To a solution of
2-[1-(benzenesulfonyl)pyrrolo[2,3-b]pyridin-6-yl]thiazole (1.8 g,
5.27 mmol, 1 eq) in THF (18 mL) was added NBS (0.94 g, 5.27 mmol, 1
eq). The mixture was stirred at 25.degree. C. for 12 h under
N.sub.2 atmosphere. The mixture was poured into water (80 mL) and
extracted with EtOAc (30 mL.times.2). The organic layers were
separated, washed with brine (150 mL), dried over Na.sub.2SO.sub.4
and concentrated to give a residue. The residue was purified by
column chromatography (SiO.sub.2, Petroleum ether/Ethyl
acetate=10/1 to 3/1) to afford the title compound (1.5 g) as a
yellow solid.
Step 3:
2-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
-yl)pyrrolo[2,3-b]pyridin-6-yl]thiazole
##STR00136##
[0344] A mixture of
2-[1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridin-6-yl]thiazole
(0.95 g, 2.26 mmol, 1 eq),
4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-
-dioxaborolane (1.15 g, 4.52 mmol, 2 eq), Pd(dppf)Cl.sub.2 (165.39
mg, 226.03 umol, 0.1 eq) and KOAc (443.65 mg, 4.52 mmol, 2 eq) in
dioxane (10 mL) was degassed and purged with N.sub.2 for 3 times,
and then the mixture was stirred at 100.degree. C. for 2 h under
N.sub.2 atmosphere. The mixture was poured into water (50 mL) and
extracted with EtOAc (20 mL.times.2). The organic layers were
separated, washed with brine (150 mL), dried over Na.sub.2SO.sub.4
and concentrated to give a residue. It was purified by column
chromatography (SiO.sub.2, Petroleum ether/Ethyl acetate=10/1 to
4/1) to afford the title compound (0.63 g, 1.15 mmol, 50% yield,
85% purity) as a yellow solid.
Step 4:
2-[1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyri-
midin-4-yl]pyrrolo[2,3-b]pyridin-6-yl]thiazole
##STR00137##
[0346] A mixture of
2-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyr-
rolo[2,3-b]pyridin-6-yl]thiazole (0.05 M, 26.96 mL, 1 eq),
4-chloro-2-methylsulfanyl-5-(trifluoromethyl)pyrimidine (462.27 mg,
2.02 mmol, 1.5 eq), Na.sub.2CO.sub.3 (285.74 mg, 2.70 mmol, 2 eq)
and ditert-butyl(cyclopentyl)phosphane:dichloropalladium:iron
(87.85 mg, 134.80 umol, 0.1 eq) in THF (20 mL) and H.sub.2O (2 mL)
was degassed and purged with N.sub.2 for 3 times, and then the
mixture was stirred at 80.degree. C. for 1 h under N.sub.2
atmosphere. The reaction mixture was diluted with addition H.sub.2O
(100 mL), and extracted with EtOAc (50 mL.times.3), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, Petroleum ether/Ethyl acetate=10/1 to
3/1) to afford the title compound (300 mg) as a yellow solid.
[0347] (Note: The reaction was combined with another reaction in 50
mg scale for work up)
Step 5:
2-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyri-
midin-4-yl]pyrrolo[2,3-b]pyridin-6-yl]thiazole
##STR00138##
[0349] A mixture of
2-[1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-
-yl]pyrrolo[2,3-b]pyridin-6-yl]thiazole (0.28 g, 524.77 umol, 1 eq)
and m-CPBA (266.35 mg, 1.31 mmol, 85% purity, 2.5 eq) in DCM (3 mL)
was stirred at 15.degree. C. for 3 h. The solution was washed with
sat.Na.sub.2SO.sub.3 (30 mL), Sat.NaHCO.sub.3 (30 mL) and brine (50
mL). The organic layers were dried over Na.sub.2SO.sub.4 and
concentrated. The residue was washed with PE/EtOAc=3/1 (4 mL), and
filtered to afford the title compound (180 mg) as a yellow
solid.
[0350] (Note: The reaction was combined with another reaction in 20
mg scale for work up).
Step 6: Tert-butyl
(5S)-5-[[4-[1-(benzenesulfonyl)-6-thiazol-2-yl-pyrrolo[2,3-b]pyridin-3-yl-
]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-3,3-dimethyl-piperidine-1-carbo-
xylate
##STR00139##
[0352] A mixture of
2-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-
-yl]pyrrolo[2,3-b]pyridin-6-yl]thiazole (0.16 g, 282.90 umol, 1
eq), tert-butyl (5S)-5-amino-3,3-dimethyl-piperidine-1-carboxylate
(96.89 mg, 424.35 umol, 1.5 eq) and DIEA (109.69 mg, 848.71 umol,
147.83 uL, 3 eq) in THF (2 mL) was stirred at 15.degree. C. for 1
h. The reaction mixture was concentrated to afford the title
compound (300 mg) as a yellow solid.
[0353] (Note: The reaction was combined with another reaction in 10
mg scale for work up)
Step 7: Tert-butyl
(5S)-3,3-dimethyl-5-[[4-(6-thiazol-2-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-5--
(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
##STR00140##
[0355] A mixture of tert-butyl
(5S)-5-[[4-[1-(benzenesulfonyl)-6-thiazol-2-yl-pyrrolo[2,3-b]pyridin-3-yl-
]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-3,3-dimethyl-piperidine-1-carbo-
xylate (0.25 g, 350.24 umol, 1 eq) and NaOH (2 M, 1.75 mL, 10 eq)
in MeOH (3 mL) was stirred at 50.degree. C. for 0.5 h. It was
diluted with H.sub.2O (30 mL) and extracted with EtOAc (15
mL.times.3). The combined organic layers were washed with brine (30
mL.times.2), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. It was purified by column
chromatography (SiO.sub.2, Petroleum ether/Ethyl acetate=10/1 to
3/1) to afford the title compound (160 mg) as a yellow solid.
(Note: The reaction was combined with another reaction in 50 mg
scale for work up.)
Step 8:
N-[(3S)-5,5-dimethyl-3-piperidyl]-4-(6-thiazol-2-yl-1H-pyrrolo[2,3-
-b]pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
##STR00141##
[0357] A mixture of tert-butyl
(5S)-3,3-dimethyl-5-[[4-(6-thiazol-2-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-5--
(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate (150
mg, 261.49 umol, 1 eq) and HCl/EtOAc (4 M, 2 mL) was stirred at
15.degree. C. for 0.5 h. The reaction mixture was concentrated to
give a residue. The residue was purified by prep-HPLC (HCl
condition) to afford the title compound (21.7 mg, HCl) as a yellow
solid.
[0358] (Note: The reaction was combined with another reaction in 10
mg scale for purification.)
Example 32. Synthesis of
4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3-
S)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
139)
Step 1: N'-hydroxy-1H-pyrrolo[2,3-b]pyridine-6-carboxamidine
##STR00142##
[0360] To a solution of 1H-pyrrolo[2,3-b]pyridine-6-carbonitrile (1
g, 6.99 mmol, 1 eq) in EtOH (30 mL) was added NH.sub.2OH.HCl (4.85
g, 69.86 mmol, 10 eq) and Na.sub.2CO.sub.3 (8.89 g, 83.83 mmol, 12
eq). The mixture was stirred at 30.degree. C. for 12 h. The
suspension was filtered and concentrated under reduced pressure to
afford the title compound (1.3 g, crude) as yellow solid, which was
used into the next step without further purification.
Step 2:
5-methyl-3-(1H-pyrrolo[2,3-b]pyridin-6-yl)-1,2,4-oxadiazole
##STR00143##
[0362] To a solution of
N'-hydroxy-1H-pyrrolo[2,3-b]pyridine-6-carboxamidine (1.2 g, 6.81
mmol, 1 eq) in AcOH (12 mL) was added Ac.sub.2O (1.39 g, 13.62
mmol, 1.28 mL, 2 eq). The mixture was stirred at 90.degree. C. for
1 h. The solution was poured into H.sub.2O (100 mL) and the yellow
solid was formed. It was filtered and dried to afford the title
compound (0.85 g, crude) as yellow solid, which was used into the
next step without further purification.
Step 3:
3-(3-bromo-1H-pyrrolo[2,3-b]pyridin-6-yl)-5-methyl-1,2,4-oxadiazol-
e
##STR00144##
[0364] To a solution of
5-methyl-3-(1H-pyrrolo[2,3-b]pyridin-6-yl)-1,2,4-oxadiazole (0.8 g,
4.00 mmol, 1 eq) in THF (80 mL) was added NBS (711.24 mg, 4.00
mmol, 1 eq). The mixture was stirred at 15.degree. C. for 2 h. The
solution was poured into H.sub.2O (100 mL), while the yellow solid
was formed. It was filtered to give a residue. It was purified by
column chromatography (SiO.sub.2, PE/EtOAc=1/1 to 0/1) to afford
the title compound (0.4 g, 1.15 mmol, 28% yield, 80% purity) as
yellow solid. (Note: The reaction was combined with another
reaction in 150 mg scale for work up)
Step 4:
3-[1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridin-6-yl]-5-methy-
l-1,2,4-oxadiazole
##STR00145##
[0366] To a solution of
3-(3-bromo-1H-pyrrolo[2,3-b]pyridin-6-yl)-5-methyl-1,2,4-oxadiazole
(0.3 g, 1.07 mmol, 1 eq) in DMF (3 mL) was added NaH (64.49 mg,
1.61 mmol, 60% purity, 1.5 eq) at 0.degree. C. The reaction mixture
was stirred at 0.degree. C. for 0.5 h. Then the benzenesulfonyl
chloride (227.82 mg, 1.29 mmol, 165.09 uL, 1.2 eq) was added to the
above mixture, and stirred at 15.degree. C. for another 1.5 h.
After the material was converted completely, the solution was
poured into H.sub.2O (20 mL), while the yellow solid formed. It was
filtered to give the crude product. It was purified by column
chromatography (SiO.sub.2, PE/EtOAc=10/1 to 1/1) to afford the
title compound (0.2 g, 429.34 umol, 39% yield, 90% purity) as
yellow solid.
Step 5: 3-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1, 3
2-dioxaborolan-2-yl)pyrrolo[2,3-b]pyridin-6-yl]-5-methyl-1,2,4-oxadiazole
##STR00146##
[0368] A mixture of
3-[1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridin-6-yl]-5-methyl-1,2,4-
-oxadiazole (0.2 g, 477.04 umol, 1 eq), BPD (181.71 mg, 715.56
umol, 1.5 eq), Pd(dppf)Cl.sub.2 (34.91 mg, 47.70 umol, 0.1 eq),
KOAc (93.64 mg, 954.08 umol, 2 eq) in dioxane (2 mL) was degassed
and purged with N.sub.2 for 3 times, and then the mixture was
stirred at 100.degree. C. for 1 h under N.sub.2 atmosphere. The
solution was poured into H.sub.2O (20 mL) and extracted with EtOAc
(20 mL.times.2). The combined organic layers were washed with brine
(40 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by column chromatography (SiO.sub.2, PE/EtOAc=10/1 to
2/1) to afford the title compound (140 mg) as brown solid.
Step 6: Tert-butyl
(3S)-3-[[4-[1-(benzenesulfonyl)-6-(5-methyl-1,2,4-oxadiazol-3-yl)pyrrolo[-
2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]aminopiperidine-1-ca-
rboxylate
##STR00147##
[0370] A mixture of
3-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyr-
rolo[2,3-b]pyridin-6-yl]-5-methyl-1,2,4-oxadiazole (0.12 g, 257.34
umol, 1 eq), tert-butyl
(3S)-3-[[4-chloro-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-ca-
rboxylate (97.99 mg, 257.34 umol, 1 eq), Pd(dppf)Cl.sub.2 (18.83
mg, 25.73 umol, 0.1 eq), Na.sub.2CO.sub.3 (54.55 mg, 514.67 umol, 2
eq) in dioxane (1.5 mL) and H.sub.2O (0.3 mL) was degassed and
purged with N.sub.2 for 3 times, and then the mixture was stirred
at 100.degree. C. for 2 h under N.sub.2 atmosphere. The solution
was poured into H.sub.2O (20 mL), and extracted with EtOAc (20
mL.times.3). The combined organic layers were washed by brine (60
mL.times.2), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give the residue. The residue was
purified by column chromatography (SiO.sub.2, PE/EtOAc=3/1 to 1/1)
to afford the title compound (60 mg) as yellow solid.
[0371] (Note: The reaction mixture was combined with another
reaction in 20 mg scale for work-up and purification).
Step 7: Tert-butyl
(3S)-3-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-3--
yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
##STR00148##
[0373] To a solution of tert-butyl
(3S)-3-[[4-[1-(benzenesulfonyl)-6-(5-methyl-1,2,4-oxadiazol-3-yl)pyrrolo[-
2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-c-
arboxylate (0.04 g, 58.42 umol, 1 eq) in MeOH (0.5 mL) was added
NaOH (2 M, 0.5 mL, 17.12 eq). The mixture was stirred at 60.degree.
C. for 1 h. The solution was concentrated under reduced pressure to
afford the title compound (40 mg, crude) as yellow solid, which was
used into the next step without further purification.
[0374] (Note: The reaction was combined with another reaction in 20
mg scale for work up)
Step 8:
4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl-
]-N-[(3S)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00149##
[0376] A solution of tert-butyl
(3S)-3-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-3--
yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
(30 mg, 55.09 umol, 1 eq) in HCl/EtOAc (4 M, 1.50 mL, 108.91 eq)
was stirred at 15.degree. C. for 1 h. The solution was concentrated
under reduced pressure to give a residue. The residue was purified
by prep-HPLC to afford the title compound (8.2 mg, FA salt, 98.14%
purity) as a white solid
[0377] (Note: The reaction was combined with another reaction in 10
mg scale for work up)
Example 33. Synthesis of
4-[6-(3,5-dimethyl-1,2,4-triazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[-
(3S)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
140)
Step 1:
6-(3,5-dimethyl-1,2,4-triazol-4-yl)-1H-pyrrolo[2,3-b]pyridine
##STR00150##
[0379] A mixture of 1H-pyrrolo[2,3-b]pyridin-6-amine (2 g, 15.02
mmol, 1 eq), 1,1-dimethoxy-N,N-dimethyl-ethanamine (18.22 g, 136.80
mmol, 20.00 mL, 9.11 eq) was stirred at 130.degree. C. for 1 h. The
mixture was cooled to 0.degree. C., conc. HCl (4 mL) was added
dropwise, followed by acetohydrazide (6.68 g, 90.12 mmol, 6 eq) and
the resulting mixture was stirred at 0.degree. C. for 30 min. Then,
the temperature was raised to 130.degree. C. and the solution was
stirred for another 2 h. The resulting solution was concentrated.
The residue was purified by column chromatography (SiO.sub.2,
DCM/MeOH=70/1 to 7/1) to afford the title compound (10.5 g, crude,
HCl) as a yellow oil.
[0380] (Note: The reaction was combined with another reaction in
200 mg scale for work up)
Step 2:
3-bromo-6-(3,5-dimethyl-1,2,4-triazol-4-yl)-1H-pyrrolo[2,3-b]pyrid-
ine
##STR00151##
[0382] To a solution of
6-(3,5-dimethyl-1,2,4-triazol-4-yl)-1H-pyrrolo[23-b]pyridine (10 g,
46.90 mmol, 1 eq) in DCM (200 mL) was added NBS (4.17 g, 23.45
mmol, 0.5 eq) at 0.degree. C. The mixture was stirred at 15.degree.
C. for 1 h. The reaction mixture was diluted with H.sub.2O (800 mL)
and extracted with EtOAc (300 mL.times.2). The combined organic
layers were washed with brine (1000 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, DCM/MeOH=50/1 to 10/1) to afford the
title compound (5.5 g, 13.74 mmol, 73% purity) as a yellow
solid.
[0383] (Note: The reaction was combined with another reaction in
500 mg scale for work up).
Step 3:
1-(benzenesulfonyl)-3-bromo-6-(3,5-dimethyl-1,2,4-triazol-4-yl)pyr-
rolo[2,3-b]pyridine
##STR00152##
[0385] To a solution of
3-bromo-6-(3,5-dimethyl-1,2,4-triazol-4-yl)-1H-pyrrolo[2,3-b]pyridine
(5 g, 17.12 mmol, 1 eq) in DMF (90 mL) and THF (10 mL) was added
NaH (821.54 mg, 20.54 mmol, 60% purity, 1.2 eq) at 0.degree. C.
batch wise. After addition, the mixture was stirred at this
temperature for 0.5 h, and then benzenesulfonyl chloride (3.93 g,
22.25 mmol, 2.85 mL, 1.3 eq) was added dropwise at 0.degree. C. The
resulting mixture was stirred at 0.degree. C. for 1 h. The
resulting mixture was diluted with H.sub.2O (500 mL) and extracted
with EtOAc (100 mL.times.3). The combined organic layers were
washed with brine (300 mL.times.2), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
It was purified by column chromatography (SiO.sub.2, DCM/MeOH=10/1)
to afford the title compound (3.6 g) as a black brown solid.
[0386] (Note: The reaction was combined with another reaction in
500 mg scale for work up)
Step 4: [1-(benzenesulfonyl)-6-(3, 5-dimethyl-1,2, 4-triazol-4-yl)
pyrrolo[2,3-b]pyridin-3-yl]boronic acid
##STR00153##
[0388] A mixture of
1-(benzenesulfonyl)-3-bromo-6-(3,5-dimethyl-1,2,4-triazol-4-yl)
pyrrolo[2,3-b]pyridine (0.8 g, 1.85 mmol, 1 eq),
4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-
-dioxaborolane (939.87 mg, 3.70 mmol, 2 eq), Pd(dppf)Cl.sub.2
(135.41 mg, 185.06 umol, 0.1 eq) and KOAc (363.23 mg, 3.70 mmol, 2
eq) in dioxane (3 mL) was degassed and purged with N.sub.2 for 3
times, and then the mixture was stirred at 80.degree. C. for 2 h
under N.sub.2 atmosphere. The solution was diluted with H.sub.2O
100 (mL) and extracted with EtOAc (30 mL.times.3). The combined
organic layers were washed with brine (100 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue to afford the title compound (3.2 g, crude) as a
brown oil.
Step 5: Tert-butyl
(3S)-3-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethyl-1,2,4-triazol-4-yl)pyrrol-
o[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-
-carboxylate
##STR00154##
[0390] A mixture of
[1-(benzenesulfonyl)-6-(3,5-dimethyl-1,2,4-triazol-4-yl)pyrrolo[2,3-b]pyr-
idin-3-yl]boronic acid (2.7 g, 6.80 mmol, 1 eq), tert-butyl
(3S)-3-[[4-chloro-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-ca-
rboxylate (864.00 mg, 2.27 mmol, 3.34 e-1 eq), Na.sub.2CO.sub.3
(1.44 g, 13.59 mmol, 2 eq) and Pd(dppf)Cl.sub.2 (497.37 mg, 679.73
umol, 0.1 eq) in dioxane (5 mL) and H.sub.2O (1 mL) was degassed
and purged with N.sub.2 for 3 times, and then the mixture was
stirred at 80.degree. C. for 1 h under N.sub.2 atmosphere. It was
diluted with H.sub.2O (100 mL) and extracted with EtOAc (30
mL.times.3). The combined organic layers were washed with brine
(100 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. It was
purified by column chromatography (SiO.sub.2, DCM/MeOH=50/1 to 7/1)
to afford the title compound (800 mg, crude) as a black brown
oil.
[0391] (Note: The reaction was combined with another reaction in
500 mg scale for work up)
Step 6: Tert-butyl
(3S)-3-[[4-[6-(3,5-dimethyl-1,2,4-triazol-4-yl)-1H-pyrrolo[2,3-b]pyridin--
3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
##STR00155##
[0393] A mixture of tert-butyl
(3S)-3-[[4-[1-(benzenesulfonyl)-6-(3,5-dimethyl-1,2,4-triazol-4-yl)pyrrol-
o[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-
-carboxylate (0.7 g, 1.00 mmol, 1 eq) and NaOH (2 M, 5.02 mL, 10
eq) in MeOH (10 mL) was stirred at 50.degree. C. for 0.5 h. The
reaction mixture was diluted with H.sub.2O 30 mL and extracted with
EtOAc (15 mL.times.3). The combined organic layers were washed with
brine (30 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue that was
purified by column chromatography (SiO.sub.2, DCM: MeOH=50/1 to
10/1) to afford the title compound (180 mg) as a black brown
oil.
[0394] (Note: The reaction was combined with another reaction in
100 mg scale for purification).
Step 7: 4-[6-(3, 5-dimethyl-1,2,
4-triazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3S)-3-piperidyl]-5-(tr-
ifluoromethyl)pyrimidin-2-amine
##STR00156##
[0396] A mixture of tert-butyl (3S)-3-[[4-[6-(3,
5-dimethyl-1,2,4-triazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluor-
omethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate (0.16 g,
286.96 umol, 1 eq) and HCl/EtOAc (4 M, 3 mL, 41.82 eq) was stirred
at 15.degree. C. for 0.5 h. The mixture was concentrated to give a
residue. The residue was purified by prep-HPLC (neutral condition)
to afford the title compound (19.9 mg, free) as a white solid.
(Note: The reaction was combined with another reaction in 20 mg
scale for purification.)
Example 34. Synthesis of
4-[6-(5-methyltetrazol-1-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3S)-3-pip-
eridyl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound 141)
Step 1:
2-[(6-bromopyrrolo[2,3-b]pyridin-1-yl)methoxy]ethyl-trimethyl-sila-
ne
##STR00157##
[0398] To a solution of 6-bromo-1H-pyrrolo[2,3-b]pyridine (1.5 g,
7.61 mmol, 1 eq) in DMF (13.5 mL) and THF (1.5 mL) was added NaH
(365.42 mg, 9.14 mmol, 60% purity, 1.2 eq) at 0.degree. C., the
mixture was stirred at 0.degree. C. for 0.5 h. Then SEM-Cl (1.65 g,
9.90 mmol, 1.75 mL, 1.3 eq) was added, the mixture was stirred at
15.degree. C. for 11.5 h. The reaction mixture was diluted with
H.sub.2O (60 mL) and extracted with EtOAc (30 mL.times.3). The
combined organic layers were washed with brine (50 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by flash silica gel
chromatography (ISCO.RTM.; 20 g SepaFlash.RTM. Silica Flash Column,
Eluent of 0.about.8% Ethyl acetate/Petroleum ether gradient @ 75
mL/min) to afford the title compound (2.1 g, 6.42 mmol, 84.28%
yield) as yellow oil.
Step 2:
trimethyl-[2-[[6-(5-methyltetrazol-1-yl)pyrrolo[2,3-b]pyridin-1-yl-
]methoxy]ethyl]silane
##STR00158##
[0400] A mixture of
2-[(6-bromopyrrolo[2,3-b]pyridin-1-yl)methoxy]ethyl-trimethyl-silane
(2.1 g, 6.42 mmol, 1 eq), 5-methyl-1H-tetrazole (539.48 mg, 6.42
mmol, 1 eq), CuI (488.79 mg, 2.57 mmol, 0.4 eq), K.sub.2CO.sub.3
(2.13 g, 15.40 mmol, 2.4 eq) and
(1S,2S)--N1,N2-dimethylcyclohexane-1,2-diamine (365.06 mg, 2.57
mmol, 0.4 eq) in DMF (20 mL) was degassed and purged with N.sub.2
for 3 times, and then the mixture was stirred at 130.degree. C. for
12 h under N.sub.2 atmosphere. It was cooled to the room
temperature. The mixture was diluted with H.sub.2O (70 mL) and
extracted with EtOAc (50 mL.times.3). The combined organic layers
were washed with brine (80 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
It was purified by flash silica gel chromatography (ISCO.RTM.; 40 g
SepaFlash.RTM. Silica Flash Column, Eluent of 0.about.35% Ethyl
acetate/Petroleum ether gradient @ 100 mL/min) to afford the title
compound (1.2 g, 3.63 mmol, 56.60% yield) as a yellow solid.
Step 3:
2-[[3-bromo-6-(5-methyltetrazol-1-yl)pyrrolo[2,3-b]pyridin-1-yl]me-
thoxy]ethyl-trimethyl-silane
##STR00159##
[0402] To a solution of
trimethyl-[2-[[6-(5-methyltetrazol-1-yl)pyrrolo[2,3-b]pyridin-1-yl]methox-
y]ethyl]silane (1.2 g, 3.63 mmol, 1 eq) in DMF (4.5 mL) was added
NBS (710.95 mg, 3.99 mmol, 1.1 eq). The mixture was stirred at
15.degree. C. for 2 h. The solution was diluted with H.sub.2O (60
mL) and extracted with EtOAc (30 mL.times.3). The combined organic
layers were washed with brine (50 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
It was purified by flash silica gel chromatography (ISCO.RTM.; 20 g
SepaFlash.RTM. Silica Flash Column, Eluent of 0.about.8% Ethyl
acetate/Petroleum ether gradient @ 75 mL/min) to afford the title
compound (1 g, 2.44 mmol, 67% yield) as a yellow solid.
Step 4:
Trimethyl-[2-[[6-(5-methyltetrazol-1-yl)-3-(4,4,5,5-tetramethyl-1,-
3,2-dioxaborolan-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
##STR00160##
[0404] A mixture of
2-[[3-bromo-6-(5-methyltetrazol-1-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]e-
thyl-trimethyl-silane (850 mg, 2.08 mmol, 1 eq), BPD (790.93 mg,
3.11 mmol, 1.5 eq), Pd(dppf)Cl.sub.2 (151.93 mg, 207.64 umol, 0.1
eq), KOAc (407.56 mg, 4.15 mmol, 2 eq) in dioxane (10 mL) was
degassed and purged with N.sub.2 for 3 times, and then the mixture
was stirred at 100.degree. C. for 2 h under N.sub.2 atmosphere. It
was cooled to the room temperature. The reaction mixture was
diluted with H.sub.2O (60 mL) and extracted with EtOAc (30
mL.times.3). The combined organic layers were washed with brine (50
mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. It was purified by flash silica
gel chromatography (ISCO.RTM.; 20 g SepaFlash.RTM. Silica Flash
Column, Eluent of 0.about.8% Ethyl acetate/Petroleum ether gradient
@ 75 mL/min) to afford the title compound (600 mg, 47% purity) as
colorless oil.
[0405] (Note: The reaction was combined with another reaction in 90
mg scale for work up)
Step 5: Tert-butyl
(3S)-3-[[4-[6-(5-methyltetrazol-1-yl)-1-(2-trimethylsilylethoxymethyl)pyr-
rolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-
e-1-carboxylate
##STR00161##
[0407] A mixture of
trimethyl-[2-[[6-(5-methyltetrazol-1-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dio-
xaborolan-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane (600
mg, 1.31 mmol, 1 eq),
tert-butyl(3S)-3-[[4-chloro-5-(trifluoromethyl)pyrimidin-2-yl]amino]piper-
idine-1-carboxylate (300.35 mg, 788.75 umol, 0.6 eq),
Pd(dppf)Cl.sub.2 (96.19 mg, 131.46 umol, 0.1 eq), Na.sub.2CO.sub.3
(278.66 mg, 2.63 mmol, 2 eq) in dioxane (6 mL) and H.sub.2O (1.2
mL) was degassed and purged with N.sub.2 for 3 times, and then the
mixture was stirred at 100.degree. C. for 12 h under N.sub.2
atmosphere. It was cooled to the room temperature. The reaction
mixture was diluted with H.sub.2O (60 mL) and extracted with EtOAc
(30 mL.times.3). The combined organic layers were washed with brine
(50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. It was purified by flash
silica gel chromatography (ISCO.RTM.; 12 g SepaFlash.RTM. Silica
Flash Column, Eluent of 0.about.30% Ethyl acetate/Petroleum ether
gradient @ 75 mL/min) to afford the title compound (200 mg, 296.39
umol, 22% yield) as a yellow oil.
Step 6:
4-[6-(5-methyltetrazol-1-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3S-
)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00162##
[0409] To a solution of tert-butyl
(3S)-3-[[4-[6-(5-methyltetrazol-1-yl)-1-(2-trimethylsilylethoxy
methyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amin-
o]piperidine-1-carboxylate (200 mg, 296.39 umol, 1 eq) in dioxane
(2 mL) was added H.sub.2SO.sub.4 (296.63 mg, 2.96 mmol, 161.21 uL,
98% purity, 10 eq). The mixture was stirred at 40.degree. C. for 2
h. It was diluted with H.sub.2O (10 mL) and adjusted to pH=12 with
aq. NaOH (4 N), then extracted with EtOAc (10 mL.times.3). The
combined organic layers were washed with brine (10 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by twice prep-HPLC (FA
condition and neutral condition) to afford the title compound (4.3
mg, free) as a white solid. (Note: The reaction was combined with
another reaction in 20 mg scale for purification and work up.)
Example 35. Synthesis of
4-[6-(3-methyl-1,2,4-thiadiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(-
3S)-3-piperidyl-5-(trifluoromethyl)pyrimidin-2-amine (Compound
142)
Step 1: 1H-pyrrolo[2,3-b]pyridine-6-carbothioamide
##STR00163##
[0411] To a solution of 1H-pyrrolo[2,3-b]pyridine-6-carbonitrile (2
g, 13.97 mmol, 1 eq) in MeOH (20 mL) and THF (10 mL) was added
ammonium sulfide (1.71 g, 25.15 mmol, 1.72 mL, 1.8 eq) at
15.degree. C. Then the solution was stirred at 15.degree. C. for 12
h under N.sub.2 atmosphere. The solution was quenched by addition
of H.sub.2O (30 mL) at 15.degree. C., and extracted with EtOAc (30
mL.times.2). The combined organic layers were washed with brine (20
mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by column chromatography (SiO.sub.2, PE/EtOAc=30/1 to
1/1) to afford the title compound (1 g, 5.64 mmol, 40.38% yield) as
a yellow solid. (Note: The reaction was combined with another
reaction (ET19050-19) in 1 g scale for purification and work
up.)
Step 2:
(NE)-N-[1-(dimethylamino)ethylidene]-1H-pyrrolo[2,3-b]pyridine-6-c-
arbothioamide
##STR00164##
[0413] To a solution of 1H-pyrrolo[2,3-b]pyridine-6-carbothioamide
(1 g, 5.64 mmol, 1 eq) in DCM (10 mL) was added 1, 1-dimethoxy-N,
N-dimethyl-ethanamine (751.52 mg, 5.64 mmol, 824.94 uL, 1 eq) at
15.degree. C. The solution was stirred at 15.degree. C. for 12 h.
It was concentrated under reduced pressure to afford the title
compound (1.3 g, crude) as a yellow solid which was used into next
step without further purification. (Note: The reaction was combined
with another reaction in 1.2 g scale for work up.)
Step 3:
3-methyl-5-(1H-pyrrolo[2,3-b]pyridin-6-yl)-1,2,4-thiadiazole
##STR00165##
[0415] To a solution of
(NE)-N-[1-(dimethylamino)ethylidene]-1H-pyrrolo[2,3-b]pyridine-6-carbo
thioamide (1.3 g, 5.28 mmol, 1 eq) in EtOH (10 mL) was added a
solution of Py (1 mL) and amino hydrogen sulfate (2.03 g, 17.96
mmol, 3.40 eq) in MeOH (10 mL). The mixture was stirred at
15.degree. C. for 3 h under N.sub.2 atmosphere. The reaction
mixture was quenched by addition H.sub.2O (20 mL) at 15.degree. C.,
and extracted with EtOAc (30 mL.times.2). The combined organic
layers were washed with brine (20 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. It was purified by column chromatography
(SiO.sub.2, PE/EtOAc=30/1 to 1/1) to afford the title compound (1
g, 4.62 mmol, 87.62% yield) as a yellow solid. (Note: The reaction
was combined with another reaction in 450 mg scale for purification
and work up.)
Step 4:
5-(3-bromo-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methyl-1,2,4-thiadiazo-
le
##STR00166##
[0417] To a solution of
3-methyl-5-(1H-pyrrolo[2,3-b]pyridin-6-yl)-1,2,4-thiadiazole (1 g,
4.62 mmol, 1 eq) in DCM (10 mL) was added Br.sub.2 (738.96 mg, 4.62
mmol, 238.37 uL, 1 eq) at 0.degree. C. The mixture was degassed and
purged with N.sub.2 for 3 times and stirred at 0.degree. C. for 1 h
under N.sub.2 atmosphere. The reaction was concentrated under
reduced pressure to give a residue. It was triturated with MTBE
(methyl tert-butyl ether) at 15.degree. C. for 10 min to afford
title compound (1.2 g, 2.03 mmol, 43.96% yield, 50% purity) as a
yellow solid. (Note: The reaction was combined with another
reaction in 290 mg scale for purification and work up.)
Step 5:
2-[[3-bromo-6-(3-methyl-1,2,4-thiadiazol-5-yl)pyrrolo[2,3-b]pyridi-
n-1-yl]methoxy]ethyl-trimethyl-silane
##STR00167##
[0419] To a solution of
5-(3-bromo-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methyl-1,2,4-thiadiazole
(1.2 g, 4.07 mmol, 1 eq) in THF (10 mL) was added NaH (325.22 mg,
8.13 mmol, 60% purity, 2 eq) and
2-(chloromethoxy)ethyl-trimethyl-silane (1.36 g, 8.13 mmol, 1.44
mL, 2 eq) at 0.degree. C. Then the mixture was stirred at 0.degree.
C. for 1 h under N.sub.2 atmosphere. The reaction mixture was
quenched by addition of H.sub.2O (20 mL) at 15.degree. C., and
extracted with EtOAc (20 mL.times.2). The combined organic layers
were washed with (20 mL.times.2), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
The residue was purified by column chromatography (SiO.sub.2,
PE/EtOAc=30/1 to 1/1) to afford the title compound (1.1 g, 2.33
mmol, 57.24% yield, 90% purity) as a yellow solid.
Step 6:
Trimethyl-[2-[[6-(3-methyl-1,2,4-thiadiazol-5-yl)-3-(4,4,5,5-tetra-
methyl-1,3,2-dioxaborolan-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]si-
lane
##STR00168##
[0421] A mixture of
2-[[3-bromo-6-(3-methyl-1,2,4-thiadiazol-5-yl)pyrrolo[2,3-b]pyridin-1-yl]-
m ethoxy]ethyl-trimethyl-silane (500 mg.times.2, 1.18 mmol, 1 eq),
4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-
-dioxaborolane (447.69 mg, 1.76 mmol, 1.5 eq), Pd(dppf)Cl.sub.2
(86.00 mg, 117.53 umol, 0.1 eq), KOAc (230.70 mg, 2.35 mmol, 2 eq)
in dioxane (8 mL) was degassed and purged with N.sub.2 for 3 times,
and then the mixture was stirred at 100.degree. C. for 1 h under
N.sub.2 atmosphere. The reaction mixture was quenched by addition
of H.sub.2O (10 mL) at 15.degree. C., and extracted with EtOAc (20
mL.times.2). The combined organic layers were washed with brine (10
mL.times.2), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. It was purified by column
chromatography (SiO.sub.2, PE/EtOAc=30/1 to 1/1) to afford the
title compound (800 mg, 541.82 umol, 23.05% yield, 32% purity) as a
black brown solid
Step 7:
Tert-butyl(3S)-3-[[4-[6-(3-methyl-1,2,4-thiadiazol-5-yl)-1-(2-trim-
ethylsilylethoxymethyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyri-
midin-2-yl]amino]piperidine-1-carboxylate
##STR00169##
[0423] A mixture of
trimethyl-[2-[[6-(3-methyl-1,2,4-thiadiazol-5-yl)-3-(4,4,5,5-tetramethyl--
1,3,2-dioxaborolan-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
(800 mg, 1.69 mmol, 1 eq),
tert-butyl(3S)-3-[[4-chloro-5-(trifluoromethyl)pyrimidin-2-yl]amino]piper-
idine-1-carboxylate (644.75 mg, 1.69 mmol, 1 eq), Na.sub.2CO.sub.3
(358.92 mg, 3.39 mmol, 2 eq), Pd(dppf)Cl.sub.2 (123.89 mg, 169.32
umol, 0.1 eq) in dioxane (10 mL) and H.sub.2O (2 mL) was degassed
and purged with N.sub.2 for 3 times, and then the mixture was
stirred at 100.degree. C. for 3 h under N.sub.2 atmosphere. The
resulting mixture was quenched by addition H.sub.2O (10 mL) at
15.degree. C. and extracted with EtOAc (20 mL.times.2). The
combined organic layers were washed with brine (15 mL.times.2),
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. It was purified by column
chromatography (SiO.sub.2, PE/EtOAc=30/1 to 5/1) to afford the
title compound (150 mg, 136.79 umol, 8.08% yield, 63% purity) as a
brown solid.
Step 8:
4-[6-(3-methyl-1,2,4-thiadiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-y-
l]-N-[(3S)-3-piperidyl-5-(trifluoromethyl)pyrimidin-2-amine
##STR00170##
[0425] To a solution of tert-butyl
(3S)-3-[[4-[6-(3-methyl-1,2,4-thiadiazol-5-yl)-1-(2-trimethylsilylethoxym-
ethyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifleoromethyl)pyrimidin-2-yl]amino]-
piperidin e-1-carboxylate (150 mg, 217.12 umol, 1 eq) in dioxane (1
mL) was added H.sub.2SO.sub.4 (217.30 mg, 2.17 mmol, 118.10 uL, 98%
purity, 10 eq). The mixture was degassed and purged with N.sub.2
for 3 times and stirred at 40.degree. C. for 3 h under N.sub.2
atmosphere. The reaction mixture was quenched by addition of
H.sub.2O (10 mL) at 15.degree. C., and extracted with EtOAc (15
mL.times.2). The organic layer was concentrated to give crude
residue, which was dissolved in H.sub.2O (5 mL). NaOH was added to
adjust pH to 12. The mixture was stirred for 10 min at 15.degree.
C. The stirred solution was extracted with EtOAc (10 mL.times.2).
The combined organic layers were washed with brine (10 mL.times.2),
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. It was purified by prep-HPLC
(HCl condition) to afford the title compound (6.7 mg, HCl salt 95%
purity) as a yellow solid.
Example 36. Synthesis of (3R,
5S)--N-methyl-5-[[4-(6-thiazol-2-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(tri-
fluoromethyl)pyrimidin-2-yl]amino]piperidine-3-carboxamide
(Compound 145)
Step 1: O1-tert-butyl
O3-methyl-(3R,5S)-5-[[4-[1-(benzenesulfonyl)-6-thiazol-2-yl-pyrrolo[2,3-b-
]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1,3-dica-
rboxylate
##STR00171##
[0427] A mixture of
2-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-
-yl]pyrrolo[2,3-b]pyridin-6-yl]thiazole (0.28 g, 495.08 umol, 1
eq), O1-tert-butyl O3-methyl
(3R)-5-aminopiperidine-1,3-dicarboxylate (191.83 mg, 742.62 umol,
1.5 eq) and DIEA (191.95 mg, 1.49 mmol, 258.70 uL, 3 eq) in THF (5
mL) was stirred at 15.degree. C. for 30 min. The reaction mixture
was diluted with H.sub.2O (50 mL) and extracted with EtOAc (20
mL.times.3). The combined organic layers were washed with brine
(100 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. It was
purified by column chromatography (SiO.sub.2, Petroleum ether/Ethyl
acetate=10/1 to 1/1) to afford the title compound (260 mg) as a
yellow solid.
[0428] (Note: The reaction was combined with another reaction in 20
mg scale for work up)
Step 2: (3R,
5S)-1-tert-butoxycarbonyl-5-[[4-(6-thiazol-2-yl-1H-pyrrolo[2,3-b]pyridin--
3-yl)-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-3-carboxylic
acid
##STR00172##
[0430] To a solution of O1-tert-butyl O3-methyl
(3R,5S)-5-[[4-[1-(benzenesulfonyl)-6-thiazol-2-yl-pyrrolo[2,3-b]pyridin-3-
-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1,3-dicarboxylate
(240 mg, 322.68 umol, 1 eq) in MeOH (5 mL) was added NaOH (2 M,
1.61 mL, 10 eq). The mixture was stirred at 15.degree. C. for 30
min. The reaction mixture was adjusted pH to 4 with aqueous HCl
(0.5 M), and extracted with EtOAc (30 mL.times.3). The combined
organic layers were washed with brine (90 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to afford the title compound (170 mg) as a yellow solid.
[0431] (Note: The reaction was combined with another reaction in 20
mg scale for work up).
Step 3: Tert-butyl (3R,
5S)-3-(methylcarbamoyl)-5-[[4-(6-thiazol-2-yl-1H-pyrrolo[2,3-b]pyridin-3--
yl)-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
##STR00173##
[0433] To a solution of
(3R,5S)-1-tert-butoxycarbonyl-5-[[4-(6-thiazol-2-yl-1H-pyrrolo[2,3-b]pyri-
din-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-3-carboxylic
acid (140 mg, 237.45 umol, 1 eq) in DMF (2 mL) was added HATU
((Dimethylamino)-N,N-dimethyl(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy)me-
thaniminium hexafluorophosphate; 135.43 mg, 356.18 umol, 1.5 eq),
DIEA (92.07 mg, 712.36 umol, 124.08 uL, 3 eq), methanamine (2 M,
356.18 uL, 3 eq). The mixture was stirred at 15.degree. C. for 1 h.
The reaction mixture was poured into saturated aqueous NaHCO.sub.3
(20 mL), stirred for 5 min. Then, the solid was formed. It was
filtered to afford the title compound (160 mg) as a yellow
solid.
[0434] (Note: The reaction was combined with another reaction in 10
mg scale for work up)
Step 4: (3R,
5S)--N-methyl-5-[[4-(6-thiazol-2-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(tri-
fluoromethyl)pyrimidin-2-yl]amino]piperidine-3-carboxamide
##STR00174##
[0436] A mixture of tert-butyl
(3R,5S)-3-(methylcarbamoyl)-5-[[4-(6-thiazol-2-yl-1H-pyrrolo[2,3-b]pyridi-
n-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
(150 mg, 248.91 umol, 1 eq) and HCl/EtOAc (4 M, 3 mL) was stirred
at 15.degree. C. for 30 min. The mixture was concentrated to give a
residue. It was purified by prep-HPLC (neutral condition) to afford
the title compound (19.4 mg) as a white solid. (Note: The reaction
was combined with another reaction in 10 mg scale for
purification).
Example 37. Synthesis of (3R, 5S)-5-[[4-[6-(3,
5-dimethylisoxazol-4-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-5-(trifluorometh-
yl)pyrimidin-2-yl]amino]piperidin-3-ol (Compound 146)
Step 1: (3R, 5S)-1-benzyl-5-[[4-[6-(3,
5-dimethylisoxazol-4-yl)-1-tetrahydropyran-2-yl-pyrazolo[3,4-b]pyridin-3--
yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-3-ol
##STR00175##
[0438] A mixture of
3,5-dimethyl-4-[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl]-1--
tetrahydropyran-2-yl-pyrazolo[3,4-b]pyridin-6-yl]isoxazole (0.25 g,
478.47 umol, 1 eq), (3R,5S)-5-amino-1-benzyl-piperidin-3-ol (98.70
mg, 478.47 umol, 1 eq), DIEA (185.51 mg, 1.44 mmol, 250.02 uL, 3
eq) in THF (5 mL) was stirred at 15.degree. C. for 1 h. The mixture
was poured into water (20 mL), and then extracted with EtOAc (10
mL.times.3). The combined organic layers were washed with brine (50
mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. The residue was purified by
column chromatography (SiO.sub.2, Petroleum ether/Ethyl
acetate=10/1 to 0/1) to afford the title compound (190 mg) as a
yellow solid
Step 2: Tert-butyl (3S, 5R)-3-[[4-[6-(3,
5-dimethylisoxazol-4-yl)-1-tetrahydropyran-2-yl-pyrazolo[3,4-b]pyridin-3--
yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-5-hydroxy-piperidine-1-carbox-
ylate
##STR00176##
[0440] To a solution of (3R,5S)-1-benzyl-5-[[4-[6-(3,
5-dimethylisoxazol-4-yl)-1-tetrahydropyran-2-yl-pyrazolo[3,4-b]pyridin-3--
yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-3-ol (0.17 g,
262.07 umol, 1 eq) tert-butoxycarbonyl tert-butyl carbonate (171.59
mg, 786.22 umol, 180.62 uL, 3 eq) in MeOH (1 mL) was added Pd/C
(10%, 0.2 g) under N.sub.2. The suspension was degassed under
vacuum and purged with H.sub.2 several times. It was stirred at
15.degree. C. for 16 h under H.sub.2 atmosphere (15 psi). The
reaction mixture was filtered and the filter was concentrated. The
residue was purified by column chromatography (SiO.sub.2, Petroleum
ether/Ethyl acetate=10/1 to 0/1) to afford the title compound (90
mg) as a white solid. (Note: The reaction was combined with another
reaction in 20 mg scale for work up.)
Step 3: (3R,
5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl]--
5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-3-ol
##STR00177##
[0442] A mixture of tert-butyl
(3S,5R)-3-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-tetrahydropyran-2-yl-pyraz-
olo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-5-hydroxy-
-piperidine-1-carboxylate (80 mg, 121.46 umol, 1 eq) in HCl/MeOH (4
M, 910.93 uL, 30 eq) was stirred at 60.degree. C. for 16 h. It was
concentrated and the residue was purified by prep-HPLC to afford
the title compound (20 mg, 99.47% purity, FA) as a white solid.
(Note: The reaction was combined with another reaction in 10 mg
scale for work up.)
Example 38. Synthesis of
(3R,5S)-5-[[4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-5-
-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-3-ol (Compound
148)
Step 1: 3-methyl-4-(1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole
##STR00178##
[0444] A mixture of 6-bromo-1H-pyrrolo[2,3-b]pyridine (1 g, 5.08
mmol, 1 eq),
3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole
(1.11 g, 5.33 mmol, 1.05 eq), Pd(dppf)Cl.sub.2--CH.sub.2Cl.sub.2
(207.24 mg, 253.77 umol, 0.05 eq), NaHCO.sub.3 (1.28 g, 15.23 mmol,
592.19 uL, 3 eq) in DME (10 mL) and H.sub.2O (4 mL) was degassed
and purged with N.sub.2 for 3 times, and then the mixture was
stirred at 100.degree. C. for 12 h under N.sub.2 atmosphere. It was
cooled to the room temperature. The reaction mixture was diluted
with H.sub.2O (30 mL) and extracted with DCM (20 mL.times.3). The
combined organic layers were washed with brine (20 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by flash silica gel
chromatography (ISCO.RTM.; 12 g SepaFlash.RTM. Silica Flash Column,
Eluent of 0.about.15% Ethyl acetate/Petroleum ether gradient @ 50
mL/min) to afford the title compound (800 mg) as a yellow solid.
(Note: The reaction was combined with another reaction
(ET12872-179) in 50 mg scale for purification and work up.)
Step 2:
4-(3-bromo-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methyl-isoxazole
##STR00179##
[0446] To a solution of
3-methyl-4-(1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole (720 mg, 3.61
mmol, 1 eq) in DMF (7 mL) was added NBS (771.93 mg, 4.34 mmol, 1.2
eq). The mixture was stirred at 15.degree. C. for 2 h. The reaction
mixture was diluted with H.sub.2O (60 mL) and extracted with EtOAc
(30 mL.times.3). The combined organic layers were washed with brine
50 (mL), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. The residue was purified
by flash silica gel chromatography (ISCO.RTM.; 12 g SepaFlash.RTM.
Silica Flash Column, Eluent of 0.about.25% Ethyl acetate/Petroleum
ether gradient @ 75 mL/min) to afford the title compound (600 mg,
2.16 mmol, 59.69% yield) as a colorless oil.
Step 3:
4-[1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridin-6-yl]-3-methy-
l-isoxazole
##STR00180##
[0448] To a solution of
4-(3-bromo-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methyl-isoxazole (600
mg, 2.16 mmol, 1 eq) in DMF (5 mL) and THF (0.5 mL) was added NaH
(129.44 mg, 3.24 mmol, 60% purity, 1.5 eq). The mixture was stirred
at 15.degree. C. for 10 min. Then benzenesulfonyl chloride (495.37
mg, 2.80 mmol, 358.96 uL, 1.3 eq) was added, the mixture was
stirred at 15.degree. C. for 2.3 h. The reaction mixture was
diluted with H.sub.2O (60 mL) and EtOAc (30 mL). Then it was
filtered. The filtrate was partitioned and the water layer was
extracted with EtOAc (30 mL.times.2). The filter cake and the
combined organic layers were concentrated under reduced pressure to
give a residue. The residue was diluted with MeOH (30 mL) and
stirred for 20 min. The reaction mixture was filtered and the
filter cake was dried in vacuum to afford the title compound (800
mg, 1.91 mmol, 88.65% yield) was obtained as a brown solid
Step 4:
4-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
-yl)pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole
##STR00181##
[0450] A mixture of
4-[1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxa-
zole (670.00 mg, 1.60 mmol, 1 eq),
4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-
-dioxaborolane (610.16 mg, 2.40 mmol, 1.5 eq), Pd(dppf)Cl.sub.2
(117.21 mg, 160.19 umol, 0.1 eq), KOAc (314.41 mg, 3.20 mmol, 2 eq)
in dioxane (8 mL) was degassed and purged with N.sub.2 for 3 times,
and then the mixture was stirred at 100.degree. C. for 1 h under
N.sub.2 atmosphere. It was cooled to the room temperature. The
mixture was diluted with dioxane (30 mL) and filtered. The filtrate
was concentrated under reduced pressure to afford the title
compound (800 mg, crude) was obtained as a brown solid, and it was
used into the next step without further purification.
Step 5:
4-[1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyri-
midin-4-yl]pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole
##STR00182##
[0452] A mixture of
4-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyr-
rolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole (800 mg, 1.03 mmol, 1
eq), 4-chloro-2-methylsulfanyl-5-(trifluoromethyl)pyrimidine
(235.83 mg, 1.03 mmol, 1 eq), Pd(dppf)Cl.sub.2 (75.48 mg, 103.15
umol, 0.1 eq), Na.sub.2CO.sub.3 (218.66 mg, 2.06 mmol, 2 eq) in
dioxane (10 mL) and H.sub.2O (2 mL) was degassed and purged with
N.sub.2 for 3 times, and then the mixture was stirred at
100.degree. C. for 12 hr under N.sub.2 atmosphere. It was cooled to
the room temperature. The reaction mixture was diluted with
H.sub.2O (60 mL) and extracted with EtOAc (30 mL.times.3). The
combined organic layers were washed with brine (60 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. It was purified by flash silica gel
chromatography (ISCO.RTM.; 20 g SepaFlash.RTM. Silica Flash Column,
Eluent of 0.about.30% Ethyl acetate/Petroleum ether gradient @ 75
mL/min) to afford the title compound (200 mg, 150.51 umol, 14.59%
yield, 40% purity) was obtained as a yellow solid. (Note: The
reaction was combined with another reaction in 120 mg scale for
purification and work up.)
Step 6:
4-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyri-
midin-4-yl]pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole
##STR00183##
[0454] To a solution of
4-[1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-
-yl]pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole (180 mg, 338.65
umol, 1 eq) in EtOAc (10 mL) was added Oxone (1.04 g, 1.69 mmol, 5
eq). The mixture was stirred at 80.degree. C. for 12 h. It was
cooled to the room temperature. The reaction mixture was filtered
and the filtrate was quenched by saturation Na.sub.2SO.sub.3 (30
mL). The water layer was extracted with EtOAc (20 mL.times.3). The
combined organic layers were washed with brine (30 mL), dried over
Na.sub.2SO.sub.4, filtrate and concentrated under reduced pressure
to give a residue. The residue was purified by flash silica gel
chromatography (ISCO.RTM.; 20 g SepaFlash.RTM. Silica Flash Column,
Eluent of 0.about.50% Ethyl acetate/Petroleum ether gradient @ 75
mL/min) to afford the title compound (40 mg) as a white solid.
Step 7:
(3R,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3-methylisoxazol-4-yl)pyrrol-
o[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-1-benzyl-pi-
peridin-3-ol
##STR00184##
[0456] To a solution of
4-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-
-yl]pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole (35 mg, 62.11
umol, 1 eq) and DIPEA (40.14 mg, 310.54 umol, 54.09 uL, 5 eq) in
THF (0.5 mL) was added (3R,5S)-5-amino-1-benzyl-piperidin-3-ol
(12.81 mg, 62.11 umol, 1 eq). The mixture was stirred at 15.degree.
C. for 3 h. The reaction mixture was concentrated under reduced
pressure to remove solvent. The residue was purified by flash
silica gel chromatography (ISCO.RTM.; 4 g SepaFlash.RTM. Silica
Flash Column, Eluent of 0-2% Dichloromethane/Methanol @ 50 mL/min)
to afford the title compound (27 mg) was obtained as a white solid.
(Note: The reaction was combined with another reaction in 5 mg
scale for purification and work up.)
Step 8: Tert-butyl
(3S,5R)-3-[[4-[1-(benzenesulfonyl)-6-(3-methylisoxazol-4-yl)pyrrolo[2,3-b-
]pyridine-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-5-hydroxy-piperid-
ine-1-carboxylate
##STR00185##
[0458] A mixture of
(3R,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3-methylisoxazol-4-yl)pyrrolo[2,3-b-
]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-1-benzyl-piperidin-
-3-ol (22 mg, 31.90 umol, 1 eq), Boc.sub.2O (10.44 mg, 47.85 umol,
10.99 uL, 1.5 eq), Pd/C (50 mg, 10% purity) in THF (0.5 mL) was
degassed and purged with H.sub.2 for 3 times, and then the mixture
was stirred at 15.degree. C. for 12 h under H.sub.2 atmosphere (15
psi). The reaction mixture was filtered and concentrated under
reduced pressure to afford the title compound (30 mg, crude) as
yellow oil, and it was used into the next step without further
purification.
Step 9: Tert-butyl
(3R,5S)-3-hydroxy-5-[[4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyrid-
in-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
##STR00186##
[0460] To a solution of tert-butyl
(3S,5R)-3-[[4-[1-(benzenesulfonyl)-6-(3-methylisoxazol-4-yl)pyrrolo[2,3-b-
]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-5-hydroxy-piperidi-
ne-1-carboxylate (30 mg, 42.88 umol, 1 eq) in MeOH (0.5 mL) was
added NaOH (2 M, 0.4 mL). The mixture was stirred at 60.degree. C.
for 0.5 h. It was cooled to the room temperature. The reaction
mixture was diluted with H.sub.2O (10 mL) and extracted with EtOAc
(10 mL.times.4). The combined organic layers were washed with brine
(20 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. The residue was purified
by flash silica gel chromatography (ISCO.RTM.; 4 g SepaFlash.RTM.
Silica Flash Column, Eluent of 0.about.8% Dichloromethane/Methanol
@ 50 mL/min) to afford the title compound (5 mg, 8.94 umol, 20.84%
yield) as a white solid.
Step 10: (3R,
5S)-5-[[4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-5-(tr-
ifluoromethyl)pyrimidin-2-yl]amino]piperidin-3-ol
##STR00187##
[0462] To a solution of tert-butyl
(3R,5S)-3-hydroxy-5-[[4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyrid-
in-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
(4 mg, 7.15 umol, 1 eq) in EtOAc (0.5 mL) was added HCl/EtOAc (4 M,
0.5 mL, 279.77 eq). The mixture was stirred at 15.degree. C. for
0.5 h. The reaction mixture was concentrated under reduced pressure
to remove solvent. The residue was purified by prep-HPLC to afford
the title compound (FA, 2 mg, 94.37% purity) was obtained as a
white solid. (Note: The reaction was combined with another reaction
in 1 mg scale for purification and work up.)
Example 39. Synthesis of
N-[(3S,6S)-6-methyl-3-piperidyl]-4-[6-(3-methyltriazol-4-yl)-1H-pyrrolo[2-
,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
150)
Step 1: 1-(benzenesulfonyl)-6-bromo-pyrrolo[2,3-b]pyridine
##STR00188##
[0464] To a solution of 6-bromo-1H-pyrrolo[2,3-b]pyridine (2 g,
10.15 mmol, 1 eq) in THF (30 mL) was added NaH (608.98 mg, 15.23
mmol, 60% purity, 1.5 eq) at -10.degree. C. The mixture was stirred
at -10.degree. C. to 0.degree. C. for 0.5 h. Then benzenesulfonyl
chloride (2.15 g, 12.18 mmol, 1.56 mL, 1.2 eq) was added to the
above and the resulting mixture was stirred at 0.degree. C. for 1
h. It was poured into water (50 mL). The aqueous phase was
extracted with EtOAc (50 mL.times.3). The combined organic phase
was washed with brine (50 mL.times.2), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue
was washed with MeOH (8 mL), filtered and the filter cake was
collected to afford the title compound (3.0 g, 7.83 mmol, 77.13%
yield, 88% purity) as a white solid.
Step 2: 1-(benzenesulfonyl)-6-(3-methyltriazol-4-yl)
pyrrolo[2,3-b]pyridine
##STR00189##
[0466] To a solution of
1-(benzenesulfonyl)-6-bromo-pyrrolo[2,3-b]pyridine (1.1 g, 3.26
mmol, 1 eq) and tributyl-(3-methyltriazol-4-yl) stannane (3.40 g,
4.57 mmol, 1.4 eq) in DMF (30 mL) was added Pd(PPh.sub.3).sub.4
(376.97 mg, 326.22 umol, 0.1 eq) and CuI (62.13 mg, 326.22 umol,
0.1 eq). The mixture was stirred at 130.degree. C. under N.sub.2
for 12 h. The reaction mixture was cooled to r.t., and poured into
water (50 mL). The aqueous phase was extracted with EtOAc (50
mL.times.3). The combined organic phase was washed with brine (100
mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuum. The residue was purified by silica gel
chromatography (100-200 mesh silica gel, Petroleum ether/EtOAc=5/1,
0/1) to afford 1 g product with 77% purity. Then the product was
washed with MeOH (5 mL), filtered and the filter cake was collected
to afford the title compound (0.9 g, 2.39 mmol, 73.16% yield, 90%
purity) as a yellow solid.
Step 3:6-(3-methyltriazol-4-yl)-1H-pyrrolo[2,3-b]pyridine
##STR00190##
[0468] To a solution of
1-(benzenesulfonyl)-6-(3-methyltriazol-4-yl) pyrrolo[2,3-b]pyridine
(0.7 g, 2.06 mmol, 1 eq) in dioxane (15 mL) was added NaOH (2 M,
5.16 mL, 5 eq). The mixture was stirred at 90.degree. C. for 1 h.
The mixture was diluted with water (10 mL), extracted with EtOAc
(15 mL.times.3). The combined organic phase was washed with brine
(10 mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuum to afford the title compound (0.5 g,
crude) as a yellow solid and used directly in next step.
Step 4:
3-bromo-6-(3-methyltriazol-4-yl)-1H-pyrrolo[2,3-b]pyridine
##STR00191##
[0470] To a solution of
6-(3-methyltriazol-4-yl)-1H-pyrrolo[2,3-b]pyridine (0.5 g, 2.51
mmol, 1 eq) in DMF (10 mL) was added NBS (446.72 mg, 2.51 mmol, 1
eq). The mixture was stirred at 15.degree. C. for 12 h. The residue
was poured into water (100 mL) and stirred for 5 min. The solid was
formed and filtered. The filter cake was collected to afford the
title compound (0.7 g, crude) as a yellow solid which was used in
next step directly.
Step 5:1-(benzenesulfonyl)-3-bromo-6-(3-methyltriazol-4-yl)
pyrrolo[2,3-b]pyridine
##STR00192##
[0472] To a solution of
3-bromo-6-(3-methyltriazol-4-yl)-1H-pyrrolo[2,3-b]pyridine (0.65 g,
2.34 mmol, 1 eq) in DMF (9 mL)/THF (1 mL) was added NaH (140.22 mg,
3.51 mmol, 60% purity, 1.5 eq) at 0.degree. C. After 0.5 h,
benzenesulfonyl chloride (619.20 mg, 3.51 mmol, 448.70 uL, 1.5 eq)
was added and the resulting mixture was stirred at 0.degree. C. for
1 h. It was poured into water (100 mL) and stirred for 5 min. The
mixture was filtered and the filter cake was collected. It was
triturated with MeOH (5 mL) and filtered to give the crude product.
It was purified by silica gel chromatography (PE:EtOAc=1:1-0:1) to
afford the title compound (1 g, crude) as a pink solid.
Step
6:1-(benzenesulfonyl)-6-(3-methyltriazol-4-yl)-3-(4,4,5,5-tetramethyl-
-1,3,2-dioxaborolan-2-yl) pyrrolo[2,3-b]pyridine
##STR00193##
[0474] To a solution of
1-(benzenesulfonyl)-3-bromo-6-(3-methyltriazol-4-yl)
pyrrolo[2,3-b]pyridine (0.7 g, 1.67 mmol, 1 eq), BPD (637.47 mg,
2.51 mmol, 1.5 eq) in dioxane (15 mL) was added KOAc (328.49 mg,
3.35 mmol, 2 eq) and Pd(dppf)Cl.sub.2 (122.46 mg, 167.36 umol, 0.1
eq). The mixture was stirred at 100.degree. C. under N.sub.2
atmosphere for 2 h. It was cooled to r.t., and poured into water
(30 mL). The aqueous phase was extracted with EtOAc (30
mL.times.3). The combined organic phase was washed with brine (30
mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuum. The residue was purified by silica gel
chromatography (100-200 mesh silica gel, PE/EtOAc=5/1, 1/1) to
afford the title compound (0.4 g, batch 1) as a white solid. Batch
2: (0.2 g) was obtained as a yellow solid which contained some
de-Br product.
Step
7:1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidi-
n-4-yl]-6-(3-methyltriazol-4-yl) pyrrolo[2,3-b]pyridine
##STR00194##
[0476] To a solution of
1-(benzenesulfonyl)-6-(3-methyltriazol-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-
-dioxaborolan-2-yl) pyrrolo[2,3-b]pyridine (0.05 M, 25.79 mL, 1
eq), 4-chloro-2-methylsulfanyl-5-(trifluoromethyl)pyrimidine
(294.79 mg, 1.29 mmol, 1 eq) in H.sub.2O (2 mL)/THF (16 mL) was
added Na.sub.2CO.sub.3 (273.32 mg, 2.58 mmol, 2 eq) and ditertbutyl
(cyclopentyl)phosphane;dichloropalladium;iron (84.04 mg, 128.94
umol, 0.1 eq). The mixture was stirred at 80.degree. C. under
N.sub.2 for 1 h. It was cooled to r.t. and poured into water (50
mL). The aqueous phase was extracted with ethyl acetate (50
mL.times.3). The combined organic phase was washed with brine (50
mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuum. The residue was purified by silica gel
chromatography (100-200 mesh silica gel, PE/EtOAc=10/1, 0/1,
contained 10% DCM) to afford the title compound (0.43 g, 647.19
umol, 50.19% yield, 80% purity) as a yellow solid.
Step
8:1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidi-
n-4-yl]-6-(3-methyltriazol-4-yl) pyrrolo[2,3-b]pyridine
##STR00195##
[0478] To a solution of
1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-yl-
]-6-(3-methyltriazol-4-yl) pyrrolo[2,3-b]pyridine (0.41 g, 771.35
umol, 1 eq) in DCM (20 mL) was added m-CPBA (391.50 mg, 1.93 mmol,
2.5 eq). The mixture was stirred at 15.degree. C. for 12 h. The
solution was quenched by sat.NaHCO.sub.3 (20 mL) and
sat.Na.sub.2SO.sub.3 (20 mL). The mixture was stirred for 5 min.
The aqueous phase was extracted with DCM (30 mL.times.3). The
combined organic phase dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuum. The residue was washed with a
mixture of Petroleum ether/EtOAc=3/1 (5 mL), filtered and the
filter cake was collected to afford the title compound (0.35 g,
crude) as a yellow solid and used directly.
Step 9:
Benzyl(2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3-methyltriazol-4-yl)p-
yrrolo[2,3-b]pyridine-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-met-
hyl-piperidine-1-carboxylate (Product 1) and
Benzyl(2R,5R)-5-[[4-[1-(benzenesulfonyl)-6-(3-methyltriazol-4-yl)pyrrolo[-
2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-pipe-
ridine-1-carboxylate (Product 2)
##STR00196##
[0480] To a solution of
1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl-
]-6-(3-methyltriazol-4-yl) pyrrolo[2,3-b]pyridine (0.35 g, 621.08
umol, 1 eq) and benzyl 5-amino-2-methyl-piperidine-1-carboxylate
(308.45 mg, 1.24 mmol, 2 eq) in THF (15 mL) was added DIEA (401.35
mg, 3.11 mmol, 540.91 uL, 5 eq). The mixture was stirred at
15.degree. C. for 12 h. The resulting mixture was poured into water
(30 mL). The aqueous phase was extracted with EtOAc (30
mL.times.2). The combined organic phase was washed with brine (30
mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuum. The residue was purified by silica gel
chromatography (100-200 mesh silica gel, Petroleum
ether/EtOAc=10/1, 1/1) to afford a racemate product (150 mg). Then
the product was separation by SFC (column: DAICEL CHIRALPAK AS-H
(250 mm*30 mm, 5 um); mobile phase:[0.1% NH.sub.3H.sub.2O MeOH]; B
%: 45%-45%, 5 min) to afford product 1 (peak 1: RT=1.71 min, 70 mg,
80.36 umol, 12.94% yield, 84% purity) as a yellow solid, and
product 2 (peak 2: 50 mg, 64.23 umol, 10.34% yield, 94% purity) as
a yellow solid.
Step 10:
(2S,5S)-2-methyl-5-[[4-[6-(3-methyltriazol-4-yl)-1H-pyrrolo[2,3-b-
]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carbox-
ylate
##STR00197##
[0482] To a solution of benzyl
(2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3-methyltriazol-4-yl)
pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-me-
thyl-piperidine-1-carboxylate (50 mg, 68.33 umol, 1 eq) in dioxane
(4 mL) was added NaOH (2 M, 170.82 uL, 5 eq). The mixture was
stirred at 90.degree. C. for 1 h. It was cooled to 15.degree. C.
and poured into water (20 mL). The aqueous phase was extracted with
EtOAc (20 mL.times.3). The combined organic phase was washed with
brine (20 mL.times.2), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuum to afford the title compound
(50 mg, crude) as a yellow solid and used directly.
[0483] (Note: The reaction was combined with another batch in 20 mg
scale for purification.)
Step 11:
N-[(3S,6S)-6-methyl-3-piperidyl]-4-[6-(3-methyltriazol-4-yl)-1H-p-
yrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00198##
[0485] To a solution of benzyl
(2S,5S)-2-methyl-5-[[4-[6-(3-methyltriazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-
-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
(50 mg, 84.52 umol, 1 eq) in DCM (5 mL) was added BBr.sub.3 (105.87
mg, 422.59 umol, 40.72 uL, 5 eq). The mixture was stirred at
15.degree. C. for 2 h. The mixture was concentrated in vacuum. The
residue was washed with a mixture solution of Petroleum
ether/EtOAc=3/1 (15 mL), filtered and the filter cake was
collected. Then the filter cake was dissolved with MeOH (1 mL) and
purified by prep-HPLC (HCl) to afford the title compound (20.6 mg,
99% purity, HCl) as a yellow solid. (Note: The reaction was
combined with another batch in 20 mg scale for purification.)
Example 40. Synthesis of
N-[(3R,6R)-6-methyl-3-piperidyl]-4-[6-(3-methyltriazol-4-yl)-1H-pyrrolo[2-
,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
151)
Step 1:
(2R,5R)-2-methyl-5-[[4-[6-(3-methyltriazol-4-yl)-1H-pyrrolo[2,3-b]-
pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxy-
late
##STR00199##
[0487] To a solution of benzyl (2R,
5R)-5-[[4-[1-(benzenesulfonyl)-6-(3-methyltriazol-4-yl)pyrrolo[2,3-b]pyri-
din-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-c-
arboxylate (50 mg, 68.33 umol, 1 eq) in dioxane (4 mL) was added
NaOH (2 M, 170.82 uL, 5 eq). The mixture was stirred at 90.degree.
C. for 1 h. The reaction mixture was cooled to r.t. and poured into
water (20 mL).The aqueous phase was extracted with EtOAc (20
mL.times.3). The combined organic phase was washed with brine (20
mL.times.2), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuum to afford the title compound (50 mg, crude)
as a yellow solid and used directly.
Step 2: N-[(3R,
6R)-6-methyl-3-piperidyl]-4-[6-(3-methyltriazol-4-yl)-1H-pyrrolo[2,3-b]py-
ridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00200##
[0489] To a solution of benzyl (2R,
5R)-2-methyl-5-[[4-[6-(3-methyltriazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-y-
l]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
(50 mg, 84.52 umol, 1 eq) in DCM (5 mL) was added BBr.sub.3 (105.87
mg, 422.59 umol, 40.72 uL, 5 eq) at 0.degree. C. The mixture was
stirred at 0.degree. C. for 2 h. The mixture was concentrated in
vacuum. The residue was washed with a mixture solution of Petroleum
ether/EtOAc=3/1 (15 mL), filtered and the filter cake was
collected. Then the filter cake was dissolved with MeOH (1 mL) and
purified by prep-HPLC (HCl) to afford the title compound (21 mg,
42.41 umol, 50.17% yield, 99.74% purity, HCl) as a yellow
solid.
Example 41. Synthesis of
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-N-[(3S,6-
S)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 152)
Step 1:
Benzyl(2S,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-tetrahydropyr-
an-2-yl-pyrazolo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]ami-
no]-2-methyl-piperidine-1-carboxylate
##STR00201##
[0491] To a solution of
3,5-dimethyl-4-[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl]-1--
tetrahydropyran-2-yl-pyrazolo[3,4-b]pyridin-6-yl]isoxazole (178.85
mg, 342.30 umol, 1 eq) in THF (1 mL) was added DIPEA (132.72 mg,
1.03 mmol, 178.86 uL, 3 eq) and benzyl
(2S,5S)-5-amino-2-methyl-piperidine-1-carboxylate (0.17 g, 684.60
umol, 2 eq). The mixture was stirred at 15.degree. C. for 1 h. The
reaction mixture concentrated under reduced pressure to give a
residue. The residue was purified by column chromatography
(SiO.sub.2, Petroleum ether/EtOAc=2/1) to afford the title compound
(150 mg) as a white solid.
Step 2:
Benzyl(2S,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrazolo[3,4-
-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperid-
ine-1-carboxylate
##STR00202##
[0493] To a solution of benzyl(2S,5S)-5-[[4-[6-(3,
5-dimethylisoxazol-4yl)-1-tetrahydropyran-2-yl-pyrazolo[3,4-b]pyridin-3-y-
l]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carboxyl-
ate (0.15 g, 217.17 umol, 1 eq) in MeOH (2 mL) was added HCl/MeOH
(4 M, 54.29 uL, 1.00 eq). The mixture was stirred at 60.degree. C.
for 12 h. The reaction mixture was concentrated under reduced
pressure to afford the title compound (120 mg) as a yellow
solid.
Step 3:
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-N-
-[(3S,6S)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00203##
[0495] To a solution of benzyl
(2S,5S)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrazolo[3,4-b]pyridin-3--
yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carboxy-
late (0.1 g, 164.85 umol, 1 eq) in DCM (0.2 mL) was added BBr.sub.3
(82.60 mg, 329.71 umol, 31.77 uL, 2 eq) at 0.degree. C. The mixture
was stirred at 0.degree. C. for 1 h. The reaction mixture was
concentrated. The residue was purified by prep-HPLC to afford the
title compound (37.6 mg, HCl salt, 100% purity) as a yellow solid.
(Note: The reaction was combined with another reaction in 20 mg
scale for work-up and purification).
Example 42. Synthesis of
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-N-[(3R,6-
R)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 153)
Step 1: benzyl
(2R,5R)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-tetrahydropyran-2-yl-pyraz-
olo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl--
piperidine-1-carboxylate
##STR00204##
[0497] To a solution of
3,5-dimethyl-4-[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl]-1--
tetrahydropyran-2-yl-pyrazolo[3,4-b]pyridin-6-yl]isoxazole (157.81
mg, 302.03 umol, 1 eq) in THF (1 mL) was added DIPEA (117.10 mg,
906.09 umol, 157.82 uL, 3 eq) and benzyl
(2R,5R)-5-amino-2-methyl-piperidine-1-carboxylate (0.15 g, 604.06
umol, 2 eq).The mixture was stirred at 15.degree. C. for 1 h. The
reaction mixture concentrated under reduced pressure to give a
residue. The residue was purified by column chromatography
(SiO.sub.2, Petroleum ether/Ethyl acetate=2/1) to afford the title
compound (140 mg) as a white solid.
Step 2: benzyl
(2R,5R)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrazolo[3,4-b]pyridin-3--
yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carboxy-
late
##STR00205##
[0499] To a solution of benzyl
(2R,5R)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1-tetrahydropyran-2-yl-pyraz-
olo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl--
piperidine-1-carboxylate (120.00 mg, 173.73 umol, 1 eq) in MeOH
(0.2 mL) was added HCl/MeOH (4 M, 1 mL). The mixture was stirred at
60.degree. C. for 12 h. It was concentrated under reduced pressure
to afford the title compound (110 mg) as a yellow solid.
Step 3:
4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-N-
-[(3R,6R)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00206##
[0501] To a solution of benzyl
(2R,5R)-5-[[4-[6-(3,5-dimethylisoxazol-4-yl)-1H-pyrazolo[3,4-b]pyridin-3--
yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carboxy-
late (0.09 g, 148.37 umol, 1 eq) in DCM (2 mL) was added BBr.sub.3
(74.34 mg, 296.74 umol, 28.59 uL, 2 eq) at 0.degree. C. The mixture
was stirred at 0.degree. C. for 1 h. The reaction mixture was
concentrated under reduced pressure to give a residue. It was
purified by prep-HPLC (HCl condition) to afford the title compound
(21.7 mg) as a yellow solid.
[0502] (Note: The reaction was combined with another reaction in 20
mg scale for work-up and purification).
Example 43. Synthesis of
N-[(3S,6S)-6-methyl-3-piperidyl]-4-[6-(5-methyl-1,3,4-thiadiazol-2-yl)-1H-
-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 154)
Step 1:
Trimethyl-[2-[[3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4--
yl]-6-(5-methyl-1,3,4-thiadiazol-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]e-
thyl]silane
##STR00207##
[0504] A mixture of trimethyl-[2-[[6-(5-methyl-1,
3,4-thiadiazol-2-yl)-3-(4,4,5, 5-tetramethyl-1,3,
2-dioxaborolan-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
(0.05 M, 22.86 mL, 1 eq),
4-chloro-2-methylsulfanyl-5-(trifluoromethyl)pyrimidine (261.29 mg,
1.14 mmol, 1 eq), Na.sub.2CO.sub.3 (242.27 mg, 2.29 mmol, 2 eq) and
ditert-butyl (cyclopentyl)phosphane:dichloropalladium:iron (74.49
mg, 114.29 umol, 0.1 eq) in THF (20 mL) and H.sub.2O (2 mL) was
degassed and purged with N.sub.2 for 3 times, and then the mixture
was stirred at 80.degree. C. for 1 h under N.sub.2 atmosphere. It
was diluted by addition H.sub.2O (100 mL), and extracted with EtOAc
(50 mL.times.3), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by column chromatography (SiO.sub.2, Petroleum
ether/Ethyl acetate=10/1 to 5/1) to afford the title compound (150
mg) as a yellow solid. (Note: The reaction was combined with
another reaction in 20 mg scale for purification.)
Step 2:
Trimethyl-[2-[[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4--
yl]-6-(5-methyl-1,3,4-thiadiazol-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]e-
thyl]silane
##STR00208##
[0506] To a solution of
trimethyl-[2-[[3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-yl]-6-(-
5-methyl-1,3,4-thiadiazol-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]si-
lane (140 mg, 259.89 umol, 1 eq) in DCM (3 mL) was added m-CPBA
(131.91 mg, 649.73 umol, 85% purity, 2.5 eq).The mixture was
stirred at 15.degree. C. for 3 h. The solution was washed with
Sat.Na.sub.2SO.sub.3 (30 mL), Sat.NaHCO.sub.3 (30 mL), brine (50
mL). The organic layers were dried over Na.sub.2SO.sub.4 and
concentrated. The residue was washed with PE/EtOAc=5/1 (5 mL) and
filtered to afford the title compound (120 mg) as a yellow solid.
(Note: The reaction was combined with another reaction in 10 mg
scale for purification.)
Step 3:
Benzyl(2S,5S)-2-methyl-5-[[4-[6-(5-methyl-1,3,4-thiadiazol-2-yl)-1-
-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromet-
hyl)pyrimidin2-yl]amino]piperidine-1-carboxylate
##STR00209##
[0508] To a solution of
trimethyl-[2-[[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl]-6-(-
5-methyl-1,3,4-thiadiazol-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]si-
lane (0.12 g, 210.27 umol, 1 eq) in THF (2 mL) was added DIEA
(81.53 mg, 630.82 umol, 109.88 uL, 3 eq), benzyl
(2S,5S)-5-amino-2-methyl-piperidine-1-carboxylate (104.43 mg,
420.55 umol, 2 eq).The mixture was stirred at 15.degree. C. for 1
h. The resulting mixture was concentrated. The residue was purified
by column chromatography (SiO.sub.2, Petroleum ether/Ethyl
acetate=10/1 to 2/1) to afford the title compound (0.09 g, 85.26
umol, 40.55% yield, 70% purity) as yellow oil.
Step 4: Benzyl (2S,5S)-2-methyl-5-[[4-[6-(5-methyl-1, 3,
4-thiadiazol-2-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyri-
midin-2-yl]amino]piperidine-1-carboxylate
##STR00210##
[0510] To a solution of benzyl
(2S,5S)-2-methyl-5-[[4-[6-(5-methyl-1,3,4-thiadiazol-2-yl)-1-(2-trimethyl-
silylethoxymethyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-
-2-yl]amino]piperidine-1-carboxylate (90 mg, 121.80 umol, 1 eq) in
dioxane (3 mL) was added H.sub.2SO.sub.4 (1.19 g, 12.18 mmol,
649.26 uL, 100 eq). The mixture was stirred at 45.degree. C. for 2
h. The mixture was adjusted pH to 8 with saturated aqueous NaOH,
and extracted with EtOAc (50 mL.times.3), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to afford the title compound (0.12 g, crude) as a yellow solid.
Step 5: N-[(3S,6S)-6-methyl-3-piperidyl]-4-[6-(5-methyl-1, 3,
4-thiadiazol-2-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyri-
midin-2-amine
##STR00211##
[0512] To a solution of
benzyl(2S,5S)-2-methyl-5-[[4-[6-(5-methyl-1,3,
4-thiadiazol-2-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyri-
midin-2-yl]amino]piperidine-1-carboxylate (0.12 g, 197.16 umol, 1
eq) in DCM (3 mL) was added BBr.sub.3 (148.18 mg, 591.49 umol,
56.99 uL, 3 eq). The mixture was stirred at 15.degree. C. for 30
min. It was concentrated. The residue was triturated with
PE/EtOAc=5/1 (5 mL) and filtered. The crude product was purified by
prep-HPLC (HCl condition) to afford the title compound (19.5 mg,
38.16 umol, 19.36% yield, 100% purity, HCl) as a yellow solid.
Example 44. Synthesis of
4-[6-(5-methyl-1,3,4-thiadiazol-2-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(-
3S)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
155)
Step 1: Trimethyl-[2-[[6-(5-methyl-1, 3, 4-thiadiazol-2-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
##STR00212##
[0514] A mixture of
[1-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridin-6-yl]boronic
acid (4 g, 13.69 mmol, 1 eq), 2-bromo-5-methyl-1,3,4-thiadiazole
(2.45 g, 13.69 mmol, 1 eq), Cs.sub.2CO.sub.3 (8.92 g, 27.38 mmol, 2
eq), Pd(PPh.sub.3).sub.4 (1.58 g, 1.37 mmol, 0.1 eq) in dioxane (50
mL) and H.sub.2O (5 mL) was degassed and purged with N.sub.2 for 3
times, and then the mixture was stirred at 100.degree. C. for 1 h
under N.sub.2 atmosphere. The reaction mixture was diluted with
H.sub.2O (300 mL) and extracted with EtOAc (100 mL.times.3). The
combined organic layers were washed with brine (300 mL.times.2),
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. The residue was purified by
column chromatography (SiO.sub.2, Petroleum ether/Ethyl
acetate=10/1 to 4/1) to afford the title compound (1.6 g, 4.39
mmol, 32.04% yield, 95% purity) as a yellow solid.
Step 2: 2-[[3-bromo-6-(5-methyl-1, 3, 4-thiadiazol-2-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl-trimethyl-silane
##STR00213##
[0516] To a solution of trimethyl-[2-[[6-(5-methyl-1, 3,
4-thiadiazol-2-yl) pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
(1.6 g, 4.62 mmol, 1 eq) in DMF (20 mL) was added NBS (821.79 mg,
4.62 mmol, 1 eq). The mixture was stirred at 15.degree. C. for 1 h.
The resulting mixture was diluted with H.sub.2O 200 mL and
extracted with EtOAc (100 mL.times.3). The combined organic layers
were washed with brine (300 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, Petroleum ether/Ethyl acetate=10/1 to
5/1) to afford the title compound (1.6 g) as a yellow solid.
Step 3: Trimethyl-[2-[[6-(5-methyl-1,
3,4-thiadiazol-2-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrro-
lo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
##STR00214##
[0518] A mixture of 2-[[3-bromo-6-(5-methyl-1, 3,
4-thiadiazol-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl-trimethyl-sila-
ne (1.6 g, 3.76 mmol, 1 eq),
4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-
-dioxaborolane (1.91 g, 7.52 mmol, 2 eq), AcOK (738.21 mg, 7.52
mmol, 2 eq) and Pd(dppf)Cl.sub.2 (275.20 mg, 376.10 umol, 0.1 eq)
in dioxane (30 mL) was degassed and purged with N.sub.2 for 3
times, and then the mixture was stirred at 100.degree. C. for 2 h
under N.sub.2 atmosphere. It was filtered and the filtrate was
concentrated. The residue was purified by column chromatography
(SiO.sub.2, Petroleum ether/Ethyl acetate=10/1 to 4:1, plate 1) to
give a residue (1.5 g). 200 mg of crude product was purified by
prep-HPLC (TFA condition). Then the eluent was concentrated to give
a residue to afford the title compound (100 mg, TFA) as yellow
oil.
Step 4:
Tert-butyl(3S)-3-[[4-[6-(5-methyl-1,3,4-thiadiazol-2-yl)-1-(2-trim-
ethylsilylethoxymethyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyri-
midin-2-yl]amino]piperidine-1-carboxylate
##STR00215##
[0520] A mixture of
trimethyl-[2-[[6-(5-methyl-1,3,4-thiadiazol-2-yl)-3-(4,4,5,5-tetramethyl--
1,3,2-dioxaborolan-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
(0.05 M, 3.41 mL, 1 eq, TFA), tert-butyl
(3S)-3-[[4-chloro-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-ca-
rboxylate (77.91 mg, 204.60 umol, 1.2 eq), Na.sub.2CO.sub.3 (36.14
mg, 341.00 umol, 2 eq) and
ditert-butyl(cyclopentyl)phosphane;dichloropalladium;iron (11.11
mg, 17.05 umol, 0.1 eq) in THF (5 mL) and H.sub.2O (0.5 mL) was
degassed and purged with N.sub.2 for 3 times, and then the mixture
was stirred at 80.degree. C. for 1 h under N.sub.2 atmosphere. The
reaction mixture was diluted with H.sub.2O (50 mL) and extracted
with EtOAc (20 mL.times.3). The combined organic layers were washed
with brine (60 mL.times.2), dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give a residue. The
residue was purified by column chromatography (SiO.sub.2, Petroleum
ether/Ethyl acetate=10/1 to 1/1) to afford the title compound (0.11
g) as a white solid.
Step 5:
4-[6-(5-methyl-1,3,4-thiadiazol-2-yl)-1H-pyrrolo[2,3-b]pyridin-3-y-
l]-N-[(3S)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00216##
[0522] To a solution of
tert-butyl(3S)-3-[[4-[6-(5-methyl-1,3,4-thiadiazol-2-yl)-1-(2-trimethylsi-
lylethoxymethyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-
-yl]amino]piperidine-1-carboxylate (0.11 g, 159.22 umol, 1 eq) in
dioxane (3 mL) was added H.sub.2SO.sub.4 (1.56 g, 15.92 mmol,
848.73 uL, 100 eq). The solution was stirred at 45.degree. C. for 2
h. The mixture was adjusted pH to 8 with saturated aqueous NaOH,
and extracted with EtOAc (50 mL.times.3), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by prep-HPLC (HCl
condition) to afford the title compound (17.4 mg, 34.89 umol,
21.91% yield, 99.65% purity, HCl) as a yellow solid.
Example 45. Synthesis of
4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3S,6S)-6--
methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
156)
Step 1:
4-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
-yl)pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole
##STR00217##
[0524] A mixture of
4-[1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxa-
zole (1.5 g, 3.59 mmol, 1 eq),
4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-
-dioxaborolane (1.37 g, 5.38 mmol, 1.5 eq), Pd(dppf)Cl.sub.2
(262.41 mg, 358.63 umol, 0.1 eq), KOAc (703.91 mg, 7.17 mmol, 2 eq)
in dioxane (90 mL) was degassed and purged with N.sub.2 for 3
times, and then the mixture was stirred at 100.degree. C. for 2 h
under N.sub.2 atmosphere. It was cooled to the room temperature and
concentrated under reduced pressure to remove solvent. The residue
was purified by column chromatography (SiO.sub.2, Petroleum
ether/Ethyl acetate=20/1 to 5/1) to afford the title compound (1.2
g) was obtained as a white solid. (Note: The reaction was combined
with another reaction in 200 mg scale for purification and work
up.)
Step 2:
4-[1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyri-
midin-4-yl]pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazol)
##STR00218##
[0526] A mixture of
4-[1-(benzenesulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyr-
rolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole (1.1 g, 2.36 mmol, 1
eq), 4-chloro-2-methylsulfanyl-5-(trifluoromethyl)pyrimidine
(540.45 mg, 2.36 mmol, 1 eq), ditert-butyl(cyclopentyl)
phosphane;dichloropalladium;iron (154.07 mg, 236.39 umol, 0.1 eq),
Na.sub.2CO.sub.3 (501.10 mg, 4.73 mmol, 2 eq) in THF (40 mL) and
H.sub.2O (4 mL) was degassed and purged with N.sub.2 for 3 times,
and then the mixture was stirred at 80.degree. C. for 12 h under
N.sub.2 atmosphere. It was cooled to the room temperature and
diluted with H.sub.2O (60 mL) and extracted with EtOAc (30
mL.times.3). The combined organic layers were washed with brine (60
mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to give a residue. The residue was purified by
flash silica gel chromatography (ISCO.RTM.; 20 g SepaFlash.RTM.
Silica Flash Column, Eluent of 0.about.30% Ethyl acetate/Petroleum
ether gradient @ 75 mL/min) to afford the title compound (800 mg,
crude) as a yellow solid.
Step 3:
4-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyri-
midin-4-yl]pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole
##STR00219##
[0528] To a solution of
4-[1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-
-yl]pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole (800 mg, 1.51
mmol, 1 eq) in EtOAc (40 mL) was added Oxone (4.63 g, 7.53 mmol, 5
eq). The mixture was stirred at 80.degree. C. for 12 h. The
reaction mixture was filtered and the filtrate was quenched by
saturation Na.sub.2SO.sub.3 (60 mL). The water layer was extracted
with EtOAc (30 mL.times.3). The combined organic layers were washed
with brine (50 mL), dried over Na.sub.2SO.sub.4, filtrate and
concentrated. The residue was purified by flash silica gel
chromatography (ISCO.RTM.; 20 g SepaFlash.RTM. Silica Flash Column,
Eluent of 0.about.50% Ethyl acetate/Petroleum ether gradient @ 75
mL/min) to afford the title compound (380 mg, 674.32 umol, 44.80%
yield) as a pink solid.
Step 4:
Benzyl(2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3-methylisoxazol-4-yl)-
pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-met-
hyl-piperidine-1-carboxylate
##STR00220##
[0530] To a solution of
4-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-
-yl]pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole (170 mg, 301.67
umol, 1 eq) and benzyl
(2S,5S)-5-amino-2-methyl-piperidine-1-carboxylate (74.91 mg, 301.67
umol, 1 eq) in THF (2 mL) was added DIPEA (194.94 mg, 1.51 mmol,
262.72 uL, 5 eq). The mixture was stirred at 15.degree. C. for 6 h.
It was concentrated under reduced pressure to remove solvent. The
residue was purified by flash silica gel chromatography (ISCO.RTM.;
4 g SepaFlash.RTM. Silica Flash Column, Eluent of 0.about.25% Ethyl
acetate/Petroleum ether gradient @ 50 mL/min) to afford the title
compound (100 mg) as a yellow solid. (Note: The reaction was
combined with another reaction in 20 mg scale for purification and
work up.)
Step 5:
Benzyl(2S,5S)-2-methyl-5-[[4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo-
[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1--
carboxylate
##STR00221##
[0532] To a solution of
benzyl(2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-(3-methylisoxazol-4-yl)pyrrolo-
[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-pip-
eridine-1-carboxylate (90 mg, 122.99 umol, 1 eq) in EtOH (3 mL) was
added NaOH (2 M, 2.5 mL). The mixture was stirred at 60.degree. C.
for 1 h. The reaction mixture was diluted with H.sub.2O (10 mL) and
extracted with EtOAc (10 mL.times.3). The combined organic layers
were washed with brine (10 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to afford the
title compound (80 mg) as a yellow solid, and it was used into the
next step without further purification. (Note: The reaction was
combined with another reaction in 10 mg scale for purification and
work up.)
Step 6:
4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3S-
,6S)-6-methyl-3-piperidyl]5-(trifluoromethyl)pyrimidin-2-amine
##STR00222##
[0534] To a solution of
benzyl(2S,5S)-2-methyl-5-[[4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo[2,3-b]-
pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxy-
late (70 mg, 118.33 umol, 1 eq) in DCM (7 mL) was added BBr.sub.3
(148.22 mg, 591.63 umol, 57.01 uL, 5 eq). The mixture was stirred
at 15.degree. C. for 2 h. The reaction mixture was quenched by
saturation NaHCO.sub.3 solution (1.5 mL), and then concentrated
under reduced pressure to give a residue. The residue was diluted
with H.sub.2O (1.5 mL) and DMF (1.5 mL), then adjusted to pH=4 with
HCl (2 N). The residue was purified by prep-HPLC (FA condition) to
afford the title compound (FA, 22.2 mg, 98.26% purity) as a white
solid.
Example 46. Synthesis of
4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3R,6R)-6--
methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
157)
Step 1: Benzyl
(2R,5R)-5-[[4-[1-(benzenesulfonyl)-6-(3-methylisoxazol-4-yl)pyrrolo[2,3-b-
]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidin-
e-1-carboxylate
##STR00223##
[0536] To a solution of
4-[1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-
-yl]pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-isoxazole (170 mg, 301.67
umol, 1 eq) and benzyl
(2R,5R)-5-amino-2-methyl-piperidine-1-carboxylate (74.91 mg, 301.67
umol, 1 eq) in THF (2 mL) was added DIPEA (194.94 mg, 1.51 mmol,
262.72 uL, 5 eq). The mixture was stirred at 15.degree. C. for 6 h.
The solution was concentrated under reduced pressure to remove
solvent. The residue was purified by flash silica gel
chromatography (ISCO.RTM.; 4 g SepaFlash.RTM. Silica Flash Column,
Eluent of 0.about.25% EtOAc/PE @ 50 mL/min) to afford the title
compound (120 mg) as a yellow solid. (Note: The reaction was
combined with another reaction in 20 mg scale for work up)
Step 2: Benzyl
(2R,5R)-2-methyl-5-[[4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridi-
n-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
##STR00224##
[0538] To a solution of benzyl
(2R,5R)-5-[[4-[1-(benzenesulfonyl)-6-(3-methylisoxazol-4-yl)pyrrolo[2,3-b-
]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidin-
e-1-carboxylate (110.15 mg, 150.53 umol, 1 eq) in EtOH (3 mL) was
added NaOH (2 M, 2.5 mL). The mixture was stirred at 60.degree. C.
for 1 h. The reaction mixture was diluted with H.sub.2O 10 mL and
extracted with EtOAc (10 mL.times.3). The combined organic layers
were washed with brine (10 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to afford the
title (100 mg) as a yellow solid which was used into the next step
without further purification.
Step 3:
4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3R-
,6R)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00225##
[0540] To a solution of benzyl
(2R,5R)-2-methyl-5-[[4-[6-(3-methylisoxazol-4-yl)-1H-pyrrolo[2,3-b]pyridi-
n-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
(90 mg, 152.13 umol, 1 eq) in DCM (9 mL) was added BBr.sub.3
(190.56 mg, 760.67 umol, 73.29 uL, 5 eq). The mixture was stirred
at 15.degree. C. for 2 h. The reaction mixture was quenched by
saturation NaHCO.sub.3 (1.5 mL), and then concentrated under
reduced pressure to give a residue. The residue was diluted with
H.sub.2O (1.5 mL) and DMF (1.5 mL), then adjusted to pH=4 with HCl
(2 N). The residue was purified by prep-HPLC (FA condition) to
afford the title compound (FA, 20.3 mg) as a white solid.
Example 47. Synthesis of
N-[(3R,6R)-6-methyl-3-piperidyl]-4-[6-(5-methyl-1,3,4-thiadiazol-2-yl)-1H-
-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 158)
Step 1: Benzyl (2R,
5R)-2-methyl-5-[[4-[6-(5-methyl-1,3,4-thiadiazol-2-yl)-1-(2-trimethylsily-
lethoxymethyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-y-
l]amino]piperidine-1-carboxylate
##STR00226##
[0542] To a solution of
trimethyl-[2-[[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl]-6-(-
5-methyl-1,3,4-thiadiazol-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]si-
lane (0.2 g, 350.46 umol, 1 eq) in THF (2 mL) was added DIEA
(135.88 mg, 1.05 mmol, 183.13 uL, 3 eq), benzyl
(2R,5R)-5-amino-2-methyl-piperidine-1-carboxylate (149.68 mg,
602.79 umol, 1.72 eq). The mixture was stirred at 15.degree. C. for
2 h. The reaction mixture was concentrated to give a residue. The
residue was purified by column chromatography (SiO.sub.2,
PE/EtOAc=10/1 to 2/1) to afford the title compound (250 mg) as
yellow oil.
Step 2:
N-[(3R,6R)-6-methyl-3-piperidyl]-4-[6-(5-methyl-1,3,4-thiadiazol-2-
-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00227##
[0544] To a solution of benzyl
(2R,5R)-2-methyl-5-[[4-[6-(5-methyl-1,3,4-thiadiazol-2-yl)-1-(2-trimethyl-
silylethoxymethyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-
-2-yl]amino]piperidine-1-carboxylate (0.25 g, 338.34 umol, 1 eq) in
dioxane (5 mL) was added H.sub.2SO.sub.4 (3.32 g, 33.83 mmol, 1.80
mL, 100 eq). The mixture was stirred at 45.degree. C. for 2 h. The
mixture was adjusted pH to 8 with saturated aqueous NaOH, and
extracted with EtOAc (50 mL.times.3), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
It was purified by prep-HPLC (HCl condition) to afford the title
compound (27.8 mg, 51.69 umol, 15.28% yield, 95% purity, HCl) as a
yellow solid.
Example 48. Synthesis of
N-[(3R,6R)-6-methyl-3-piperidyl]-4-[6-(3-methyl-1,2,4-thiadiazol-5-yl)-1H-
-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 159)
Step 1:
Trimethyl-[2-[[6-(3-methyl-1,2,4-thiadiazol-5-yl)pyrrolo[2,3-b]pyr-
idin-1-yl]methoxy]ethyl]silane
##STR00228##
[0546] To a solution of
[1-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridin-6-yl]boronic
acid (3.92 g, 13.40 mmol, 1.6 eq),
5-bromo-3-methyl-1,2,4-thiadiazole (1.5 g, 8.38 mmol, 1 eq) in
dioxane (40 mL) was added K.sub.2CO.sub.3 (2.89 g, 20.95 mmol, 2.5
eq) and Pd(dppf)Cl.sub.2.CH.sub.2Cl.sub.2 (684.19 mg, 837.81 umol,
0.1 eq) at 15.degree. C., the reaction mixture was degassed and
purged with N.sub.2 for 3 times and stirred at 100.degree. C. for 2
h. The resulting solution was filtered and concentrated to give a
residue. The residue was purified by silica gel column
(PE/EtOAc=100/1 to 25/1) to afford the title compound (1.1 g, 3.17
mmol, 37.89% yield) as a colorless oil.
Step 2:
2-[[3-bromo-6-(3-methyl-1,2,4-thiadiazol-5-yl)pyrrolo[2,3-b]pyridi-
n-1-yl]methoxy]ethyl-trimethyl-silane
##STR00229##
[0548] To a stirred solution of
trimethyl-[2-[[6-(3-methyl-1,2,4-thiadiazol-5-yl)pyrrolo[2,3-b]pyridin-1--
yl]methoxy]ethyl]silane (1.1 g, 3.17 mmol, 1 eq) in DMF (30 mL) was
added NBS (553.69 mg, 3.11 mmol, 0.98 eq), the reaction mixture was
stirred at 15.degree. C. for 2 h. Then it was diluted with EtOAc
(80 mL), washed with sub-saturated brine (50 mL.times.3) and
saturated brine (50 mL). The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated to give a residue. It was
purified by silica gel column (PE/EtOAc=50/1 to 20/1) to afford the
title compound (1.15 g, 2.70 mmol, 85.16% yield) as a white
solid.
Step 3:
Trimethyl-[2-[[6-(3-methyl-1,2,4-thiadiazol-5-yl)-3-(4,4,5,5-tetra-
methyl-1,3,2-dioxaborolan-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]si-
lane
##STR00230##
[0550] To a solution of
2-[[3-bromo-6-(3-methyl-1,2,4-thiadiazol-5-yl)pyrrolo[2,3-b]pyridin-1-yl]-
methoxy]ethyl-trimethyl-silane (1 g, 2.35 mmol, 1 eq) and
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (895.38
mg, 3.53 mmol, 1.5 eq) in dioxane (20 mL) was added KOAc (461.39
mg, 4.70 mmol, 2 eq) and Pd(dppf)Cl.sub.2.CH.sub.2Cl.sub.2 (191.96
mg, 235.06 umol, 0.1 eq) at 15.degree. C., the reaction mixture was
degassed and purged with N.sub.2 for 3 times and stirred at
100.degree. C. for 3 h. The final reaction mixture was filtered and
concentrated to give a residue. The residue was purified by silica
gel column (PE/EtOAc=50/1) to afford the title compound (1.05 g,
not pure) as a yellow solid. (Note: The reaction was combined with
another reaction in 200 mg scale for work up.)
Step 4:
Trimethyl-[2-[[3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4--
yl]-6-(3-methyl-1,2,4-thiadiazol-5-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]e-
thyl]silane
##STR00231##
[0552] To a solution of
trimethyl-[2-[[6-(3-methyl-1,2,4-thiadiazol-5-yl)-3-(4,4,5,5-tetramethyl--
1,3,2-dioxaborolan-2-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
(929.99 mg, 1.97 mmol, 1 eq) and
4-chloro-2-methylsulfanyl-5-(trifluoromethyl)pyrimidine (0.45 g,
1.97 mmol, 1 eq) in H.sub.2O (2.5 mL) and THF (12.5 mL) was added
Na.sub.2CO.sub.3 (417.24 mg, 3.94 mmol, 2 eq) and
ditert-butyl(cyclopentyl)phosphane;dichloropalladium;iron (128.28
mg, 196.83 umol, 0.1 eq) at 15.degree. C., the reaction mixture was
degassed and purged with N.sub.2 for 3 times, and stirred at
75.degree. C. for 2 h. The mixture was diluted with EtOAc (50 mL),
washed with H.sub.2O (20 mL). The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated to give a residue. It was
purified by silica gel column (PE/EtOAc=50/1 to 10/1) to afford the
title compound (550 mg) as a white solid. (Note: The reaction was
combined with another reaction in 100 mg scale for work up.)
Step 5:
Trimethyl-[2-[[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4--
yl]-6-(3-methyl-1,2,4-thiadiazol-5-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]e-
thyl]silane
##STR00232##
[0554] A mixture of
trimethyl-[2-[[3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-yl]-6-(-
3-methyl-1,2,4-thiadiazol-5-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]si-
lane (0.5 g, 928.19 umol, 1 eq) and Oxone (2.85 g, 4.64 mmol, 5 eq)
in EtOAc (20 mL) was stirred at 80.degree. C. for 12 h. It was
filtered and washed with EtOAc (30 mL). The organic layer was
washed with aq.Na.sub.2SO.sub.3 (10 mL.times.3), dried over
Na.sub.2SO.sub.4 and concentrated to give a residue. The residue
was purified by silica gel column (PE/EtOAc=15/1 to 5/1) to afford
the title compound (120 mg) as a yellow solid.
[0555] (Note: The reaction was combined with another reaction in 50
mg scale for work up)
Step 6: Benzyl
(2R,5R)-2-methyl-5-[[4-[6-(3-methyl-1,2,4-thiadiazol-5-yl)-1-(2-trimethyl-
silylethoxy
methyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amin-
o]piperidine-1-carboxylate
##STR00233##
[0557] To a solution of
trimethyl-[2-[[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl]-6-(-
3-methyl-1,2,4-thiadiazol-5-yl)pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]si-
lane (0.1 g, 175.23 umol, 1 eq) and benzyl
(2R,5R)-5-amino-2-methylpiperidine-1-carboxylate (43.51 mg, 175.23
umol, 1 eq) in THF (2.5 mL) was added DIEA (113.24 mg, 876.14 umol,
152.61 uL, 5 eq), the reaction mixture was stirred at 20.degree. C.
for 12 h. The final reaction mixture was concentrated to give a
residue. The residue was purified by silica gel column
(PE/EtOAc=20/1 to 5/1) to afford the title compound (90 mg) as a
yellow solid. (Note: The reaction was combined with another
reaction in 20 mg scale for work up.)
Step 7: Benzyl
(2S,5S)-2-methyl-5-[[4-[6-(3-methyl-1,2,4-thiadiazol-5-yl)-1H-pyrrolo[2,3-
-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carb-
oxylate
##STR00234##
[0559] To a solution of benzyl
(2S,5S)-2-methyl-5-[[4-[6-(3-methyl-1,2,4-thiadiazol-5-yl)-1-(2-trimethyl-
silylethoxymethyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-
-2-yl]amino]piperidine-1-carboxylate (0.085 g, 115.04 umol, 1 eq)
in THF (2 mL) was added TBAF (1 M, 1.73 mL, 15 eq) at 15.degree.
C., the reaction mixture was stirred at 40.degree. C. for 4 h. It
was partitioned between aq.NaHCO.sub.3 (15 mL) and EtOAc (25 mL).
The organic layer was washed with aq.NaHCO.sub.3 (15 mL.times.3),
dried over Na.sub.2SO.sub.4 and concentrated to give a residue. The
residue was purified by silica gel column (PE/EtOAc=20/1 to 5/1) to
afford the title compound (75 mg) of as a yellow solid. (Note: The
reaction was combined with another reaction in 5 mg scale for work
up.)
Step 8:
N-[(3R,6R)-6-methyl-3-piperidyl]-4-[6-(3-methyl-1,2,4-thiadiazol-5-
-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00235##
[0561] To a solution of benzyl
(2R,5R)-2-methyl-5-[[4-[6-(3-methyl-1,2,4-thiadiazol-5-yl)-1H-pyrrolo[2,3-
-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carb-
oxylate (68 mg, 111.73 umol, 1 eq) in DCM (3 mL) was added
BBr.sub.3 (139.95 mg, 558.63 umol, 53.83 uL, 5 eq) at 0.degree. C.,
the reaction mixture was stirred at 20.degree. C. for 0.33 h. The
resulting solution was quenched with H.sub.2O (0.2 mL), basified to
pH=7 with NaHCO.sub.3 and concentrated. The residue was purified by
HCl prep-HPLC to afford the title compound (14.3 mg, 94.3% purity)
as a white solid.
Example 49. Synthesis of
4-[6-(4-methyloxazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3S)-3-piper-
idyl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound 161)
Step 1: 2-[(6-bromopyrrolo[2,3-b]pyridin-1-yl)
methoxy]ethyl-trimethyl-silane
##STR00236##
[0563] To a solution of 6-bromo-1H-pyrrolo[2,3-b]pyridine (2 g,
10.15 mmol, 1 eq) in DMF (18 mL) and THF (2 mL) was added NaH
(609.04 mg, 15.23 mmol, 60% purity, 1.5 eq) at 0.degree. C. The
mixture was stirred at 0.degree. C. for 0.5 h. Then the
2-(chloromethoxy) ethyl-trimethyl-silane (2.03 g, 12.18 mmol, 2.16
mL, 1.2 eq) was added. The resulting mixture was stirred at
15.degree. C. for another 11.5 h. The solution was poured into
H.sub.2O (100 mL), extracted by EtOAc (50 mL.times.3). The combined
organic layers were washed by brine (100 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to afford the title compound (3.3 g, crude) as yellow oil, which
was used into the next step without further purification.
Step 2:
1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridine-6-carbonitrile
##STR00237##
[0565] 2-[(6-bromopyrrolo[2,3-b]pyridin-1-yl)
methoxy]ethyl-trimethyl-silane (0.3 g, 916.61 umol, 1 eq),
4-methyloxazole-5-carboxylic acid (233.00 mg, 1.83 mmol, 2 eq) and
palladium; tritert-butylphosphane (23.42 mg, 45.83 umol, 0.05 eq),
tetrabutylammonium; chloride; hydrate (271.26 mg, 916.61 umol, 1
eq), Cs.sub.2CO.sub.3 (447.98 mg, 1.37 mmol, 1.5 eq) were taken up
into a microwave tube in DMF (9 mL). The sealed tube was heated at
170.degree. C. for 15 min under microwave. The reaction mixture was
poured into H.sub.2O (100 mL) and extracted with EtOAc (50
mL.times.3). The combined organic layers were washed with brine
(150 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by column chromatography (SiO.sub.2, PE/EtOAc=10/1 to
2/1) to afford the title compound (0.35 g, 1.06 mmol, 23.18% yield,
5 batches in parallel) as yellow oil.
Step 3: 2-[[3-bromo-6-(4-methyloxazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl-trimethyl-silane
##STR00238##
[0567] To a solution of trimethyl-[2-[[6-(4-methyloxazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane (0.25 g, 758.80
umol, 1 eq) in DCM (3 mL) was added NBS (81.03 mg, 455.28 umol, 0.6
eq). The mixture was stirred at 15.degree. C. for 1 h. The solution
was poured into H.sub.2O (20 mL) and extracted with DCM (10
mL.times.3). The combined organic layers were washed with brine (30
mL.times.2), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. The residue was purified
by column chromatography (SiO.sub.2, PE/EtOAc=10/1 to 3/1) to
afford the title compound (250 mg) as yellow solid.
Step 4: trimethyl-[2-[[6-(4-methyloxazol-5-yl)-3-(4, 4, 5,
5-tetramethyl-1, 3, 2-dioxaborolan-2-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
##STR00239##
[0569] To a solution of 2-[[3-bromo-6-(4-methyloxazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl-trimethyl-silane (0.16 g,
391.81 umol, 1 eq) in dioxane (3.5 mL) was added BPD (149.24 mg,
587.71 umol, 1.5 eq), KOAc (76.91 mg, 783.61 umol, 2 eq) and
Pd(dppf)Cl.sub.2 (28.67 mg, 39.18 umol, 0.1 eq). The mixture was
stirred at 100.degree. C. for 1 h under N.sub.2. It was
concentrated under reduced pressure to give a residue. It was
purified by column chromatography (SiO.sub.2, PE/EtOAc=10/1 to 1/1)
to afford the title compound (0.2 g) as yellow oil.
Step 5: Tert-butyl
(3S)-3-[[4-[6-(4-methyloxazol-5-yl)-1-(2-trimethylsilylethoxymethyl)pyrro-
lo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine--
1-carboxylate
##STR00240##
[0571] To a solution of
trimethyl-[2-[[6-(4-methyloxazol-5-yl)-3-(4, 4, 5, 5-tetramethyl-1,
3, 2-dioxaborolan-2-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane (0.2 g, 439.15
umol, 1 eq) in THF (6 mL) and H.sub.2O (1 mL) was added tert-butyl
(3S)-3-[[4-chloro-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-ca-
rboxylate (167.22 mg, 439.15 umol, 1 eq), ditert-butyl(cyclopentyl)
phosphane:dichloropalladium:iron (28.62 mg, 43.91 umol, 0.1 eq) and
Na.sub.2CO.sub.3 (93.09 mg, 878.29 umol, 2 eq). The mixture was
stirred at 80.degree. C. for 2 h under N.sub.2. The solution was
concentrated under reduced pressure to give a residue. The residue
was purified by column chromatography (SiO.sub.2, PE/EtOAc=10/1 to
1/1) to afford the title compound (70 mg, 68% purity) as a white
solid. (Note: The reaction mixture was combined with another batch
in 20 mg scale for worked up and purification.)
Step 6:
4-[6-(4-methyloxazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3S)--
3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00241##
[0573] To a solution of tert-butyl
(3S)-3-[[4-[6-(4-methyloxazol-5-yl)-1-(2-trimethylsilylethoxymethyl)pyrro-
lo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine--
1-carboxylate (0.06 g, 89.05 umol, 1 eq) in dioxane (1 mL) was
added H.sub.2SO.sub.4 (87.34 mg, 890.47 umol, 47.47 uL, 10 eq). The
mixture was stirred at 40.degree. C. for 2 h. It was diluted with
H.sub.2O (10 mL) and adjusted pH to 12 with NaOH. Then the solution
was extracted with EtOAc (10 mL.times.3). The combined organic
layers were washed with brine (30 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
It was purified by prep-HPLC (FA condition) to afford the title
compound (5.5 mg, FA salt, 100% purity) as a white solid. (Note:
The reaction mixture was combined with another batch in 10 mg scale
for work-up and purification.)
Example 50. Synthesis of
4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3-
S,6S)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 162)
Step 1: 1H-pyrrolo[2,3-b]pyridine-6-carbonitrile
##STR00242##
[0575] A mixture of 6-bromo-1H-pyrrolo[2,3-b]pyridine (10 g, 50.75
mmol, 1 eq), Zn (331.88 mg, 5.08 mmol, 0.1 eq), Zn(CN).sub.2 (4.17
g, 35.53 mmol, 2.26 mL, 0.7 eq) and
Pd(dppf)Cl.sub.2.CH.sub.2Cl.sub.2 (2.07 g, 2.54 mmol, 0.05 eq) in
DMF (100 mL) was degassed and purged with N.sub.2 for 3 times, and
then the mixture was stirred at 140.degree. C. for 5 h under
N.sub.2 atmosphere. It was diluted with water (500 mL) and EtOAc
(300 mL), and then filtered to remove the solid, the solid was
washed with EtOAc (200 mL.times.2). The combined organic layers
were washed with brine (500 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
The residue was purified by column chromatography (SiO.sub.2,
PE/EtOAc=5/1 to 2/1) to afford the title compound (5.3 g, 89%
purity) as a green solid.
Step 2: 3-bromo-1H-pyrrolo[2,3-b]pyridine-6-carbonitrile
##STR00243##
[0577] To a solution of 1H-pyrrolo[2,3-b]pyridine-6-carbonitrile (4
g, 25.15 mmol, 1 eq) in DCM (40 mL) was added NBS (5.37 g, 30.18
mmol, 1.2 eq) at 0.degree. C. The mixture was stirred at 15.degree.
C. for 1 h. It was concentrated and the residue was washed with PE
(500 mL) and the solid was filtered to afford the title compound (5
g, crude) as a yellow solid.
Step 3:
1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridine-6-carbonitrile
##STR00244##
[0579] To a solution of
3-bromo-1H-pyrrolo[2,3-b]pyridine-6-carbonitrile (5 g, 22.52 mmol,
1 eq) in THF (50 mL) was added NaH (900.65 mg, 22.52 mmol, 60%
purity, 1 eq) at 0.degree. C. The mixture was stirred at 0.degree.
C. for 0.5 h. And then benzenesulfonyl chloride (5.17 g, 29.27
mmol, 3.75 mL, 1.3 eq) was added into the mixture was stirred at
15.degree. C. for 1 h. It was quenched by addition water (200 mL)
at 15.degree. C., and then extracted with EtOAc (100 mL.times.2).
The combined organic layers were washed with brine (200 mL), dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to give a residue. The residue was purified by column
chromatography (SiO.sub.2, Petroleum ether/Ethyl acetate=7/1 to
2/1) to afford a crude product (4.5 g). Then it was triturated with
EtOAc (5 mL) and PE (50 mL), the solid was filtered and dried to
afford the title compound (4 g, 95% purity) as a white solid.
Step 4: 1-(benzenesulfonyl)-3-(4, 4, 5, 5-tetramethyl-1, 3,
2-dioxaborolan-2-yl)pyrrolo[2,3-b]pyridine-6-carbonitrile
##STR00245##
[0581] A mixture of
1-(benzenesulfonyl)-3-bromo-pyrrolo[2,3-b]pyridine-6-carbonitrile
(4 g, 11.04 mmol, 1 eq), BPD (4.21 g, 16.57 mmol, 1.5 eq), KOAc
(2.17 g, 22.09 mmol, 2 eq), Pd(dppf)Cl.sub.2 (1.21 g, 1.66 mmol,
0.15 eq) in dioxane (40 mL) was degassed and purged with N.sub.2
for 5 times, and then the mixture was stirred at 100.degree. C. for
2 h under N.sub.2 atmosphere. It was filtered, and washed by EtOAc
(200 mL.times.3). The combined organic layers were washed with
brine (300 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was combined with the solid and purified by column chromatography
(SiO.sub.2, Petroleum ether/Ethyl acetate=6/1 to 3/1) to afford the
title compound (4 g, 85% purity) as an orange solid.
Step 5:
1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimid-
in-4-yl]pyrrolo[2,3-b]pyridine-6-carbonitrile
##STR00246##
[0583] A mixture of 1-(benzenesulfonyl)-3-(4, 4, 5,
5-tetramethyl-1, 3,
2-dioxaborolan-2-yl)pyrrolo[2,3-b]pyridine-6-carbonitrile (2 g,
4.89 mmol, 1 eq),
4-chloro-2-methylsulfanyl-5-(trifluoromethyl)pyrimidine (1.68 g,
7.33 mmol, 1.5 eq), ditert-butyl (cyclopentyl)
phosphane:dichloropalladium:iron (318.50 mg, 488.68 umol, 0.1 eq),
Na.sub.2CO.sub.3 (1.04 g, 9.77 mmol, 2 eq) in THF (60 mL) and
H.sub.2O (6 mL) was degassed and purged with N.sub.2 for 5 times,
and then the mixture was stirred at 80.degree. C. for 1 h under
N.sub.2 atmosphere. It was filtered to remove the solid, and washed
with EtOAc (250 mL.times.2). The filtrate was washed with brine
(500 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to give a residue. The residue was purified
by column chromatography (SiO.sub.2, PE/EtOAc=6/1 to 2/1) to afford
the title compound (0.8 g) as a white solid. (Note: The reaction
was combined with another reaction in 100 mg scale for
work-up.)
Step 6:
1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimid-
in-4-yl]pyrrolo[2,3-b]pyridine-6-carbonitrile
##STR00247##
[0585] To a solution of
1-(benzenesulfonyl)-3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-yl-
]pyrrolo[2,3-b]pyridine-6-carbonitrile (0.06 g, 115.05 umol, 1 eq,
FA) in DCM (4 mL) was added m-CPBA (51.39 mg, 253.12 umol, 85%
purity, 2.2 eq) and Na.sub.2SO.sub.4 (18.23 mg, 128.38 umol, 13.02
uL, 1.12 eq). The mixture was stirred at 15.degree. C. for 2 h. The
reaction mixture was poured into the saturated Na.sub.2SO.sub.3 and
NaHCO.sub.3 solution (200 mL), then extracted with DCM (100
mL.times.3), the combined organic layers were washed with brine
(300 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure to afford the title compound (900 mg, crude)
as a white solid which was used directly into the next step without
purification. (Note: The reaction was combined with another
reaction in 800 mg scale for work-up.)
Step 7: Benzyl
(2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-cyano-pyrrolo[2,3-b]pyridin-3-yl]-5--
(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carboxylate
##STR00248##
[0587] To a solution of
1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl-
]pyrrolo[2,3-b]pyridine-6-carbonitrile (0.42 g, 827.64 umol, 1 eq)
in THF (4.6 mL) was added DIPEA (213.93 mg, 1.66 mmol, 288.31 uL, 2
eq) and benzyl (2S,5S)-5-amino-2-methyl-piperidine-1-carboxylate
(246.62 mg, 993.17 umol, 1.2 eq). The mixture was stirred at
15.degree. C. for 1 h. It was concentrated under reduced pressure
to remove solvent. The residue was diluted with water (20 mL) and
extracted with EtOAc (20 mL.times.2). The combined organic layers
were washed with brine (40 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
The residue was purified by column chromatography (SiO.sub.2,
PE/EtOAc=4/1 to 2/1) to afford the title compound (300 mg, 80%
purity) as a white solid. (Note. The reaction was combined with
another reaction in 40 mg scale for work-up.)
Step 8: Benzyl
(2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-[(E)-N'-hydroxycarbamimidoyl]pyrrolo-
[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-pip-
eridine-1-carboxylate
##STR00249##
[0589] To a solution of benzyl
(2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-cyano-pyrrolo[2,3-b]pyridin-3-yl]-5--
(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carboxylate
(300.00 mg, 444.00 umol, 1 eq) in EtOH (20 mL) was added
hydroxylamine; hydrochloride (308.54 mg, 4.44 mmol, 10 eq) and
Na.sub.2CO.sub.3 (564.71 mg, 5.33 mmol, 12 eq). The mixture was
stirred at 30.degree. C. for 16 h. It was filtered, and the
filtrate was concentrated under reduced pressure to afford the
title compound (400 mg, crude) as a white solid which was used
directly into the next step without purification.
Step 9: Benzyl (2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-(5-methyl-1,
2,4-oxadiazol-3-yl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimid-
in-2-yl]amino]-2-methyl-piperidine-1-carboxylate
##STR00250##
[0591] Benzyl
(2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-[(E)-N'-hydroxycarbamimidoyl]pyrrolo-
[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-pip-
eridine-1-carboxylate (0.36 g, 507.97 umol, 1 eq) in 1, 1,
1-triethoxyethane (0.4 mL) was added TFA (2.90 mg, 25.40 umol, 1.88
uL, 0.05 eq). The mixture was stirred at 15.degree. C. for 10 min.
Then the mixture was stirred at 60.degree. C. for 30 min. The
solution was concentrated under reduced pressure to give a residue.
The residue was purified by column chromatography (SiO.sub.2,
PE/EtOAc=2/1) to afford the title compound (220 mg, 96% purity) as
a white solid.
Step 10: Benzyl
(2S,5S)-2-methyl-5-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3--
b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carbo-
xylate
##STR00251##
[0593] To a solution of benzyl
(2S,5S)-5-[[4-[1-(benzenesulfonyl)-6-(5-methyl-1,2,4-oxadiazol-3-yl)pyrro-
lo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-p-
iperidine-1-carboxylate (0.2 g, 272.95 umol, 1 eq) in MeOH (24 mL)
was added NaOH (2 M, 4.09 mL, 30 eq). The mixture was stirred at
60.degree. C. for 0.5 h. The reaction mixture was concentrated
under reduced pressure to remove solvent. It was diluted with water
30 mL, adjusted pH to 7 by added HCl (1 M), and then extracted with
EtOAc (30 mL). The organic layers were washed with brine (30 mL),
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to afford the title compound (180 mg, crude) as a
white solid. The crude product was used directly into the next step
without purification. (Note: The reaction was combined with another
reaction in 20 mg scale for work-up.)
Step 11: 4-[6-(5-methyl-1,
2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3S,6S)-6-methyl-3--
piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00252##
[0594] To a solution of benzyl
(2S,5S)-2-methyl-5-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3--
b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carbo-
xylate (160 mg, 270.01 umol, 1 eq) in DCM (3.2 mL) was added
BBr.sub.3 (676.43 mg, 2.70 mmol, 260.17 uL, 10 eq) at 0.degree. C.,
and then the mixture was stirred at this temperature for 1 h. It
was concentrated under reduced pressure to give a residue, and then
it was washed with PE (160 mL) and collected the cake. The crude
product was purified by prep-HPLC (FA condition) to afford the
title compound (26.7 mg, FA, 100% purity, 100% ee value) as a white
solid. (Note: The reaction was combined with another reaction in 20
mg scale for work-up.)
Example 51. Synthesis of
4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3-
R, 6R)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 163)
Step 1: Benzyl (2R,
5R)-5-[[4-[1-(benzenesulfonyl)-6-cyano-pyrrolo[2,3-b]pyridin-3-yl]-5-(tri-
fluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carboxylate
##STR00253##
[0596] To a solution of
1-(benzenesulfonyl)-3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl-
]pyrrolo[2,3-b]pyridine-6-carbonitrile (400 mg, 788.23 umol, 1 eq)
in THF (4 mL) was added DIPEA (203.75 mg, 1.58 mmol, 274.59 uL, 2
eq) and benzyl (2R, 5R)-5-amino-2-methyl-piperidine-1-carboxylate
(234.88 mg, 945.88 umol, 1.2 eq). The mixture was stirred at
15.degree. C. for 1 h. It was concentrated under reduced pressure
to remove solvent. The residue was diluted with water 20 mL and
extracted with EtOAc (20 mL.times.2). The combined organic layers
were washed with brine (40 mL), dried over Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to give a residue.
It was purified by column chromatography (SiO.sub.2, PE/EtOAc=4/1
to 2/1) to afford the title compound (350 mg, 70% purity) as a
white solid.
Step 2: Benzyl
(2R,5R)-5-[[4-[1-(benzenesulfonyl)-6-[(E)-N'-hydroxycarbamimidoyl]pyrrolo-
[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-pip-
eridine-1-carboxylate
##STR00254##
[0598] To a solution of benzyl
(2R,5R)-5-[[4-[1-(benzenesulfonyl)-6-cyano-pyrrolo[2,3-b]pyridin-3-yl]-5--
(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carboxylate
(350.00 mg, 518.00 umol, 1 eq) in EtOH (23 mL) was added
hydroxylamine; hydrochloride (359.96 mg, 5.18 mmol, 10 eq) and
Na.sub.2CO.sub.3 (658.82 mg, 6.22 mmol, 12 eq). The mixture was
stirred at 30.degree. C. for 16 h. It was filtered, and the
filtrate was concentrated under reduced pressure to afford the
title compound (400 mg, crude) as a white solid which was used
directly into the next step without purification.
Step 3: Benzyl (2R,
5R)-5-[[4-[1-(benzenesulfonyl)-6-(5-methyl-1,2,4-oxadiazol-3-yl)pyrrolo[2-
,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piper-
idine-1-carboxylate
##STR00255##
[0600] To a solution of benzyl (2R,
5R)-5-[[4-[1-(benzenesulfonyl)-6-[(E)-N'-hydroxycarbamimidoyl]pyrrolo[2,3-
-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperid-
ine-1-carboxylate (400.00 mg, 564.41 umol, 1 eq) in AcOH (4 mL) was
added Ac.sub.2O (69.14 mg, 677.29 umol, 63.44 uL, 1.2 eq). The
mixture was stirred at 90.degree. C. for 16 h. It was residue was
diluted with water 30 mL. The solid was formed and filtered to
collect the cake. The crude product was purified by column
chromatography (SiO.sub.2, Petroleum ether/Ethyl acetate=5/1 to
3/1) to afford the title compound (180 mg, 94% purity) as a white
solid.
Step 4: Benzyl (2R,
5R)-2-methyl-5-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3-b]py-
ridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxyla-
te
##STR00256##
[0602] To a solution of benzyl (2R,
5R)-5-[[4-[1-(benzenesulfonyl)-6-(5-methyl-1,2,4-oxadiazol-3-yl)pyrrolo[2-
,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piper-
idine-1-carboxylate (0.1 g, 136.48 umol, 1 eq) in MeOH (12 mL) was
added NaOH (2 M, 2.05 mL, 30 eq). The mixture was stirred at
60.degree. C. for 0.5 h. It was concentrated under reduced pressure
to remove solvent. The residue was diluted with water 30 mL,
adjusted pH to 7 by added HCl (1 M), and then extracted with EtOAc
(30 mL). The organic layers were washed with brine (30 mL), dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to afford the title compound (110 mg, crude) as a white
solid. The crude product was used directly into the next step
without purification. (Note: The reaction was combined with another
reaction in 20 mg scale for work-up and purification.)
Step 5: 4-[6-(5-methyl-1,
2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-N-[(3R,
6R)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00257##
[0604] To a solution of benzyl (2R,
5R)-2-methyl-5-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrrolo[2,3-b]py-
ridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxyla-
te (80 mg, 135.01 umol, 1 eq) in DCM (1.6 mL) was added BBr.sub.3
(338.22 mg, 1.35 mmol, 130.08 uL, 10 eq) at 0.degree. C., and then
the mixture was stirred at this temperature for 1 h. The reaction
mixture was concentrated under reduced pressure to give a residue,
and then the residue was triturated with PE (60 mL), filtered and
dried. The crude product was purified by prep-HPLC (FA condition)
to afford the title compound (17.3 mg, FA, 100% purity, 100% ee
value) as a white solid. (Note: The reaction mixture was combined
with another batch in 15 mg scale for worked up and
purification.)
Example 52. Synthesis of N-[(3R,
6R)-6-methyl-3-piperidyl]-4-[6-(1-methyltetrazol-5-yl)-1H-pyrrolo[2,3-b]p-
yridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
164)
Step 1: Benzyl (2R,
5R)-2-methyl-5-[[4-[6-(1-methyltetrazol-5-yl)-1-(2-trimethylsilylethoxyme-
thyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]p-
iperidine-1-carboxylate
##STR00258##
[0606] To a solution of
trimethyl-[2-[[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl]-6-(-
1-methyltetrazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane (0.27 g, 486.82
umol, 1 eq), benzyl (2R,
5R)-5-amino-2-methyl-piperidine-1-carboxylate (181.33 mg, 730.23
umol, 1.5 eq) in THF (5 mL) was added DIEA (188.75 mg, 1.46 mmol,
254.38 uL, 3 eq). The mixture was stirred at 20.degree. C. for 2 h.
The reaction mixture was concentrated to give a residue. It was
purified by column chromatography (SiO.sub.2, PE/EtOAc=10/1 to 2/1)
to afford the title compound (0.22 g, 295.23 umol, 60.64% yield,
97% purity) as a yellow solid.
Step 2: N-[(3R,
6R)-6-methyl-3-piperidyl]-4-[6-(1-methyltetrazol-5-yl)-1H-pyrrolo[2,3-b]p-
yridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00259##
[0608] To a solution of benzyl (2R,
5R)-2-methyl-5-[[4-[6-(1-methyltetrazol-5-yl)-1-(2-trimethylsilylethoxyme-
thyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]p-
iperidine-1-carboxylate (0.22 g, 304.36 umol, 1 eq) in dioxane (5
mL) was added H.sub.2SO.sub.4 (4.48 g, 45.65 mmol, 2.43 mL, 150
eq). The mixture was stirred at 45.degree. C. for 2 h. The mixture
was adjusted pH to 8 with saturated aqueous NaOH, and extracted
with EtOAc (50 mL.times.3), dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give a residue. It was
purified by prep-HPLC (FA condition) to afford the title compound
(31.4 mg, 56.64 umol, 18.610% yield, 910% purity, FA) as a white
solid.
Example 53. Synthesis of
N-[(3S,6S)-6-methyl-3-piperidyl]-4-[6-(1-methyltetrazol-5-yl)-1H-pyrrolo[-
2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
165)
Step 1: 5-iodo-1-methyl-tetrazole
##STR00260##
[0610] To a solution of 1-methyltetrazole (3 g, 35.68 mmol, 1 eq)
in THF (90 mL) was added dropwise n-BuLi (2.5 M, 18.55 mL, 1.3 eq)
at -78.degree. C. under N.sub.2 atmosphere. After addition, the
mixture was stirred at this temperature for 30 min, and then
molecular iodine (9.06 g, 35.68 mmol, 7.19 mL, 1 eq) was added at
-78.degree. C. The resulting mixture was stirred at 15.degree. C.
for 1 h. The mixture was diluted with H.sub.2O (200 mL) and
extracted with EtOAc (100 mL.times.3). The combined organic layers
were washed with brine (300 mL.times.2), dried over
Na.sub.2SO.sub.4 filtered and concentrated under reduced pressure
to afford the title compound (3 g, crude) as brown oil.
Step 2: Trimethyl-[2-[[6-(1-methyltetrazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
##STR00261##
[0612] A mixture of
[1-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridin-6-yl]boronic
acid (3.96 g, 13.55 mmol, 1 eq), 5-iodo-1-methyl-tetrazole (3.7 g,
17.62 mmol, 1.3 eq), Cs.sub.2CO.sub.3 (8.83 g, 27.11 mmol, 2 eq),
Pd(PPh.sub.3).sub.4 (1.57 g, 1.36 mmol, 0.1 eq) in dioxane (70 mL)
and H.sub.2O (7 mL) was degassed and purged with N.sub.2 for 3
times, and then the mixture was stirred at 100.degree. C. for 1 h
under N.sub.2 atmosphere. It was diluted with H.sub.2O 300 mL and
extracted with EtOAc (100 mL.times.3). The combined organic layers
were washed with brine (300 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue that was purified by column chromatography
(SiO.sub.2, PE/EtOAc=10/1 to 4/1) to afford the title compound (2
g) as a yellow solid. (Note: The reaction was combined with another
reaction in 500 mg scale for work up.)
Step 3: 2-[[3-bromo-6-(1-methyltetrazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl-trimethyl-silane
##STR00262##
[0614] To a solution of trimethyl-[2-[[6-(1-methyltetrazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane (1.7 g, 5.14 mmol,
1 eq) in DMF (40 mL) was added NBS (915.59 mg, 5.14 mmol, 1 eq).
The mixture was stirred at 15.degree. C. for 1 h. The mixture was
diluted with H.sub.2O 200 mL and extracted with EtOAc (100
mL.times.3). The combined organic layers were washed with brine
(300 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. It was
washed with MTBE (8 mL) and filtered to afford the title compound
(1.9 g) as a red solid. (Note: The reaction was combined with
another reaction in 300 mg scale for work up.)
Step 4: Trimethyl-[2-[[6-(1-methyltetrazol-5-yl)-3-(4, 4, 5,
5-tetramethyl-1, 3, 2-dioxaborolan-2-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
##STR00263##
[0616] A mixture of 2-[[3-bromo-6-(1-methyltetrazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl-trimethyl-silane (1.8 g,
4.40 mmol, 1 eq), 4, 4, 5, 5-tetramethyl-2-(4, 4, 5,
5-tetramethyl-1, 3, 2-dioxaborolan-2-yl)-1, 3, 2-dioxaborolane
(2.23 g, 8.79 mmol, 2 eq), AcOK (863.07 mg, 8.79 mmol, 2 eq) and
Pd(dppf)Cl.sub.2 (321.74 mg, 439.72 umol, 0.1 eq) in dioxane (40
mL) was degassed and purged with N.sub.2 for 3 times, and then the
mixture was stirred at 100.degree. C. for 2 h under N.sub.2
atmosphere. The solution was filtered and the filtrate was
concentrated to give a residue. It was purified by column
chromatography (SiO.sub.2, PE/EtOAc=10/1 to 5/1) to give a residue.
Then the residue was purified by reverse column. The eluent
solution was adjusted pH to 8 with NaHCO.sub.3 and extracted with
EtOAc (60 mL.times.3), The combined organic layers were washed with
brine (200 mL.times.2), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to afford the title compound
(700 mg) as a white solid.
Step 5:
Trimethyl-[2-[[3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4--
yl]-6-(1-methyltetrazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
##STR00264##
[0618] A mixture of trimethyl-[2-[[6-(1-methyltetrazol-5-yl)-3-(4,
4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane (0.05 M, 28.48 mL,
1 eq), 4-chloro-2-methylsulfanyl-5-(trifluoromethyl)pyrimidine
(488.38 mg, 2.14 mmol, 1.5 eq), Na.sub.2CO.sub.3 (301.89 mg, 2.85
mmol, 2 eq) and ditert-butyl (cyclopentyl)
phosphane:dichloropalladium:iron (92.82 mg, 142.41 umol, 0.1 eq) in
THF (30 mL) and H.sub.2O (3 mL) was degassed and purged with
N.sub.2 for 3 times, and then the mixture was stirred at 80.degree.
C. for 1 h under N.sub.2 atmosphere. It was diluted with H.sub.2O
80 mL and extracted with EtOAc (30 mL.times.3). The combined
organic layers were washed with brine (60 mL.times.2), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, PE/EtOAc=10/1 to 5/1) to afford the
title compound (660 mg) as yellow oil. (Note: The reaction was
combined with another reaction in 50 mg scale for work up.)
Step 6:
Trimethyl-[2-[[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4--
yl]-6-(1-methyltetrazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane
##STR00265##
[0620] To a solution of
trimethyl-[2-[[3-[2-methylsulfanyl-5-(trifluoromethyl)pyrimidin-4-yl]-6-(-
1-methyltetrazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane (0.66 g, 1.26 mmol,
1 eq) in DCM (10 mL) was added m-CPBA (640.98 mg, 3.16 mmol, 85%
purity, 2.5 eq). The mixture was stirred at 15.degree. C. for 3 h.
The mixture was washed with Sat.Na.sub.2SO.sub.3 (30 mL),
Sat.NaHCO.sub.3 (30 mL), brine (50 mL). The organic layers were
dried over Na.sub.2SO.sub.4 and concentrated to give a residue. The
residue was washed with PE/EtOAc=5/1 (8 mL) and filtered to afford
the title compound (0.54 g, crude) as a yellow solid.
Step 7: Benzyl
(2S,5S)-2-methyl-5-[[4-[6-(1-methyltetrazol-5-yl)-1-(2-trimethylsilyletho-
xymethyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]ami-
no]piperidine-1-carboxylate
##STR00266##
[0622] To a solution of
trimethyl-[2-[[3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl]-6-(-
1-methyltetrazol-5-yl)
pyrrolo[2,3-b]pyridin-1-yl]methoxy]ethyl]silane (0.27 g, 486.82
umol, 1 eq), benzyl
(2S,5S)-5-amino-2-methyl-piperidine-1-carboxylate (145.06 mg,
584.18 umol, 1.2 eq) in THF (5 mL) was added DIEA (188.75 mg, 1.46
mmol, 254.38 uL, 3 eq). The mixture was stirred at 20.degree. C.
for 2 h. The resulting solution was concentrated to give a residue.
It was purified by column chromatography (SiO.sub.2, PE/EtOAc=10/1
to 2/1) to afford the title compound (0.26 g, 348.90 umol, 71.67%
yield, 97% purity) as a yellow solid.
Step 8:
N-[(3S,6S)-6-methyl-3-piperidyl]-4-[6-(1-methyltetrazol-5-yl)-1H-p-
yrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00267##
[0624] To a solution of benzyl
(2S,5S)-2-methyl-5-[[4-[6-(1-methyltetrazol-5-yl)-1-(2-trimethylsilyletho-
xymethyl)pyrrolo[2,3-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]ami-
no]piperidine-1-carboxylate (0.25 g, 345.86 umol, 1 eq) in dioxane
(5 mL) was added H.sub.2SO.sub.4 (5.09 g, 51.88 mmol, 2.77 mL, 150
eq). The mixture was stirred at 45.degree. C. for 2 h. The solution
was adjusted pH to 8 with saturated aqueous NaOH, and extracted
with EtOAc (50 mL.times.3), dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to give a residue. The
residue was purified by prep-HPLC (FA condition) to afford the
title compound (30.9 mg, FA) as a white solid. (Note: The reaction
was combined with another reaction in 10 mg scale for
purification).
Example 54. Synthesis of
4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-N-[(-
3S,6S)-6-methyl-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 167)
Step 1: Benzyl
(2S,5S)-5-[[4-(6-cyano-1-tetrahydropyran-2-yl-pyrazolo[3,4-b]pyridin-3-yl-
)-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carboxyla-
te
##STR00268##
[0626] To a solution of
3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl]-1-tetrahydropyran--
2-yl-pyrazolo[3,4-b]pyridine-6-carbonitrile (150 mg, 331.56 umol, 1
eq) in THF (2 mL) was added benzyl
(2S,5S)-5-amino-2-methyl-piperidine-1-carboxylate (98.80 mg, 397.87
umol, 1.2 eq) and DIEA (428.52 mg, 3.32 mmol, 577.51 uL, 10 eq).
The mixture was stirred at 15.degree. C. for 1 h. It was poured
into water (5 mL) and extracted with EtOAc (10 mL.times.3). The
combined organic layer was washed with brine (10 mL.times.3) and
sat. NaHCO.sub.3 (20 mL), dried over Na.sub.2SO.sub.4, filtered.
The filtrate was concentrated under reduced pressure to give a
residue. The residue was purified by MPLC (SiO.sub.2, PE/EtOAc=3/1
to 1/1) to afford the title compound (85 mg, crude) as white
solid.
Step 2: Benzyl
(2S,5S)-5-[[4-[6-[(Z)--N'-hydroxycarbamimidoyl]-1-tetrahydropyran-2-yl-py-
razolo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-meth-
yl-piperidine-1-carboxylate
##STR00269##
[0628] To a mixture of benzyl
(2S,5S)-5-[[4-(6-cyano-1-tetrahydropyran-2-yl-pyrazolo[3,4-b]pyridin-3-yl-
)-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-methyl-piperidine-1-carboxyla-
te (85.00 mg, 136.96 umol, 1 eq) in EtOH (2 mL) was added
Na.sub.2CO.sub.3 (174.19 mg, 1.64 mmol, 12 eq) and NH.sub.2OH.HCl
(95.17 mg, 1.37 mmol, 10 eq). The mixture was stirred at 30.degree.
C. for 12 h. It was 10 filtered and concentrated under reduced
pressure to afford the title compound (100 mg, crude) as yellow
solid which was used next step without further purification. (Note:
The reaction was combined with another reaction in 200 mg scale for
work up.)
Step 3: Benzyl
(2S,5S)-2-methyl-5-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1-tetrahydropyr-
an-2-yl-pyrazolo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]ami-
no]piperidine-1-carboxylate
##STR00270##
[0630] To a solution of benzyl
(2S,5S)-5-[[4-[6-[(Z)--N'-hydroxycarbamimidoyl]-1-tetrahydropyran-2-yl-py-
razolo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-2-meth-
yl-piperidine-1-carboxylate (100 mg, 152.99 umol, 1 eq) in 1, 1,
1-triethoxyethane (885.00 mg, 5.46 mmol, 1 mL, 35.66 eq) was added
TFA (3.49 mg, 30.60 umol, 2.27 uL, 0.2 eq) at 15.degree. C. The
mixture was stirred at 15.degree. C. for 10 min, then the mixture
was stirred at 60.degree. C. for 50 min. The mixture was
concentrated to obtain the residue. It was purified by MPLC
(SiO.sub.2, PE/EtOAc=5/1 to 1/1) to afford the title compound (50
mg, 67.51 umol, 44.13% yield, 91.5% purity) as brown solid.
Step 4: Benzyl
(2S,5S)-2-methyl-5-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrazolo[3,4-
-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carb-
oxylate
##STR00271##
[0632] To a solution of benzyl
(2S,5S)-2-methyl-5-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1-tetrahydropyr-
an-2-yl-pyrazolo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]ami-
no]piperidine-1-carboxylate (50.00 mg, 73.78 umol, 1 eq) in
HCl/MeOH (2 mL) was stirred at 60.degree. C. for 12 h. The reaction
mixture was concentrated under reduced pressure to afford the title
compound (50 mg, crude) as yellow solid.
Step 5: 4-[6-(5-methyl-1,
2,4-oxadiazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-N-[(3S,6S)-6-methyl-3-
-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine
##STR00272##
[0634] To a solution of benzyl
(2S,5S)-2-methyl-5-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrazolo[3,4-
-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carb-
oxylate (50.00 mg, 84.24 umol, 1 eq) in DCM (2 mL) was added
BBr.sub.3 (105.52 mg, 421.19 umol, 40.58 uL, 5 eq) at 0.degree. C.
The mixture was stirred at 0.degree. C. for 1 h. The reaction
mixture was concentrated under reduced pressure to give a residue.
It was purified by prep-HPLC (HCl condition) to afford the title
compound (12.7 mg, 23.76 umol, 28.21% yield, 92.78% purity, HCl) as
white solid.
Example 55. Synthesis of
4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-N-[(-
3S)-3-piperidyl]-5-(trifluoromethyl)pyrimidin-2-amine (Compound
168)
Step 1: Tert-butyl
(3S)-3-[[4-(6-cyano-1-tetrahydropyran-2-yl-pyrazolo[3,4-b]pyridin-3-yl)-5-
-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
##STR00273##
[0636] To a solution of
3-[2-methylsulfonyl-5-(trifluoromethyl)pyrimidin-4-yl]-1-tetrahydropyran--
2-yl-pyrazolo[3,4-b]pyridine-6-carbonitrile (160 mg, 353.66 umol, 1
eq) in THF (1.6 mL) was added tert-butyl
(3S)-3-aminopiperidine-1-carboxylate (77.91 mg, 389.03 umol, 1.1
eq), DIEA (228.54 mg, 1.77 mmol, 308.01 uL, 5 eq) at 15.degree. C.
The mixture was stirred at 15.degree. C. for 1 h. It was poured
into water (10 mL) and extracted with EtOAc (10 mL.times.3). The
combined organic layer was washed with brine (10 mL.times.3), dried
over Na.sub.2SO.sub.4, filtered. The filtrate was concentrated
under reduced pressure to give a residue. It was purified by MPLC
(SiO.sub.2, PE/EtOAc=3/1 to 1/1) to afford the title compound (200
mg) as yellow solid.
Step 2: Tert-butyl
(3S)-3-[[4-[6-[(Z)--N'-hydroxycarbamimidoyl]-1-tetrahydropyran-2-yl-pyraz-
olo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-
-1-carboxylate
##STR00274##
[0638] To a solution of tert-butyl
(3S)-3-[[4-(6-cyano-1-tetrahydropyran-2-yl-pyrazolo[3,4-b]pyridin-3-yl)-5-
-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-1-carboxylate
(200 mg, 349.30 umol, 1 eq) in EtOH (5 mL) was added NH.sub.2OH.HCl
(242.73 mg, 3.49 mmol, 10 eq) and Na.sub.2CO.sub.3 (444.26 mg, 4.19
mmol, 12 eq). The mixture was stirred at 30.degree. C. for 12 h. It
was filtered and concentrated under reduced pressure to afford the
title compound (200 mg, crude) as yellow solid which was used next
step directly. (Note: The reaction was combined with another
reaction in 200 mg scale for work up.)
Step 3: Tert-butyl
(3S)-3-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1-tetrahydropyran-2-yl-pyra-
zolo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-
e-1-carboxylate
##STR00275##
[0640] To a mixture of tert-butyl
(3S)-3-[[4-[6-[(Z)--N'-hydroxycarbamimidoyl]-1-tetrahydropyran-2-yl-pyraz-
olo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidine-
-1-carboxylate (180 mg, 297.22 umol, 1 eq) and 1, 1,
1-triethoxyethane (964.36 mg, 5.94 mmol, 1.09 mL, 20 eq) was added
TFA (3.39 mg, 29.72 umol, 2.20 uL, 0.1 eq). The mixture was stirred
at 15.degree. C. for 10 min, and then the mixture was stirred at
60.degree. C. for 50 min. It was concentrated. The residue was
purified by MPLC (SiO.sub.2, PE/EtOAc=4/1 to 1/1) to afford the
title compound (100 mg, 93.7% purity) as white solid.
Step 4: 4-[6-(5-methyl-1,
2,4-oxadiazol-3-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-N-[(3S)-3-piperidyl]--
5-(trifluoromethyl)pyrimidin-2-amine
##STR00276##
[0642] A solution of tert-butyl
(3S)-3-[[4-[6-(5-methyl-1,2,4-oxadiazol-3-yl)-1-tetrahydropyran-2-yl-pyra-
zolo[3,4-b]pyridin-3-yl]-5-(trifluoromethyl)pyrimidin-2-yl]amino]piperidin-
e-1-carboxylate (100 mg, 158.82 umol, 1 eq) in HCl/MeOH (3 mL) was
stirred at 60.degree. C. for 12 h. It was concentrated under
reduced pressure to give a residue. It was purified by prep-HPLC
(HCl condition) to afford the title compound (42 mg, 86.40 umol,
54.40% yield, 99.13% purity, HCl) as white solid.
Example 56. Synthesis of
4-(6-(3,5-dimethylisoxazol-4-yl)-5-fluoro-1H-indol-3-yl)-N--((S)-piperidi-
n-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (Compound 134)
Step 1:
6-bromo-3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-5-fluoro-1H-
-indole
##STR00277##
[0644] To a andrt solution of
2,4-dichloro-5-trifluoromethylpyrimidine (945 uL, 7.01 mmol) in
anh. (anhydrous) DCE (3 mL) under nitrogen, was added aluminum
trichloride (472 mg, 3.51 mmol), and the resulting mixture was
stirred at 80.degree. C. for 30 min. After this time, the reaction
mixture was cooled to RT, and 6-bromo-5-fluoro indole (500 mg, 2.34
mmol) was added. The resulting mixture was stirred at 80.degree. C.
for 3 h, where the solution became reddish overtime. The mixture
was then poured into crushed ice and EtOAc. Layers were separated,
and organic layer was then dried over anh. Na.sub.2SO.sub.4,
filtered, and concentrated under reduced pressure. The residue was
purified by reverse phase chromatography (C18, ACN in aq. 10 mM
ammonium formate, pH 3.8, 45 to 85% gradient) to afford the title
compound (431 mg, 1.09 mmol, 47% yield) as a beige solid.
Step 2: (S)-tert-butyl
3-((4-(6-bromo-5-fluoro-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)-
amino)piperidine-1-carboxylate
##STR00278##
[0646] To a solution of
6-bromo-3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-5-fluoro-1H-indole
(410 mg, 1.04 mmol) and (S)-tert-butyl
3-aminopiperidine-1-carboxylate (234 mg, 1.14 mmol) in anh. NMP
(2.1 mL), was added DIPEA (0.55 mL, 3.12 mmol). The reaction
mixture was stirred at 70.degree. C. for 16 h. The mixture was then
diluted with EtOAc, washed with brine (2.times.20 mL), dried over
anh. Na.sub.2SO.sub.4, filtered, and concentrated under reduced
pressure. The residue was purified by column chromatography on
silica gel (EtOAc in hexanes, 0 to 100% gradient) to afford the
title compound (355 mg, 0.635 mmol, 61% yield) as a pale yellow oil
that crystallized overtime.
Step 3: (3S)-tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-5-fluoro-1H-indol-3-yl)-5-(trifluoro-
methyl)pyrimidin-2-yl)amino)piperidine-1-carboxylate
##STR00279##
[0648] (S)-tert-Butyl
3-((4-(6-bromo-5-fluoro-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)-
amino)piperidine-1-carboxylate (100 mg, 0.179 mmol) and
3,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole
(80 mg, 0.358 mmol) were dissolved in a previously degassed mixture
of dioxane (3.0 mL) and water (1.5 mL). Cesium carbonate (118 mg,
0.358 mmol) and Pd(PPh.sub.3).sub.4 (21 mg, 0.018 mmol) were then
added to the reaction mixture subsequently. The resulting mixture
was stirred at 100.degree. C. for 3 h. After full conversion of
starting material, the mixture was cooled to RT, diluted with
water, and extracted with EtOAc (3.times.20 mL). Organic layers
were combined, dried over anh. Na.sub.2SO.sub.4, filtered, and
concentrated under reduced pressure. The residue was purified by
column chromatography on silica gel (EtOAc in Hexanes, 0 to 100%
gradient) to provide the title compound (88 mg, 0.153 mmol, 86%
yield) as a pale yellow oil.
Step 4:
4-(6-(3,5-dimethylisoxazol-4-yl)-5-fluoro-1H-indol-3-yl)-N--((S)-p-
iperidin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
##STR00280##
[0650] To a solution of (3S)-tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-5-fluoro-1H-indol-3-yl)-5-(trifluoro-
methyl)pyrimidin-2-yl)amino)piperidine-1-carboxylate (88 mg, 0.153
mmol) in DCM (2.25 mL), was added TFA (0.6 mL, 7.67 mmol). After 1
h of stirring at RT, the reaction mixture was concentrated to
dryness in vacuo. The crude was then re-dissolved in MeTHF and
washed twice with aq. sat. NaHCO.sub.3. Organic layer was
concentrated, and the obtained residue was purified by reverse
phase chromatography (C18, ACN in aq. 10 mM ammonium formate, pH
3.8, 0 to 55% gradient) to afford the title compound (29.6 mg,
0.062 mmol, 41% yield) as a white solid after lyophilization.
Example 57. Synthesis of (3S)-tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-5-fluoro-1H-pyrrolo[2,3-b]pyridin-3--
yl)-5-(trifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carboxylate
(Compound 138)
Step 1:
4-(5-fluoro-1H-pyrrolo[2,3-b]pyridin-6-yl)-3,5-dimethylisoxazole
##STR00281##
[0652] 6-Bromo-5-fluoro-1H-pyrrolo[2,3-b]pyridine (592 mg, 2.67
mmol) and 3,5-dimethylisoxazol-4-yl-4-boronic acid (792 mg, 5.34
mmol) were dissolved in degassed 2:1 mixture of dioxane and water
(30/15 mL). Cesium carbonate (1.75 g, 5.34 mmol) was added, and the
mixture was degassed for 5 min. Pd(Ph.sub.3).sub.4 (312 mg, 267
umol) was then added, and the reaction mixture was stirred at
100.degree. C. for 3 h. The mixture was cooled to RT and diluted
with water, the crude product was extracted from aq. phase with
MeTHF (3 times). Organics were combined, dried over anh.
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. The residue
was purified by silica gel column chromatography (EtOAc in hexanes,
0 to 100% gradient) to afford the title compound (500 mg, 2.16
mmol, 81% yield) as a yellowish solid.
Step 2:
4-(3-bromo-5-fluoro-1H-pyrrolo[2,3-b]pyridin-6-yl)-3,5-dimethyliso-
xazole
##STR00282##
[0654] To a solution of
4-(5-fluoro-1H-pyrrolo[2,3-b]pyridin-6-yl)-3,5-dimethylisoxazole
(500 mg, 2.16 m mol) in DCM (22 mL), was added NBS (388 mg, 2.16
mmol), and the reaction mixture was stirred at RT for 16 h. The
mixture was quenched by the addition of Na.sub.2S.sub.2O.sub.3 (aq.
sat.) and stirred vigorously for 30 min. DCM was concentrated, and
the crude product was extracted from aq. phase with EtOAc
(3.times.30 mL). The combined organic phase was again washed with
aq. sat. NaS.sub.2SO.sub.3 (20 mL), water (2.times.10 mL), brine
(2.times.20 mL), then dried over anh. Na.sub.2SO.sub.4, filtered,
and concentrated in vacuo to afford the title compound as an
orange-brown solid (666 mg, 2.15 mmol, quant. yield), which was
used in the next step without further purification.
Step 3:
4-(3-bromo-5-fluoro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6--
yl)-3,5-dimethylisoxazole
##STR00283##
[0656] A solution of
4-(3-bromo-5-fluoro-1H-pyrrolo[2,3-b]pyridin-6-yl)-3,5-dimethylisoxazole
(666 mg, 2.15 mmol) in THF (21.5 mL) was cooled to 0.degree. C. and
then treated with sodium tert-butoxide (248 mg, 2.58 mmol). After
stirring for 30 min at this temperature, benzenesulfonyl chloride
(303 uL, 2.25 mmol) was added, and the reaction mixture was stirred
at RT until full conversion. The mixture was poured into water (50
mL), and then THF was concentrated under reduced pressure. The
formed solid was filtered, washed with water, and dried in vacuo to
afford the title compound as a tan solid (919 mg, 2.04 mmol, 95%
yield), which was used in the next step without further
purification.
Step 4:
4-(5-fluoro-1-(phenylsulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxab-
orolan-2-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3,5-dimethylisoxazole
##STR00284##
[0658] A mixture of
4-(3-bromo-5-fluoro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3,5-
-dimethylisoxazole (455 mg, 1.01 mmol), potassium acetate (501 mg,
5.05 mmol) and B.sub.2Pin.sub.2 (bis(pinacolato)diboron; 655 mg,
2.53 mmol) in dioxane (10 mL) was degassed for 30 min by bubbling
with nitrogen. Pd(dppf)Cl.sub.2 (37.5 mg, 50.5 umol) was then
added, and the reaction mixture was stirred in a sealed vial at
100.degree. C. until full conversion (30 min). The mixture was
cooled to RT, diluted with EtOAc, and filtered through Celite pad.
The organic phase was washed with water (2.times.10 mL), brine
(2.times.10 mL), dried over anh. Na.sub.2SO.sub.4, and concentrated
in vacuo to afford the title compound as a yellowish oil (503 mg,
1.01 mmol, quant. yield), which was used in the next step without
further purification.
Step 5: (3S)-tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-5-fluoro-1-(phenylsulfonyl)-1H-pyrro-
lo[2,3-b]pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)amino)piperidine--
1-carboxylate
##STR00285##
[0660] A solution of
4-(5-fluoro-1-(phenylsulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan--
2-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3,5-dimethylisoxazole (503 mg,
1.01 mmol), (S)-tert-butyl
3-((4-chloro-5-(trifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carboxy-
late [prepared as in PCT/US2014/015256] (424 mg, 1.11 mmol) and
cesium carbonate (664 mg, 2.02 mmol) in dioxane and water (20 mL/10
mL) was degassed for 30 min by bubbling with nitrogen.
Pd(PPh.sub.3).sub.4 (119 mg, 0.101 mmol) was then added, and the
reaction mixture was stirred at 100.degree. C. for 2 h. The mixture
was cooled to RT and concentrated under reduced pressure to remove
dioxane. After re-dissolving in EtOAc (30 mL) and water (15 mL),
the aqueous layer was extracted with EtOAc (2.times.15 mL). The
collected organic phase was washed with water (2.times.15 mL),
brine (2.times.15 mL), dried over anh. Na.sub.2SO.sub.4, filtered,
and concentrated. The residue was purified by flash silica gel
column chromatography (EtOAc in hexanes, 0 to 100% gradient) to
provide the title compound (410 mg, 0.573 mmol, 57% yield) as a
pale yellow solid.
Step 6: (3S)-tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-5-fluoro-1H-pyrrolo[2,3-b]pyridin-3--
yl)-5-(trifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carboxylate
##STR00286##
[0662] To a solution of (3S)-tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-5-fluoro-1-(phenylsulfonyl)-1H-pyrro-
lo[2,3-b]pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)amino)piperidine--
1-carboxylate (410 mg, 0.573 mmol) in dioxane (5 mL), was added aq.
5 M solution of sodium hydroxide (2.29 mL, 11.5 mmol), and the
reaction mixture was stirred at 70.degree. C. until full conversion
(30 min). Cooled to RT, the mixture was concentrated, then
re-dissolved in MeTHF, and water (20 mL) was added. The mixture was
acidified to pH 3 with aq. 1 M HCl, then aq. layer was extracted
with MeTHF (2.times.15 mL). The combined organic phase was washed
with water (2.times.15 mL), brine (2.times.15 mL), dried over anh.
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo to afford the
title compound (319 mg, 0.554 mmol, 97% yield) as a pale yellow
solid, which was used in the next step without further
purification.
Example 58. Synthesis of (S)-tert-butyl
3-((4-(6-(3,5-dimethylisothiazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(-
trifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carboxylate
(Compound 166)
Step 1:
(S)-3-(2-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-5-(trifluo-
romethyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridine 7-oxide
##STR00287##
[0664] To a suspension of (S)-tert-butyl
3-((4-(1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)a-
mino)piperidine-1-carboxylate [prepared from
1-(phenylsulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyr-
rolo[2,3-b]pyridine and (S)-tert-butyl
3-((4-chloro-5-(trifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carboxy-
late as in Example 57] (3.20 g, 6.92 mmol) in DCM (68 mL) at
0.degree. C., was added m-CPBA, 77%, portion wise (4.65 g, 20.76
mmol, 3 eq in total) over 16 h, stirring the reaction mixture at RT
and monitoring conversion by LCMS. Then, water (50 mL) was added,
and the crude product was extracted with DCM (2.times.100 mL). The
combined organic phase was washed with sat. aq. NaHCO.sub.3 (50
mL), brine (2.times.50 mL), dried over anh. Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. The residue was purified by
reverse phase column chromatography (C18, ACN in aq. 10 mM ammonium
formate, pH 3.8, 0 to 100% gradient) to afford the title compound
(1.2 g, 2.5 mmol, 36% yield) as a red solid.
Step 2: (S)-Methyl
3-(2-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-5-(trifluoromethyl)py-
rimidin-4-yl)-6-chloro-1H-pyrrolo[2,3-b]pyridine-1-carboxylate
##STR00288##
[0666] To a suspension of
(S)-3-(2-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-5-(trifluoromethy-
l)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridine 7-oxide (1.09 g, 2.27
mmol) in THF (12 mL), was added HMDS
(1,1,1,3,3,3-Hexamethyldisilazane; 517 uL, 2.50 mmol), and the
mixture was stirred under nitrogen at 0.degree. C. Then, methyl
chloroformate (443 uL, 5.68 mmol) was added, and the reaction
mixture was stirred at RT until full conversion (4 h). The mixture
was diluted with EtOAc, washed with aq. sat. NaHCO.sub.3, water and
brine, then dried over anh. Na.sub.2SO.sub.4, filtered, and
concentrated under reduced pressure. The residue was purified by
flash column silica gel chromatography (EtOAc in hexanes, 0 to 100%
gradient) to provide the title compound (567 mg, 1.02 mmol, 45%
yield) as a yellow foam.
Step 3: (S)-tert-butyl
3-((4-(6-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(trifluoromethyl)pyrimid-
in-2-yl)amino)piperidine-1-carboxylate
##STR00289##
[0668] (S)-Methyl
3-(2-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-5-(trifluoromethyl)py-
rimidin-4-yl)-6-chloro-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (567
mg, 1.02 mmol) was dissolved in THF (5 mL), and 2 M aq. sodium
hydroxide (1 mL, 2.04 mmol) was added. The reaction mixture was
stirred at RT until full conversion (1 h). The mixture was then
diluted with MeTHF and water, and the organic layer was separated.
The organic phase was dried over anh. Na.sub.2SO.sub.4, filtered,
and concentrated in vacuo to afford the title compound (507 mg,
quant. yield), which was used in the next step without further
purification.
Step 4:
(S)-(3-(2-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-5-(triflu-
oromethyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)boronic
acid
##STR00290##
[0670] A flask was charged with (S)-tert-butyl
3-((4-(6-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(trifluoromethyl)pyrimid-
in-2-yl)amino)piperidine-1-carboxylate (507 mg, 1.02 mmol),
bis(pinacolato)diboron (317 mg, 1.22 mmol), potassium acetate (506
mg, 5.10 mmol), and degassed dioxane (13 mL). The mixture was
degassed for another 5 min, and then Pd(dppf)Cl.sub.2.times.DCM
complex (85 mg, 102 umol) was added. The reaction mixture was
stirred at 100.degree. C. for 16 h. The reaction mixture was cooled
to RT, diluted with EtOAc, and filtered through Celite pad. The
organic phase was washed with water and brine, dried over anh.
Na.sub.2SO.sub.4, and concentrated in vacuo. The obtained
dark-brown crude was triturated with the mixture of hexane/ether to
afford the title compound as a grey solid, which was used in the
next step without further purification.
Step 5: (S)-tert-butyl
3-((4-(6-(3,5-dimethylisothiazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(-
trifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carboxylate
##STR00291##
[0672]
(S)-(3-(2-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-5-(trifluo-
romethyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)boronic acid
(crude from the previous step, 1.02 mmol) and
4-bromo-3,5-dimethylisothiazole (274 mg, 1.43 mmol) were dissolved
in degassed 2:1 mixture of dioxane and water (14/7 mL). Cesium
carbonate (669 mg, 2.04 mmol) was added, and after degassing for 5
min, Pd(PPh.sub.3).sub.4 (120 mg, 102 umol) was added. The reaction
mixture was stirred at 100.degree. C. for 1 h. Reaction mixture was
cooled to RT and diluted with water, the product was extracted 2
times with EtOAc. Organics were combined, dried over anh.
Na.sub.2SO.sub.4, filtered, and concentrated. Residue was purified
by reverse phase chromatography (C18, ACN in aq. 10 mM ammonium
formate, pH 3.8, 0 to 100% gradient) to provide the title compound
(132 mg, 0.23 mmol, 22% yield over 2 steps) as a brown foam.
Example 59. Synthesis of
(S)--N-(6,6-dimethylpiperidin-3-yl)-4-(6-(3-methylisoxazol-4-yl)-1H-pyraz-
olo[3,4-b]pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
(Compound 160)
Step 1: 3-methyl-4-(1H-pyrazolo[3,4-b]pyridin-6-yl)isoxazole
##STR00292##
[0674] A mixture of 6-chloro-1H-pyrazolo[3,4-b]pyridine (565 mg,
3.68 mmol),
3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole
(1.10 g, 5.15 mmol), Pd(dppf)Cl.sub.2 (150 mg, 184 umol), sodium
bicarbonate (927 mg, 11.0 mmol) in dioxane (10 mL) and water (3 mL)
was degassed with nitrogen (5 min), and then the reaction mixture
was stirred at 100.degree. C. for 1.5 h. The mixture was cooled
down to RT, diluted with ethyl acetate (20 mL) and filtered through
Celite pad. The organic phase was dried over anh. Na.sub.2SO.sub.4,
filtered, concentrated, and the crude material was triturated in
20% DCM/Hexanes to afford the title compound (620 mg, 3.10 mmol,
84% yield) as a pale brown solid, which was used in the next step
without further purification.
Step 2:
4-(3-iodo-1H-pyrazolo[3,4-b]pyridin-6-yl)-3-methylisoxazole
##STR00293##
[0676] To a solution of
3-methyl-4-(1H-pyrazolo[3,4-b]pyridin-6-yl)isoxazole (500 mg, 2.50
mmol) in DMF (5 mL), was added N-iodosuccinimide (710 mg, 3.00
mmol), and the resulting mixture was stirred at RT for 3 h. The
reaction mixture was then quenched with 10% aqueous sodium
thiosulfate (25 mL), and the resulting suspension was stirred for
10 minutes. The suspension was then filtered to afford the title
compound (757 mg, 2.32 mmol, 93% yield) as a brown solid, which was
used in the next step without further purification.
Step 3:
4-(3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-b]pyridin-6-
-yl)-3-methylisoxazole
##STR00294##
[0678] To a solution of
4-(3-iodo-1H-pyrazolo[3,4-b]pyridin-6-yl)-3-methylisoxazole (757
mg, 2.32 mmol) in DCM (10 mL), was added 3,4-dihydro-2H-pyran (240
uL, 2.55 mmol) followed by p-toluenesulfonic acid monohydrate (44.8
mg, 232 umol), and the solution was stirred at RT for 12 h. The
reaction mixture was then diluted with ethyl acetate (20 mL), and
the organic phase was 25 washed with aq. sat. sodium bicarbonate
(2.times.10 mL). The organic phase was dried over anh.
Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure
to afford the title compound (952 mg, 2.32 mmol, quant. yield),
which was used in the next step without further purification.
Step 4:
3-methyl-4-(3-(2-(methylthio)-5-(trifluoromethyl)pyrimidin-4-yl)-1-
-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl)isoxazole
##STR00295##
[0680] To a mixture of
4-(3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-3--
methylisoxazole (952 mg, 2.32 mmol),
4-chloro-2-(methylthio)-5-(trifluoromethyl)pyrimidine (637 mg, 2.78
mmol), and hexamethylditin (486 uL, 2.32 mmol) in toluene (13 mL),
purged with nitrogen (5 min), was added Pd(PPh.sub.3).sub.4 (268
mg, 232 umol), and the reaction mixture was stirred at 105.degree.
C. until full conversion (24 h). The reaction mixture was poured
into a freshly prepared solution of aqueous potassium fluoride (1.0
g in 50 mL), and stirred for 30 min. The resulting mixture was
filtered on Celite pad, and the filtrate was extracted with ethyl
acetate (3.times.30 mL). The organic layers were combined, washed
with brine (30 mL), dried over anh. Na.sub.2SO.sub.4, filtered, and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography (EtOAc in hexanes, 10 to 70%
gradient) to afford the title compound (529 mg, 1.11 mmol, 48%
yield).
Step 5:
3-methyl-4-(3-(2-(methylsulfonyl)-5-(trifluoromethyl)pyrimidin-4-y-
l)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl)isoxazole
##STR00296##
[0682] To a solution of
3-methyl-4-(3-(2-(methylthio)-5-(trifluoromethyl)pyrimidin-4-yl)-1-(tetra-
hydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl)isoxazole (500
mg, 1.05 mmol) in DCM (19 mL), was added m-CPBA (495 mg, 2.15
mmol), and the reaction mixture was stirred at RT, monitoring the
conversion by LCMS. After 2.5 h with the conversion of 47%, another
0.7 eq of m-CPBA were added, and the reaction mixture was stirred
at RT for 1 h. The reaction was then quenched with 10% aq. sodium
thiosulfate (10 mL) and sat. aq. NaHCO.sub.3 (10 mL). The resulting
mixture was stirred for 15 minutes, then diluted with EtOAc (100
mL). The biphasic mixture was separated, and the organic phase was
washed with sat. aq. NaHCO.sub.3 (2.times.10 mL). The organic phase
was dried over anh. Na.sub.2SO.sub.4, filtered, and concentrated
under reduced pressure to afford the title compound (534 mg, quant.
yield), which was used in the next step without further
purification.
Step 6:
N--((S)-6,6-dimethylpiperidin-3-yl)-4-(6-(3-methylisoxazol-4-yl)-1-
-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl)-5-(trifluorome-
thyl)pyrimidin-2-amine
##STR00297##
[0684] To a solution of
3-methyl-4-(3-(2-(methylsulfonyl)-5-(trifluoromethyl)pyrimidin-4-yl)-1-(t-
etrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl)isoxazole
(534 mg, 1.05 mmol) in THF (6.4 mL), was added
(S)-6,6-dimethylpiperidin-3-amine (202 mg, 1.58 mmol) followed by
DIPEA (554 uL, 3.15 mmol), and the reaction mixture was stirred at
RT for 1 h. The mixture was concentrated under reduced pressure,
and the residue was purified by reverse phase chromatography (C18,
ACN in aq. 10 mM ammonium bicarbonate buffer, pH 10, 0 to 100%
gradient) to afford the title compound (143 mg, 0.26 mmol, 24%
yield over two steps).
Step 7:
(S)--N-(6,6-dimethylpiperidin-3-yl)-4-(6-(3-methylisoxazol-4-yl)-1-
H-pyrazolo[3,4-b]pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine
##STR00298##
[0686]
N--((S)-6,6-dimethylpiperidin-3-yl)-4-(6-(3-methylisoxazol-4-yl)-1--
(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-b]pyridin-3-yl)-5-(trifluoromet-
hyl)pyrimidin-2-amine (111 mg, 0.20 mmol) was dissolved in 4 N HCl
in dioxane (2 mL), and the mixture was stirred at 60.degree. C. for
2 h. The solution was then concentrated under reduced pressure, and
the residue was purified by reverse phase chromatography (C18, ACN
in aq. 0.1% formic acid buffer, 10 to 50% gradient) to afford the
title compound (47 mg, 0.10 mmol, 50% yield) as a white HCl salt
after lyophilization.
Example 60.
3-methyl-4-(3-(2-(methylthio)-5-(trifluoromethyl)pyrimidin-4-yl)-1-(pheny-
lsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole (Compound
147)
Step 1:
4-(3-iodo-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylisoxazole
##STR00299##
[0688] To a solution of
3-methyl-4-(1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole [prepared from
3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole
and 6-bromo-1H-pyrrolo[2,3-b]pyridine as in Example 59] (550 mg,
2.76 mmol) in DMF (2 mL), iodine (702 mg, 2.76 mmol) and KOH (94
mg, 5.52 mmol) were added in one portion. The reaction mixture was
stirred at 75.degree. C. for 1 h. The mixture was then cooled down
to RT and quenched by addition of 10% HCl until pH was neutral,
followed by the addition of aq. sat. sodium sulfite (3 mL) at
15.degree. C. The resulting mixture was poured into water (20 mL),
and the obtained solid was filtered to afford the title compound
(847 mg, 2.60 mmol, 94% yield) as a pale orange solid, which was
used in the next step without further purification.
Step 2:
4-(3-iodo-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-meth-
ylisoxazole
##STR00300##
[0690] 4-(3-Iodo-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylisoxazole
(847 mg, 2.60 mmol) was dissolved in THF (10 mL), and the resulting
solution was cooled to 0.degree. C. To the mixture, was added
sodium tert-butoxide (325 mg, 3.38 mmol) portion wise, and the
mixture was stirred for 10 min. Benzenesulfonyl chloride (332 uL,
2.60 mmol) was then added dropwise, and the reaction mixture was
allowed to reach RT and stirred for 1 h. The reaction was diluted
with water (60 mL), and the resulting suspension was filtered and
washed with water (15 mL) to afford the title compound (1.12 g,
2.42 mmol, 93% yield) as a pale orange solid, which was used in the
next step without further purification.
Step 3:
3-methyl-4-(3-(2-(methylthio)-5-(trifluoromethyl)pyrimidin-4-yl)-1-
-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole
##STR00301##
[0692] To a degassed mixture of
4-(3-iodo-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-methylisoxa-
zole (1.28 g, 2.75 mmol) and
dichlorobis(triphenylphosphine)palladium(II) (96.5 mg, 138 umol) in
dioxane (25.6 mL), was added triethylamine (2.30 mL, 16.5 mmol)
followed by HBP in (4,4,5,5-Tetramethyl-1,3,2-dioxaborolane; 599
uL, 4.13 mmol). The resulting yellow suspension was stirred at
80.degree. C. in a sealed vessel for 2 h. The reaction mixture was
then cooled down to RT, and
4-chloro-2-(methylthio)-5-(trifluoromethyl)pyrimidine (629 uL, 2.75
mmol) and degassed water (600 uL) were added followed by the sodium
carbonate (874 mg, 8.25 mmol). The resulting mixture was stirred at
100.degree. C. for 12 h. The mixture was then cooled down to RT,
diluted with EtOAc (25 mL), filtered on Celite pad, and
concentrated under reduced pressure. The crude solid was triturated
with 50% MTBE in hexanes, and the suspension was filtered to afford
the title compound (510 mg, 0.96 mmol, 35% yield) as a pale beige
solid, which was used in the next step without further
purification.
Example 61. (3S,4S)-tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]-
pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-4-methylpiperidine--
1-carboxylate (Compound 143)
##STR00302##
[0694] To a solution of
3,5-dimethyl-4-(3-(2-(methylsulfonyl)-5-(trifluoromethyl)pyrimidin-4-yl)--
1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole
[prepared from (3,5-dimethylisoxazol-4-yl)boronic acid and
6-bromo-1H-pyrrolo[2,3-b]pyridine, following Examples 59 and 60]
(220 mg, 0.38 mmol) and racemic cis tert-butyl
3-amino-4-methylpiperidine-1-carboxylate (122 mg, 0.57 mmol) in
anh. THF (3.8 mL), was added anh. DIPEA (200 uL, 1.14 mmol) at RT.
The reaction mixture was stirred at 70.degree. C. for 16 h. The
mixture was diluted with EtOAc, washed with brine, dried over anh.
Na.sub.2SO.sub.4, filtered, and concentrated under reduced
pressure. The residue was purified by column silica gel
chromatography (EtOAc in hexanes, 0 to 100% gradient) to provide
the racemic product (137 mg, 0.19 mmol, 51% yield) as a pale yellow
solid. The racemic compound was then separated by chiral HPLC using
a ChiralPak IA column and 6:6:88 mixture of MeOH/DCM/Hexanes for
elution. Two enantiomers were obtained: Peak 1 (tentatively
assigned as (3R,4R), 36 mg) and Peak 2 (tentatively assigned as
(3S,4S), 31 mg).
Example 62. (3S,4S)-tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl)-5-(trifluoromethyl)py-
rimidin-2-yl)amino)-4-methylpiperidine-1-carboxylate (Compound
135)
Step 1: racemic cis tert-butyl
3-((4-(6-bromo-1-(phenylsulfonyl)-1H-indol-3-yl)-5-(trifluoromethyl)pyrim-
idin-2-yl)amino)-4-methylpiperidine-1-carboxylate
##STR00303##
[0696]
6-Bromo-3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-1-(phenylsul-
fonyl)-1H-indole [prepared from 6-bromo-1H-indole and
2,4-dichloro-5-(trifluoromethyl)pyrimidine as in Example 56 and
then protected as in Example 57] (360 mg, 0.70 mmol), racemic cis
tert-butyl 3-amino-4-methylpiperidine-1-carboxylate (180 mg, 0.84
mmol) and DIPEA (183 uL, 1.05 mmol) were dissolved in anh. THF (7
mL). The reaction mixture was stirred at 70.degree. C. until full
conversion (4 h). The mixture was then diluted with EtOAc, washed
with sat. aq. NaHCO.sub.3, water, and brine. Organic phase was
dried over anh. Na.sub.2SO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by normal phase silica gel
chromatography (EtOAc in DCM, 0 to 100% gradient) to afford the
title compound (473 mg, 0.68 mmol, 97% yield) as an off-white
solid.
Step 2: (3S,4S)-tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl)-5-(trifluoromethyl)py-
rimidin-2-yl)amino)-4-methylpiperidine-1-carboxylate
##STR00304##
[0698] Racemic cis tert-butyl
3-((4-(6-(3,5-dimethylisoxazol-4-yl)-1H-indol-3-yl)-5-(trifluoromethyl)py-
rimidin-2-yl)amino)-4-methylpiperidine-1-carboxylate (292 mg,
yellow solid) was separated by Chiral SFC using ChiralPak IC
column, 10.times.250 mm 5 um, 10% MeOH, 10 mL/min, 150 bar, column
T: 40.degree. C., run time (min): 27. Two enantiomers were
obtained: Peak 1 (tentatively assigned as (3S,4S), 43 mg, white
solid, ee=99.7%) and Peak 2 (tentatively assigned as (3R,4R), 38
mg, white solid, ee=99.15%).
[0699] The table of FIG. 3 provides additional details of the
synthesis of certain compounds of the invention, as well as their
NMR (nuclear magnetic resonance) and MS (mass spectroscopy)
values.
Example 63. Inhibition of CDK Kinase Activity
[0700] Compounds described herein were assayed for inhibition of
CDK7, CDK9, CDK12, and CDK2 activity at Biortus Biosciences
(Jiangyin, Jiangsu Province, P.R. of China) using kinase assays for
each CDK developed with a Caliper/LabChip EZ Reader (Perkin Elmer,
Waltham, Mass.). These assays measure the amount of phosphorylated
peptide substrate produced as a fraction of the total peptide
following an incubation period at 27.degree. C. with the following
components: test compounds (variable concentrations from 10 .mu.M
down to 0.508 nM in a series of 3-fold serial dilutions), active
CDK kinase protein (with the indicated cyclin, listed below for
each CDK), ATP (2 mM), and substrate peptide (listed below) in the
buffer 2-(N-morpholino)ethanesulfonate (MES buffer, 20 mM), pH
6.75, 0.01% (v/v) Tween 20 detergent, 0.05 mg/mL bovine serum
albumin (BSA).
[0701] Specifically, the CDK7 inhibition assay used CDK7/Cyclin
H/MAT1 complex (6 nM) and "5-FAM-CDK7tide" peptide substrate (2
.mu.M, synthesized fluorophore-labeled peptide with the following
sequence: 5-FAM-YSPTSPSYSPTSPSYSPTSPSKKKK (SEQ ID NO:1), where
"5-FAM" means 5-carboxyfluorescein) with 6 mM MgCl.sub.2 in the
buffer composition listed above. Furthermore, the CDK9 inhibition
assay used CDK9/Cyclin T1 complex (8 nM) and "5-FAM-CDK9tide"
peptide substrate (2 .mu.M, synthesized fluorophore-labeled peptide
with the following sequence: 5-FAM-GSRTPMY-NH.sub.2 (SEQ ID NO:2)
where 5-FAM is defined above and NH.sub.2 signifies a C-terminal
amide) with 10 mM MgCl.sub.2 in the buffer composition listed
above. The CDK12 inhibition assay used CDK12 (aa686-1082)/Cyclin K
complex (50 nM) and "5-FAM-CDK9tide" (2 .mu.M) as defined above,
with 2 mM MgCl.sub.2 in the buffer composition above. Additionally,
the CDK2 inhibition assay used CDK2/Cyclin E1 complex (0.5 nM) and
"5-FAM-CDK7tide" (2 .mu.M; defined above) with 2 mM MgCl.sub.2 in
the buffer composition above.
[0702] The incubation period at 27.degree. C. for each CDK
inhibition assay was chosen such that the fraction of
phosphorylated peptide product produced in each assay, relative to
the total peptide concentration, was approximately 20% (.+-.5%) for
the uninhibited kinase (35 min. for CDK7, 35 min. for CDK2, 3 hr.
for CDK12, 15 min. for CDK9). In cases where the compound
titrations were tested and resulted in inhibition of peptide
product formation, these data were fit to produce best-fit
IC.sub.50 values. The results of these assays are shown below in
Table 1 where "A" represents a calculated IC.sub.50 of less than 30
nM; "B" represents a calculated IC.sub.50 of between 30 nM and less
than 100 nM; "C" represents a calculated IC.sub.50 of between 100
nM and less than 500 nM; "D" represents a calculated IC.sub.50 of
greater than or equal to 500 nM, and "NT" represents that the
specified compound was not tested in the specified assay.
[0703] The inhibitory activity of selected compounds against CDK2,
CDK7, CDK9, and CDK12 was found to be as follows:
TABLE-US-00001 CDK CDK CDK CDK Compound 12 2 7 9 100 D D A D 101 C
D A D 102 D D A D 103 C D A D 104 D D A D 105 D D A D 106 D D A D
107 D D A D 108 D D A D 109 D D A D 110 D D A D 111 D D A D 112 D D
A D 113 D D A D 114 D D A D 115 C C A D 116 D D A D 117 D D A D 118
D D A D 119 D D A D 120 D D A D 121 D D A D 122 D D A D 123 D D A D
124 D D A D 125 D D A D 126 D D A D 127 D D A D 128 D D A D 129 D D
A D 130 D D A D 131 D D A D 132 D D A D 133 D D A D 134 D D A D 135
D D B D 136 D D A D 137 D D A D 138 D D A D 139 D D A D 140 D D A D
141 D D A D 142 D D A D 143 D D A D 144 D D A D 145 D D A D 146 D D
A D 147 D D A D 148 D D A D 149 D D A D 150 D D A D 151 D D A D 152
D D A D 153 D D A D 154 D D A D 155 D D A D 156 D D A D 157 D D A D
158 D D A D 159 D D A D 160 D D A D 161 D D A D 162 D D A D 163 D D
A D 164 D D A D 165 D D B D 166 D D A D 167 D D A D 168 D D A D
Example 64. Inhibition of Cell Proliferation
[0704] HCC70 and MB453 cells are cell lines derived from human
triple negative breast cancer. COV318 and COV504 cells are cell
lines derived from human ovarian cancer. Representative compounds
of the invention were tested at different concentrations (from 4
.mu.M to 126.4 .mu.M; 0.5 log serial dilutions) for their ability
to inhibit the proliferation of each of these cell lines. Known CDK
inhibitors dinaciclib or N-((1S,3R)-3-((5-chloro-4-(1H-indol-3-yl)
pyrimidin-2-yl)amino)cyclohexyl)-5-((E)-4-(dimethylamino)but-2-enamido)pi-
colinamide and triptolide were used as positive controls. Cells
were seeded at approximately 2,000 cells/well and grown in
ATCC-formulated RPMI-1640 Medium (ATCC 30-2001)+10% FBS. The cells
were cultured at 37.degree. C. in a humidified chamber in the
presence of 5% CO.sub.2. Proliferation assays were conducted over a
72-hour time period. CyQUANT.RTM. Direct Cell Proliferation Assay
(Life Technologies, Chicago, Ill. USA) was used to assess the
anti-proliferative effects of the compounds following
manufacturer's directions and utilizing the reagents supplied with
the CyQUANT.RTM. Direct Cell kit. The results of the assay are
shown below in Table 2 where "A" represents a calculated IC.sub.50
of less than 100 nM; "B" represents a calculated IC.sub.50 of
between 100 nM and less than 500 nM; "C" represents a calculated
IC.sub.50 of between 500 nM and less than 1 .mu.M; "D" represents a
calculated IC.sub.50 of greater than 1 .mu.M. Cell proliferation
inhibition activity of selected compounds against HCC70, MB453,
COV318 and COV504 cells was found to be as follows:
TABLE-US-00002 Compound HCC70 COV318 COV504 MB453 100 A B A A 101 A
A A A 102 A B A A 103 B B A A 104 A B A A 105 A C B B 106 A B A A
107 B B B B 108 A A A A 109 B A A B 110 A B A A 111 B B A B 112 B D
B B 113 C D C B 114 A A A A 115 A A A A 116 A A A A 117 B C A A 118
A B A A 119 A D A A 120 B C B A 121 B B A A 122 B D A A 123 B B A A
124 A A A A 125 B B A A 126 A B A A 127 D B A A 128 A B A A 129 A A
A A 130 B B B B 131 B B A A 132 A A A A 133 A B A A 134 A A A A 135
B D A A 136 A D A A 137 A A A A 138 A A A A 139 A D A A 140 D D B C
141 B B A B 142 A C A A 143 A D A A 144 A D A A 145 B D B B 146 C D
B B 147 A D A A 148 D D C B 149 A A A A 150 B B A A 151 D D D C 152
D D C B 153 A A A A 154 B D B D 155 A A A A 156 C D B B 157 A A A A
158 A B A A 159 A D A A 160 D D B B 161 A B A A 162 B D A C 163 A D
A A 164 B C A A 165 D D C C 166 A A A A 167 A B A B 168 A A A A
[0705] In the claims, articles such as "a," "an," and "the" may
mean one or more than one unless indicated to the contrary or
otherwise evident from the context. Claims or descriptions that
include "or" between one or more members of a group are considered
satisfied if one, more than one, or all of the group members are
present in, employed in, or otherwise relevant to a given product
or process unless indicated to the contrary or otherwise evident
from the context (for example, where it is evident from the context
that "A or B" can mean only "A" or, alternatively, only "B"). The
invention includes embodiments in which exactly one member of the
group is present in, employed in, or otherwise relevant to a given
product or process as well as embodiments in which more than one,
or all of the group members are present in, employed in, or
otherwise relevant to a given product or process.
[0706] Furthermore, the invention encompasses all variations,
combinations, and permutations in which one or more limitations,
elements, clauses, and descriptive terms from one or more of the
listed claims are introduced into another claim. For example, any
claim that is dependent on another claim can be modified to include
one or more limitations found in any other claim that is dependent
on the same base claim. Where elements are presented as lists every
possible subgroup of the elements is also disclosed, and any
element(s) can be removed from the group. In general, where the
invention, or aspects or embodiments of the invention, is/are
referred to as comprising or including particular elements and/or
features, other aspects or embodiments of the invention consist of,
or consist essentially of, such elements and/or features. For
practicality and simplicity, not all of those aspects or
embodiments have been specifically set forth in haec verba herein
but are nevertheless within the scope of the present invention.
Where ranges are given, endpoints are included. Furthermore, unless
otherwise indicated or otherwise evident from the context, values
that are expressed as ranges can assume any specific value or
sub-range within the stated ranges in different embodiments of the
invention, to the tenth of the unit of the lower limit of the
range, and any value may be as stated or "about" the stated
value.
[0707] One of ordinary skill in the art will recognize or be able
to ascertain using no more than routine experimentation, many
equivalents to the specific embodiments of the compositions and
methods described and claimed herein. Such equivalents are intended
to be encompassed by the following claims.
* * * * *