U.S. patent application number 15/553870 was filed with the patent office on 2018-08-02 for pyrimidine derivatives as kinase inhibitors and their therapeutical applications.
The applicant listed for this patent is NantBioScience, Inc.. Invention is credited to Forrest Arp, David Ho, Laxman Nallan, Tulay Polat, Chunlin Tao, Qinwei Wang, Paul Weingarten.
Application Number | 20180215734 15/553870 |
Document ID | / |
Family ID | 56789187 |
Filed Date | 2018-08-02 |
United States Patent
Application |
20180215734 |
Kind Code |
A1 |
Tao; Chunlin ; et
al. |
August 2, 2018 |
PYRIMIDINE DERIVATIVES AS KINASE INHIBITORS AND THEIR THERAPEUTICAL
APPLICATIONS
Abstract
The present invention provides kinase inhibitors with
anti-proliferative activity comprising substituted pyrimidine
derivatives and pharmaceutically-acceptable formulations thereof.
In addition, the invention provides methods for making novel
compounds and methods for using the compounds.
Inventors: |
Tao; Chunlin; (Newport
Coast, CA) ; Wang; Qinwei; (Alhambra, CA) ;
Nallan; Laxman; (Rancho Mission Viejo, CA) ; Ho;
David; (Monterey Park, CA) ; Polat; Tulay;
(Tustin, CA) ; Arp; Forrest; (Irvine, CA) ;
Weingarten; Paul; (Anaheim, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
NantBioScience, Inc. |
Culver City |
CA |
US |
|
|
Family ID: |
56789187 |
Appl. No.: |
15/553870 |
Filed: |
February 29, 2016 |
PCT Filed: |
February 29, 2016 |
PCT NO: |
PCT/US16/20095 |
371 Date: |
August 25, 2017 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 401/14 20130101;
A61K 31/506 20130101; A61P 29/00 20180101; A61P 37/00 20180101;
C07D 403/14 20130101; A61K 31/5377 20130101; A61P 35/00 20180101;
C07D 405/14 20130101; C07D 417/14 20130101; A61K 45/06 20130101;
C07D 403/12 20130101; C07D 487/04 20130101 |
International
Class: |
C07D 403/12 20060101
C07D403/12; C07D 403/14 20060101 C07D403/14 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 27, 2016 |
US |
PCT/US2015/018085 |
Claims
1. A compound of the formula ##STR00278## or a pharmaceutically
acceptable salt thereof, wherein: W is selected from the group
consisting of: Cl, Br, I, CN, C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.6 alkoxy, C.sub.2-C.sub.6 alkenyl, CF.sub.3,
CF.sub.2H, CFH.sub.2, C.sub.2-C.sub.6 alkynyl, CON(R1)R2; R1 and R2
represent hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, alkylthio,
aryl, arylalkyl; Ar represents heteroaryl or aryl, each of which is
substituted with from 0 to 4 substituents independently chosen from
the group consisting of: (1) halogen, hydroxy, amino, amide, cyano,
--COOH, --SO.sub.2NH.sub.2, oxo, nitro or alkoxycarbonyl; (2) NR1;
and (3) groups of the formula (Ia): ##STR00279## wherein: R.sub.4
represents hydrogen, C.sub.1-C.sub.4 alkyl, or oxo; X--R.sub.3 is
CH; or X--R.sub.3 is O; or X is N, and R.sub.3 represents hydrogen,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.10 aryl or heteroaryl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.6
haloalkyl, C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkylthio,
C.sub.2-C.sub.6 alkanoyl, C.sub.1-C.sub.6 alkoxycarbonyl,
C.sub.2-C.sub.6 alkanoyloxy, mono- and di-(C.sub.3-C.sub.8
cycloalkyl)aminoC.sub.0-C.sub.4alkyl, (4- to 7-membered
heterocycle)C.sub.0-C.sub.4alkyl, C.sub.1-C.sub.6 alkylsulfonyl,
mono- and di-(C.sub.1-C.sub.6 alkyl) sulfonamido, and mono- and
di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, each of which is
substituted with from 0 to 4 substituents independently chosen from
halogen, hydroxy, cyano, amino, --COOH or oxo; R.sub.5 and R.sub.6
are independently selected from the comprising of: hydrogen, F, Cl,
Br, CN, C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.6 alkoxy; and
R.sub.7, R.sub.8 and R.sub.9 are independently selected from the
group consisting of: hydrogen, C.sub.1-C.sub.4 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.10
aryl or heteroaryl, C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6
alkylthio, C.sub.2-C.sub.6 alkanoyl, C.sub.1-C.sub.6
alkoxycarbonyl, and C.sub.2-C.sub.6 alkanoyloxy.
2. A compound of claim 1, having the formula ##STR00280## or a
pharmaceutically acceptable salt thereof.
3. A compound of claim 1, having the formula ##STR00281## or a
pharmaceutically acceptable salt thereof.
4. A compound of claim 1, having the formula ##STR00282## or a
pharmaceutically acceptable salt thereof.
5. A compound of claim 1, having the formula ##STR00283## or a
pharmaceutically acceptable salt thereof.
6. A compound of claim 1, having the formula ##STR00284## or a
pharmaceutically acceptable salt thereof.
7. A compound of claim 1, having the formula ##STR00285## or a
pharmaceutically acceptable salt thereof.
8. A compound of claim 1, having the formula ##STR00286## or a
pharmaceutically acceptable salt thereof.
9. A method of treating an animal suffering from a cellular
proliferative disorder comprising: a. Preparing, or causing to be
prepared, the compound or pharmaceutically acceptable salt of claim
1; b. optionally, formulating, or causing to be formulated, the
compound or pharmaceutically acceptable salt of step a with a
pharmaceutically acceptable carrier; and c. administering, or
causing to be administered, the compound or pharmaceutically
acceptable salt of step a or the formulation of step b to the
animal suffering from the proliferative disorder.
10. The method of claim 9, wherein the animal is a mammal.
11. The method of claim 10, wherein the animal is a human.
12. The method of claim 9, wherein the cellular proliferative
disorder is a cancer, a precancerous state, a benign tumor,
autoimmune disorder, transplant rejection, graft versus host
disease, response to an infection, response to an environmental
insult or a genetic disorder.
13. The method of claim 12, wherein the cellular proliferative
disorder is a cancer.
14. The method of claim 13, wherein the cancer is a skin cancer,
breast cancer, uterine cancer, ovarian cancer, testicular cancer,
prostate cancer, nasopharyngeal cancer, lung cancer, esophageal
cancer, gastric cancer, hepatic cancer, biliary cancer, pancreatic
cancer, lymphoma, lung cancer, renal cancer, bladder cancer,
esophageal cancers, myeloid leukemia, lymphocytic leukemia,
myleoproliferative disorder, myelodysplastic syndrome, lymphoma,
neuroendocrine cancer, sarcoma or brain tumor.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of PCT/US2015/018085,
filed Feb. 27, 2015, which is incorporated by reference in its
entirety.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH AND
DEVELOPMENT
[0002] This invention was made without Government support.
INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ELECTRONICALLY
[0003] None
STATEMENT REGARDING PRIOR DISCLOSURES BY THE INVENTOR OR A JOINT
INVENTOR
[0004] There have been no prior disclosures of this invention.
FIELD OF THE INVENTION
[0005] The present invention relates generally to the use of
compounds to treat a variety of disorders, diseases and pathologic
conditions and more specifically to the use of substituted
pyrimidine derivatives to modulate protein kinases and for treating
protein kinase-mediated diseases.
BACKGROUND OF THE INVENTION
[0006] Protein kinases constitute large families of structurally
related proteinaceous enzymes that are responsible for the control
of numerous signal transduction pathways in the eukaryotic cell.
Protein kinases, containing a similar 250-300 amino acid catalytic
domain, catalyze the phosphorylation of target protein substrates.
As such protein kinases are among the most promising small molecule
drug targets.
[0007] The kinases may be categorized into families by the
substrates in the phosphorylate (e.g., protein-tyrosine,
protein-serine/threonine, lipids, etc.). Tyrosine phosphorylation
is a central event in the regulation of a variety of biological
processes such as cell proliferation, migration, differentiation
and survival. Several families of receptor and non-receptor
tyrosine kinases control these events by catalyzing the transfer of
phosphate from ATP to a tyrosine residue of specific cell protein
targets. Sequence motifs have been identified that generally
correspond to each of these kinase families [Hanks et al., FASEB
J., (1995), 9, 576-596; Knighton et al., Science, (1991), 253,
407-414; Garcia-Bustos et al., EMBO J., (1994), 13:2352-2361).
Examples of kinases in the protein kinases include (without
limitation): abl, Akt, bcr-abl, Blk, Brk, Btk, c-kit, c-Met, c-src,
c-fms, CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10,
cRaf1, CSF1R, CSK, EGFR, ErbB2, ErbB3, ErbB4, Erk, Fak, fes, FGFR1,
FGFR2, FGFR3, FGFR4, FGFR5, Fgr, fit-1, Fps, Frk, Fyn, Hck, IGF-1R,
INS-R, Jak, KDR, Lck, Lyn, MEK, p38, PDGFR, PIK, PKC, PYK2, ros,
Tie, Tie-2, TRK, Yes, and Zap70.
[0008] Studies indicated that protein kinases play a central role
in the regulation and maintenance of a wide variety of cellular
processes. For example, kinase activity acts as molecular switches
regulating cell proliferation, activation, and/or differentiation.
Uncontrolled or excessive kinase activity, whether from mutant
kinases or wild type kinases has been observed in many disease
states including benign and malignant proliferation disorders, as
well as diseases resulting from inappropriate activation of the
immune system (autoimmune disorders), allograft rejection, and
graft vs host disease.
[0009] It is reported that many diseases are associated with
abnormal cellular responses triggered by protein kinase-mediated
events. These diseases include autoimmune diseases, inflammatory
diseases, bone diseases, metabolic diseases, neurological and
neurodegenerative diseases, cancer, cardiovascular diseases,
allergies and asthma, Alzheimer's disease and hormone-related
diseases. In addition, endothelial cell specific receptor PTKs,
such as VEGF-2 and Tie-2, mediate the angiogenic process and are
involved in supporting the progression of cancers and other
diseases involving uncontrolled vascularization. Accordingly, there
has been a substantial effort in medicinal chemistry to find
protein kinase inhibitors that are effective as therapeutic
agents.
[0010] Many cancers are characterized by disruptions in cellular
signaling pathways that lead to uncontrolled growth and
proliferation of cancerous cells. Receptor tyrosine kinases (RTKs)
play a crucial role in these signaling pathways, transmitting
extracellular molecular signals into cytoplasm and/or nucleus of a
cell. RTKs are transmembrane proteins that generally include an
extracellular ligand-binding domain, a membrane-spanning domain and
a catalytic cytoplasmic tyrosine kinase domain. The binding of
ligand to the extracellular potion is believed to promote
dimerization, resulting in trans-phosphorylation and activation of
the intracellular tyrosine kinase domain (Schlessinger et al.
Neuron 1992; 9:383-391).
[0011] Considering the lack of currently available treatment
options for the majority of the conditions associated with protein
kinases, there is still a great need for new therapeutic agents
that inhibit protein kinases. In particular, there is a need for
highly active kinase inhibitors that are also non-toxic, and
specific to certain protein kinases.
SUMMARY OF THE INVENTION
[0012] A compound of the formula
##STR00001##
[0013] or a pharmaceutically acceptable salt thereof, wherein:
[0014] W is selected from: F, Cl, Br, I, CN, C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.6 alkoxy, C.sub.2-C.sub.6 alkenyl, CF.sub.3,
CF.sub.2H, CFH.sub.2, C.sub.2-C.sub.6 alkynyl, CON(R1)R2.
[0015] R1 and R2 represent hydrogen, alkyl, cycloalkyl, alkenyl,
alkynyl, alkylthio, aryl, arylalkyl.
[0016] Ar represents heteroaryl or aryl, each of which is
substituted with from 0 to 4 substituents independently chosen
from:
[0017] (1) halogen, hydroxy, amino, amide, cyano, --COOH,
--SO.sub.2NH.sub.2, oxo, nitro and alkoxycarbonyl; and
[0018] (2) NR1
[0019] (3) groups of the formula (Ia):
##STR00002##
[0020] wherein:
[0021] R.sub.4 represents hydrogen, C.sub.1-C.sub.4 alkyl, oxo;
[0022] X is CH, when R.sub.3 is hydrogen; or X--R.sub.3 is O; or X
is N, R.sub.3 represents groups of hydrogen, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.10
aryl or heteroaryl, (C.sub.3-C.sub.7cycloalkyl)C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 alkoxy,
C.sub.1-C.sub.6 alkylthio, C.sub.2-C.sub.6 alkanoyl,
C.sub.1-C.sub.6 alkoxycarbonyl, C.sub.2-C.sub.6 alkanoyloxy, mono-
and di-(C.sub.3-C.sub.5 cycloalkyl)aminoC.sub.0-C.sub.4alkyl, (4-
to 7-membered heterocycle)C.sub.0-C.sub.4alkyl, C.sub.1-C.sub.6
alkylsulfonyl, mono- and di-(C.sub.1-C.sub.6 alkyl) sulfonamido,
and mono- and di-(C.sub.1-C.sub.6 alkyl)aminocarbonyl, each of
which is substituted with from 0 to 4 substituents independently
chosen from halogen, hydroxy, cyano, amino, --COOH and oxo.
[0023] Substituents on indole are as the following: R.sub.5 and
R.sub.6 are independently selected from: Hydrogen, F, Cl, Br, CN,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.6 alkoxy.
[0024] R.sub.7, R.sub.8 and R.sub.9 are independently selected from
Hydrogene, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.10 aryl or heteroaryl,
C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkylthio, C.sub.2-C.sub.6
alkanoyl, C.sub.1-C.sub.6 alkoxycarbonyl, C.sub.2-C.sub.6
alkanoyloxy.
[0025] A pharmaceutical composition comprising at least one
compound of claim 1 or its pharmaceutically acceptable salts,
hydrates, solvates, crystal forms salts and individual
diastereomers thereof, and a pharmaceutically acceptable
carrier.
[0026] Accordingly, it is an objective of the present invention to
provide an antitumor agent comprising substituted pyrimidine
derivatives as described in formula (I),
pharmaceutically-acceptable formulations thereof, methods for
making novel compounds and compositions for using the compounds.
The compounds and compositions comprising the compounds in formula
(I) have utility in treatment of a variety of diseases.
[0027] The combination therapy described herein may be provided by
the preparation of the substituted pyrimidine derivatives of
formula (I) and the other therapeutic agent as separate
pharmaceutical formulations followed by the administration thereof
to a patient simultaneously, semi-simultaneously, separately or
over regular intervals.
[0028] The present invention provides methods of use for certain
chemical compounds such as kinase inhibitors for treatment of
various diseases, disorders, and pathologies, for example, cancer,
and vascular disorders, such as myocardial infarction (MI), stroke,
or ischemia. The compounds described in this invention may block
the enzymatic activity of some or many of the members of the FGFR
kinase family, in addition to blocking
[0029] the activity of other receptor and non-receptor kinases.
BRIEF DESCRIPTION OF THE FIGURES
[0030] FIG. 1 depicts a dose response curve for Compound 19.
[0031] FIG. 2 depicts Compound 19's anti-tumor activity.
[0032] FIG. 3 depicts Compound 19's toxicity in rats based on
weight loss.
DETAILED DESCRIPTION OF THE INVENTION
[0033] The present invention is related to compounds having general
Formula (I)
##STR00003##
or a pharmaceutically acceptable salt thereof, wherein:
[0034] W is selected from: F, Cl, Br, I, CN, C1-C4 alkyl, C1-C6
alkoxy, C2-C6 alkenyl, CF3, CF2H, CFH2, C2-C6 alkynyl,
CON(R1)R2.
[0035] R1 and R2 represent hydrogen, alkyl, cycloalkyl, alkenyl,
alkynyl, alkylthio, aryl, arylalkyl.
[0036] Ar represents heteroaryl or aryl, each of which is
substituted with from 0 to 4 substituents independently chosen
from:
[0037] (1) halogen, hydroxy, amino, amide, cyano, --COOH,
--SO.sub.2NH.sub.2, Oxo, nitro and alkoxycarbonyl; and
[0038] (2) NR1
[0039] (3) groups of the formula (Ia):
##STR00004##
[0040] A compound of the formula,
[0041] or a pharmaceutically acceptable salt thereof, wherein:
##STR00005##
[0042] W is selected from: F, Cl, Br, I, CN, C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.6 alkoxy, C.sub.2-C.sub.6 alkenyl, CF.sub.3,
CF.sub.2H, CFH.sub.2, C.sub.2-C.sub.6 alkynyl, CON(R1)R2.
1. R1 and R2 represent hydrogen, alkyl, cycloalkyl, alkenyl,
alkynyl, alkylthio, aryl, arylalkyl. 2. Ar represents heteroaryl or
aryl, each of which is substituted with from 0 to 4 substituents
independently chosen from:
[0043] (1) halogen, hydroxy, amino, amide, cyano, --COOH,
--SO.sub.2NH.sub.2, oxo, nitro and alkoxycarbonyl; and
[0044] (2) NR1
[0045] (3) groups of the formula (Ia):
##STR00006##
[0046] wherein:
[0047] R.sub.4 represents hydrogen, C.sub.1-C.sub.4 alkyl, oxo;
[0048] X is CH, when R.sub.3 is hydrogen; or X--R.sub.3 is O; or X
is N, R.sub.3 represents groups of hydrogen, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.10
aryl or heteroaryl, (C.sub.3-C.sub.7cycloalkyl)C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 alkoxy,
C.sub.1-C.sub.6 alkylthio, C.sub.2-C.sub.6 alkanoyl,
C.sub.1-C.sub.6 alkoxycarbonyl, C.sub.2-C.sub.6 alkanoyloxy, mono-
and di-(C.sub.3-C.sub.8 cycloalkyl)aminoC.sub.0-C.sub.4alkyl, (4-
to 7-membered heterocycle)C.sub.0-C.sub.4alkyl, C.sub.1-C.sub.6
alkylsulfonyl, mono- and di-(C.sub.1-C.sub.6 alkyl) sulfonamido,
and mono- and di-(C.sub.1-C.sub.6 alkyl)aminocarbonyl, each of
which is substituted with from 0 to 4 substituents independently
chosen from halogen, hydroxy, cyano, amino, --COOH and oxo.
3. Substituents on indole are as the following:
[0049] R.sub.5 and R.sub.6 are independently selected from:
Hydrogen, F, Cl, Br, CN, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.6
alkoxy.
4. R.sub.7, R.sub.8 and R.sub.9 are independently selected from
Hydrogene, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.10 aryl or heteroaryl,
C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkylthio, C.sub.2-C.sub.6
alkanoyl, C.sub.1-C.sub.6 alkoxycarbonyl, C.sub.2-C.sub.6
alkanoyloxy. 5. A pharmaceutical composition comprising at least
one compound of claim 1 or its pharmaceutically acceptable salts,
hydrates, solvates, crystal forms salts and individual
diastereomers thereof, and a pharmaceutically acceptable carrier.
wherein:
[0050] R.sub.4 represents hydrogen, C.sub.1-C.sub.4 alkyl, oxo;
[0051] X is CH, when R.sub.3 is hydrogen; or X--R.sub.3 is O; or X
is N, R.sub.3 represents groups of hydrogen, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.10
aryl or heteroaryl, (C.sub.3-C.sub.7cycloalkyl)C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 alkoxy,
C.sub.1-C.sub.6 alkylthio, C.sub.2-C.sub.6 alkanoyl,
C.sub.1-C.sub.6 alkoxycarbonyl, C.sub.2-C.sub.6 alkanoyloxy, mono-
and di-(C.sub.3-C.sub.5 cycloalkyl)aminoC.sub.0-C.sub.4alkyl, (4-
to 7-membered heterocycle)C.sub.0-C.sub.4alkyl, C.sub.1-C.sub.6
alkylsulfonyl, mono- and di-(C.sub.1-C.sub.6 alkyl) sulfonamido,
and mono- and di-(C.sub.1-C.sub.6 alkyl)aminocarbonyl, each of
which is substituted with from 0 to 4 substituents independently
chosen from halogen, hydroxy, cyano, amino, --COOH and oxo.
[0052] R.sub.5 and R.sub.6 are independently selected from:
hydrogen, F, Cl, Br, CN, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.6
alkoxy.
[0053] R.sub.7, R.sub.8 and R.sub.9 are independently selected from
Hydrogene, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.10 aryl or heteroaryl,
C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkylthio, C.sub.2-C.sub.6
alkanoyl, C.sub.1-C.sub.6 alkoxycarbonyl, C.sub.2-C.sub.6
alkanoyloxy.
[0054] The term "halo" or "halogen" refers to fluorine, chlorine,
bromine or iodine.
[0055] The term "alkyl" herein alone or as part of another group
refers to a monovalent alkane (hydrocarbon) derived radical
containing from 1 to 12 carbon atoms unless otherwise defined.
Alkyl groups may be substituted at any available point of
attachment. An alkyl group substituted with another alkyl group is
also referred to as a "branched alkyl group". Exemplary alkyl
groups include methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl,
isobutyl, pentyl, hexyl, isohexyl, heptyl, dimethylpentyl, octyl,
2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, and the
like. Exemplary substituents include but are not limited to one or
more of the following groups: alkyl, aryl, halo (such as F, Cl, Br,
I), haloalkyl (such as CCl.sub.3 or CF.sub.3), alkoxy, alkylthio,
hydroxy, carboxy (--COOH), alkyloxycarbonyl (--C(O)R),
alkylcarbonyloxy (--OCOR), amino (--NH.sub.2), carbamoyl
(--NHCOOR-- or --OCONHR--), urea (--NHCONHR--) or thiol (--SH). In
some preferred embodiments of the present invention, alkyl groups
are substituted with, for example, amino, heterocycloalkyl, such as
morpholine, piperazine, piperidine, azetidine, hydroxyl, methoxy,
or heteroaryl groups such as pyrrolidine,
[0056] The term "cycloalkyl" herein alone or as part of another
group refers to fully saturated and partially unsaturated
hydrocarbon rings of 3 to 9, preferably 3 to 7 carbon atoms. The
examples include cyclopropyl, cyclobutyl, cyclopentyl and
cyclohexyl, and like. Further, a cycloalkyl may be substituted. A
substituted cycloalkyl refers to such rings having one, two, or
three substituents, selected from the group consisting of halo,
alkyl, substituted alkyl, alkenyl, alkynyl, nitro, cyano, oxo
(.dbd.O), hydroxy, alkoxy, thioalkyl, --CO.sub.2H, --C(.dbd.O)H,
CO.sub.2-alkyl, --C(.dbd.O)alkyl, keto, .dbd.N--OH,
.dbd.N--O-alkyl, aryl, heteroaryl, heterocyclo, --NR'R'',
--C(.dbd.O)NR'R'', --CO.sub.2NR'R'', --C(.dbd.O)NR'R'',
--NR'CO.sub.2R'', --NR'C(.dbd.O)R'', --SO.sub.2NR'R'', and
--NR'SO.sub.2R'', wherein each of R' and R'' are independently
selected from hydrogen, alkyl, substituted alkyl, and cycloalkyl,
or R' and R'' together form a heterocyclo or heteroaryl ring.
[0057] The term "alkenyl" herein alone or as part of another group
refers to a hydrocarbon radical straight, branched or cyclic
containing from 2 to 12 carbon atoms and at least one carbon to
carbon double bond. Examples of such groups include the vinyl,
allyl, 1-propenyl, isopropenyl, 2-methyl-1-propenyl, 1-butenyl,
2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl,
4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl,
1-heptenyl, and like. Alkenyl groups may also be substituted at any
available point of attachment. Exemplary substituents for alkenyl
groups include those listed above for alkyl groups, and especially
include C3 to C7 cycloalkyl groups such as cyclopropyl, cyclopentyl
and cyclohexyl, which may be further substituted with, for example,
amino, oxo, hydroxyl, etc.
[0058] The term "alkynyl" refers to straight or branched chain
alkyne groups, which have one or more unsaturated carbon-carbon
bonds, at least one of which is a triple bond. Alkynyl groups
include C2-C8 alkynyl, C.sub.2-C.sub.6 alkynyl and C.sub.2-C.sub.4
alkynyl groups, which have from 2 to 8, 2 to 6 or 2 to 4 carbon
atoms, respectively. Illustrative of the alkynyl group include
ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, pentenyl, and
hexenyl. Alkynyl groups may also be substituted at any available
point of attachment. Exemplary substituents for alkynyl groups
include those listed above for alkyl groups such as amino,
alkylamino, etc. The numbers in the subscript after the symbol "C"
define the number of carbon atoms a particular group can
contain.
[0059] The term "alkoxy" alone or as part of another group denotes
an alkyl group as described above bonded through an oxygen linkage
(--O--). Preferred alkoxy groups have from 1 to 8 carbon atoms.
Examples of such groups include the methoxy, ethoxy, n-propoxy,
isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy,
n-pentyloxy, isopentyloxy, n-hexyloxy, cyclohexyloxy, n-heptyloxy,
n-octyloxy and 2-ethylhexyloxy.
[0060] The term "alkylthio" refers to an alkyl group as described
above attached via a sulfur bridge. Preferred alkoxy and alkylthio
groups are those in which an alkyl group is attached via the
heteroatom bridge. Preferred alkylthio groups have from 1 to 8
carbon atoms. Examples of such groups include the methylthio,
ethylthio, n-propythiol, n-butylthiol, and like.
[0061] The term "oxo," as used herein, refers to a keto (C.dbd.O)
group. An oxo group that is a substituent of a nonaromatic carbon
atom results in a conversion of --CH.sub.2-- to --C(.dbd.O)--.
[0062] The term "alkoxycarbonyl" herein alone or as part of another
group denotes an alkoxy group bonded through a carbonyl group. An
alkoxycarbonyl radical is represented by the formula: --C(O)OR,
where the R group is a straight or branched C.sub.1-C.sub.6 alkyl
group, cycloalkyl, aryl, or heteroaryl.
[0063] The term "arylalkyl" herein alone or as part of another
group denotes an aromatic ring bonded through an alkyl group (such
as benzyl) as described above.
[0064] The term "aryl" herein alone or as part of another group
refers to monocyclic or bicyclic aromatic rings, e.g. phenyl,
substituted phenyl and the like, as well as groups which are fused,
e.g., napthyl, phenanthrenyl and the like. An aryl group thus
contains at least one ring having at least 6 atoms, with up to five
such rings being present, containing up to 20 atoms therein, with
alternating (resonating) double bonds between adjacent carbon atoms
or suitable heteroatoms. Aryl groups may optionally be substituted
with one or more groups including, but not limited to halogen such
as I, Br, F, or C1; alkyl, such as methyl, ethyl, propyl, alkoxy,
such as methoxy or ethoxy, hydroxy, carboxy, carbamoyl,
alkyloxycarbonyl, nitro, alkenyloxy, trifluoromethyl, amino,
cycloalkyl, aryl, heteroaryl, cyano, alkyl S(O)m (m=0, 1, 2), or
thiol.
[0065] The term "aromatic" refers to a cyclically conjugated
molecular entity with a stability, due to delocalization,
significantly greater than that of a hypothetical localized
structure, such as the Kekule structure.
[0066] The term "amino" herein alone or as part of another group
refers to --NH.sub.2. An "amino" may optionally be substituted with
one or two substituents, which may be the same or different, such
as alkyl, aryl, arylalkyl, alkenyl, alkynyl, heteroaryl,
heteroarylalkyl, cycloheteroalkyl, cycloheteroalkylalkyl,
cycloalkyl, cycloalkylalkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl,
thioalkyl, carbonyl or carboxyl. These substituents may be further
substituted with a carboxylic acid, any of the alkyl or aryl
substituents set out herein. In some embodiments, the amino groups
are substituted with carboxyl or carbonyl to form N-acyl or
N-carbamoyl derivatives.
[0067] The term "alkylsulfonyl" refers to groups of the formula
(SO.sub.2)-alkyl, in which the sulfur atom is the point of
attachment. Preferably, alkylsulfonyl groups include
C.sub.1-C.sub.6 alkylsulfonyl groups, which have from 1 to 6 carbon
atoms. Methylsulfonyl is one representative alkylsulfonyl
group.
[0068] The term "heteroatom" refers to any atom other than carbon,
for example, N, O, or S.
[0069] The term "heteroaryl" herein alone or as part of another
group refers to substituted and unsubstituted aromatic 5 or 6
membered monocyclic groups, 9 or 10 membered bicyclic groups, and
11 to 14 membered tricyclic groups which have at least one
heteroatom (O, S or N) in at least one of the rings. Each ring of
the heteroaryl group containing a heteroatom can contain one or two
oxygen or sulfur atoms and/or from one to four nitrogen atoms
provided that the total number of heteroatoms in each ring is four
or less and each ring has at least one carbon atom.
[0070] The term "heterocycle" or "heterocycloalkyl" herein alone or
as part of another group refers to a cycloalkyl group (nonaromatic)
in which one of the carbon atoms in the ring is replaced by a
heteroatom selected from O, S or N. The "heterocycle" has from 1 to
3 fused, pendant or spiro rings, at least one of which is a
heterocyclic ring (i.e., one or more ring atoms is a heteroatom,
with the remaining ring atoms being carbon). The heterocyclic ring
may be optionally substituted which means that the heterocyclic
ring may be substituted at one or more substitutable ring positions
by one or more groups independently selected from alkyl (preferably
lower alkyl), heterocycloalkyl, heteroaryl, alkoxy (preferably
lower alkoxy), nitro, monoalkylamino (preferably a lower
alkylamino), dialkylamino (preferably a alkylamino), cyano, halo,
haloalkyl (preferably trifluoromethyl), alkanoyl, aminocarbonyl,
monoalkylaminocarbonyl, dialkylaminocarbonyl, alkyl amido
(preferably lower alkyl amido), alkoxyalkyl (preferably a lower
alkoxy; lower alkyl), alkoxycarbonyl (preferably a lower
alkoxycarbonyl), alkylcarbonyloxy (preferably a lower
alkylcarbonyloxy) and aryl (preferably phenyl), said aryl being
optionally substituted by halo, lower alkyl and lower alkoxy
groups. A heterocyclic group may generally be linked via any ring
or substituent atom, provided that a stable compound results.
N-linked heterocyclic groups are linked via a component nitrogen
atom.
[0071] Typically, a heterocyclic ring comprises 1-4 heteroatoms;
within certain embodiments each heterocyclic ring has 1 or 2
heteroatoms per ring. Each heterocyclic ring generally contains
from 3 to 8 ring members (rings having from to 7 ring members are
recited in certain embodiments), and heterocycles comprising fused,
pendant or spiro rings typically contain from 9 to 14 ring members
which consists of carbon atoms and contains one, two, or three
heteroatoms selected from nitrogen, oxygen and/or sulfur.
[0072] Examples of "heterocycle" or "heterocycloalkyl groups
include piperazine, piperidine, morpholine, thiomorpholine,
pyrrolidine, imidazolidine and thiazolide.
[0073] The term "substituent," as used herein, refers to a
molecular moiety that is covalently bonded to an atom within a
molecule of interest. For example, a "ring substituent" may be a
moiety such as a halogen, alkyl group, haloalkyl group or other
group discussed herein that is covalently bonded to an atom
(preferably a carbon or nitrogen atom) that is a ring member.
[0074] The term "optionally substituted" as it refers that the aryl
or heterocyclyl or other group may be substituted at one or more
substitutable positions by one or more groups independently
selected from alkyl (preferably lower alkyl), alkoxy (preferably
lower alkoxy), nitro, monoalkylamino (preferably with one to six
carbons), dialkylamino (preferably with one to six carbons), cyano,
halo, haloalkyl (preferably trifluoromethyl), alkanoyl,
aminocarbonyl, monoalkylaminocarbonyl, dialkylaminocarbonyl, alkyl
amido (preferably lower alkyl amido), alkoxyalkyl (preferably a
lower alkoxy and lower alkyl), alkoxycarbonyl (preferably a lower
alkoxycarbonyl), alkylcarbonyloxy (preferably a lower
alkylcarbonyloxy) and aryl (preferably phenyl), said aryl being
optionally substituted by halo, lower alkyl and lower alkoxy
groups. Optional substitution is also indicated by the phrase
"substituted with from 0 to X substituents," where X is the maximum
number of possible substituents. Certain optionally substituted
groups are substituted with from 0 to 2, 3 or 4 independently
selected substituents.
[0075] A dash ("-") that is not between two letters or symbols is
used to indicate a point of t attachment for a substituent. For
example, --CONH.sub.2 is attached through the carbon atom.
[0076] A dashed cycle that locates inside of a heterocyle ring is
used to indicate a conjugated system. The bonds between two atoms
may be single bond or double bond. The term "kinase" refers to any
enzyme that catalyzes the addition of phosphate groups to a protein
residue; for example, serine and threonine kinases catalyze the
addition of phosphate groups to serine and threonine residues.
[0077] The term "therapeutically effective amount" refers to the
amount of the compound or pharmaceutical composition that will
elicit the biological or medical response of a tissue, system,
animal or human that is being sought by the researcher,
veterinarian, medical doctor or other clinician, e.g., restoration
or maintenance of vasculostasis or prevention of the compromise or
loss or vasculostasis; reduction of tumor burden; reduction of
morbidity and/or mortality.
[0078] The term "pharmaceutically acceptable" refers to the fact
that the carrier, diluent or excipient is compatible with the other
ingredients of the formulation and not deleterious to the recipient
thereof.
[0079] The terms "administration of a compound" or "administering a
compound" refer to the act of providing a compound of the invention
or pharmaceutical composition to the subject in need of
treatment.
[0080] The term "protected" refers that the group is in modified
form to preclude undesired side reactions at the protected site.
Suitable protecting groups for the compounds of the present
invention will be recognized from the present application taking
into account the level of skill in the art, and with reference to
standard textbooks, such as Greene, T. W. et al., Protective Groups
in Organic Synthesis, John Wiley & Sons, New York (1999).
[0081] The term "pharmaceutically acceptable salt" of a compound
recited herein is an acid or base salt that is suitable for use in
contact with the tissues of human beings or animals without
excessive toxicity or carcinogenicity, and preferably without
irritation, allergic response, or other problem or complication.
Such salts include mineral and organic acid salts of basic residues
such as amines, as well as alkali or organic salts of acidic
residues such as carboxylic acids. Specific pharmaceutical salts
include, but are not limited to, salts of acids such as
hydrochloric, phosphoric, hydrobromic, malic, glycolic, fumaric,
sulfuric, sulfamic, sulfanilic, formic, toluenesulfonic,
methanesulfonic, benzene sulfonic, ethane disulfonic,
2-hydroxyethylsulfonic, nitric, benzoic, 2-acetoxybenzoic, citric,
tartaric, lactic, stearic, salicylic, glutamic, ascorbic, pamoic,
succinic, fumaric, maleic, propionic, hydroxymaleic, hydroiodic,
phenylacetic, alkanoic such as acetic, HOOC-- (CH.sub.2)n-COOH
where n is 0-4, and the like. Similarly, pharmaceutically
acceptable cations include, but are not limited to sodium,
potassium, calcium, aluminum, lithium and ammonium. Those of
ordinary skill in the art will recognize further pharmaceutically
acceptable salts for the compounds provided herein. In general, a
pharmaceutically acceptable acid or base salt can be synthesized
from a parent compound that contains a basic or acidic moiety by
any conventional chemical method. Briefly, such salts can be
prepared by reacting the free acid or base forms of these compounds
with a stoichiometric amount of the appropriate base or acid in
water or in an organic solvent, or in a mixture of the two;
generally, the use of nonaqueous media, such as ether, ethyl
acetate, ethanol, isopropanol or acetonitrile, is preferred. It
will be apparent that each compound of Formula I may, but need not,
be formulated as a hydrate, solvate or non-covalent complex. In
addition, the various crystal forms and polymorphs are within the
scope of the present invention. Also provided herein are prodrugs
of the compounds of Formula I.
[0082] Preferred W groups of formula (I) are: F, Cl, Br, CN,
CF.sub.3, CF.sub.2H, CFH.sub.2, CH.sub.3, OCH.sub.3, NH.sub.2 and
the list below:
##STR00007##
[0083] Preferred substituted indole groups of formula (I) are
listed below:
##STR00008## ##STR00009## ##STR00010## ##STR00011## ##STR00012##
##STR00013## ##STR00014## ##STR00015## ##STR00016## ##STR00017##
##STR00018## ##STR00019## ##STR00020## ##STR00021## ##STR00022##
##STR00023## ##STR00024##
[0084] Preferred Ar groups of formula (I) are as below:
##STR00025## ##STR00026## ##STR00027## ##STR00028## ##STR00029##
##STR00030## ##STR00031## ##STR00032##
[0085] Examples of specific compounds of the present invention are
those compounds defined in the following:
##STR00033## ##STR00034## ##STR00035## ##STR00036## ##STR00037##
##STR00038## ##STR00039## ##STR00040## ##STR00041## ##STR00042##
##STR00043## ##STR00044## ##STR00045## ##STR00046## ##STR00047##
##STR00048## ##STR00049## ##STR00050## ##STR00051## ##STR00052##
##STR00053## ##STR00054## ##STR00055## ##STR00056## ##STR00057##
##STR00058## ##STR00059## ##STR00060## ##STR00061## ##STR00062##
##STR00063## ##STR00064## ##STR00065## ##STR00066## ##STR00067##
##STR00068## ##STR00069## ##STR00070## ##STR00071## ##STR00072##
##STR00073## ##STR00074## ##STR00075## ##STR00076## ##STR00077##
##STR00078## ##STR00079## ##STR00080## ##STR00081## ##STR00082##
##STR00083## ##STR00084## ##STR00085## ##STR00086## ##STR00087##
##STR00088## ##STR00089## ##STR00090## ##STR00091## ##STR00092##
##STR00093## ##STR00094## ##STR00095## ##STR00096## ##STR00097##
##STR00098## ##STR00099## ##STR00100## ##STR00101## ##STR00102##
##STR00103## ##STR00104## ##STR00105## ##STR00106## ##STR00107##
##STR00108## ##STR00109## ##STR00110## ##STR00111## ##STR00112##
##STR00113## ##STR00114## ##STR00115## ##STR00116## ##STR00117##
##STR00118## ##STR00119## ##STR00120## ##STR00121## ##STR00122##
##STR00123## ##STR00124## ##STR00125## ##STR00126## ##STR00127##
##STR00128## ##STR00129## ##STR00130## ##STR00131## ##STR00132##
##STR00133## ##STR00134## ##STR00135## ##STR00136## ##STR00137##
##STR00138## ##STR00139## ##STR00140## ##STR00141## ##STR00142##
##STR00143## ##STR00144## ##STR00145## ##STR00146## ##STR00147##
##STR00148## ##STR00149## ##STR00150## ##STR00151## ##STR00152##
##STR00153## ##STR00154## ##STR00155## ##STR00156## ##STR00157##
##STR00158## ##STR00159## ##STR00160## ##STR00161## ##STR00162##
##STR00163## ##STR00164## ##STR00165## ##STR00166## ##STR00167##
##STR00168## ##STR00169## ##STR00170## ##STR00171## ##STR00172##
##STR00173## ##STR00174## ##STR00175## ##STR00176## ##STR00177##
##STR00178## ##STR00179## ##STR00180## ##STR00181## ##STR00182##
##STR00183## ##STR00184## ##STR00185## ##STR00186## ##STR00187##
##STR00188## ##STR00189## ##STR00190## ##STR00191## ##STR00192##
##STR00193## ##STR00194## ##STR00195## ##STR00196## ##STR00197##
##STR00198## ##STR00199## ##STR00200## ##STR00201## ##STR00202##
##STR00203## ##STR00204## ##STR00205## ##STR00206##
##STR00207##
[0086] In another embodiment, a method of preparing the inventive
compounds is provided. The compounds of the present invention can
be generally prepared using 4, 6-dichloro-pyrimidine, with various
substituents on position "5". Compound (I) may contain various
stereoisomers, geometric isomers, tautomeric isomers, and the like.
All of possible isomers and their mixtures are included in the
present invention, and the mixing ratio is not particularly
limited.
[0087] The pyrimidine derivative compounds of Formula (I) in this
invention can be synthesized from commercially available precursors
using established protocols. By way of example, a synthetic route
similar to that shown in any of the following Schemes may be used,
together with synthetic methods known in the art of synthetic
organic chemistry, or variations thereon as appreciated by those
skilled in the art. Each variable in the following schemes refers
to any group consistent with the description of the compounds
provided herein.
[0088] In the Schemes that follow the term "reduction" refers to
the process of reducing a nitro functionality to an amino
functionality, or the process of transforming an ester
functionality to an alcohol. The reduction of a nitro group can be
carried out in a number of ways well known to those skilled in the
art of organic synthesis including, but not limited to, catalytic
hydrogenation, reduction with SnCI2 and reduction with titanium
bichloride. In the Schemes that follow, the term "hydrolyze" refers
to the reaction of a substrate or reactant with water. More
specifically, "hydrolyze" refers to the conversion of an ester or
nitrite functionality into a carboxylic acid. This process can be
catalyzed by a variety of acids or bases well known to those
skilled in the art of organic synthesis.
[0089] The compounds of Formula (I) may be prepared by use of known
chemical reactions and procedures. The following general
preparative methods are presented to aid one of skill in the art in
synthesizing the inhibitors, with more detailed examples being
presented in the experimental section describing the working
examples.
[0090] Propenyl-pyrazol amine as defined in formula (III) is not
commercially available. It can be prepared by several methods as
described earlier (see, e.g., U.S. provisional application No.
61/555,738).
##STR00208##
[0091] Precursors of substituted indol-5-ol as defined in formula
(III) can be purchased from suppliers, or synthesized from
commercially available precursors using established protocols. (WO
2004/009542, P33-38; Journal of Medicinal Chemistry, 2006, 49, No.
7, P2143-2146; Org. Lett. Vol 10, No 12, 2008, P 2369-2372; WO
00/47212, P245-250; WO 2009036055 A1, P57).
[0092] Especially, precursor 47-difluoroindol-5-ol as defined in
formula (IIIa) was not reported before and can be prepared by
several methods as described earlier (WO2014145403 A1).
##STR00209##
[0093] Precursors of 5-substituted 4, 6-dichloro-pyrimidines as
defined in formula (IV) can be purchased from suppliers.
Especially, precursor as defined in formula (IVa) can be
synthesized from commercially available precursors using
established protocols (PCT Int. Appl., 2010141406, 9 Dec. 2010,
Compound 310F).
##STR00210##
[0094] Generally, precursors of ArNH.sub.2 can be purchased from
suppliers. Precursors of ArNH.sub.2 as defined in formula (V) can
be purchased from suppliers, or synthesized from commercially
available precursors using established protocols. (J. Med. Chem.
2010, 53, 7938-7957, specifically, P7949).
##STR00211##
[0095] The preparation of the compounds of formula (I) in this
invention can be carried out by methods listed in scheme 1.
##STR00212##
[0096] As shown in scheme 1, the pyrimidine derivative (I) can be
synthesized by the reaction of 5-substituted 4,6-dichloropyrimidine
with a sequence of substituted idole-5-ol to give
monochlororopyrimidine intermediate of compound b, which can react
with ArNH.sub.2 to produce the final compound (I). The reaction can
be stepwise or in one pot. Alternative sequence can also be used to
make pyrimidine derivatives.
##STR00213## [0097] As shown in scheme 2, the final compounds as
defined in (I-b), can be synthesized from the corresponding
precursors, where W is "CN".
##STR00214##
[0098] The reaction is preferably conducted in the presence of an
inert solvent. There is no particular restriction on the nature of
the solvent to be employed, provided that it has no adverse effect
on the reaction or on the reagents involved and that it can
dissolve the reagents, at least to some extent. Examples of
suitable solvents include: aliphatic hydrocarbons, such as hexane,
heptane, ligroin and petroleum ether; aromatic hydrocarbons, such
as benzene, toluene and xylene; halogenated hydrocarbons,
especially aromatic and aliphatic hydrocarbons, such as methylene
chloride, chloroform, carbon tetrachloride, dichloroethane,
chlorobenzene and the dichlorobenzenes; esters, such as ethyl
formate, ethyl acetate, propyl acetate, butyl acetate and diethyl
carbonate; ethers, such as diethyl ether, diisopropyl ether,
tetrahydrofuran, dioxane, dimethoxyethane and diethylene glycol
dimethyl ether; ketones, such as acetone, methyl ethyl ketone,
methyl isobutyl ketone, isophorone and cyclohexanone; nitro
compounds, which may be nitroalkanes or nitroaranes, such as
nitroethane and nitrobenzene; nitriles, such as acetonitrile and
isobutyronitrile; amides, which may be fatty acid amides, such as
formamide, dimethylformamide, dimethylacetamide and
hexamethylphosphoric triamide; and sulphoxides, such as dimethyl
sulphoxide and sulpholane.
[0099] The reaction can take place over a wide range of
temperatures, and the precise reaction temperature is not critical
to the invention. In general, we find it convenient to carry out
the reaction at a temperature of from -50.degree. C. to 100.degree.
C.
[0100] The present invention provides compositions of matter that
are formulations of one or more active drugs and a
pharmaceutically-acceptable carrier. In this regard, the invention
provides a composition for administration to a mammalian subject,
which may include a compound of formula I, or its pharmaceutically
acceptable salts.
[0101] Pharmaceutically acceptable salts of the compounds of this
invention include those derived from pharmaceutically acceptable
inorganic and organic acids and bases. Examples of suitable acid
salts include acetate, adipate, alginate, aspartate, benzoate,
benzenesulfonate, bisulfate, butyrate, citrate, camphorate,
camphorsulfonate, cyclopentanepropionate, digluconate,
dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptanoate, glycerophosphate, glycolate, hemisulfate,
heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide,
2-hydroxyethanesulfonate, lactate, maleate, malonate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,
oxalate, palmoate, pectinate, persulfate, 3-phenylpropionate,
phosphate, picrate, pivalate, propionate, salicylate, succinate,
sulfate, tartrate, thiocyanate, tosylate and undecanoate. Other
acids, such as oxalic, while not in themselves pharmaceutically
acceptable, may be employed in the preparation of salts useful as
intermediates in obtaining the compounds of the invention and their
pharmaceutically acceptable acid addition salts.
[0102] Salts derived from appropriate bases include alkali metal
(e.g., sodium and potassium), alkaline earth metal (e.g.,
magnesium), ammonium and N+(C1-4 alkyl)4 salts. This invention also
envisions the quaternization of any basic nitrogen-containing
groups of the compounds disclosed herein. Water or oil-soluble or
dispersible products may be obtained by such quaternization.
[0103] The compositions of the present invention may be
administered orally, parenterally, by inhalation spray, topically,
rectally, nasally, buccally, vaginally or via an implanted
reservoir. The term "parenteral" as used herein includes
subcutaneous, intravenous, intramuscular, intra-articular,
intra-synovial, intrasternal, intrathecal, intrahepatic,
intralesional and intracranial injection or infusion techniques.
Preferably, the compositions are administered orally,
intraperitoneally or intravenously.
[0104] The pharmaceutically acceptable compositions of this
invention may be orally administered in any orally acceptable
dosage form including, but not limited to, capsules, tablets,
troches, elixirs, suspensions, syrups, wafers, chewing gums,
aqueous suspensions or solutions.
[0105] The oral compositions may contain additional ingredients
such as: a binder such as microcrystalline cellulose, gum
tragacanth or gelatin; an excipient such as starch or lactose, a
disintegrating agent such as alginic acid, corn starch and the
like; a lubricant such as magnesium stearate; a glidant such as
colloidal silicon dioxide; and a sweetening agent such as sucrose
or saccharin or flavoring agent such as peppermint, methyl
salicylate, or orange flavoring. When the dosage unit form is a
capsule, it may additionally contain a liquid carrier such as a
fatty oil. Other dosage unit forms may contain other various
materials which modify the physical form of the dosage unit, such
as, for example, a coating. Thus, tablets or pills may be coated
with sugar, shellac, or other enteric coating agents. A syrup may
contain, in addition to the active ingredients, sucrose as a
sweetening agent and certain preservatives, dyes and colorings and
flavors. Materials used in preparing these various compositions
should be pharmaceutically or veterinarally pure and non-toxic in
the amounts used.
[0106] For the purposes of parenteral therapeutic administration,
the active ingredient may be incorporated into a solution or
suspension. The solutions or suspensions may also include the
following components: a sterile diluent such as water for
injection, saline solution, fixed oils, polyethylene glycols,
glycerine, propylene glycol or other synthetic solvents;
antibacterial agents such as benzyl alcohol or methyl parabens;
antioxidants such as ascorbic acid or sodium bisulfite; chelating
agents such as ethylenediaminetetraacetic acid; buffers such as
acetates, citrates or phosphates and agents for the adjustment of
tonicity such as sodium chloride or dextrose. The parenteral
preparation can be enclosed in ampoules, disposable syringes or
multiple dose vials made of glass or plastic.
[0107] The pharmaceutical forms suitable for injectable use include
sterile solutions, dispersions, emulsions, and sterile powders. The
final form should be stable under conditions of manufacture and
storage. Furthermore, the final pharmaceutical form should be
protected against contamination and should, therefore, be able to
inhibit the growth of microorganisms such as bacteria or fungi. A
single intravenous or intraperitoneal dose can be administered.
Alternatively, a slow long-term infusion or multiple short-term
daily infusions may be utilized, typically lasting from 1 to 8
days. Alternate day dosing or dosing once every several days may
also be utilized.
[0108] Sterile, injectable solutions may be prepared by
incorporating a compound in the required amount into one or more
appropriate solvents to which other ingredients, listed above or
known to those skilled in the art, may be added as required.
Sterile injectable solutions may be prepared by incorporating the
compound in the required amount in the appropriate solvent with
various other ingredients as required. Sterilizing procedures, such
as filtration, may then follow. Typically, dispersions are made by
incorporating the compound into a sterile vehicle which also
contains the dispersion medium and the required other ingredients
as indicated above. In the case of a sterile powder, the preferred
methods include vacuum drying or freeze drying to which any
required ingredients are added.
[0109] Suitable pharmaceutical carriers include sterile water;
saline, dextrose; dextrose in water or saline; condensation
products of castor oil and ethylene oxide combining about 30 to
about 35 moles of ethylene oxide per mole of castor oil; liquid
acid; lower alkanols; oils such as corn oil; peanut oil, sesame oil
and the like, with emulsifiers such as mono- or di-glyceride of a
fatty acid, or a phosphatide, e.g., lecithin, and the like;
glycols; polyalkylene glycols; aqueous media in the presence of a
suspending agent, for example, sodium carboxymethylcellulose;
sodium alginate; poly(vinylpyrolidone); and the like, alone, or
with suitable dispensing agents such as lecithin; polyoxyethylene
stearate; and the like. The carrier may also contain adjuvants such
as preserving stabilizing, wetting, emulsifying agents and the like
together with the penetration enhancer. In all cases, the final
form, as noted, must be sterile and should also be able to pass
readily through an injection device such as a hollow needle. The
proper viscosity may be achieved and maintained by the proper
choice of solvents or excipients. Moreover, the use of molecular or
particulate coatings such as lecithin, the proper selection of
particle size in dispersions, or the use of materials with
surfactant properties may be utilized.
[0110] In accordance with the invention, there are provided
compositions containing pyrimidine derivatives and methods useful
for the in vivo delivery of pyrimidine derivatives in the form of
nanoparticles, which are suitable for any of the aforesaid routes
of administration.
[0111] U.S. Pat. Nos. 5,916,596, 6,506,405 and 6,537,579 teach the
preparation of nanoparticles from the biocompatible polymers, such
as albumin. Thus, in accordance with the present invention, there
are provided methods for the formation of nanoparticles of the
present invention by a solvent evaporation technique from an
oil-in-water emulsion prepared under conditions of high shear
forces (e.g., sonication, high pressure homogenization, or the
like).
[0112] Alternatively, the pharmaceutically acceptable compositions
of this invention may be administered in the form of suppositories
for rectal administration. These can be prepared by mixing the
agent with a suitable non-irritating excipient that is solid at
room temperature but liquid at rectal temperature and therefore
will melt in the rectum to release the drug. Such materials include
cocoa butter, beeswax and polyethylene glycols.
[0113] The pharmaceutically acceptable compositions of this
invention may also be administered topically, especially when the
target of treatment includes areas or organs readily accessible by
topical application, including diseases of the eye, the skin, or
the lower intestinal tract. Suitable topical formulations are
readily prepared for each of these areas or organs.
[0114] Topical application for the lower intestinal tract can be
effected in a rectal suppository formulation (see above) or in a
suitable enema formulation. Topically-transdermal patches may also
be used.
[0115] For topical applications, the pharmaceutically acceptable
compositions may be formulated in a suitable ointment containing
the active component suspended or dissolved in one or more
carriers. Carriers for topical administration of the compounds of
this invention include, but are not limited to, mineral oil, liquid
petrolatum, white petrolatum, propylene glycol, polyoxyethylene,
polyoxypropylene compound, emulsifying wax and water.
Alternatively, the pharmaceutically acceptable compositions can be
formulated in a suitable lotion or cream containing the active
components suspended or dissolved in one or more pharmaceutically
acceptable carriers. Suitable carriers include, but are not limited
to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl
esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and
water.
[0116] For ophthalmic use, the pharmaceutically acceptable
compositions may be formulated as micronized suspensions in
isotonic, pH adjusted sterile saline, or, preferably, as solutions
in isotonic, pH adjusted sterile saline, either with or without a
preservative such as benzylalkonium chloride. Alternatively, for
ophthalmic uses, the pharmaceutically acceptable compositions may
be formulated in an ointment such as petrolatum.
[0117] The pharmaceutically acceptable compositions of this
invention may also be administered by nasal aerosol or inhalation.
Such compositions are prepared according to techniques well-known
in the art of pharmaceutical formulation and may be prepared as
solutions in saline, employing benzyl alcohol or other suitable
preservatives, absorption promoters to enhance bioavailability,
fluorocarbons, and/or other conventional solubilizing or dispersing
agents.
[0118] Most preferably, the pharmaceutically acceptable
compositions of this invention are formulated for oral
administration.
[0119] In accordance with the invention, the compounds of the
invention may be used to treat diseases associated with cellular
proliferation or hyperproliferation, such as cancers which include
but are not limited to tumors of the nasal cavity, paranasal
sinuses, nasopharynx, oral cavity, oropharynx, larynx, hypopharynx,
salivary glands, and paragangliomas. The compounds of the invention
may also be used to treat cancers of the liver and biliary tree
(particularly hepatocellular carcinoma), intestinal cancers,
particularly colorectal cancer, ovarian cancer, small cell and
non-small cell lung cancer, breast cancer, sarcomas (including
fibrosarcoma, malignant fibrous histiocytoma, embryonal
rhabdomysocarcoma, leiomysosarcoma, neuro-fibrosarcoma,
osteosarcoma, synovial sarcoma, liposarcoma, and alveolar soft part
sarcoma), neoplasms of the central nervous systems (particularly
brain cancer), and lymphomas (including Hodgkin's lymphoma,
lymphoplasmacytoid lymphoma, follicular lymphoma, mucosa-associated
lymphoid tissue lymphoma, mantle cell lymphoma, B-lineage large
cell lymphoma, Burkitt's lymphoma, and T-cell anaplastic large cell
lymphoma).
[0120] The compounds and methods of the present invention, either
when administered alone or in combination with other agents (e.g.,
chemotherapeutic agents or protein therapeutic agents described
below) are also useful in treating a variety of disorders,
including but not limited to, for example: stroke, cardiovascular
disease, myocardial infarction, congestive heart failure,
cardiomyopathy, myocarditis, ischemic heart disease, coronary
artery disease, cardiogenic shock, vascular shock, pulmonary
hypertension, pulmonary edema (including cardiogenic pulmonary
edema), pleural effusions, rheumatoid arthritis, diabetic
retinopathy, retinitis pigmentosa, and retinopathies, including
diabetic retinopathy and retinopathy of prematurity, inflammatory
diseases, restenosis, asthma, acute or adult respiratory distress
syndrome (ARDS), lupus, vascular leakage, protection from ischemic
or reperfusion injury such as ischemic or reperfusion injury
incurred during organ transplantation, transplantation tolerance
induction; ischemic or reperfusion injury following angioplasty;
arthritis (such as rheumatoid arthritis, psoriatic arthritis or
osteoarthritis); multiple sclerosis; inflammatory bowel disease,
including ulcerative colitis and Crohn's disease; lupus (systemic
lupus crythematosis); graft vs. host diseases; T-cell mediated
hypersensitivity diseases, including contact hypersensitivity,
delayed-type hypersensitivity, and gluten-sensitive enteropathy
(Celiac disease); Type 1 diabetes; psoriasis; contact dermatitis
(including that due to poison ivy); Hashimoto's thyroiditis;
Sjogren's syndrome; Autoimmune Hyperthyroidism, such as Graves'
disease; Addison's disease (autoimmune disease of the adrenal
glands); autoimmune polyglandular disease (also known as autoimmune
polyglandular syndrome); autoimmune alopecia; pernicious anemia;
vitiligo; autoimmune hypopituatarism; Guillain-Barre syndrome;
other autoimmune diseases; cancers, including those where kineses
such as Src-family kineses are activated or overexpressed, such as
colon carcinoma and thymoma, or cancers where kinase activity
facilitates tumor growth or survival; glomerulonephritis, serum
sickness; uticaria; allergic diseases such as respiratory allergies
(asthma, hayfever, allergic rhinitis) or skin allergies; mycosis
fungoides; acute inflammatory responses (such as acute or adult
respiratory distress syndrome and ischemialreperfusion injury);
dermatomyositis; alopecia areata; chronic actinic dermatitis;
eczema; Behcet's disease; Pustulosis palmoplanteris; Pyoderma
gangrenum; Sezary's syndrome; atopic dermatitis; systemic
schlerosis; morphea; peripheral limb ischemia and ischemic limb
disease; bone disease such as osteoporosis, osteomalacia,
hyperparathyroidism, Paget's disease, and renal osteodystrophy;
vascular leak syndromes, including vascular leak syndromes induced
by chemotherapies or immunomodulators such as IL-2; spinal cord and
brain injury or trauma; glaucoma; retinal diseases, including
macular degeneration; vitreoretinal disease; pancreatitis;
vasculatides, including vasculitis, Kawasaki disease,
thromboangiitis obliterans, Wegener s granulomatosis, and Behcet's
disease; scleroderma; preeclampsia; thalassemia; Kaposi's sarcoma;
von Hippel Lindau disease; and the like.
[0121] In accordance with the invention, the compounds of the
invention may be used to treat diseases associated with undesired
cellular proliferation or hyperproliferation comprising identifying
the animal afflicted with said disease or condition and
administering to said afflicted animal a composition comprising the
compound of formula (I), wherein the disease or condition is
associated with a kinase.
[0122] The invention also provides methods of treating an animal
afflicted with the above diseases and conditions. The amount of the
compounds of the present invention that may be combined with the
carrier materials to produce a composition in a single dosage form
will vary depending upon the host treated, the particular mode of
administration. Preferably, the compositions should be formulated
so that a dosage of between 0.01-100 mg/kg body weight/day of the
inhibitor can be administered to a patient receiving these
compositions.
[0123] In one aspect, the invention compounds are administered in
combination with chemotherapeutic agent, an anti-inflammatory
agent, antihistamines, chemotherapeutic agent, immunomodulator,
therapeutic antibody or a protein kinase inhibitor, e.g., a
tyrosine kinase inhibitor, to a subject in need of such
treatment.
[0124] The method includes administering one or more of the
inventive compounds to the afflicted mammal. The method may further
include the administration of a second active agent, such as a
cytotoxic agent, including alkylating agents, tumor necrosis
factors, intercalators, microtubulin inhibitors, and topoisomerase
inhibitors. The second active agent may be co-administered in the
same composition or in a second composition. Examples of suitable
second active agents include, but are not limited to, a cytotoxic
drug such as Acivicin; Aclarubicin; Acodazole Hydrochloride;
AcrQnine; Adozelesin; Aldesleukin; Altretamine; Ambomycin;
Ametantrone Acetate; Aminoglutethimide; Amsacrine; Anastrozole;
Anthramycin; Asparaginase; Asperlin; Azacitidine; Azetepa;
Azotomycin; Batimastat; Benzodepa; Bicalutamide; Bisantrene
Hydrochloride; Bisnafide Dimesylate; Bizelesin; Bleomycin Sulfate;
Brequinar Sodium; Bropirimine; Busulfan; Cactinomycin; Calusterone;
Caracemide; Carbetimer; Carboplatin; Carmustine; Carubicin
Hydrochloride; Carzelesin; Cedefingol; Chlorambucil; Cirolemycin;
Cisplatin; Cladribine; Crisnatol Mesylate; Cyclophosphamide;
Cytarabine; Dacarbazine; Dactinomycin; Daunorubicin Hydrochloride;
Decitabine; Dexormaplatin; Dezaguanine; Dezaguanine Mesylate;
Diaziquone; Docetaxel; Doxorubicin; Doxorubicin Hydrochloride;
Droloxifene; Droloxifene Citrate; Dromostanolone Propionate;
Duazomycin; Edatrexate; Eflomithine Hydrochloride; Elsamitrucin;
Enloplatin; Enpromate; Epipropidine; Epirubicin Hydrochloride;
Erbulozole; Esorubicin Hydrochloride; Estramustine; Estramustine
Phosphate Sodium; Etanidazole; Ethiodized Oil 131; Etoposide;
Etoposide Phosphate; Etoprine; Fadrozole Hydrochloride; Fazarabine;
Fenretinide; Floxuridine; Fludarabine Phosphate; Fluorouracil;
Flurocitabine; Fosquidone; Fostriecin Sodium; Gemcitabine;
Gemcitabine Hydrochloride; Gold Au 198; Hydroxyurea; Idarubicin
Hydrochloride; Ifosfamide; Ilmofosine; Interferon Alfa-2a;
Interferon Alfa-2b; Interferon Alfa-n1; Interferon Alfa-n3;
Interferon Beta-.quadrature.a; Interferon Gamma-Ib; Iproplatin;
Irinotecan Hydrochloride; Lanreotide Acetate; Letrozole; Leuprolide
Acetate; Liarozole Hydrochloride; Lometrexol Sodium; Lomustine;
Losoxantrone Hydrochloride; Masoprocol; Maytansine; Mechlorethamine
Hydrochloride; Megestrol Acetate; Melengestrol Acetate; Melphalan;
Menogaril; Mercaptopurine; Methotrexate; Methotrexate Sodium;
Metoprine; Meturedepa; Mitindomide; Mitocarcin; Mitocromin;
Mitogillin; Mitomalcin; Mitomycin; Mitosper; Mitotane; Mitoxantrone
Hydrochloride; Mycophenolic Acid; Nocodazole; Nogalamycin;
Ormaplatin; Oxisuran; Paclitaxel; Pegaspargase; Peliomycin;
Pentamustine; Peplomycin Sulfate; Perfosfamide; Pipobroman;
Piposulfan; Piroxantrone Hydrochloride; Plicamycin; Plomestane;
Porfimer Sodium; Porfiromycin; Prednimustine; Procarbazine
Hydrochloride; Puromycin; Puromycin Hydrochloride; Pyrazofurin;
Riboprine; Rogletimide; Safmgol; Safingol Hydrochloride; Semustine;
Simtrazene; Sparfosate Sodium; Sparsomycin; Spirogermanium
Hydrochloride; Spiromustine; Spiroplatin; Streptonigrin;
Streptozocin; Strontium Chloride Sr 89; Sulofenur; Talisomycin;
Taxane; Taxoid; Tecogalan Sodium; Tegafur; Teloxantrone
Hydrochloride; Temoporfin; Teniposide; Teroxirone; Testolactone;
Thiamiprine; Thioguanine; Thiotepa; Tiazofurin; Tirapazamine;
Topotecan Hydrochloride; Toremifene Citrate; Trestolone Acetate;
Triciribine Phosphate; Trimetrexate; Trimetrexate Glucuronate;
Triptorelin; Tubulozole Hydrochloride; Uracil Mustard; Uredepa;
Vapreotide; Verteporfin; Vinblastine Sulfate; Vincristine Sulfate;
Vindesine; Vindesine Sulfate; Vinepidine Sulfate; Vinglycinate
Sulfate; Vinleurosine Sulfate; Vinorelbine Tartrate; Vinrosidine
Sulfate; Vinzolidine Sulfate; Vorozole; Zeniplatin; Zinostatin; and
Zorubicin Hydrochloride.
[0125] In accordance with the invention, the compounds and
compositions may be used at sub-cytotoxic levels in combination
with other agents in order to achieve highly selective activity in
the treatment of non-neoplastic disorders, such as heart disease,
stroke and neurodegenerative diseases (Whitesell et al., Curr
Cancer Drug Targets (2003), 3(5), 349-58).
[0126] The exemplary therapeutical agents that may be administered
in combination with invention compounds include EGFR inhibitors,
such as gefitinib, erlotinib, and cetuximab. Her2 inhibitors
include canertinib, EKB-569, and GW-572016. Also included are Src
inhibitors, dasatinib, as well as Casodex (bicalutamide),
Tamoxifen, MEK-1 kinase inhibitors, MARK kinase inhibitors, PI3
inhibitors, and PDGF inhibitors, such as imatinib, Hsp90
inhibitors, such as 17-AAG and 17-DMAG. Also included are
anti-angiogenic and antivascular agents which, by interrupting
blood flow to solid tumors, render cancer cells quiescent by
depriving them of nutrition. Castration, which also renders
androgen dependent carcinomas non-proliferative, may also be
utilized. Also included are IGF 1R inhibitors, inhibitors of
non-receptor and receptor tyrosine kineses, and inhibitors of
integrin.
[0127] The pharmaceutical composition and method of the present
invention may further combine other protein therapeutic agents such
as cytokines, immunomodulatory agents and antibodies. As used
herein the term "cytokine" encompasses chemokines, interleukins,
lymphokines, monokines, colony stimulating factors, and receptor
associated proteins, and functional fragments thereof. As used
herein, the term "functional fragment" refers to a polypeptide or
peptide which possesses biological function or activity that is
identified through a defined functional assay. The cytokines
include endothelial monocyte activating polypeptide II (EMAP-II),
granulocyte-macrophage-CSF (GM-CSF), granulocyte-CSF (G-CSF),
macrophage-CSF (M-CSF), IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-12,
and IL-13, interferons, and the like and which is associated with a
particular biologic, morphologic, or phenotypic alteration in a
cell or cell mechanism.
[0128] Other therapeutic agents for the combinatory therapy include
cyclosporins (e.g., cyclosporin A), CTLA4-Ig, antibodies such as
ICAM-3, anti-IL-2 receptor (Anti-Tac), anti-CD45RB, anti-CD2,
anti-CD3 (OKT-3), anti-CD4, anti-CD80, anti-CD86, agents blocking
the interaction between CD40 and gp39, such as antibodies specific
for CD40 and for gpn39 (i.e., CD154), fusion proteins constructed
from CD40 and gp39 (CD40Ig and CD8gp39), inhibitors, such as
nuclear translocation inhibitors, of NF-kappa B function, such as
deoxyspergualin (DSG), cholesterol biosynthesis inhibitors such as
HM:G CoA reductase inhibitors (lovastatin and simvastatin),
non-steroidal antiinflammatory drugs (NSAIDs) such as ibuprofen and
cyclooxygenase inhibitors such as rofecoxib, steroids such as
prednisone or dexamethasone, gold compounds, antiproliferative
agents such as methotrexate, FK506 (tacrolimus, Prograf),
mycophenolate mofetil, cytotoxic drugs such as azathioprine and
cyclophosphamide, TNF-.alpha. inhibitors such as tenidap, anti-TNF
antibodies or soluble TNF receptor, and rapamycin (sirolimus or
Rapamune) or derivatives thereof.
[0129] When other therapeutic agents are employed in combination
with the compounds of the present invention they may be used for
example in amounts as noted in the Physician Desk Reference (PDR)
or as otherwise determined by one having ordinary skill in the
art.
[0130] The following demonstrates the chemical structure and
synthesis of 2 "intermediates" useful in the synthesis of 61
"compounds." The 61 compounds will be tested for their ability to
inhibit a range of kinases.
[0131] All synthesis were performed under anhydrous conditions
(i.e. dry solvents) in an atmosphere of argon, except where stated,
using oven-dried apparatus and employing standard techniques in
handling air-sensitive materials. Aqueous solutions of sodium
bicarbonate (NaHCO3) and sodium chloride (brine) were
saturated.
[0132] Analytical thin layer chromatography (TLC) was carried out
on Merck Kiesel gel 60 F254 plates with visualization by
ultraviolet and/or anisaldehyde, potassium permanganate or
phosphomolybdic acid dips.
NMR spectra: 1H Nuclear magnetic resonance spectra were recorded at
400 MHz. Data are presented as follows: chemical shift,
multiplicity (s=singlet, d=doublet, t=triplet, q=quartet,
qn=quintet, dd=doublet of doublets, m=multiplet, bs=broad singlet),
coupling constant (J/Hz) and integration. Coupling constants were
taken and calculated directly from the spectra and are uncorrected.
Low resolution mass spectra: Electrospray (ES+) ionization was
used. The protonated parent ion (M+H) or parent sodium ion (M+Na)
or fragment of highest mass is quoted. Analytical gradient
consisted of 10% ACN in water ramping up to 100% ACN over 5 minutes
unless otherwise stated.
[0133] High performance liquid chromatography (HPLC) was use to
analyze the purity of derivatives. HPLC was performed on a
Phenomenex Synergi Polar-RP, 4u, 80 A, 150.times.4.6 mm column
using a Shimadzu system equipted with SPD-M10A Phosphodiode Array
Detector. Mobile phase A was water and mobile phase B was
acetonitrile with a gradient from 20% to 80% B over 60 minutes and
re-equilibrate at A/B (80:20) for 10 minutes. UV detection was at
220 and 54 nm.
The following demonstrates the chemical structure and synthesis of
two intermediate compounds ("intermediate 1" and "intermediate 2")
known by those of ordinary skill in art to have been useful in
synthesizing pyrimidine derivatives which can have kinase
inhibitory activity.
##STR00215##
To a solution of 4-fluoro-2-methyl-1H-indol-5-ol (200 mg, 1.21
mmol) in a mixture of acetonitrile (4 mL) and N,
N-dimethylformamide (1 mL) was added potassium carbonate (200 mg,
1.45 mmol). The reaction mixture was stirred for 1 h at room
temperature before a suspension of 4,
6-dichloropyrimidine-5-carbonitrile (221 mg, 1.27 mmol) in 3 mL of
acetonitrile was added. This mixture was stirred at room
temperature for 1 h. TLC was checked and the reaction was
completed. The mixture was diluted with water and ethyl acetate.
The layers were separated and the aqueous phase was extracted twice
with ethyl acetate. The combined organic phases were washed once
with water, then with brine, dried over sodium sulfate, filtered,
and the filtrate was concentrated in vacuo to give the desired
product as brown solids (365 mg, 99% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.46 (br, 1H), 8.83 (s, 1H), 7.17 (d, J=8.8
Hz, 1H), 7.00 (t, J=7.6 Hz, 1H), 6.27 (s, 1H), 2.41 (s, 3H);
ESI-MS: calcd for (C14H8ClFN4O) 302, found 303 (MH.sup.+).
##STR00216##
[0134] To a solution of 4,7-difluoro-2-methyl-1H-indol-5-ol (500
mg, 2.73 mmol) in a mixture of acetonitrile (9 mL) and N,
N-dimethylformamide (1 mL) was added potassium carbonate (453 mg,
3.28 mmol). The reaction mixture was stirred for 30 min. at room
temperature before a suspension of 2, 4-dichloro5-cyanopyrimidine
(499 mg, 2.87 mmol) in acetonitrile/DMF (2.5 mL/2.5 mL) was added.
This mixture was stirred at 0.degree. C. for 2 h. TLC was checked
and the reaction was completed. The mixture was diluted with
water/brine and ethyl acetate. The layers were separated and the
aqueous phase was extracted twice with ethyl acetate. The combined
organic phases were washed once with water/brine three times, dried
over sodium sulfate, filtered, and the filtrate was concentrated in
vacuo to give the deired compound as purple sticky solids (890 mg,
100% yield, contained some DMF). The product was used without
further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
11.91 (br, 1H), 8.84 (s, 1H), 7.03 (dd, J=5.6 Hz, J=10.4 Hz, 1H),
6.35 (br, 1H), 2.39 (s, 3H); ESI-MS: calcd for (C14H7ClF2N4O) 320,
found 321 (MH.sup.+).
[0135] The following demonstrate the chemical structure and a
method of synthesizing 61 pyrimidine derivatives ("compound 1" to
"compound 61") which are species the generic compound disclosed
herein.
##STR00217##
[0136] A mixture of Intermediate 1 (100 mg, 0.33 mmol),
2-methoxy-4-(4-methylpiperazin-1-yl)aniline (73 mg, 0.33 mmol), and
DIPEA (0.08 ml, 0.49 mmol) in DMSO (5 ml) was stirred at room
temperature for 30 min. After checking the TLC, the mixture was
added to water (100 ml). After cooled with ice-bath, the solids
were collected by filtration, washed by water. The crude product
was purified by column chromatography (silica gel, 0-15% MeOH in
DCM) to give the desired product as yellow solids (64 mg, 40%
yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.34 (br, 1H),
9.45 (br s, 1H), 8.15 (s, 1H), 7.13 (m, 2H), 6.93 (m, 1H), 6.65 (m,
1H), 6.50 (m, 1H), 6.23 (s, 1H), 3.77 (s, 3H), 3.18 (m, 4H), 2.46
(m, 4H), 2.39 (s, 3H), 2.23 (s, 3H); ESI-MS: calcd for C26H26FN7O2
487, found 488 (MH.sup.+). HPLC: retention time: 18.63 min. purity:
96%.
##STR00218##
[0137] A mixture Intermediate 1 (100 mg, 0.33 mmol),
3-fluoro-4-(4-methylpiperazin-1-yl)aniline (69 mg, 0.33 mmol), and
DIPEA (0.15 ml, 0.82 mmol) in DMSO (5 ml) was stirred at room
temperature for 30 min. After checking the TLC, the mixture was
added to water (100 ml). After cooled with ice-bath, the solids
were collected by filtration, washed by water. The crude product
was purified by column chromatography (silica gel, 0-15% MeOH in
DCM) to give the desired product as light brown solids (90 mg, 57%
yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.36 (br, 1H),
10.0 (s, 1H), 8.30 (s, 1H), 7.41 (m, 1H), 7.26 (m, 1H), 7.13 (m,
1H), 6.98 (m, 2H), 6.24 (br s, 1H), 3.00 (m, 4H), 2.50 (m, 4H),
2.40 (s, 3H), 2.23 (s, 3H); ESI-MS: calcd for C25H23F2N7O 475,
found 476 (MH.sup.+). HPLC: retention time: 19.15 min. purity:
95%.
##STR00219##
[0138] A mixture Intermediate 1 (100 mg, 0.33 mmol),
4-((4-methylpiperazin-1-yl)methyl)aniline (68 mg, 0.33 mmol), and
DIPEA (0.15 ml, 0.82 mmol) in DMSO (5 ml) was stirred at room
temperature for 30 min. After checking the TLC, the mixture was
added to water (100 ml). After cooled with ice-bath, the solids
were collected by filtration, washed by water. The crude product
was purified by column chromatography (silica gel, 0-15% MeOH in
DCM) to give the desired product as off white solids (41 mg, 26%
yield).sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.34 (br s, 1H),
8.24 (m, 1H), 7.12 (m, 1H), 6.92 (m, 1H), 6.22 (s, 1H), 3.95 (m,
4H), 2.41 s (m, 4H), 2.39 (s, 3H), 2.24 (s, 3H); ESI-MS: calcd for
C19H19FN6O 366, found 367 (MH.sup.+). HPLC: retention time: 12.36
min. purity: 98%.
##STR00220##
[0139] A mixture Intermediate 1 (100 mg, 0.33 mmol),
4-((4-ethylpiperazin-1-yl)methyl)aniline (72 mg, 0.33 mmol), and
DIPEA (0.15 ml, 0.82 mmol) in DMSO (5 ml) was stirred at room
temperature for 30 min. After checking the TLC, the mixture was
added to water (100 ml). After cooled with ice-bath, the solids
were collected by filtration, washed by water. The crude product
was purified by column chromatography (silica gel, 0-15% MeOH in
DCM) to give the desired product as off white solids (18 mg, 11%
yield).sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.34 (br s, 1H),
8.24 (s, 1H), 7.12 (m, 1H), 6.92 (m, 1H), 6.23 (s, 1H), 3.96 (m,
4H), 2.52 (m, 3H), 2.45 (m, 4H), 1.23 (m, 2H), 1.050 (m, 3H);
ESI-MS: calcd for C20H21FN6O 380, found 381 (MH.sup.+). HPLC:
retention time: 13.77 min. purity: 93%.
##STR00221##
[0140] Step 1: To a solution of flouroindole (415 mg, 2.51 mmol)
and 4,6-dichloro-5-(difluoromethyl)pyridine (500 mg, 2.51 mmol) in
DMSO (3 ml) was added potassium carbonate (695 mg, 5.03 mmol) at
room temperature and the mixture was heated at room temperature for
overnight. TLC was checked and the starting material was consumed.
The reaction mixture was added to a flask with water/brine (50
ml/50 ml) and the mixture was stirred at room temperature for 1
hour then colled with ice bath. The solids were collected by
filtration and washed by water to obtain the desired product as
yellow solids (803 mg, 98% yield). No further purification was
performed and the product was used for the next step reaction.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.39 (br, 1H), 8.71
(s, 1H), 7.51 (t, J=52.4 Hz, 1H), 7.15 (d, J=8.8 Hz, 1H), 6.96 (t,
J=7.6 Hz, 1H), 6.25 (s, 1H), 2.40 (s, 3H); ESI-MS: calcd for
(C14H9ClF3N3O) 327, found 328 (MH.sup.+).
[0141] Step 2: A mixture of above intermediate (125 mg, 0.61 mmol),
4-(4-Ethylpiperazine-1-yl))aniline (200 mg, 0.61 mmol), and DIPEA
(0.27 ml, 1.52 mmol) in DMSO (3.0 ml) was stirred at 100.degree. C.
for 2 h, then at room temperature for overnight. TLC was checked
and the reaction was completed. The mixture was added to water/sat.
NH.sub.4Cl (50 ml/50 ml) and stirred at room temperature for 30
min. The pH of the mixture was adjusted to .about.6 using 2N HCl.
Cooled at 4.degree. C. and the solids were collected by filtration,
washed by water to give the sticky fine crude product. The crude
product was dissolved into DCM/MeOH (2 ml/2 ml), dried over sodium
sulfate and concentrated. The crude product was purified on column
(0-10% MeOH in DCM) to give the desired product as yellow solids
(103 mg, 34% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
11.32 (br, 1H), 10.49 (br, 1H), 8.93 (s, 1H), 8.12 (s, 1H), 7.36
(d, J=8.8 Hz, 2H), 7.10 (d, J=8.8 Hz, 1H), 7.00 (d, J=9.2 Hz, 2H),
6.86 (t, J=7.6 Hz, 1H), 6.21 (s, 1H), 3.80 (br, 2H), 3.55 (br, 2H),
3.10 (m, 6H), 2.39 (s, 3H), 1.28 (t, J=7.2 Hz, 3H); ESI-MS: calcd
for (C26H27F3N6O) 496, found 497 (MH.sup.+).
##STR00222##
[0142] A flask was charged with intermediate 1 (150 mg, 0.5 mmol),
1-(4-aminobenzoyl)-4-methyl piperazine (109 mg, 0.5 mmol), TFA (50
uL), isopropanol (3 mL). The reaction was heated to 100.degree. C.
for 4 h. The reaction mixture was basified with a saturated aqueous
sodium bicarbonate solution and then was extracted with DCM/(10
ml.times.3). The combined organic was washed by brine, dried over
sodium sulfate and concentrated. The crude product was purified
with flash chromatography (0-10% MeOH in DCM) to afford the desired
product as light yellow solids (160 mg, 66% yield). .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 11.36 (br, 1H), 10.17 (br, 1H),
8.34 (s, 1H), 7.63 (d, J=8.8 Hz, 2H), 7.39 (d, J=8.8 Hz, 2H), 7.13
(d, J=8.4 Hz, 1H), 6.96 (t, J=7.6 Hz, 1H), 6.24 (s, 1H), 3.80-3.40
(br, 4H), 2.40 (s, 3H), 2.36-2.24 (br, 4H), 2.19 (s, 3H); ESI-MS:
calcd for (C26H24FN7O2) 485, found 486 (MH.sup.+).
##STR00223##
[0143] A flask was charged with intermediate 1 (100 mg, 0.33 mmol),
4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)aniline (86 mg, 0.33
mmol), TFA (50 uL), isopropanol (3 mL). The reaction was heated to
100.degree. C. for 4 h. The reaction mixture was basified with a
saturated aqueous sodium bicarbonate solution and then was
extracted with DCM/(10 ml.times.3). The combined organic was washed
by brine, dried over sodium sulfate and concentrated. The crude
product was purified with flash chromatography (0-10% MeOH-in DCM)
to afford the desired product as light yellow solids (103 mg, 60%
yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.36 (br, 1H),
10.18 (br, 1H), 8.33 (s, 1H), 7.90-7.80 (m, 2H), 7.55 (d, J=8.8 Hz,
1H), 7.13 (d, J=8.8 Hz, 1H), 6.95 (t, J=7.2 Hz, 1H), 6.24 (s, 1H),
2.85 (m, 4H), 2.49-2.32 (m, 7H), 2.23 (s, 3H); ESI-MS: calcd for
(C26H23F4N7O) 525, found 526 (MH.sup.+).
##STR00224##
[0144] A flask was charged with intermediate 1 (100 mg, 0.33 mmol),
4-(4-methylpiperazin-1-yl)-3-fluoroaniline (74 mg, 0.33 mmol), TFA
(50 uL), isopropanol (3 mL). The reaction was heated to 100.degree.
C. for 4 h. The reaction mixture was basified with a saturated
aqueous sodium bicarbonate solution and then was extracted with
DCM/(10 ml.times.3). The combined organic was washed by brine,
dried over sodium sulfate and concentrated. The crude product was
purified with flash chromatography (0-10% MeOH-in DCM) to afford
the desired product as light yellow solids (120 mg, 74% yield).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.36 (br, 1H), 9.99
(br, 1H), 8.29 (s, 1H), 7.39 (dd, J=2.0 Hz, J=14.4 Hz, 1H), 7.26
(d, J=8.8 Hz, 1H), 7.13 (d, J=8.4 Hz, 1H), 7.01 (t, J=9.2 Hz, 1H),
6.95 (t, J=8.0 Hz, 1H), 6.24 (s, 1H), 3.00 (m, 4H), 2.60-2.45 (m,
4H), 2.40 (m, 5H), 1.02 (t, J=7.2 Hz, 3H); ESI-MS: calcd for
(C26H25F2N7O) 489, found 490 (MH.sup.+).
##STR00225##
[0145] A mixture intermediate 1 (100 mg, 0.33 mmol), tert-butyl
4-(4-amino-2-fluorophenyl)piperazine-1-carboxylate (97 mg, 0.33
mmol), and DIPEA (0.15 ml, 0.82 mmol) in DMSO (5 ml) was stirred at
room temperature for 30 min. After checking the TLC, the mixture
was added to water (100 ml). After cooled with ice-bath, the solids
were collected by filtration, washed by water. After air-drying at
room temperature overnight, the solids were suspended into DCM/MeOH
(10/1, 5 mL) and 1 ml of TFA was added. The mixture was stirred at
room temperature for overnight. After concentrated, the residue was
dissolved into DCM/MeOH (8/2, 15 ml) and sat. Sodium bicarbonate
solution was added to pH about 7. The organic was dried over sodium
sulfate and concentrated. The crude product was purified by column
chromatography (silica gel, 0-15% MeOH in DCM) to give the desired
product as off white solids (60 mg, 39% yield). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 11.36 (br, 1H), 8.30 (s, 1H), 7.43 (m,
1H), 7.28 (m, 1H), 7.14 (m, 1H), 7.02 (m, 1H), 6.94 (m, 1H), 6.24
(s, 1H), 2.97 (m, 8H), 2.40 (m, 3H); ESI-MS: calcd for C24H21F2N7O
461, found 462 (MH.sup.+). HPLC: retention time: NA
##STR00226##
[0146] A mixture intermediate 1 (100 mg, 0.33 mmol), tert-butyl
4-(4-aminobenzyl)piperazine-1-carboxylate (96 mg, 0.33 mmol), and
DIPEA (0.15 ml, 0.82 mmol) in DMSO (5 ml) was stirred at room
temperature for 30 min. After checking the TLC, the mixture was
added to water (100 ml). After cooled with ice-bath, the solids
were collected by filtration, washed by water. After air-drying at
room temperature overnight, the solids were suspended into DCM/MeOH
(10/1, 5 mL) and 1 ml of TFA was added. The mixture was stirred at
room temperature for overnight. After concentrated, the residue was
dissolved into DCM/MeOH (8/2, 15 ml) and sat. Sodium bicarbonate
solution was added to pH about 7. The organic was dried over sodium
sulfate and concentrated. The crude product was purified by column
chromatography (silica gel, 0-15% MeOH in DCM) to give the desired
product as light yellow solids (15 mg, 10% yield). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 11.48 (br, 1H), 10.06 (s, 1H), 8.90 (m,
2H), 8.28 (m, 1H), 7.48 (m, 2H), 7.29 (m, 2H), 7.13 (m, 1H), 6.94
(m, 1H), 6.23 (s, 1H), 3.53 (m, 2H), 3.09 (m, 4H), 2.54 (m, 4H),
2.40 (m, 3H); ESI-MS: calcd for C25H24FN7O 457, found 458
(MH.sup.+). HPLC: retention time: 13.75 min. purity: 91%.
##STR00227##
[0147] Step 1: A mixture of 1-fluoro-4-nitrobenzene (3.00 g, 21.3
mmol), N,N-dimethylpiperidin-4-amine dihydrochloride (4.70 g, 23.4
mmol) and DIPEA (15 mL, 86.1 mmol) was stirred at 95.degree. C. for
18 h. The mixture was cooled to room temperature, diluted with 1:1
EtOAc/hexanes (100 mL), washed twice with aq. calcium gluconate
(100 mL each, 50% saturation), and the organic layer was separated
and dried over anhydrous Na.sub.2SO.sub.4 and concentrated to yield
the desired product as a red oil (4.1 g, 77% yield). .sup.1H NMR
(DMSO-d6, 400 MHz) .delta. 8.03 (d, J=9.6 Hz, 1H), 7.01 (d, J=9.6
Hz, 1H), 4.04-4.01 (m, 2H), 3.02-2.95 (m, 2H), 2.39-2.32 (m, 1H),
2.17 (s, 6H), 1.85-1.81 (m, 2H), 1.44-1.34 (m, 2H); MS (ESI): calcd
for C13H19N3O2: 249, found: 250 (MH.sup.+).
[0148] Step 2: A mixture of
N,N-dimethyl-1-(4-nitrophenyl)piperidin-4-amine (1.5 g, 6.0 mmol),
tin (II) chloride dihydrate (6.8 g, 30 mmol) and methanol (100 mL)
was stirred at 70.degree. C. for 19 h. The mixture was cooled to
room temperature, diluted with 4:1 EtOAc/hexanes (200 mL), washed
with aq. 5M NaOH (200 mL), and the organic layer was separated and
dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The
resulting residue was purified by flash column chromatography on
silica gel using 0-20% MeOH in DCM (v/v) as eluent to afford the
desired product 1-(4-aminophenyl)-N,N-dimethylpiperidin-4-amine as
a light yellow solid (340 mg, 26% yield). .sup.1H NMR (DMSO-d6, 400
MHz) .delta. 6.67 (d, J=8.8 Hz, 2H), 6.47 (d, J=8.8 Hz, 2H), 4.53
(br s, 2H), 3.37-3.34 (m, 2H), 2.48-2.42 (m, 2H), 2.18 (s, 6H),
2.11-2.05 (m, 1H), 1.80-1.77 (m, 2H), 1.51-1.41 (m, 2H); MS (ESI):
calcd for C13H21N3: 219, found: 220 (MH.sup.+).
[0149] Step 3: To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (200 mg, 0.661 mmol) and
1-(4-aminophenyl)-N,N-dimethylpiperidin-4-amine (142 mg, 0.647
mmol) in anhydrous DMSO (2.0 mL) was added TEA (0.28 mL; 1.98
mmol), and the resulting biphasic mixture was efficiently stirred
at room temperature under argon atmosphere for 19 h. The resulting
mixture was diluted with 4:1 EtOAc/hexanes (100 mL) and washed with
aq. NH.sub.4Cl (ca. 100 mL each; 50% NH.sub.4Cl saturation)
followed by brine (100 mL; 50% saturation). The organic layer was
separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by crystallization
out of EtOAc to afford the desired product as a yellow solid (254
mg, 81% yield). .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.35 (br s,
1H), 9.83 (br s, 1H), 8.21 (s, 1H), 7.30-7.28 (m, 2H), 7.13-7.11
(m, 1H), 6.95-6.91 (m, 2H), 6.23 (s, 1H), 3.71-3.68 (m, 2H),
2.69-2.62 (m, 2H), 2.40 (s, 3H), 2.24-2.21 (m, 1H), 2.19 (s, 6H),
1.84-1.81 (m, 2H), 1.52-1.42 (m, 2H); MS (ESI): calcd for
C27H28FN7O: 485, found: 486 (MH.sup.+).
##STR00228##
[0150] A mixture of intermediate 1 (150 mg, 0.50 mmol),
4-imidazole-1-yl-phenylamine (91 mg, 0.57 mmol), and DIPEA (0.22
ml, 1.245 mmol) in DMSO (3.0 ml) was stirred at room temperature
for overnight. TLC was checked and the reaction was completed. The
mixture was added to sat. NH.sub.4Cl/water (25 ml/50 ml) and
stirred at room temperature for 30 min. The pH of the mixture was
adjusted to about 6 using 2N HCl. After cooled with ice for 1 h,
the solids were collected by filtration, washed by water to give
the crude product. The crude product was purified by column on
silica gel (0-10% MeOH in DCM) to give the desired product as
yellow solids (140 mg, 66% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.36 (br, 1H), 10.16 (s, 1H), 8.31 (s, 1H),
8.24 (s, 1H), 7.66 (m, 5H), 7.14 (m, 2H), 6.95 (t, J=7.6 Hz, 1H),
6.24 (s, 1H), 2.40 (s, 3H); ESI-MS: calcd for (C23H16FN7O) 425,
found 426 (MH.sup.+).
##STR00229##
[0151] A mixture of intermediate 1 (150 mg, 0.50 mmol),
4-(2-methylimidazole-1-yl-phenylamine (99 mg, 0.57 mmol), and DIPEA
(0.22 ml, 1.245 mmol) in DMSO (3.0 ml) was stirred at room
temperature for overnight. TLC was checked and the reaction was
completed. The mixture was added to sat. NH.sub.4Cl/water (25 ml/50
ml) and stirred at room temperature for 30 min. The pH of the
mixture was adjusted to about 6 using 2N HCl. After cooled with ice
for 1 h, the solids were collected by filtration, washed by water
to give the crude product. The crude product was purified by column
on silica gel (0-10% MeOH in DCM) to give the desired product as
yellow solids (108 mg, 50% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.36 (br, 1H), 10.16 (br, 1H), 8.33 (s, 1H),
7.70 (d, J=8.8 Hz, 2H), 7.42 (d, J=8.8 Hz, 2H), 7.28 (br, 1H), 7.14
(d, J=8.4 Hz, 1H), 6.96 (t, J=7.6 Hz, 1H), 6.25 (s, 1H), 2.41 (s,
3H), 2.30 (s, 3H); ESI-MS: calcd for (C24H18FN7O) 439, found 440
(MH.sup.+).
##STR00230##
[0152] A mixture of intermediate 2 (150 mg, 0.47 mmol), tert-butyl
4-(4-aminophenyl)piperazine-1-carboxylate (149 mg, 0.54 mmol), and
DIPEA (0.21 ml, 1.17 mmol) in DMSO (3.0 ml) was stirred at room
temperature for overnight. TLC was checked and the reaction was
completed. The mixture was added to sat. NH.sub.4Cl/water (25 ml/50
ml) and stirred at room temperature for 30 min. The pH of the
mixture was adjusted to about 6 using 2N HCl. After cooled with ice
for 1 h, the solids were collected by filtration, washed by water
to give the crude product. The crude product was suspended in DCM
(10 ml) and 1 ml of TFA was added (the mixture become clear
solution). The mixture was stirred at room temperature for
overnight. Potassium phosphate in water was added to the mixture
(pH about 8), and extracted with DCM/MeOH. The combined organic was
washed by brine, concentrated and purified by column on silica gel
(5-15% MeOH in DCM) to give the desired product as yellow solids
(97 mg, 45% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
11.83 (br, 1H), 9.90 (br, 1H), 8.24 (s, 1H), 7.32 (d, J=8.8 Hz,
2H), 7.00 (dd, J=5.6 Hz, J=10.4 Hz, 1H), 6.94 (d, J=8.8 Hz, 2H),
6.34 (s, 1H), 3.14 (m, 4H), 2.99 (m, 4H), 2.41 (s, 3H) (NH may be
berried under solvent peak); ESI-MS: calcd for (C24H21F2N7O) 461,
found 462 (MH.sup.+).
##STR00231##
[0153] A flask was charged with intermediate 1 (100 mg, 0.33 mmol),
5-Amino-2-methylbenzenesulfonamide (77 mg, 0.41 mmol), TFA (50 uL),
isopropanol (3 mL). The reaction was heated to 100.degree. C. for 4
h. The reaction mixture was basified with a saturated aqueous
sodium bicarbonate solution and then was extracted with DCM/(10
ml.times.3). The combined organic was washed by brine, dried over
sodium sulfate and concentrated. The crude product was purified
with flash chromatography (0-10% MeOH-in DCM) to afford the desired
product as light yellow solids (79 mg, 53% yield). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 11.36 (br, 1H), 10.25 (br, 1H), 8.30 (s,
1H), 8.07 (br, 1H), 7.69 (dd, J=2.4 Hz, J=8.4 Hz, 1H), 7.39 (br,
2H), 7.37 (d, J=8.4 Hz, 1H), 7.13 (d, J=8.8 Hz, 1H), 6.95 (t, J=7.6
Hz, 1H), 6.24 (s, 1H), 2.57 (s, 3H), 2.40 (s, 3H); ESI-MS: calcd
for (C21H17FN6O3S) 452, found 453 (MH.sup.+).
##STR00232##
[0154] A flask was charged with intermediate 1 (100 mg, 0.33 mmol),
4-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-fluoroaniline (92 mg, 0.41
mmol), TFA (50 uL), isopropanol (3 mL). The reaction was heated to
100.degree. C. for 4 h. The reaction mixture was basified with a
saturated aqueous sodium bicarbonate solution and then was
extracted with DCM/(10 ml.times.3). The combined organic was washed
by brine, dried over sodium sulfate and concentrated. The crude
product was purified with flash chromatography (0-10% MeOH-in DCM)
to afford the desired product as yellow solids (118 mg, 73% yield).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.35 (br, 1H), 9.98
(br, 1H), 8.29 (s, 1H), 7.40 (dd, J=2.4 Hz, J=14.4 Hz, 1H), 7.26
(dd, J=2.0 Hz, J=8.4 Hz, 1H), 7.12 (d, J=8.8 Hz, 1H), 6.95 (m, 2H),
6.24 (s, 1H), 3.17 (d, J=9.6 Hz, 2H), 2.91 (m, 2H), 2.40 (s, 3H),
2.19 (m, 3H), 1.00 (s, 3H), 0.98 (s, 3H); ESI-MS: calcd for
(C26H25F2N7O) 489, found 490 (MH.sup.+).
##STR00233##
[0155] A flask was charged with intermediate 1 (100 mg, 0.33 mmol),
4-((3S,5R)-3,5-dimethylpiperazin-1-yl)aniline (85 mg, 0.41 mmol),
TFA (50 uL), isopropanol (5 mL). The reaction was heated to
100.degree. C. for 4 h. The reaction mixture was basified with a
saturated aqueous sodium bicarbonate solution and then was
extracted with DCM/(10 ml.times.3). The combined organic was washed
by brine, dried over sodium sulfate and concentrated. The crude
product was purified with flash chromatography (0-10% MeOH-in DCM)
to afford the desired product as yellow solids (122 mg, 78% yield).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.36 (br, 1H), 9.84
(br, 1H), 8.21 (s, 1H), 7.28 (d, J=8.8 Hz, 2H), 7.12 (d, J=8.8 Hz,
1H), 6.92 (m, 3H), 6.24 (s, 1H), 3.51 (m, 2H), 2.84 (m, 2H), 2.40
(s, 3H), 2.11 (m, 3H), 1.03 (s, 3H), 1.01 (s, 3H); ESI-MS: calcd
for (C26H26FN7O) 471, found 472 (MH.sup.+).
##STR00234##
[0156] A flask was charged with intermediate 1 (100 mg, 0.33 mmol),
4-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)aniline (86 mg, 0.33
mmol), TFA (50 uL), isopropanol (5 mL). The reaction was heated to
100.degree. C. for 4 h. The reaction mixture was basified with a
saturated aqueous sodium bicarbonate solution and then was
extracted with DCM/(10 ml.times.3). The combined organic was washed
by brine, dried over sodium sulfate and concentrated. The crude
product was purified with flash chromatography (0-10% EtOAc in DCM)
to afford the desired product as yellow solids (73 mg, 42% yield).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.36 (br, 1H), 9.86
(br, 1H), 8.21 (s, 1H), 7.31 (d, J=8.8 Hz, 2H), 7.12 (d, J=8.4 Hz,
1H), 6.94 (m, 3H), 6.24 (s, 1H), 3.23 (q, J=10 Hz, 2H), 3.13 (m,
4H), 2.76 (m, 4H), 2.34 (s, 3H); ESI-MS: calcd for (C26H23F4N7O)
525, found 526 (MH.sup.+).
##STR00235##
[0157] A flask was charged with intermediate 2 (100 mg, 0.31 mmol),
4-((3 S,5R)-3,5-dimethylpiperazin-1-yl)aniline (80 mg, 0.39 mmol),
TFA (50 uL), isopropanol (5 mL). The reaction was heated to
100.degree. C. for 4 h. The reaction mixture was basified with a
saturated aqueous sodium bicarbonate solution and then was
extracted with DCM/(10 ml.times.3). The combined organic was washed
by brine, dried over sodium sulfate and concentrated. The crude
product was purified with flash chromatography (0-10% MeOH-in DCM)
to afford the desired product as yellow solids (146 mg, 95% yield).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.83 (br, 1H), 9.86
(br, 1H), 8.23 (s, 1H), 7.30 (d, J=8.4 Hz, 2H), 7.00 (dd, J=5.6 Hz,
J=10.4 Hz, 1H), 6.90 (d, J=8.8 Hz, 2H), 6.34 (s, 1H), 3.51 (m, 2H),
2.84 (m, 2H), 2.41 (s, 3H), 2.11 (m, 2H), 1.02 (d, J=6.4 Hz, 6H);
ESI-MS: calcd for (C26H25F2N7O) 489, found 490 (MH.sup.+).
##STR00236##
[0158] A flask was charged with intermediate 2 (100 mg, 0.31 mmol),
4-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)aniline (101 mg, 0.39
mmol), TFA (50 uL), isopropanol (5 mL). The reaction was heated to
100.degree. C. for 4 h. The reaction mixture was basified with a
saturated aqueous sodium bicarbonate solution and then was
extracted with DCM/(10 ml.times.3). The combined organic was washed
by brine, dried over sodium sulfate and concentrated. The crude
product was purified with flash chromatography (0-10% EtOAc in DCM)
to afford the desired product as yellow solids (101 mg, 60% yield).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.83 (br, 1H), 9.88
(br, 1H), 8.23 (s, 1H), 7.30 (d, J=8.8 Hz, 2H), 7.00 (dd, J=5.6 Hz,
J=10.8 Hz, 1H), 6.94 (d, J=9.2 Hz, 2H), 6.34 (s, 1H), 3.25 (q,
J=10.0 Hz, 2H), 3.14 (m, 4H), 2.76 (m, 4H), 2.41 (s, 3H); ESI-MS:
calcd for (C26H22F5N7O) 543, found 544 (MH.sup.+).
##STR00237##
[0159] To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (200 mg, 0.661 mmol) and 3-morpholinoaniline (124 mg, 0.694 mmol)
in anhydrous DMSO (2.0 mL) was added TEA (0.28 mL; 1.98 mmol), and
the resulting biphasic mixture was efficiently stirred at room
temperature under argon atmosphere for 18 h. The resulting mixture
was diluted with EtOAc (100 mL) and washed with aq. NH.sub.4Cl (ca.
100 mL each; 50% NH.sub.4Cl saturation) followed by brine (100 mL;
50% saturation). The organic layer was separated and dried over
anhydrous Na.sub.2SO.sub.4 and concentrated. The resulting residue
was purified by crystallization out of EtOAc to afford the desired
product as a brown solid (123 mg, 42% yield). .sup.1H NMR (DMSO-d6,
400 MHz) .delta. 11.36 (br s, 1H), 9.89 (br s, 1H), 8.28 (s, 1H),
7.23-7.19 (m, 1H), 7.14-7.10 (m, 2H), 7.05-7.03 (m, 1H), 6.96-6.92
(m, 1H), 6.79-6.77 (m, 1H), 3.75-3.73 (m, 4H), 3.11-3.09 (m, 4H),
2.40 (s, 3H); MS (ESI): calcd for C24H21FN6O2: 444, found: 445
(MH.sup.+).
##STR00238##
[0160] To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (200 mg, 0.661 mmol) and 3-(4-ethylpiperazin-1-yl)aniline (193
mg, 0.694 mmol) in anhydrous DMSO (2.0 mL) was added TEA (0.28 mL;
1.98 mmol), and the resulting biphasic mixture was efficiently
stirred at room temperature under argon atmosphere for 18 h. The
resulting mixture was diluted with EtOAc (100 mL) and washed with
aq. NH.sub.4Cl (ca. 100 mL each; 50% NH.sub.4Cl saturation)
followed by brine (100 mL; 50% saturation). The organic layer was
separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by crystallization
out of EtOAc to afford the desired product as a brown solid (221
mg, 69% yield). .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.36 (br s,
1H), 9.86 (br s, 1H), 8.28 (s, 1H), 7.20-7.08 (m, 3H), 7.02-6.92
(m, 2H), 6.78-6.75 (m, 1H), 3.14-3.12 (4H), 2.48 (m, 4H); obscured
by DMSO signal), 2.40 (s, 3H), 2.37 (q, J=7.2 Hz, 2H), 1.03 (t,
J=7.2 Hz, 3H); MS (ESI): calcd for C26H26FN7O: 471, found: 472
(MH.sup.+).
##STR00239##
[0161] To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (200 mg, 0.330 mmol) and aniline (33 .mu.L, 0.363 mmol) in
anhydrous DMSO (2.0 mL) was added TEA (0.14 mL; 0.99 mmol), and the
resulting biphasic mixture was efficiently stirred at room
temperature under argon atmosphere for 18 h. The resulting mixture
was diluted with EtOAc (100 mL) and washed with aq. NH.sub.4Cl (ca.
100 mL each; 50% NH.sub.4Cl saturation) followed by brine (100 mL;
50% saturation). The organic layer was separated and dried over
anhydrous Na.sub.2SO.sub.4 and concentrated. The resulting residue
was purified by flash column chromatography on silica gel using
0-100% EtOAc in hexanes (v/v) to afford the desired product as an
off-white solid (93 mg, 78% yield). .sup.1H NMR (DMSO-d6, 400 MHz)
.delta. 11.36 (br s, 1H), 10.04 (br s, 1H), 8.29 (s, 1H), 7.53 (d,
J=7.6 Hz, 2H), 7.39-7.35 (m, 2H), 7.20-7.12 (m, 2H), 6.97-6.93 (m,
1H), 6.24 (s, 1H), 2.40 (s, 3H); MS (ESI): calcd for C20H14FN5O:
359, found: 360 (MH.sup.+).
##STR00240##
[0162] To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (100 mg, 0.330 mmol) and aniline (33 .mu.L, 0.363 mmol) in
anhydrous DMSO (2.0 mL) was added TEA (0.14 mL; 0.99 mmol), and the
resulting biphasic mixture was efficiently stirred at room
temperature under argon atmosphere for 18 h. The resulting mixture
was diluted with EtOAc (100 mL) and washed with aq. NH.sub.4Cl (ca.
100 mL each; 50% NH.sub.4Cl saturation) followed by brine (100 mL;
50% saturation). The organic layer was separated and dried over
anhydrous Na.sub.2SO.sub.4 and concentrated. The resulting residue
was purified by flash column chromatography on silica gel using
0-100% EtOAc in hexanes (v/v) to afford the desired product as an
off-white solid (93 mg, 78% yield). .sup.1H NMR (DMSO-d6, 400 MHz)
.delta. 11.35 (br s, 1H), 9.82 (br s, 1H), 8.21 (d, J=0.8 Hz, 1H),
7.29 (d, J=8.0 Hz, 2H), 7.12 (d, J=8.8 Hz, 1H), 6.95-6.90 (m, 3H),
6.23 (d, J=0.8 Hz, 1H), 3.14-3.11 (m, 4H), 2.40 (s, 3H), 1.65-1.51
(m, 6H); MS (ESI): calcd for C25H23FN6O: 442, found: 443
(MH.sup.+).
##STR00241##
[0163] To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (100 mg, 0.330 mmol) and N-(4-aminophenyl)acetamide (54 mg, 0.36
mmol) in anhydrous DMSO (2.0 mL) was added TEA (0.14 mL; 0.99
mmol), and the resulting biphasic mixture was efficiently stirred
at room temperature under argon atmosphere for 18 h. The resulting
mixture was diluted with EtOAc (100 mL) and washed with aq.
NH.sub.4Cl (ca. 100 mL each; 50% NH.sub.4Cl saturation) followed by
brine (100 mL; 50% saturation). The organic layer was separated and
dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The
resulting residue was purified by crystallization out of EtOAc to
afford the desired product
N-(4-((5-cyano-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidin-4-yl)ami-
no)phenyl)acetamide as an off-white solid (88 mg, 64% yield).
.sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.37 (br s, 1H), 9.99 (br
s, 1H), 9.98 (br s, 1H), 8.25 (s, 1H), 7.57-7.55 (m, 2H), 7.42-7.40
(m, 2H), 7.14-7.11 (m, 1H), 6.96-6.92 (m, 1H), 6.24 (s, 1H), 2.40
(s, 3H), 2.04 (s, 3H); MS (ESI): calcd for C22H17FN6O2: 416, found:
417 (MH.sup.+).
##STR00242##
[0164] To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (100 mg, 0.330 mmol) and N-(4-aminophenyl)methanesulfonamide (68
mg, 0.36 mmol) in anhydrous isopropanol (5.0 mL) was added TFA
(0.05 mL; 0.65 mmol), and the resulting mixture was efficiently
stirred at 80.degree. C. under argon atmosphere for 22 h. The
resulting mixture was diluted with EtOAc (100 mL) and washed with
aq. NaHCO.sub.3 (ca. 100 mL each; 50% NaHCO.sub.3 saturation). The
organic layer was separated and dried over anhydrous
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified by crystallization out of EtOAc afford the desired product
as an off-white solid (97 mg, 65% yield). .sup.1H NMR (DMSO-d6, 400
MHz) .delta. 11.35 (br s, 1H), 10.02 (br s, 1H), 9.71 (br s, 1H),
8.27 (s, 1H), 7.49-7.46 (m, 2H), 7.21-7.11 (m, 3H), 6.96-6.92 (m,
1H), 6.24 (s, 1H), 2.98 (s, 3H), 2.40 (s, 3H); MS (ESI): calcd for
C21H17FN6O3S: 452, found: 453 (MH.sup.+).
##STR00243##
[0165] To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (100 mg, 0.330 mmol) and 4-(4-aminophenyl)piperazin-2-one (69 mg,
0.36 mmol) in anhydrous DMSO (3.0 mL) was added TEA (0.14 mL; 0.99
mmol), and the resulting biphasic mixture was efficiently stirred
at room temperature under argon atmosphere for 27 h. The resulting
mixture was diluted with EtOAc (100 mL) and washed with aq.
NaHCO.sub.3 (ca. 100 mL each; 50% NaHCO.sub.3 saturation). The
organic layer was separated and dried over anhydrous
Na.sub.2SO.sub.4 and concentrated, whereupon a precipitate formed.
The precipitate was filtered and washed with EtOAc to afford the
desired product as a yellow solid (88 mg, 64% yield). .sup.1H NMR
(DMSO-d6, 400 MHz) .delta. 11.34 (br s, 1H), 9.86 (br s, 1H), 8.22
(s, 1H), 8.04 (br s, 1H), 7.35-7.33 (m, 2H), 7.12 (d, J=8.8 Hz,
1H), 6.95-6.91 (m, 2H), 6.24-6.23 (m, 1H), 3.71 (s, 2H), 3.41-3.38
(m, 2H), 3.32-3.29 (m, 2H; obscured by water signal), 2.40 (s, 3H);
MS (ESI): calcd for C24H20FN7O2: 457, found: 458 (MH.sup.+).
##STR00244##
[0166] To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (100 mg, 0.330 mmol) and 2-(4-aminophenyl)isothiazolidine
1,1-dioxide (77 mg, 0.36 mmol) in anhydrous isopropanol (5.0 mL)
was added TFA (0.05 mL; 0.65 mmol), and the resulting mixture was
stirred at 80.degree. C. under argon atmosphere for 22 h. The
resulting mixture was diluted with EtOAc (100 mL) and washed with
aq. NaHCO.sub.3 (ca. 100 mL each; 50% NaHCO.sub.3 saturation). The
organic layer was separated and dried over anhydrous
Na.sub.2SO.sub.4 and concentrated, whereupon a precipitate formed.
The precipitate was filtered and washed with EtOAc to afford the
desired product as an off-white solid (97 mg, 65% yield). .sup.1H
NMR (DMSO-d6, 400 MHz) .delta. 11.37 (br s, 1H), 10.06 (br s, 1H),
8.26 (s, 1H), 7.52-7.49 (m, 2H), 7.22-7.19 (m, 2H), 7.12 (d, J=8.4
Hz, 1H), 6.97-6.93 (m, 1H), 3.74 (t, J=6.4 Hz, 2H), 3.51 (t, J=7.2
Hz, 2H), 2.44-2.37 (m, 2H), 2.40 (s, 3H); MS (ESI): calcd for
C23H19FN6O3S: 478, found: 479 (MH.sup.+).
##STR00245##
[0167] To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (100 mg, 0.330 mmol) and 4-ethoxyaniline (47 .mu.L, 0.36 mmol) in
anhydrous DMSO (3.0 mL) was added TEA (0.14 mL; 0.99 mmol), and the
resulting biphasic mixture was efficiently stirred at room
temperature under argon atmosphere for 23 h. The resulting mixture
was diluted with EtOAc (100 mL) and washed with aq. NaHCO.sub.3
(ca. 100 mL each; 50% NaHCO.sub.3 saturation). The organic layer
was separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by flash column
chromatography on silica gel using 0-100% EtOAc in hexanes (v/v) to
afford the desired product as a yellow solid (111 mg, 83% yield).
.sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.35 (br s, 1H), 9.90 (br
s, 1H), 8.22 (s, 1H), 7.37 (d, J=9.2 Hz, 1H), 7.12 (d, J=8.8 Hz,
1H), 6.95-6.90 (m, 3H), 6.23 (s, 1H), 4.02 (q, J=6.8 Hz, 2H), 2.40
(s, 3H), 1.33 (t, J=6.8 Hz, 3H); MS (ESI): calcd for C22H18FN5O2:
403, found: 404 (MH.sup.+).
##STR00246##
[0168] Step 1: To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (200 mg, 0.661 mmol) and tert-butyl
4-(4-aminophenyl)piperidine-1-carboxylate (201 mg, 0.727 mmol) in
anhydrous DMSO (3.0 mL) was added TEA (0.28 mL; 1.98 mmol), and the
resulting biphasic mixture was efficiently stirred at room
temperature under argon atmosphere for 20 h. The resulting mixture
was diluted with EtOAc (100 mL) and washed with aq. NaHCO.sub.3
(ca. 100 mL each; 50% NaHCO.sub.3 saturation). The organic layer
was separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by flash column
chromatography on silica gel using 0-100% EtOAc in hexanes (v/v) to
afford the desired product as a yellow solid (322 mg, 90% yield).
.sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.36 (br s, 1H), 9.98 (br
s, 1H), 8.25 (s, 1H), 7.42 (d, J=8.4 Hz, 2H), 7.23 (d, J=8.8 Hz,
2H), 7.12 (d, J=8.4 Hz, 1H), 6.96-6.92 (m, 1H), 6.24-6.23 (m, 1H),
4.11-4.06 (m, 1H), 2.84-2.78 (m, 2H), 2.70-2.64 (m, 2H), 2.40 (s,
3H), 1.77-1.74 (m, 2H), 1.54-1.41 (m, 2H), 1.42 (s, 9H); MS (ESI):
calcd for C30H31FN6O3: 542, found: 442 (M-BOC+H.sup.+).
[0169] Step 2: A mixture of tert-butyl
4-(4-((5-cyano-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)
pyrimidin-4-yl)amino)phenyl)piperidine-1-carboxylate (200 mg, 0.369
mmol) was stirred into 8% TFA in DCM (10 mL) and the resulting
mixture was stirred at room temperature for 17 h. The resulting
mixture was diluted with EtOAc (100 mL) and washed with aq.
NaHCO.sub.3 (ca. 100 mL each; 50% NaHCO.sub.3 saturation) whereupon
a precipitate formed. The precipitate was filtered and washed with
EtOAc. The mother liquid was concentrated to afford the desired
product as a glassy, white solid (60 mg, 40% yield). .sup.1H NMR
(DMSO-d6, 400 MHz) .delta. 11.37 (br s, 1H), 9.99 (br s, 1H), 8.26
(s, 1H), 7.44-7.42 (m, 2H), 7.23-7.20 (m, 2H), 7.13 (d, J=8.8 Hz,
1H), 6.96-6.92 (m, 1H), 6.24-6.23 (m, 1H), 3.13-3.10 (m, 2H),
2.72-2.60 (m, 3H), 2.40 (s, 3H), 1.77-1.72 (m, 2H), 1.62-1.52 (m,
2H); MS (ESI): calcd for C25H23FN6O: 442, found: 443
(MH.sup.+).
##STR00247##
[0170] Step 1: To a solution of 4-fluoronitrobenzene (2.0 g, 14.16
mmol) in AcN (15 mL), 2-(piperazin-1-yl)ethanol (1.85 g, 14.17
mmol) and DIEA (2.97 mL, 17.01 mmol) were added. The mixture was
refluxed for 15 h (in a sealed tube). After cooling, the resulting
mixture was poured to water (300 ml). The mixture was stirred at
room temperature for 30 min. The solids were collected by
filtration and washed with water to afford the desired product as
off white solids (2.74 g, 72% yield). ESI-MS calcd for (C12H17N3O3)
251, found 252 [M+H].sup.+.
[0171] Step 2: A solution of the above product (2.74 g) in methanol
(50 mL) was hydrogenated in the presence of 10% Pd/C (270 mg) by
using an H.sub.2 balloon. After 16 h, the reaction mixture was
filtered through a pad of Celite and rinsed with methanol
(3.times.15 mL). The filtrate was concentrated to the desired
product (2.70 g) as light yellow solids. The product was used
directly in the next step without further purification. ESI-MS:
calcd for (C13H21N3) 219, found 220 (MH.sup.+).
[0172] Step 3: A flask was charged with intermediate 2 (50 mg, 0.16
mmol), 2-(4-(4-aminophenyl)piperazin-1-yl)ethanol (30.4 mg, 0.16
mmol), DIPEA (60 uL), DMSO (2 mL). The reaction was stirred at room
temperature for over. The crude product was purified with flash
chromatography (0-10% MeOH-in DCM) to afford the desired product as
light yellow solids (42 mg, 54% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.83 (br, 1H), 9.86 (br, 1H), 8.23 (s, 1H),
7.30 (d, J=8.4 Hz, 2H), 7.00 (dd, J=5.6 Hz, J=10.4 Hz, 1H), 6.90
(d, J=8.8 Hz, 2H), 6.34 (s, 1H), 3.51 (m, 2H), 2.84 (m, 2H), 2.41
(s, 3H), 2.11 (m, 2H), 1.03 (s, 3H), 1.01 (s, 3H); ESI-MS: calcd
for (C26H25F2N7O) 489, found 490 (MH.sup.+).
##STR00248##
[0173] A flask was charged with intermediate 2 (50 mg, 0.16 mmol),
2-(4-(4-aminophenyl)piperazin-1-yl)ethanol (30.4 mg, 0.16 mmol),
DIPEA (60 uL), DMSO (2 mL). The reaction was stirred at room
temperature for over. The crude product was purified with flash
chromatography (0-10% MeOH-in DCM) to afford the desired product as
light yellow solids (42 mg, 54% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.83 (br, 1H), 9.86 (br, 1H), 8.23 (s, 1H),
7.30 (d, J=8.4 Hz, 2H), 7.00 (dd, J=5.6 Hz, J=10.4 Hz, 1H), 6.90
(d, J=8.8 Hz, 2H), 6.34 (s, 1H), 3.51 (m, 2H), 2.84 (m, 2H), 2.41
(s, 3H), 2.11 (m, 2H), 1.03 (s, 3H), 1.01 (s, 3H); ESI-MS: calcd
for (C26H25F2N7O) 489, found 490 (MH.sup.+).
##STR00249##
[0174] Step 1: To a solution of 4-fluoronitrobenzene (0.5 g, 3.54
mmol) in AcN (30 mL), (S)-tert-butyl
2-methylpiperazine-1-carboxylate (0.71 g, 3.54 mmol) and DIEA (0.74
mL, 4.25 mmol) were added. The mixture was refluxed for 15 h (in a
sealed tube). After cooling, the resulting mixture was poured to
water (300 ml). The mixture was stirred at room temperature for 30
min. The resulting reaction mixture was extracted with EtOAc
(3.times.30 mL) and anhydrous Na.sub.2SO.sub.4 and concentrated in
vacuum. The resulting crude product was purified by Teledyne-Isco
flash system by using EtOAc/Hex, 0 to 30% of ethylacetate in hexane
to provide compound tert-butyl
(S)-4-(4-nitrophenyl)-2-methylpiperazine-1-carboxylate as light
yellow solids (710 mg, 62%) as off white solids. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 8.01 (d, J=9.6 Hz, 2H), 6.97 (d, J=9.2
Hz, 2H), 3.84 (m, 2H), 2.95-2.50 (m, 4H), 2.40 (m, 1H), 2.30 (br,
1H), 1.00 (d, J=6.0 Hz, 3H); ESI-MS: calcd for (C11H15N3O2) 221,
found 222 (MH.sup.+).
[0175] Step 2: A solution of tert-butyl
(S)-4-(4-nitrophenyl)-2-methylpiperazine-1-carboxylatein methanol
(30 mL) was hydrogenated in the presence of 10% Pd/C (70 mg) by
using an H.sub.2 balloon. After 16 h, the reaction mixture was
filtered through a pad of Celite and rinsed with methanol
(3.times.15 mL). The filtrate was concentrated to afford the title
compound tert-butyl
(S)-4-(4-aminophenyl)-2-methylpiperazine-1-carboxylate (0.62 g,
97%) as light yellow solids. The product was used directly in the
next step without further purification. ESI-MS: calcd for
(C13H21N3) 219, found 220 (MH.sup.+).
[0176] Step 3: A mixture of Intermediate 2 (150 mg, 0.47 mmol),
(S)-tert-butyl 2-methylpiperazine-1-carboxylate (LN927, 0.05 g,
0.016 mmol), and DIPEA (60 uL, 0.037 mmol) in DMSO (3.0 ml) was
stirred at room temperature for overnight. TLC was checked and the
reaction was completed. The mixture was added to sat.
NH.sub.4Cl/water (25 ml/50 ml) and stirred at room temperature for
30 min. The pH of the mixture was adjusted to about 6 using 2N HCl.
After cooled with ice for 1 h, the solids were collected by
filtration, washed by water to give the crude product. The crude
product was suspended in DCM (10 ml) and 1 ml of TFA was added (the
mixture become clear solution). The mixture was stirred at room
temperature for overnight. Potassium phosphate in water was added
to the mixture (pH about 8), and extracted with DCM/MeOH. The
combined organic was washed by brine, concentrated and purified by
column on silica gel (5-15% MeOH in DCM) to give the desired
product as yellow solids (97 mg, 45% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.83 (br, 1H), 9.90 (br, 1H), 8.24 (s, 1H),
7.32 (d, J=8.8 Hz, 2H), 7.00 (dd, J=5.6 Hz, J=10.4 Hz, 1H), 6.94
(d, J=8.8 Hz, 2H), 6.34 (s, 1H), 3.14 (m, 4H), 2.99 (m, 4H), 2.41
(s, 3H) (NH may be berried under solvent peak); ESI-MS: calcd for
(C24H21F2N7O) 461, found 462 (MH.sup.+).
##STR00250##
[0177] Step 1: To a solution of 4-fluoronitrobenzene (0.5 g, 3.54
mmol) in AcN (30 mL), (R)-tert-butyl
2-methylpiperazine-1-carboxylate (0.71 g, 3.54 mmol) and DIEA (0.74
mL, 4.25 mmol) were added. The mixture was refluxed for 15 h (in a
sealed tube). After cooling, the resulting mixture was poured to
water (300 ml). The mixture was stirred at room temperature for 30
min. The resulting reaction mixture was extracted with EtOAc
(3.times.30 mL) and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated in vacuum. The resulting crude product was purified by
Teledyne-Isco flash system by using EtOAc/Hex, 0 to 30% of
ethylacetate in hexane to provide the desired product tert-butyl
(S)-2-methyl-4-(4-nitrophenyl)piperazine-1-carboxylate as light
yellow solids (720 mg, 63%) as off white solids. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 8.01 (d, J=9.6 Hz, 2H), 6.97 (d, J=9.2
Hz, 2H), 3.84 (m, 2H), 2.95-2.50 (m, 4H), 2.40 (m, 1H), 2.30 (br,
1H), 1.00 (d, J=6.0 Hz, 3H); ESI-MS: calcd for (C11H15N3O2) 221,
found 222 (MH.sup.+).
[0178] Step 2: A solution of tert-butyl
(S)-2-methyl-4-(4-nitrophenyl)piperazine-1-carboxylate (0.7 g) in
methanol (30 mL) was hydrogenated in the presence of 10% Pd/C (70
mg) by using an H.sub.2 balloon. After 16 h, the reaction mixture
was filtered through a pad of Celite and rinsed with methanol
(3.times.15 mL). The filtrate was concentrated to afford the title
compound tert-butyl
(S)-4-(4-aminophenyl)-2-methylpiperazine-1-carboxylate (0.61 g,
95%) as light yellow solids. The product was used directly in the
next step without further purification. ESI-MS: calcd for
(C13H21N3) 219, found 220 (MH.sup.+).
[0179] Step 3: A mixture of intermediate 3 (150 mg, 0.47 mmol),
tert-butyl (S)-4-(4-aminophenyl)-2-methylpiperazine-1-carboxylate
(0.05 g, 0.016 mmol), and DIPEA (60 uL, 0.037 mmol) in DMSO (3.0
ml) was stirred at room temperature for overnight. TLC was checked
and the reaction was completed. The mixture was added to sat.
NH.sub.4Cl/water (25 ml/50 ml) and stirred at room temperature for
30 min. The pH of the mixture was adjusted to about 6 using 2N HCl.
After cooled with ice for 1 h, the solids were collected by
filtration, washed by water to give the crude product. The crude
product was suspended in DCM (10 ml) and 1 ml of TFA was added (the
mixture become clear solution). The mixture was stirred at room
temperature for overnight. Potassium phosphate in water was added
to the mixture (pH about 8), and extracted with DCM/MeOH. The
combined organic was washed by brine, concentrated and purified by
column on silica gel (5-15% MeOH in DCM) to give the desired
product as yellow solids (97 mg, 45% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.83 (br, 1H), 9.90 (br, 1H), 8.24 (s, 1H),
7.32 (d, J=8.8 Hz, 2H), 7.00 (dd, J=5.6 Hz, J=10.4 Hz, 1H), 6.94
(d, J=8.8 Hz, 2H), 6.34 (s, 1H), 3.14 (m, 4H), 2.99 (m, 4H), 2.41
(s, 3H) (NH may be berried under solvent peak); ESI-MS: calcd for
(C24H21F2N7O) 461, found 462 (MH.sup.+).
##STR00251##
[0180] Step 1: To a mixture of 4-Nitroaniline (1.0 g, 7.24 mmol)
and cyclopropyl carbonylchloride (0.83 g, 7.96 mmol) were dissolved
in Dry THF (30 mL). DIPEA (3.16 mL, 18.10 mmol) was added to above
reaction mixture and stirred at the room temperature for over
night. The mixture was stirred at room temperature for 30 min. The
resulting crude mixture was extracted with EtOAc (3.times.30 mL)
and dried over anhydrous Na.sub.2SO.sub.4 and concentrated in
vacuum. The resulting crude product was purified by Teledyne-Isco
flash system by using EtOAc/Hex, 0 to 30% of ethylacetate in hexane
to provide the desired product
(4-nitrophenyl)cyclopropanecarboxamide (880 mg, 62%) as off white
solids. ESI-MS: calcd for (C10H10N2O3) 206, found 207
(MH.sup.+).
[0181] Step 2: A solution (4-nitrophenyl)cyclopropanecarboxamide
(0.7 g) in methanol (30 mL) was hydrogenated in the presence of 10%
Pd/C (70 mg) by using an H.sub.2 balloon. After 16 h, the reaction
mixture was filtered through a pad of Celite and rinsed with
methanol (3.times.15 mL). The filtrate was concentrated to afford
the title compound (4-aminophenyl)cyclopropanecarboxamide
[0182] as light yellow solids. The product was used directly in the
next step without further purification.
[0183] Step 3: A flask was charged with FA425_1 (100 mg, 0.312
mmol), N-(4-aminophenyl)cyclopropanecarboxamide (LN933_1, 0.055 g,
0.312 mmol), DIPEA (60 uL), DMSO (2 mL). The reaction was stirred
at room temperature for over. The crude product was purified with
flash chromatography (0-10% MeOH-in DCM) to afford the desired
product as light yellow solids (85 mg, 54% yield). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 11.83 (br, 1H), 9.86 (br, 1H), 8.23 (s,
1H), 7.30 (d, J=8.4 Hz, 2H), 7.00 (dd, J=5.6 Hz, J=10.4 Hz, 1H),
6.90 (d, J=8.8 Hz, 2H), 6.34 (s, 1H), 3.51 (m, 2H), 2.84 (m, 2H),
2.41 (s, 3H), 2.11 (m, 2H), 1.03 (s, 3H), 1.01 (s, 3H); ESI-MS:
calcd for (C26H25F2N7O) 489, found 490 (MH.sup.+).
##STR00252##
[0184] To a mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (100 mg, 0.330 mmol) and 4-aminobenzamide (47 mg, 0.34 mmol) in
anhydrous DMSO (1.5 mL) was added TEA (0.13 mL; 0.93 mmol), and the
resulting biphasic mixture was efficiently stirred at room
temperature under argon atmosphere for ca. 17 h. The resulting
mixture was diluted with EtOAc (100 mL) and washed with aq.
NaHCO.sub.3 (ca. 100 mL each; 50% NaHCO.sub.3 saturation). The
organic layer was separated and dried over anhydrous
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified by flash column chromatography on silica gel using 0.5-15%
MeOH in DCM (v/v) to afford the desired product as a white solid
(42 mg, 32% yield). .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.85
(br s, 1H), 10.22 (br s, 1H), 8.35 (s, 1H), 7.91 (br s, 1H), 7.86
(d, J=8.8 Hz, 2H), 7.62 (d, J=8.4 Hz, 2H), 7.29 (br s, 1H),
7.04-7.00 (m, 1H), 6.35 (s, 1H), 2.42 (s, 3H); MS (ESI): calcd for
C21H14F2N6O2: 420, found: 421 (MH.sup.+).
##STR00253##
[0185] To a mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (80 mg, 0.25 mmol) and 4-amino-N-methylbenzamide (45 mg, 0.30
mmol) in anhydrous DMSO (1.5 mL) was added TEA (0.122 mL; 0.875
mmol), and the resulting biphasic mixture was efficiently stirred
at room temperature for ca. 3 days. The resulting mixture was
diluted with EtOAc (100 mL) and washed with aq. NaHCO.sub.3 (ca.
100 mL each; 50% NaHCO.sub.3 saturation). The organic layer was
separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by flash column
chromatography on silica gel using 0.5-15% MeOH in DCM (v/v) to
afford the desired product as a white solid (18 mg, 17% yield).
.sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.85 (br s, 1H), 10.23 (br
s, 1H), 8.37-8.34 (m, 2H), 7.81 (d, J=8.8 Hz, 2H), 7.64-7.62 (m,
2H), 7.04-7.00 (m, 1H), 6.35 (s, 1H), 2.78 (d, J=4.0 Hz, 3H), 2.42
(s, 3H); MS (ESI): calcd for C22H16F2N6O2: 434, found: 435
(MH.sup.+).
##STR00254##
[0186] To a mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (80 mg, 0.25 mmol) and 4-amino-N,N-dimethylbenzamide (49 mg,
0.30 mmol) in anhydrous DMSO (1.5 mL) was added TEA (0.122 mL;
0.875 mmol), and the resulting biphasic mixture was efficiently
stirred at room temperature for ca. 3 days. The resulting mixture
was diluted with EtOAc (100 mL) and washed with aq. NaHCO.sub.3
(ca. 100 mL each; 50% NaHCO.sub.3 saturation). The organic layer
was separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by crystallization
(DCM) to afford the desired product as a white solid (49 mg, 44%
yield). .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.85 (br s, 1H),
10.20 (br s, 1H), 8.33 (s, 1H), 7.60 (d, J=8.4 Hz, 2H), 7.41 (d,
J=8.4 Hz, 2H), 7.04-7.00 (m, 1H), 6.35 (s, 1H), 2.97 (s, 6H), 2.42
(s, 3H); MS (ESI): calcd for C23H18F2N6O2: 448, found: 449
(MH.sup.+).
##STR00255##
[0187] To a mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (80 mg, 0.25 mmol) and 4-fluoroaniline (33 mg, 0.30 mmol) in
anhydrous DMSO (1.5 mL) was added TEA (0.122 mL; 0.875 mmol), and
the resulting biphasic mixture was efficiently stirred at room
temperature for ca. 3 days. The resulting mixture was diluted with
EtOAc (100 mL) and washed with aq. NaHCO.sub.3 (ca. 100 mL each;
50% NaHCO.sub.3 saturation). The organic layer was separated and
dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The
resulting residue was purified by crystallization (DCM) to afford
the desired product as a white solid (65 mg, 66% yield). .sup.1H
NMR (DMSO-d6, 400 MHz) .delta. 11.84 (br s, 1H), 10.10 (br s, 1H),
8.29 (s, 1H), 7.54-7.51 (m, 2H), 7.23-7.29 (m, 2H), 7.03-6.99 (m,
1H), 6.34 (s, 1H), 2.42 (s, 3H); MS (ESI): calcd for C20H12F3N5O:
395, found: 396 (MH.sup.+).
##STR00256##
[0188] To a mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (80 mg, 0.25 mmol) and 4-chloroaniline (33 mg, 0.30 mmol) in
anhydrous DMSO (1.5 mL) was added TEA (0.122 mL; 0.875 mmol), and
the resulting biphasic mixture was efficiently stirred at room
temperature for ca. 3 days. The resulting mixture was diluted with
EtOAc (100 mL) and washed with aq. NaHCO.sub.3 (ca. 100 mL each;
50% NaHCO.sub.3 saturation). The organic layer was separated and
dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The
resulting residue was purified by crystallization (DCM) to afford
the desired product as a white solid (65 mg, 44% yield). .sup.1H
NMR (DMSO-d6, 400 MHz) .delta. 11.85 (br s, 1H), 10.15 (br s, 1H),
8.34 (d, J=1.2 Hz, 1H), 7.59-7.57 (m, 2H), 7.43 (d, J=8.0 Hz, 2H),
7.04-7.00 (m, 1H), 6.35 (s, 1H), 2.42 (s, 3H); MS (ESI): calcd for
C20H12ClF2N5O: 411, found: 412 (MH.sup.+).
##STR00257##
[0189] To a mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (80 mg, 0.25 mmol) and 1-(4-aminophenyl)pyrrolidin-2-one (53
mg, 0.30 mmol) in anhydrous DMSO (1.5 mL) was added TEA (0.122 mL;
0.875 mmol), and the resulting biphasic mixture was efficiently
stirred at room temperature for ca. 3 days. The resulting mixture
was diluted with EtOAc (100 mL) and washed with aq. NaHCO.sub.3
(ca. 100 mL each; 50% NaHCO.sub.3 saturation). The organic layer
was separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by crystallization
(DCM) to afford the desired product as a white solid (56 mg, 49%
yield). .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.34 (br s, 1H),
10.07 (br s, 1H), 8.26 (s, 1H), 7.64 (d, J=8.8 Hz, 2H), 7.48 (d,
J=8.8 Hz, 2H), 7.03-6.99 (m, 1H), 6.34 (s, 1H), 3.84 (t, J=6.8 Hz,
2H), 2.50-2.48 (m, 2H; obscured by DMSO signal), 2.42 (s, 3H),
2.11-2.03 (m, 2H); MS (ESI): calcd for C24H18F2N6O2: 460, found:
461 (MH.sup.+).
##STR00258##
[0190] To a mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (80 mg, 0.25 mmol) and 1H-indol-5-amine (40 mg, 0.30 mmol) in
anhydrous DMSO (1.5 mL) was added TEA (0.122 mL; 0.875 mmol), and
the resulting biphasic mixture was efficiently stirred at room
temperature for ca. 3 days. The resulting mixture was diluted with
EtOAc (100 mL) and washed with aq. NaHCO.sub.3 (ca. 100 mL each;
50% NaHCO.sub.3 saturation). The organic layer was separated and
dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The
resulting residue was purified by flash column chromatography on
silica gel using 0.5-15% MeOH in DCM (v/v) to afford the desired
product as a white solid (32 mg, 33% yield). .sup.1H NMR (DMSO-d6,
400 MHz) .delta. 11.83 (br s, 1H), 11.14 (br s, 1H), 10.00 (br s,
1H), 8.21 (s, 1H), 7.58 (s, 1H), 7.39-7.36 (m, 2H), 7.14-7.11 (m,
1H), 7.03-6.99 (m, 1H), 6.44-6.42 (m, 1H), 6.34 (s, 1H), 5.76 (d,
J=0.8 Hz, 1H), 2.41 (s, 3H); MS (ESI): calcd for C22H14F2N6O: 416,
found: 417 (MH.sup.+).
##STR00259##
[0191] To a mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (80 mg, 0.25 mmol) and 1H-indol-5-amine (40 mg, 0.30 mmol) in
anhydrous DMSO (1.5 mL) was added TEA (0.122 mL; 0.875 mmol), and
the resulting biphasic mixture was efficiently stirred at room
temperature for ca. 3 days. The resulting mixture was diluted with
EtOAc (100 mL) and washed with aq. NaHCO.sub.3 (ca. 100 mL each;
50% NaHCO.sub.3 saturation). The organic layer was separated and
dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The
resulting residue was purified by flash column chromatography on
silica gel using 0.5-15% MeOH in DCM (v/v) to afford the desired
product as a white solid (32 mg, 33% yield). .sup.1H NMR (DMSO-d6,
400 MHz) .delta. 11.84 (br s, 1H), 9.76 (br s, 1H), 9.57 (br s,
1H), 8.24 (s, 1H), 7.40 (d, J=8.0 Hz, 1H), 7.13-7.09 (m, 1H),
7.02-6.98 (m, 1H), 6.91 (d, J=8.0 Hz, 1H), 6.84-6.80 (m, 1H), 6.34
(d, J=3.2 Hz, 1H), 2.41 (s, 3H); MS (ESI): calcd for C20H13F2N502:
393, found: 394 (MH.sup.+).
##STR00260##
[0192] A mixture of
4-chloro-6-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carbonitril-
e (100 mg, 0.31 mmol), 4-nitroaniline (47 mg, 0.34 mmol),
Pd(OAc).sub.2 (10 mg, 0.045 mmol), xantphos (45 mg, 0.078 mmol),
K.sub.2CO.sub.3 (150 mg, 1.09 mmol) and anhydrous dioxane (12 mL)
was sealed in a microwavable tube and degassed with argon for 10
min. The mixture was then heated to 120.degree. C. for 20 min under
microwave irradiation. The mixture was cooled to room temperature
and partitioned between EtOAc and aq. NaHCO.sub.3 (ca. 100 mL each;
50% saturated NaHCO.sub.3), and the organic layer was separated,
dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The
resulting residue was taken up into EtOAc, filtered, and washed
with additional EtOAc to yield the desired product as a light
yellow solid (54 mg, 41% yield). .sup.1H NMR (DMSO-d6, 400 MHz)
.delta. 11.86 (br s, 1H), 10.59 (br s, 1H), 8.47 (s, 1H), 8.26-8.23
(m, 2H), 7.92-7.88 (m, 2H), 7.06-7.02 (m, 1H), 6.36 (s, 1H), 2.42
(s, 3H); MS (ESI): calcd for C20H12F2N6O3: 422, found: 423
(MH.sup.+).
##STR00261##
[0193] Step 1: To a solution of 4-fluoronitrobenzene (3 g, 21.26
mmol) in AcN (60 mL), cis-2,6-dimethylpiperazine (2.55 g, 22.32
mmol) and DIEA (3.90 mL, 22.32 mmol) were added. The mixture was
stirred and refluxed for 18 h. The resulting mixture was cooled
until room temperature and evaporated to afford. EtOAc and water
was added. The mixture was extracted with EtOAc three times. The
organic was washed with brine, dried over sodium sulfate and
concentrated. The reissue was crystallized from EtOAc to give the
yellow solids of reversed connection of the side product (300 mg).
The mother liquid was concentrated to minimum amount of solvents
and then Hexanes was added to form the yellow precipitate. The
solids were collected by filtration and washed with hexanes to
afford the desire compound
(3R,5S)-3,5-dimethyl-1-(4-nitrophenyl)piperazine as light yellow
solids (2.60 g, 52% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.02 (d, J=9.6 Hz, 2H), 7.02 (d, J=9.2 Hz, 2H), 3.90 (m,
2H), 2.78 (m, 2H), 2.40 (m, 2H), 1.03 (d, J=5.6 Hz, 6H); ESI-MS:
calcd for (C12H17N3O2) 235, found 236 (MH.sup.+).
[0194] Step 2: To a solution of
(3R,5S)-3,5-dimethyl-1-(4-nitrophenyl)piperazine (600 mg, 2.55
mmol) in DMF (6 mL) was added sodium hydride (60%, 122 mg g, 3.06
mmol) potion wise and the mixture was stirred at room temperature
for 30 min. then idomethan (0.19 ml g, 3.06 mmol) was added. The
reaction mixture was stirred at room temperature for overnight. TLC
was checked and the starting material was consumed. The mixture was
poured to cold water portionwise and the mixture was extracted with
EtOAc (3.times.50 ml). The combined organic phase was washed with
water, dried over sodium sulfate and concentrated to give
4-((3R,5S)-3,4,5-trimethylpiperazin-1-yl)nitrobenzine as a yellow
solid (100 mg, 15% yield). No further purification was conducted.
ESI-MS: calcd for (C13H19N3O2) 249, found 250 (MH.sup.+).
[0195] Step 3: A solution of
(3R,5S)-3,5-dimethyl-1-(4-nitrophenyl)piperazine (.about.100 mg) in
methanol (20 mL) was hydrogenated in the presence of 10% Pd/C (10
mg) using an H.sub.2 balloon. After 16 h, the reaction mixture was
filtered through a pad of Celite and rinsed with methanol
(3.times.15 mL). The filtrate was concentrated to afford the
desired product 4-((3R,5S)-3,4,5-trimethylpiperazin-1-yl)aniline
(51 mg, 9% yield for 2 steps) as red solids. The product was used
directly for the next step reaction without further purification.
ESI-MS: calcd for (C13H21N3) 219, found 220 (MH.sup.+).
[0196] Step 4: A flask was charged with Intermediate 2 (50 mg, 0.16
mmol), 4-((3R,5S)-3,4,5-trimethylpiperazin-1-yl)aniline (38 mg,
0.17 mmol), TFA (50 uL), isopropanol (5 mL). The reaction was
heated to 80.degree. C. for overnight. The reaction mixture was
basified with a saturated aqueous potassium phosphate solution and
then was extracted with DCM/(10 ml.times.3). The combined organic
was dried over sodium sulfate and concentrated. The crude product
was purified with flash chromatography (0-10% MeOH-in DCM) to
afford the desired product as yellow solids (51 mg, 65% yield).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.84 (br, 1H), 9.87
(br, 1H), 8.23 (s, 1H), 7.30 (d, J=8.8 Hz, 2H), 7.00 (dd, J=5.6 Hz,
J=10.4 Hz, 1H), 6.91 (d, J=9.2 Hz, 2H), 6.34 (s, 1H), 3.54 (d,
J=11.2 Hz, 2H), 2.41 (s, 3H), 2.34 (d, J=11.6 Hz, 2H), 2.24 (m,
2H), 2.19 (s, 3H), 1.07 (d, J=6.0 Hz, 6H); ESI-MS: calcd for
(C27H27F2N7O) 503, found 504 (MH.sup.+).
##STR00262##
[0197] Step 1: To a solution of 4-fluoronitrobenzene (1.80 g, 12.76
mmol) in AcN (18 mL), 2-methylpiperazine (3.19 g, 31.89 mmol) and
DIEA (3.34 mL, 19.14 mmol) were added. The mixture was stirred at
75.degree. C. for 3 h (in a sealed tube). The resulting mixture was
cooled until room temperature and then transferred to water (300
ml). The mixture was stirred at room temperature for 30 min. then
cooled with ice. The solids were collected by filtration and washed
with water to afford the crude product, which was purified by
column chromatography (0-10% MeOH in DCM) to afford the desired
compound of 3-methyl-1-(4-nitrophenyl)piperazine as yellow solids
(2.03 g, 72% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
8.01 (d, J=9.6 Hz, 2H), 6.97 (d, J=9.2 Hz, 2H), 3.84 (m, 2H),
2.95-2.50 (m, 4H), 2.40 (m, 1H), 2.30 (br, 1H), 1.00 (d, J=6.0 Hz,
3H); ESI-MS: calcd for (C11H15N3O2) 221, found 222 (MH.sup.+).
[0198] Step 2: To a solution of
3-methyl-1-(4-nitrophenyl)piperazine (600 mg, 2.71 mmol) in DMF (6
mL) was added sodium hydride (60%, 130 mg g, 3.25 mmol) potion wise
and the mixture was stirred at room temperature for 30 min. then
idomethan (0.20 ml g, 3.25 mmol) was added. The reaction mixture
was stirred at room temperature for overnight. TLC was checked and
the starting material was consumed. The mixture was poured to cold
water portionwise and the mixture was extracted with EtOAc
(3.times.50 ml). The combined organic phase was washed with water,
dried over sodium sulfate and concentrated to give the crude
product which was purified by column chromatography (0-10% MeOH in
DCM) to give the yellow oil. ESI-MS: calcd for (C12H17N3O2) 235,
found 236 (MH.sup.+).
[0199] Step 3: A solution of
4-(3,4-dimethylpiperazin-1-yl)nitrobenzine in methanol (.about.20
mL) was hydrogenated in the presence of Pd/C (25 mg) using an
H.sub.2 balloon. After 16 h, the reaction mixture was filtered
through a pad of Celite and rinsed with methanol (3.times.15 mL).
The filtrate was concentrated to the desired product
4-(3,4-dimethylpiperazin-1-yl)aniline (398 mg, 72% yield for 2
steps) as brown solids. The product was used directly for the next
step reaction without further purification. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 6.65 (d, J=8.4 Hz, 2H), 6.45 (d, J=8.0 Hz,
2H), 4.51 (br, 2H), 3.17 (m, 2H), 2.74 (m, 1H), 2.57 (m, 1H),
2.30-2.00 (m, 5H), 0.98 (d, J=6.0 Hz, 3H); ESI-MS: calcd for
(C12H19N3) 205, found 206 (MH.sup.+).
[0200] Step 4: A flask was charged with intermediate 2 (50 mg, 0.16
mmol), 4-(3,4-dimethylpiperazin-1-yl)aniline (35 mg, 0.17 mmol),
TFA (50 uL), isopropanol (5 mL). The reaction was heated to
80.degree. C. for overnight. The reaction mixture was basified with
a saturated aqueous potassium phosphate solution and then was
extracted with DCM/(10 ml.times.3). The combined organic was dried
over sodium sulfate and concentrated. The crude product was
purified with flash chromatography (0-10% MeOH-in DCM) to afford
the desired product as yellow solids (60 mg, 79% yield). .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 11.83 (br, 1H), 9.88 (br, 1H),
8.23 (s, 1H), 7.30 (d, J=8.8 Hz, 2H), 7.00 (dd, J=5.6 Hz, J=10.4
Hz, 1H), 6.90 (d, J=8.8 Hz, 2H), 6.34 (s, 1H), 3.51 (m, 2H),
2.83-2.70 (m, 2H), 2.41 (s, 3H), 2.36 (t, J=10.4 Hz, 1H), 2.30-2.05
(m, 5H), 1.03 (d, J=6.4 Hz, 3H); ESI-MS: calcd for (C26H25F2N7O)
489, found 490 (MH.sup.+).
##STR00263##
[0201] Step 1: To a solution of 4-fluoronitrobenzene (3 g, 21.26
mmol) in AcN (60 mL), cis-2,6-dimethylpiperazine (2.55 g, 22.32
mmol) and DIEA (3.90 mL, 22.32 mmol) were added. The mixture was
stirred and refluxed for 18 h. The resulting mixture was cooled
until room temperature and evaporated to afford. EtOAc and water
was added. The mixture was extracted with EtOAc three times. The
organic was washed with brine, dried over sodium sulfate and
concentrated. The reissue was crystallized from EtOAc to give the
yellow solids of reversed connection of the side product (300 mg).
The mother liquid was concentrated to minimum amount of solvents
and then Hexanes was added to form the yellow precipitate. The
solids were collected by filtration and washed with hexanes to
afford the desire compound
(3R,5S)-3,5-dimethyl-1-(4-nitrophenyl)piperazine as light yellow
solids (2.60 g, 52% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.02 (d, J=9.6 Hz, 2H), 7.02 (d, J=9.2 Hz, 2H), 3.90 (m,
2H), 2.78 (m, 2H), 2.40 (m, 2H), 1.03 (d, J=5.6 Hz, 6H); ESI-MS:
calcd for (C12H17N3O2) 235, found 236 (MH.sup.+).
[0202] Step 2: To a solution of
(3R,5S)-3,5-dimethyl-1-(4-nitrophenyl)piperazine (650 mg, 2.76
mmol) and iodoethane (453 mg, 2.90 mmol) in DMF (6 mL) was added
potassium carbonate (573 mg g, 4.15 mmol) and the mixture was
stirred at room temperature for overnight. TLC was checked and the
starting material was consumed. The mixture was poured to cold
water and extracted with DCM (3.times.15 ml). The combined organic
phase was dried over sodium sulfate and concentrated to give the
crude product which was purified by column chromatography (0-10%
MeOH in DCM) to give the yellow oil. ESI-MS: calcd for (C14H21N3O2)
263, found 264 (MH.sup.+).
[0203] Step 3: A solution of above prepared nitro benzine in
methanol (.about.30 mL) was hydrogenated in the presence of Pd/C
(50 mg) using an H.sub.2 balloon. After 16 h, the reaction mixture
was filtered through a pad of Celite and rinsed with methanol
(3.times.15 mL). The filtrate was concentrated to afford the
desired product
4-((3R,5S)-4-ethyl-3,5-dimethylpiperazin-1-yl)aniline (520 mg, 81%
yield for 2 steps) as brown solids. The product was used directly
for the next step reaction without further purification. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 6.65 (d, J=7.2 Hz, 2H), 6.47
(d, J=7.6 Hz, 2H), 4.52 (br, 2H), 3.20 (d, J=10.8 Hz, 2H), 2.81 (q,
J=7.2 Hz, 2H), 2.66 (m, 2H), 2.18 (t, J=10.4 Hz, 2H), 1.00 (d,
J=6.4 Hz, 6H), 0.85 (t, J=7.2 Hz, 3H); ESI-MS: calcd for (C14H23N3)
233, found 234 (MH.sup.+).
[0204] Step 4: A flask was charged with intermediate 2 (50 mg, 0.16
mmol), 4-((3R,5S)-4-ethyl-3,5-dimethylpiperazin-1-yl)aniline (40
mg, 0.17 mmol), TFA (50 uL), isopropanol (5 mL). The reaction was
heated to 80.degree. C. for overnight. The reaction mixture was
basified with a saturated aqueous potassium phosphate solution and
then was extracted with DCM/(10 ml.times.3). The combined organic
was dried over sodium sulfate and concentrated. The crude product
was purified with flash chromatography (0-10% MeOH-in DCM) to
afford the desired product as yellow solids (65 mg, 80% yield).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.84 (br, 1H), 9.87
(br, 1H), 8.23 (s, 1H), 7.30 (d, J=8.8 Hz, 2H), 7.00 (dd, J=5.6 Hz,
J=10.4 Hz, 1H), 6.90 (d, J=8.8 Hz, 2H), 6.34 (s, 1H), 3.54 (d,
J=11.6 Hz, 2H), 2.85 (q, J=7.2 Hz, 2H), 2.67 (m, 2H), 2.41 (s, 3H),
2.33 (t, J=10.8 Hz, 2H), 1.05 (d, J=6.0 Hz, 6H), 0.86 (t, J=7.2 Hz,
3H); ESI-MS: calcd for (C28H29F2N7O) 517, found 518 (MH.sup.+).
##STR00264##
[0205] Step 1: To a solution of 4-fluoronitrobenzene (1.80 g, 12.76
mmol) in AcN (18 mL), 2-methylpiperazine (3.19 g, 31.89 mmol) and
DIEA (3.34 mL, 19.14 mmol) were added. The mixture was stirred at
75.degree. C. for 3 h (in a sealed tube). The resulting mixture was
cooled until room temperature and then transferred to water (300
ml). The mixture was stirred at room temperature for 30 min. then
cooled with ice. The solids were collected by filtration and washed
with water to afford the crude product, which was purified by
column chromatography (0-10% MeOH in DCM) to afford the desired
compound of 3-methyl-1-(4-nitrophenyl)piperazine as yellow solids
(2.03 g, 72% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
8.01 (d, J=9.6 Hz, 2H), 6.97 (d, J=9.2 Hz, 2H), 3.84 (m, 2H),
2.95-2.50 (m, 4H), 2.40 (m, 1H), 2.30 (br, 1H), 1.00 (d, J=6.0 Hz,
3H); ESI-MS: calcd for (C11H15N3O2) 221, found 222 (MH.sup.+).
[0206] Step 2: To a solution of
3-methyl-1-(4-nitrophenyl)piperazine (610 mg, 2.76 mmol) and
iodoethane (452 mg, 2.89 mmol) in DMF (6 mL) was added potassium
carbonate (572 mg g, 4.14 mmol) and the mixture was stirred at room
temperature for overnight. TLC was checked and the starting
material was consumed. The mixture was poured to cold water and
extracted with DCM (3.times.15 ml). The combined organic phase was
dried over sodium sulfate and concentrated to give the crude
product which was purified by column chromatography (0-10% MeOH in
DCM) to give 1-ethyl-2-methyl-4-(4-nitrophenyl)piperazine as yellow
oil. ESI-MS: calcd for (C13H19N3O2) 249, found 250 (MH.sup.+).
[0207] Step 3: A solution of
1-ethyl-2-methyl-4-(4-nitrophenyl)piperazine in methanol (.about.30
mL) was hydrogenated in the presence of Pd/C (50 mg) using an
H.sub.2 balloon. After 16 h, the reaction mixture was filtered
through a pad of Celite and rinsed with methanol (3.times.15 mL).
The filtrate was concentrated to afford the desired product (560
mg, 92% yield for 2 steps) as brown oil. The product was used
directly for the next step reaction without further purification.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 6.66 (d, J=8.4 Hz, 2H),
6.48 (d, J=8.0 Hz, 2H), 4.53 (br, 2H), 3.15 (m, 2H), 2.85-2.70 (m,
2H), 2.60 (m, 1H), 2.48-2.20 (m, 4H), 1.00 (d, J=6.0 Hz, 3H), 0.97
(t, J=7.2 Hz, 3H); ESI-MS: calcd for (C13H21N3) 219, found 220
(MH.sup.+).
[0208] Step 4: A flask was charged with intermediate 2 (50 mg, 0.16
mmol), amine (38 mg, 0.17 mmol), TFA (50 uL), isopropanol (5 mL).
The reaction was heated to 80.degree. C. for overnight. The
reaction mixture was basified with a saturated aqueous potassium
phosphate solution and then was extracted with DCM/(10 ml.times.3).
The combined organic was dried over sodium sulfate and
concentrated. The crude product was purified with flash
chromatography (0-10% MeOH-in DCM) to afford the desired product as
yellow solids (63 mg, 80% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.83 (br, 1H), 9.88 (br, 1H), 8.23 (s, 1H),
7.30 (d, J=8.8 Hz, 2H), 7.00 (dd, J=5.6 Hz, J=10.4 Hz, 1H), 6.91
(d, J=8.8 Hz, 2H), 6.34 (s, 1H), 3.44 (m, 2H), 3.00-2.70 (m, 3H),
2.50-2.20 (m, 7H), 1.05 (d, J=5.2 Hz, 3H), 0.98 (t, J=7.2 Hz, 3H);
ESI-MS: calcd for (C27H27F2N7O) 503, found 504 (MH.sup.+).
##STR00265##
[0209] Step 1: A solution of
(2S,6R)-2,6-dimethyl-1-(4-nitrophenyl)piperazine (290 mg, 1.23
mmol) in methanol (.about.20 mL) was hydrogenated in the presence
of Pd/C (28 mg) using an H.sub.2 balloon. After 16 h, the reaction
mixture was filtered through a pad of Celite and rinsed with
methanol (3.times.15 mL). The filtrate was concentrated to afford
the desired product (265 mg, 100% yield) as purple solids. The
product was used directly for the next step reaction without
further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
6.72 (d, J=7.2 Hz, 2H), 6.50 (d, J=7.2 Hz, 2H), 4.60 (br, 2H),
3.50-3.20 (m, 6H), 2.55 (m, 1H), 1.26 (d, J=6.0 Hz, 3H); ESI-MS:
calcd for (C12H19N3) 205, found 206 (MH.sup.+).
[0210] Step 2: A flask was charged with intermediate 2 (50 mg, 0.16
mmol), 4-((2S,6R)-2,6-dimethylpiperazin-1-yl)aniline (35 mg, 0.17
mmol), TFA (50 uL), isopropanol (5 mL). The reaction was heated to
100.degree. C. for overnight. The reaction mixture was basified
with a saturated aqueous potassium phosphate solution and then was
extracted with DCM/(10 ml.times.3). The combined organic was dried
over sodium sulfate and concentrated. The crude product was
purified with flash chromatography (0-10% MeOH-in DCM) to afford
the desired product as yellow solids (50 mg, 65% yield). .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 11.83 (br, 1H), 9.87 (br, 1H),
8.23 (s, 1H), 7.30 (d, J=8.8 Hz, 2H), 7.00 (dd, J=5.6 Hz, J=10.4
Hz, 1H), 6.91 (d, J=8.8 Hz, 2H), 6.34 (s, 1H), 3.51 (d, J=9.6 Hz,
2H), 2.84 (m, 2H), 2.41 (s, 3H), 2.11 (t, J=10.8 Hz, 2H), 1.02 (d,
J=6.0 Hz, 6H), (NH, missing); ESI-MS: calcd for (C26H25F2N7O) 489,
found 490 (MH.sup.+).
##STR00266##
[0211] Step 1: To a solution of 4-fluoronitrobenzene (0.61 ml, 5.71
mmol) in AcN (6 mL), Octahydropyrrolo[1,2-a]pyrazine (600 mg, 4.75
mmol) and DIEA (1.2 mL, 7.13 mmol) were added. The mixture was
stirred at 75.degree. C. for overnight (in a sealed tube). The
resulting mixture was cooled to room temperature and then
concentrated. The crude product was purified by column
chromatography (0-5% MeOH in DCM) to afford the desired compound as
yellow oil (1.09 g, 92% yield). ESI-MS: calcd for (C13H17N3O2) 247,
found 248 (MH.sup.+).
[0212] Step 2: A solution of above prepared nitrobenzine (1.09 g,
4.41 mmol) in methanol (30 mL) was hydrogenated in the presence of
10% Pd/C (0.08 g) using an H.sub.2 balloon. After 16 h, the
reaction mixture was filtered through a pad of Celite and rinsed
with methanol (3.times.15 mL). The filtrate was concentrated to
afford the desired product (0.93 g, 4.28 mmol, 92%) as red solids.
The product was used directly for the next step reaction without
further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
6.69 (d, J=6.8 Hz, 2H), 6.47 (d, J=6.8 Hz, 2H), 4.53 (br, 2H), 3.41
(d, J=10.8 Hz, 1H), 3.27 (d, J=10.8 Hz, 1H), 2.98 (m, 2H), 2.58 (t,
J=10.8 Hz, 1H), 2.30-2.00 (m, 4H), 1.85-1.65 (m, 3H), 1.32 (m, 1H);
ESI-MS: calcd for (C13H19N3) 217, found 218 (MH.sup.+).
[0213] Step 3: A flask was charged with Intermediate 2 (50 mg, 0.16
mmol), aniline (39 mg, 0.18 mmol), TFA (25 uL), isopropanol (3 mL).
The reaction was heated to 100.degree. C. for overnight. The
reaction mixture was basified with a saturated aqueous potassium
phosphate solution and then was extracted with DCM/(10 ml.times.3).
The combined organic was dried over sodium sulfate and
concentrated. The crude product was purified with flash
chromatography (0-10% MeOH-in DCM) to afford the desired product as
yellow solids (56 mg, 72% yield).
[0214] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.83 (br, 1H),
9.88 (br, 1H), 8.23 (s, 1H), 7.30 (d, J=8.4 Hz, 2H), 7.00 (dd,
J=5.6 Hz, J=10.4 Hz, 1H), 6.93 (d, J=8.8 Hz, 2H), 6.34 (s, 1H),
3.76 (d, J=10.8 Hz, 1H), 3.62 (d, J=11.6 Hz, 1H), 3.03 (m, 2H),
2.73 (t, J=11.6 Hz, 1H), 2.41 (s, 3H), 2.40-2.30 (m, 1H), 2.22 (t,
J=8.0 Hz, 1H), 2.12-2.08 (m, 2H), 1.90-1.60 (m, 3H), 1.42-1.32 (m,
1H); ESI-MS: calcd for (C27H25F2N7O) 501, found 502 (MH.sup.+).
##STR00267##
[0215] Step 1: To a solution of 4-fluoronitrobenzene (0.55 ml, 5.13
mmol) in AcN (6 mL), Octahydro-1H-pyrido[1,2-a]pyrazine (600 mg,
4.28 mmol) and DIEA (1.1 mL, 6.42 mmol) were added. The mixture was
stirred at 75.degree. C. for overnight (in a sealed tube). The
resulting mixture was cooled to room temperature and then
concentrated. The crude product was purified by column
chromatography (0-5% MeOH in DCM) to afford the desired product as
yellow oil (1.03 g, 92% yield). ESI-MS: calcd for (C14H19N3O2) 261,
found 262 (MH.sup.+).
[0216] Step 2: A solution of the above nitrobenzine (1.03 g, 3.94
mmol) in methanol (30 mL) was hydrogenated in the presence of 10%
Pd/C (0.08 g) using an H.sub.2 balloon. After 16 h, the reaction
mixture was filtered through a pad of Celite and rinsed with
methanol (3.times.15 mL). The filtrate was concentrated to afford
the desired product (0.89 g, 7.85 mmol, 92%) as red solids. The
product was used directly for the next step reaction without
further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
6.65 (d, J=6.8 Hz, 2H), 6.47 (d, J=6.8 Hz, 2H), 4.52 (br, 2H), 3.23
(d, J=10.8 Hz, 1H), 3.16 (d, J=10.8 Hz, 1H), 2.72 (m, 2H), 2.56 (t,
J=11.2 Hz, 1H), 2.19 (m, 2H), 1.93 (t, J=10.8 Hz, 2H), 1.75-1.35
(m, 4H), 1.30-1.00 (m, 2H); ESI-MS: calcd for (C14H21N3) 231, found
232 (MH.sup.+).
[0217] Step 3: A flask was charged with intermediate 2 (50 mg, 0.16
mmol), the above prepared aniline (42 mg, 0.18 mmol), TFA (25 uL),
isopropanol (3 mL). The reaction was heated to 100.degree. C. for
overnight. The reaction mixture was basified with a saturated
aqueous potassium phosphate solution and then was extracted with
DCM/(10 ml.times.3). The combined organic was dried over sodium
sulfate and concentrated. The crude product was purified with flash
chromatography (0-10% MeOH-in DCM) to afford the desired product as
yellow solids (62 mg, 77% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 11.83 (br, 1H), 9.87 (br, 1H), 8.23 (s, 1H),
7.30 (d, J=8.0 Hz, 2H), 7.00 (dd, J=5.6 Hz, J=10.4 Hz, 1H), 6.91
(d, J=7.6 Hz, 2H), 6.33 (s, 1H), 3.70-3.40 (m, 2H), 2.90-2.60 (m,
3H), 2.41 (s, 3H), 2.40-2.10 (m, 2H), 2.00-1.90 (m, 2H), 1.80-1.10
(m, 5H); ESI-MS: calcd for (C28H27F2N7O) 515, found 516
(MH.sup.+).
##STR00268##
[0218] A mixture of QW823 (crude, 70 mg, 0.22 mmol),
N-(3-Aminophenyl)propanamide (43 mg, 0.26 mmol), and DIPEA (0.08
ml, 0.44 mmol) in DMSO (2 ml) was stirred at room temperature for 2
hours. TLC was checked and the reaction was completed. Ethylacetate
(15 ml) was added, followed by NH4Cl (20 mL). After separation, the
aqueous was extracted with EtOAc (15 ml.times.1). The combined
organic phase was dried over Na.sub.2SO.sub.4 and concentrated. The
crude product was purified by column on silica gel (0-10% MeOH in
DCM) to give the desired product as yellow solids (57 mg, 58%
yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.84 (br, 1H),
10.10 (br, 1H), 9.91 (s, 1H), 8.31 (s, 1H), 7.84 (s, 1H), 7.37 (d,
J=8.4 Hz, 1H), 7.27 (t, J=8.0 Hz, 1H), 7.18 (d, J=8.0 Hz, 1H), 7.00
(dd, J=5.2 Hz, J=10.4 Hz, 1H), 6.34 (s, 1H), 2.42 (s, 3H), 2.32 (q,
J=7.6 Hz, 2H), 1.08 (t, J=7.6 Hz, 3H); ESI-MS: calcd for
(C23H18F2N6O2) 448, found 449 (MH.sup.+).
##STR00269##
[0219] Step 1: To a solution of 3-nitroaniline (5.00 g, 36.2 mmol)
in dry THF (50 mL) was added TEA (7.50 mL, 54.3 mmol). The mixture
was stirred at r.t. for 10 min followed by dropwise addition of
acryloyl chloride (7.38 mL, 90.50 mmol) at 0.degree. C. The mixture
was then stirred at r.t. for 4 hr. The resulting mixture was
quenched by sodium bicarbonate and extracted with EtOAc three
times. The combined organic was washed with brine, dried over
sodium sulfate and concentrated in vacuo. The crude product was
crystallized from EtOAc/Hexanes to give the desired product as
yellow solids (3.37 g, 48% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 10.65 (br, 1H), 8.71 (s, 1H), 8.00-7.90 (m,
2H), 7.63 (t, J=8.0 Hz, 1H), 6.50-6.30 (m, 2H), 5.84 (d, J=10.0 Hz,
1H); ESI-MS: calcd for (C9H8N2O3) 192, found 193 (MH.sup.+).
[0220] Step 2: To a solution of N-(3-nitrophenyl)acrylamide (2.06
g, 10.72 mmol) in a mixture of MeOH (40 mL) and THF (40 mL) was
added Tin(II) chloride dihydrate (12.09 g, 53.60 mmol). The mixture
was stirred at r.t. overnight and then concentrated. The residue
was treated with saturated aq. Na2CO3 to pH=10-11. The mixture was
extracted with EtOAc. The combined organic layers were washed with
brine, dried over sodium sulfate and concentrated. The residue was
used directly used for the next step reaction without further
purification. (yellow oil, 1.49 g, 85% yield). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 9.81 (br, 1H), 6.99 (s, 1H), 6.95 (t,
J=9.2 Hz, 1H), 6.76 (d, J=7.6 Hz, 1H), 6.41 (m, 1H), 6.30-6.10 (m,
2H), 5.6 (d, J=10.4 Hz, 1H), 5.07 (br, 2H); ESI-MS: calcd for
(C9H10N20) 162, found 163 (MH.sup.+).
[0221] Step 3: A mixture of intermediate 2 (crude, 70 mg, 0.22
mmol), aniline (43 mg, 0.26 mmol), and DIPEA (0.08 ml, 0.44 mmol)
in DMSO (2 ml) was stirred at room temperature for 2 hours. TLC was
checked and the reaction was completed. Ethylacetate (15 ml) was
added, followed by NH4Cl (20 mL). After separation, the aqueous was
extracted with EtOAc (15 ml.times.1). The combined organic phase
was dried over Na.sub.2SO.sub.4 and concentrated. The crude product
was purified by column on silica gel (0-10% MeOH in DCM) to give
the desired product as yellow solids (50 mg, 51% yield). .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 11.84 (br, 1H), 10.20 (br, 1H),
10.14 (s, 1H), 8.32 (s, 1H), 7.93 (s, 1H), 7.46 (d, J=8.0 Hz, 1H),
7.31 (t, J=8.0 Hz, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.00 (dd, J=5.2 Hz,
J=10.4 Hz, 1H), 6.46 (dd, J=10.0 Hz, J=17.2 Hz, 1H), 6.34 (s, 1H),
6.26 (d, J=16.8 Hz, 1H), 5.76 (d, J=10.0 Hz, 1H), 2.42 (s, 3H);
ESI-MS: calcd for (C23H16F2N6O2) 446, found 447 (MH.sup.+).
##STR00270##
[0222] A mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (100 mg, 0.31 mmol), methyl 4-aminobenzoate (47 mg, 0.34
mmol), Pd(OAc).sub.2 (10 mg, 0.045 mmol), xantphos (45 mg, 0.078
mmol), K.sub.2CO.sub.3 (150 mg, 1.09 mmol) and anhydrous dioxane
(12 mL) was sealed in a microwavable tube and degassed with argon
for 10 min. The mixture was then heated to 120.degree. C. for 20
min under microwave irradiation. The mixture was cooled to room
temperature and partitioned between EtOAc and aq. NaHCO.sub.3 (ca.
100 mL each; 50% NaHCO.sub.3 saturation). The organic layer was
separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated whereupon a precipitate formed. The precipitate was
filtered and washed with EtOAc to afford the desired product as a
white solid (68 mg, 50% yield). .sup.1H NMR (DMSO-d6, 400 MHz)
.delta. 11.85 (br s, 1H), 10.39 (br s, 1H), 8.38-8.37 (m, 1H),
7.95-7.93 (m, 2H), 7.75-7.73 (m, 2H), 7.04-7.00 (m, 1H), 6.35 (s,
1H), 3.84 (s, 3H), 2.42 (s, 3H); MS (ESI): calcd for C22H15F2N503:
435, found: 436 (MH.sup.+).
##STR00271##
[0223] A mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (100 mg, 0.31 mmol), 4-aminobenzonitrile (40 mg, 0.34 mmol),
Pd(OAc).sub.2 (10 mg, 0.045 mmol), xantphos (45 mg, 0.078 mmol),
K.sub.2CO.sub.3 (150 mg, 1.09 mmol) and anhydrous dioxane (12 mL)
was sealed in a microwavable tube and degassed with argon for 10
min. The mixture was then heated to 120.degree. C. for 20 min under
microwave irradiation. The mixture was cooled to room temperature
and partitioned between EtOAc and aq. NaHCO.sub.3 (ca. 100 mL each;
50% NaHCO.sub.3 saturation). The organic layer was separated and
dried over anhydrous Na.sub.2SO.sub.4 and concentrated whereupon a
precipitate formed. The precipitate was filtered and washed with
EtOAc to afford the desired product as a white solid (58 mg, 45%
yield). .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.85 (br s, 1H),
10.48 (br s, 1H), 8.38 (s, 1H), 7.81-7.77 (m, 4H), 7.04-7.00 (m,
1H), 6.35 (d, J=0.8 Hz, 1H), 2.42 (s, 3H); MS (ESI): calcd for
C21H12F2N6O: 402, found: 403 (MH.sup.+).
##STR00272##
[0224] Step 1: To a mixture of 4-nitroaniline (3.0 g, 22 mmol) and
TEA (9.2 mL, 66 mmol) in THF (100 mL) was added acryloyl chloride
(1.9 mL; 24 mmol), and the mixture was allowed to warm to room
temperature and stirred for 17 h. Then additional TEA (9.2 mL, 66
mmol) and acryloyl chloride (1.9 mL; 24 mmol) was added and the
mixture was stirred for an additional 2 h. The mixture was
partitioned between EtOAc and aq. NaHCO.sub.3 (ca. 100 mL each; 50%
NaHCO.sub.3 saturation), and the organic layer was separated and
dried over anhydrous Na.sub.2SO.sub.4 and concentrated whereupon a
precipitate formed. The precipitate was filtered and washed with
EtOAc to afford the desired as a yellow solid (1.9 g, 45% yield).
.sup.1H NMR (DMSO-d6, 400 MHz) .delta. 10.76 (br s, 1H), 8.25-8.23
(m, 2H), 7.93-7.91 (m, 2H), 6.51-6.44 (m, 1H), 6.37-6.32 (m, 1H),
5.88-5.85 (m, 1H); MS (ESI): calcd for C9H8N2O3: 192, found: 193
(MH.sup.+).
[0225] Step 2: To a suspension of N-(4-nitrophenyl)acrylamide (800
mg, 4.16 mmol) in 5:1 EtOH/water (21 mL) was added iron powder (469
mg, 8.40 mmol) and saturated aq. NH4Cl (2.1 mL), and the mixture
was stirred at 80.degree. C. for 3 h. Then additional added iron
powder (500 mg, 8.95 mmol) and NH4Cl powder (700 mg, 13.1 mmol) was
added and the mixture was stirred for an additional 17 h. The
mixture was partitioned between EtOAc and aq. NaHCO3 (100 mL each),
and the organic layer was separated and dried over anhydrous
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified by flash column chromatography on silica gel using 0.5-15%
MeOH in DCM (v/v) to afford the desired product as a yellow solid
(216 mg, 32% yield). .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 9.71
(br s, 1H), 7.30 (d, J=8.4 Hz, 2H), 6.51 (d, J=8.0 Hz, 2H),
6.40-6.33 (m, 1H), 6.18-6.13 (m, 1H), 5.66-5.63 (m, 1H); MS (ESI):
calcd for C9H10N20: 162, found: 163 (MH.sup.+).
[0226] Step 3: To a mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (230 mg, 0.71 mmol) and N-(4-aminophenyl)acrylamide (120 mg,
1.0 mmol) in anhydrous DMSO (5.0 mL) was added DIPEA (0.43 mL; 2.5
mmol), and the resulting biphasic mixture was efficiently stirred
at room temperature for 20 h. The resulting mixture was diluted
with 1:9 MeOH/EtOAc (100 mL) and washed with aq. NH.sub.4Cl (ca.
100 mL; 50% NH.sub.4Cl saturation) then brine (100 mL). The organic
layer was separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated whereupon a precipitate formed. The precipitate was
filtered and washed with EtOAc to afford the desired product as an
off-white solid (219 mg, 69% yield). .sup.1H NMR (DMSO-d6, 400 MHz)
.delta. 11.84 (br s, 1H), 10.18 (br s, 1H), 10.04 (br s, 1H), 8.29
(d, J=0.4 Hz, 1H), 7.66 (d, J=8.8 Hz, 2H), 7.46 (d, J=8.8 Hz, 2H),
7.03-6.99 (m, 1H), 6.47-6.41 (m, 1H), 6.34-6.24 (m, 2H), 5.77-5.74
(m, 1H), 2.42 (s, 3H); MS (ESI): calcd for C23H16F2N6O2: 446,
found: 447 (MH.sup.+).
##STR00273##
[0227] Step 1: To a solution of N-(4-nitrophenyl)propionamide (1.00
g, 5.15 mmol) in MeOH (100 mL) was added 10% Pd/C (100 mg), and the
resulting mixture was stirred under H.sub.2 (1 atm) for 21 h. The
mixture was then filtered over Celite and concentrated to afford
the desired product as an orange oil (858 mg, ca. 100% yield).
.sup.1H NMR (DMSO-d6, 400 MHz) .delta. 9.39 (br s, 1H), 7.20 (d,
J=8.8 Hz, 2H), 6.48 (d, J=8.4 Hz, 2H), 4.79 (br s, 2H), 2.22 (q,
J=7.6 Hz, 2H), 1.05 (t, J=7.6 Hz, 3H).
[0228] Step 2: To a mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (80 mg, 0.25 mmol) and N-(4-aminophenyl)propionamide (69 mg,
0.42 mmol) in anhydrous DMSO (1.5 mL) was added TEA (0.10 mL; 0.75
mmol), and the resulting biphasic mixture was efficiently stirred
at room temperature for 20 h. The resulting mixture was diluted
with 1:9 MeOH/EtOAc (100 mL) and washed with aq. NH.sub.4Cl (ca. 10
mL; 50% NH.sub.4Cl saturation) then brine (10 mL). The organic
layer was separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated whereupon a precipitate formed. The precipitate was
filtered and washed with EtOAc to afford the desired product as an
off-white solid (69 mg, 62% yield). .sup.1H NMR (DMSO-d6, 400 MHz)
.delta. 11.83 (br s, 1H), 10.00 (br s, 1H), 9.89 (br s, 1H), 8.27
(d, J=0.4 Hz, 1H), 7.58 (d, J=8.8 Hz, 2H), 7.41 (d, J=8.8 Hz, 2H),
7.03-6.99 (m, 1H), 6.34 (s, 1H), 2.41 (s, 3H), 2.32 (q, J=7.6 Hz,
2H), 1.09 (t, J=7.6 Hz, 3H); MS (ESI): calcd for C23H18F2N6O2: 448,
found: 449 (MH.sup.+).
##STR00274##
[0229] Step 1: To a mixture of 4-nitroaniline (5.0 g, 36 mmol) and
TEA (5.0 mL, 36 mmol) in THF (100 mL) stirred at 0.degree. C. was
added 2-chloroacetyl chloride (1.9 mL; 24 mmol), and the mixture
was allowed to warm to room temperature and stirred for 17 h. The
mixture was partitioned between 1:9 MeOH/EtOAc and aq. NaHCO.sub.3
(ca. 100 mL each; 50% NaHCO.sub.3 saturation), and the organic
layer was washed with aq. brine (ca. 100 mL; 50% saturation),
separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated whereupon a precipitate formed. The precipitate was
filtered and washed with EtOAc to afford the desired product
2-chloro-N-(4-nitrophenyl)acetamide as a yellow solid (6.17 g, 80%
yield). .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 10.90 (br s, 1H),
8.25 (d, J=8.4 Hz, 2H), 7.84 (d, J=9.2 Hz, 2H), 4.34 (s, 2H); MS
(ESI): calcd for C8H7ClN2O3: 214, found: weak signal.
[0230] Step 2: A mixture of 2-chloro-N-(4-nitrophenyl)acetamide
(500 mg, 2.33 mmol) and morpholine (2.0 mL, 23 mmol) in isopropanol
(ca. 5 mL) was stirred at 80.degree. C. for ca. 17 h. The mixture
was partitioned between EtOAc and aq. NaHCO.sub.3 (ca. 10 mL each;
50% NaHCO.sub.3 saturation), and the organic layer was separated
and dried over anhydrous Na.sub.2SO.sub.4 to afford the desired
product as a yellow solid (323 mg, 52% yield). .sup.1H NMR
(DMSO-d6, 400 MHz) .delta. 10.36 (br s, 1H), 8.24-8.21 (m, 2H),
7.92-7.89 (m, 2H), 3.63 (d, J=3.2 Hz, 4H), 3.31 (br s, 4H), 3.21
(s, 2H); MS (ESI): calcd for C12H15N3O4: 265, found: 266
(MH.sup.+).
[0231] Step 3: To a solution of
2-morpholino-N-(4-nitrophenyl)acetamide (320 mg, 1.21 mmol) in MeOH
(70 mL) was added 10% Pd/C (80 mg), and the resulting mixture was
stirred under H.sub.2 (1 atm) for 23 h. The mixture was then
filtered over Celite and concentrated to afford the desired product
as an orange oil (314 mg, ca. 100% yield). .sup.1H NMR (DMSO-d6,
400 MHz) .delta. 9.28 (br s, 1H), 7.22 (d, J=8.4 Hz, 2H), 6.50 (d,
J=8.8 Hz, 2H), 4.86 (br s, 2H), 3.64-3.62 (m, 4H), 3.04 (s, 2H),
2.51-2.47 (m, 4H); MS (ESI): calcd for C12H17N3O2: 235, found: 236
(MH.sup.+).
[0232] Step 4: To a mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (80 mg, 0.25 mmol) and
N-(4-aminophenyl)-2-morpholinoacetamide (68 mg, 0.29 mmol) in
anhydrous DMSO (1.5 mL) was added DIPEA (0.15 mL; 0.87 mmol), and
the resulting biphasic mixture was efficiently stirred at room
temperature for 1.5 days. The resulting mixture was diluted with
1:9 MeOH/EtOAc (10 mL) and washed with aq. NH.sub.4Cl (ca. 10 mL;
50% NH.sub.4Cl saturation) then brine (10 mL). The organic layer
was separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated to afford the desired product as an off-white solid
(142 mg, ca. 100% yield). .sup.1H NMR (DMSO-d6, 400 MHz) .delta.
11.84 (s, 1H), 10.03 (s, 1H), 9.77 (s, 1H), 8.28 (s, 1H), 7.63-7.61
(m, 2H), 7.44 (d, J=8.8 Hz, 2H), 7.03-6.99 (m, 1H), 6.34 (br s,
1H), 3.65-3.63 (m, 4H), 2.54-2.52 (m, 4H; obscured by DMSO signal),
2.41 (s, 3H); MS (ESI): calcd for C26H23F2N7O3: 519, found: 520
(MH.sup.+).
##STR00275##
[0233] Step 1: A mixture of 2-chloro-N-(4-nitrophenyl)acetamide
(500 mg, 2.33 mmol) and N-methylpiperazine (2.6 mL, 23 mmol) in
isopropanol (ca. 5 mL) was stirred at 80.degree. C. for ca. 17 h.
The mixture was partitioned between EtOAc and aq. NaHCO.sub.3 (ca.
10 mL each; 50% NaHCO.sub.3 saturation), and the organic layer was
separated and dried over anhydrous Na.sub.2SO.sub.4 to afford the
desired product as a yellow solid (513 mg, 79% yield). .sup.1H NMR
(DMSO-d6, 400 MHz) .delta. 10.32 (br s, 1H), 8.23-8.21 (m, 2H),
7.91-7.89 (m, 2H), 3.31 (s, 2H), 2.51 (br s, 4H), 2.33 (br s, 4H),
2.17 (s, 3H); MS (ESI): calcd for C13H18N4O3: 278, found: 279
(MH.sup.+).
[0234] Step 2: To a solution of
2-(4-methylpiperazin-1-yl)-N-(4-nitrophenyl)acetamide (500 mg, 1.80
mmol) in MeOH (70 mL) was added 10% Pd/C (75 mg), and the resulting
mixture was stirred under H.sub.2 (1 atm) for 19 h. The mixture was
then filtered over Celite and concentrated to afford the desired
product as an off-white solid (456 mg, ca. 100% yield). .sup.1H NMR
(DMSO-d6, 400 MHz) .delta. 9.21 (br s, 1H), 7.22 (d, J=8.8 Hz, 2H),
6.49 (d, J=8.4 Hz, 2H), 4.86 (br s, 2H), 3.01 (s, 2H), 2.50 (br s,
4H), 2.36 (br s, 4H), 2.16 (s, 3H); MS (ESI): calcd for C13H20N4O:
248, found: 249 (MH.sup.+).
[0235] Step 3: To a mixture of
4-chloro-6-((4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimidine-5-carboni-
trile (80 mg, 0.25 mmol) and
N-(4-aminophenyl)-2-(4-methylpiperazin-1-yl)acetamide (71 mg, 0.29
mmol) in anhydrous DMSO (1.5 mL) was added DIPEA (0.15 mL; 0.87
mmol), and the resulting biphasic mixture was efficiently stirred
at room temperature for 18 h. The resulting mixture was diluted
with 1:9 MeOH/EtOAc (10 mL) and washed with aq. NH.sub.4Cl (ca. 10
mL; 50% NH.sub.4Cl saturation) then brine (10 mL). The organic
layer was separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated, and The resulting residue was purified by flash
column chromatography on silica gel using 0-20% MeOH in DCM (v/v)
to afford the desired product as an off-white solid (54 mg, 41%
yield). .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.84 (s, 1H), 10.03
(s, 1H), 9.71 (s, 1H), 8.28 (s, 1H), 7.63-7.60 (m, 2H), 7.45-7.42
(m, 2H), 7.03-6.99 (m, 1H), 6.35 (br s, 1H), 2.55-2.52 (m, 4H;
obscured by DMSO signal), 2.41 (s, 3H), 2.40-2.37 (m, 4H), 2.17 (s,
3H); MS (ESI): calcd for C27H26F2N8O2: 532, found: 533
(MH.sup.+).
##STR00276##
[0236] To a mixture of
4-((4-aminophenyl)amino-6-4,7-difluoro-2-methyl-1H-indol-5-yl)oxy)pyrimid-
ine-5-carbonitrile (64 mg, 0.16 mmol) and pyridine (39 mg, 0.49
mmol) in DCM (3.0 mL) was added crotonoyl chloride (51 mg, 0.49
mmol). The resulting mixture was efficiently stirred at room
temperature for 1 h, whereupon it became homogeneous. The resulting
mixture was diluted with EtOAc (10 mL) and washed wit aq.
NaHCO.sub.3 (ca. 10 mL; 50% NaHCO.sub.3 saturation). The organic
layer was separated and dried over anhydrous Na.sub.2SO.sub.4 and
concentrated to afford the desired product as an off-white solid
(51 mg, 68% yield). .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.84
(br s, 1H), 10.03 (s, 1H), 9.82 (s, 1H), 8.29 (s, 1H), 7.68-7.65
(m, 2H), 7.46-7.43 (m, 2H), 7.03-6.99 (m, 1H), 6.34 (d, J=0.8 Hz,
1H), 5.80 (s, 1H), 5.52 (s, 1H), 2.42 (s, 3H), 1.96 (s, 3H); MS
(ESI): calcd for C27H26F2N8O2: 532, found: 533 (MH.sup.+).
##STR00277##
[0237] Step 1: To a solution of 2-Chloro-5-nitropyridine (10.00 g,
63.07 mmol) and Cis-2,6-dimethylpiperazine (9.00 g, 78.84 mmol) in
DMSO (50 ml) was added potassium carbonate (10.90 g, 78.84 mmol).
The mixture was stirred at 50.degree. C. for 18 h. The resulting
mixture was cooled until room temperature and added to a flsk
containing water/brine (600 ml). The mixture was stirred at room
temperature for 30 min. and then cooled to 0.degree. C. The solids
were collected by filtration, washed with water (100 ml.times.3).
The solids were triturated with hexane, collected by filtration and
further dried on vac line to give the product as yellow solids
(13.82 g, 92% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
8.93 (d, J=2.8 Hz, 1H), 8.17 (dd, J=2.8 Hz, J=9.6 Hz, 1H), 6.94 (d,
J=9.6 Hz, 1H), 4.40 (br, 2H), 2.69 (m, 2H), 2.49-2.30 (m, 3H), 1.02
(d, J=6.4 Hz, 3H); ESI-MS: calcd for (C11H16N4O2) 236, found 237
(MH.sup.+).
[0238] Step 2: A solution of QW910 (13.72 g, 58.07 mmol) in
methanol (350 mL) was hydrogenated in the presence of 10% Pd/C
(0.60 g) using an H.sub.2 balloon (3.times.). After 48 h, the
reaction mixture was filtered through a pad of celite and rinsed
with methanol (250 mL). The filtrate was concentrated to afford the
desired product (12.00 g, 100% yield) as purple solids. The product
was used directly for the next step reaction without further
purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.58 (d,
J=2.8 Hz, 1H), 6.88 (dd, J=8.8 Hz, J=2.8 Hz, 1H), 6.58 (d, J=8.8
Hz, 1H), 4.50 (br, 2H), 3.80 (m, 2H), 2.75 (m, 2H), 2.02 (m, 3H),
0.98 (d, J=6.0 Hz, 3H); ESI-MS: calcd for (C11H18N4) 206, found 207
(MH.sup.+).
[0239] Step 3: To a solution of aniline prepared in step 2 (141 mg,
0.68 mmol) in isopropanol (3 mL) was added TFA (50 uL) and shaked
thoroughly. A solution of intermediate 2 (200 mg, 0.62 mmol in
isopropanol (5 ml) was added. The reaction was heated to 85.degree.
C. for 16 h. After cooling to room temperature, hexanes (.about.8
ml) was added, cooled with ice and filtered to give the purple
slods. The solids were suspended in water (50 ml)/MeOH (5 ml) and
sat. NaHCO.sub.3 (.about.20 ml) was added (pH>8). The mixture
was stirred at room temperature for 45 min., then cooled with ice.
The solid were collected by filtration, washed with water (.about.5
ml) and hexanes (10 ml). The product was obtained as purple solids
(263 mg, 86%). HPLC was 95% and no further purification was
performed. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 11.85 (br,
1H), 9.93 (br, 1H), 8.24 (s, 1H), 8.17 (d, J=2.4 Hz, 1H), 7.62 (dd,
J=2.4 Hz, J=9.2 Hz, 1H), 7.00 (dd, J=5.2 Hz, J=10.4 Hz, 1H), 6.87
(d, J=9.2 Hz, 1H), 6.34 (s, 1H), 4.22 (d, J=12 Hz, 2H), 2.90 (br,
2H), 2.46-2.00 (m, 6H), 1.11 (d=6.4 Hz, 6H); ESI-MS: calcd for
(C25H24F2N80) 490, found 491 (MH.sup.+).
EXAMPLES
[0240] The following examples are provided to further illustrate
the present invention but, of course, should not be construed as in
any way limiting its scope.
Example 1
[0241] This example tests exemplary compounds from among Compound 1
to Compound 61 disclosed above for kinase inhibitory activity. A
significant number of these compounds have kinase inhibitory
activity collectively, over a broad spectrum of kinases.
[0242] Kinase assay protocols well known to those of ordinary skill
in the art were used. Specifically, the buffer composition was as
follows: 20 mM MOPS, 1 mM EDTA, 0.01% Brij-35, 5% Glycerol, 0.1%
.beta.-mercaptoethanol, 1 mg/mL BSA. Test compounds were initially
dissolved in DMSO at the desired concentration, then serially
diluted to the kinase assay buffer. In a final reaction volume of
25 .mu.L FGFR1(h) (5-10 mU) and KDR(h) (5-10 mU) are incubated with
8 mM MOPS pH 7.0, 0.2 mM EDTA, 200 .mu.M LRRASLG (Kemptide), 10 mM
magnesium acetate and [.gamma..sup.33P-ATP]. The reaction was
initiated by the addition of the MgATP mix. After incubation for 40
minutes at room temperature, the reaction was stopped by addition
of 5 .mu.L of a 3% phosphoric acid solution. Ten L of the reaction
was then spotted onto a P30 filter mat and washed three times for 5
minutes in 50 mM phosphoric acid and once in methanol prior to
drying and scintillation counting. Wells containing substrate but
no kinase and wells containing a phosphopeptide control were used
to set 0% and 100% phosphorylation value, respectively.
[0243] Table 1 shows representative data for the inhibition of
kinase by compounds of this invention. FGFR1 and KDR kinases would
be recognized by those of ordinary skill in the art as associated
with cancer.
TABLE-US-00001 TABLE 1 Inhibition of kinase activity in two kinases
associated with cancer. % Inhibition at 1 .mu.M Example No.
FGFR1(h) KDR(h) 1 99 96 2 100 96 3 1 4 4 -- -- 5 26 9 6 85 95 7 49
94 8 97 96 9 100 96 10 34 23 11 100 96 12 56 67 13 48 85 14 99 93
15 94 96 16 100 96 17 100 96 18 76 92 19 100 86 20 70 42 21 100 96
22 100 96 23 98 96 24 86 95 25 99 96 26 97 96 27 99 97 28 99 96 29
91 95 30 91 94 31 102 95 32 92 69 33 101 89 34 100 89 35 100 95 36
99 67 37 99 70 38 90 73 39 91 36 40 78 20 41 101 96 42 101 96 43 97
76 44 41 14 45 101 73 46 101 90 47 101 74 48 100 84 49 101 82 50
101 88 51 101 80 52 101 91 53 100 93 54 89 17 55 55 6 56 101 94
Example 2
[0244] A number of studies were performed to analyze the
consequences of tyrosine kinase inhibition in cell lines. To do
this, 1000 cells are seeded in 27 .mu.l/well in 384-well
microplates, which are then placed in a humidified CO.sub.2
incubator at 37.degree. C. overnight. The next day, 3 .mu.l/well of
10.times. concentrated drug is added and the plates are returned to
the incubator for 72 hr. After 72 hr incubation, plates are removed
and 6 .mu.l/well CellTiterblue (Promega) viability reagent is
added. Plates are returned to the incubator for 3 hrs, after which
fluorescence measurements are read on the Victor X3 plate reader
(Perkin Elmer). Data are analyzed using Excel (Microsoft), and
GI.sub.50 values are determined using Prism (Graphpad).
[0245] For the phospho-FGFR, the following assay Protocol was used.
25,000 cells are seeded in 90 .mu.l/well in 96-well microplates,
which are then placed in a humidified CO.sub.2 incubator at
37.degree. C. overnight. 96-well ELISA plates (Mesoscale Discovery)
are coated with capture antibody (R&D Systems Duo-Set) at 4
.mu.g/ml, 30 .mu.l/well. The next day, 10 .mu.l/well of 10.times.
concentrated drug is added and the plates are returned to the
incubator for 20 min. ELISA plates are washed using an automated
plate washer (BioTek Instruments). After 30 min, cells are inverted
and gently tapped to remove excess medium, and immediately placed
on ice. 30 1 mPer cell lysis reagent (Thermo Scientific) with
protease and phosphatase inhibitors is added/well. After 15 min on
ice, lysates are mixed and 30 .mu.l transferred to the ELISA plate.
Plates are incubated for 2 hr, washed, and 30 .mu.l/well detection
antibody is added. After 1 hr, plates are washed and 30 .mu.l
"SulfoTag" (MesoScale Discovery) detection reagent is added. After
1 hr, plates are washed and 150 .mu.l/well reading solution is
added. Electrochemiluminescence is determined on the Mesoscale
Discovery Sector Imager 2000. Data are analyzed using Excel
(Microsoft), and EC.sub.50 values are determined using Prism
(Graphpad).
[0246] Table 2 shows representative GI50 data for the inhibition of
selected cancer cell lines. Those of ordinary skill in the art
appreciate that each cell line is a surrogate for a particular kind
of cancer. This example confirms that protein kinase inhibitors can
have effects on cell proliferation. Those of ordinary skill would
be surprised by the specificity of the kinase inhibition. All of
the kinase inhibitors tested herein have been associated with
cancer.
TABLE-US-00002 TABLE 2 Inhibition of Proliferation GI50 (nM)
Compound No. KG1a SNU16 Kato III RT112 1 49 -- -- -- 2 62 -- -- --
8 250 -- -- -- 9 314 -- -- -- 11 25 -- -- 14 1.6 23 137 76 16 15 --
-- 17 0.74 20 113 83 19 0.2 12 79 66 22 196 -- -- -- 25 30 -- -- --
28 226 -- -- -- 31 0.35 32 205 171 33 0.28 12 88 126 34 0.28 22 98
52 35 78 -- -- -- 36 182 -- -- -- 41 180 -- -- -- 42 35 -- -- -- 45
0.43 15 79 37 46 1.37 22 111 48 47 0.5 22 123 48 48 0.55 35 152 57
49 0.6 24 86 48 50 2.91 52 174 49 51 1.62 66 208 73 52 174 -- -- 53
3.59 28 152 94 56 5.59 24 106 134 57 15 6 -- -- 58 101 122 -- -- 59
31 79 -- -- 60 101 92 -- -- 61 4 75 -- --
Example 3
[0247] In this Example the antitumor activity of compounds 14, 17,
19, 45, and 48 of the current invention are tested using an
art-recognized xenograft model of AML. ("T/C" refers to the ratio
of the tumor's size in treated animals versus the tumor size in
control untreated animals. "BWC" refers to "body weight
change").
TABLE-US-00003 TABLE 3 Antitumor Efficacy of Lead Compounds on TG1a
Human AML Xenograft Com- Dosing pound Dose Sched- T/C (%) BWC Group
No. ID (mg/kg) ule Route (day14) (%) A 4 Vehicle / Qdx10 IP / -7.2
B 4 Cpd 19 5 Qdx10 IP -69.7 -4.1 C 4 Cpd 19 10 Qdx10 IP -83.7 -6.3
D 4 Cpd 19 20 Qdx10 IP -68.7 -26.2 (day7) (d7) E 4 Cpd 14 20 Qdx10
IP -60.6 -4.0 F 4 Cpd 17 20 Qdx10 IP -79.5 -5.7 G 4 Cpd 45 20 Qdx10
IP 4.9 -11.7 H 4 Cpd 48 20 Qdx10 IP -72.8 -3.8 "Qdx10", once a day
for 10 days
Example 4
[0248] This Example demonstrates the biologic activity and
pharmaceutical suitability of an embodiment of the invention,
Compound 19. This Example is offered as exemplary of the
characteristics of the inventive compounds disclosed by the current
application. However, this Example is in no way meant to be
limiting the scope of protection obtained.
[0249] Of all of the steps in developing a drug, the hardest is
finding promising compounds. (See, e.g., Malo et al., "Statistical
practice in high-throughput screening data analysis," Nature
Biotechnology, 2006, 24, 167-75.) Kinase inhibitors often also have
an anti-proliferative effect. This characteristic attraction of
kinase inhibitors, like the compounds of Example 2, is their
ability to inhibit mutant forms of the kinase which have been shown
to be important in the transformation of normal cells into cancer
cells. The kinase inhibitory activity Compound 19 was tested using
the kinase assay described in Example 3. Tables 4 and 5 demonstrate
the kinase inhibitory activity of the Compound 19. Unpredictably,
Compound 19 is highly active against wild type and mutant forms of
fibroblast growth factor receptor kinases. These mutations in the
tested kinases are thought to play an important role in
transformation of some cancers. In addition, Table 4 also
demonstrates that Compound 19 is reasonably selective for
fibroblast growth factor receptor kinases.
[0250] Of all of the steps in developing a drug, the hardest is the
finding promising Compounds.
TABLE-US-00004 TABLE 4 PHD Kinase IC50 (nM) FGFR1(h) 0.9
FGFR1(V561M)(h) 134 FGFR2(h) 11 FGFR2(N549H)(h) 7 FGFR3(h) 12
FGFR4(h) 12 KDR(h) 485 Flt4(h) 116 Lck(h) >1,000 ABL(h), ALK(h),
Aurora-A (h), Axl(h), cSRC(h), Flt3(h), >1,000 IGF-1R(h),
AJK2(h), MEK1(h), Mer(h), Met(h), Ret(h), Tie2(h)
[0251] FIG. 1 demonstrates the kinase inhibitory dose-response
curve for compound 19 against specific kinases in a model system
that uses BaF3 cells engineered for the expression of specific
kinases. In FIG. 1 these kinases are from the fibroblast growth
factor receptor class of kinases (FGFR1-FGFR4). Compound 19 kinase
inhibitory activity FGFR1-FGFR4 are also depicted in Table 5.
Overall, Compound 19 demonstrates significant kinase inhibitory
activity.
TABLE-US-00005 TABLE 5 Compound 19 Activity Against Fibroblast
Growth Factor Receptor Kinases Compound Assay IC5o(nM) Cpd 19
Parental 1,890 FGFR1 11.4 FGFR2 17.6 FGFR3 32.6 FGFR4 92.1
[0252] Compound 19 also has significant anti-proliferation
properties. Table 6 summarizes results with Compound 19 tested by
the proliferation assay described in Example 2.
[0253] As shown in Table 6, Compound 19 has significant
anti-proliferation activity against some, but not all the cell
lines tested. Notably, a common feature of susceptible cells is
their expression of one of the fibroblast growth factor receptors.
In this assay, Compound 19 is especially effective in KG1a which is
a cell line made from a form of acute myeloid leukemia ("AML").
[0254] The pharmacokinetic profile of Compound 19 in rats is
presented in Table 7. Those of ordinary skill in the art would
appreciate that, while not perfectly predictive, these results
would be similar in other animals including, e.g., human patients.
Compound 19's pharmacokinetic profile is consistent with Compound
19 being used as a therapeutic agent.
TABLE-US-00006 TABLE 7 Pharmacokinetic Profile of Compound 19 in
Rats Results Study Compound 19 PK IV (1 Terminal Half 6.81 (Rat)
mg/Kg) Life (hr.) Vz(L/kg) 9.54 Cl 17.57 (mL/min/kg) PO (5 Oral 76
mg/Kg) Bioavailability (% F)
[0255] The metabolic half-life for Compound 19 was determined in
human, rat, and mouse model systems his presented in Table 7. Those
of ordinary skill in the art will recognize that these results
indicate that Compound 19 can be quite stable in vivo.
TABLE-US-00007 TABLE 8 The Metabolic-Stability of Compound 19
Metabolic Half-Life (min) Human Liver Rat Liver Mouse Liver Test
Article Microsomes Microsomes Microsomes DASATINIB 7.5 11.9 9.2
COMPOUND >60 >60 >60 19
[0256] Table 9 discloses toxicity data for compound 19 from various
toxicity tests well known to those of ordinary skill. These results
present nothing that would suggest against the institution of the
Phase I trial for compound 19.
TABLE-US-00008 TABLE 9 Compound 19 Toxicity Assessment Toxicity
Test Results Cellular HepG2 Cell line (CC.sub.50) 1.9 .mu.M Primary
human Hepatocyte (CC.sub.50) 20 .mu.M In Vivo Acute MTD (PO, rat)
100 mg/kg Cardiovascular hERG (IC.sub.50) >30 .mu.M CYP450 1A2
(IC.sub.50) 72.0 .mu.M 2C8 (IC.sub.50) 10.6 .mu.M 2C9 (IC.sub.50)
32.9 .mu.M 2D6 (IC.sub.50) >100 .mu.M 3A4 (IC.sub.50) 67.3
.mu.M
[0257] Surprisingly it was found that Compound 19 has significant
antitumor activity in a nude mouse model of AML. FIG. 2
demonstrates that both of the compound 19 dosages test resulted in
near complete suppression of the tumor cells expansion. FIG. 3
demonstrates that animal weight, used as a surrogate marker for
toxicity, showed no difference between the control and compound 19
treated animals. These results are confirmed in the dose-response
study in the same animal model depicted in FIGS. 4 and 5. (Of note,
the highest dose of Compound 19 (20 mg/kg) results in significant
weight loss suggesting the presence of generalized toxicity, rather
than specific effect on cancer cells.)
[0258] Overall these data on the behavior of compound 19 are
perfectly consistent with this compound being developed into an
anti-proliferation/antitumor agent.
[0259] All references, including publications, patent applications,
and patents, cited herein are hereby incorporated by reference to
the same extent as if each reference were individually and
specifically indicated to be incorporated by reference and were set
forth in its entirety herein.
[0260] The use of the terms "a" and "an" and "the" and "at least
one" and similar references in the context of describing the
invention (especially in the context of the following claims) are
to be construed to cover both the singular and the plural, unless
otherwise indicated herein or clearly contradicted by context. The
use of the term "at least one" followed by a list of one or more
items (for example, "at least one of A and B") is to be construed
to mean one item selected from the listed items (A or B) or any
combination of two or more of the listed items (A and B), unless
otherwise indicated herein or clearly contradicted by context. The
terms "comprising," "having," "including," and "containing" are to
be construed as open-ended terms (i.e., meaning "including, but not
limited to,") unless otherwise noted. Recitation of ranges of
values herein are merely intended to serve as a shorthand method of
referring individually to each separate value falling within the
range, unless otherwise indicated herein, and each separate value
is incorporated into the specification as if it were individually
recited herein. All methods described herein can be performed in
any suitable order unless otherwise indicated herein or otherwise
clearly contradicted by context. The use of any and all examples,
or exemplary language (e.g., "such as") provided herein, is
intended merely to better illuminate the invention and does not
pose a limitation on the scope of the invention unless otherwise
claimed. No language in the specification should be construed as
indicating any non-claimed element as essential to the practice of
the invention.
[0261] Preferred embodiments of this invention are described
herein, including the best mode known to the inventors for carrying
out the invention. Variations of those preferred embodiments may
become apparent to those of ordinary skill in the art upon reading
the foregoing description. The inventors expect skilled artisans to
employ such variations as appropriate, and the inventors intend for
the invention to be practiced in additional ways, not expressly
taught herein. Accordingly, this invention includes all
modifications and equivalents of the subject matter recited in the
claims appended hereto as permitted by applicable law. Moreover,
any combination of the above-described elements in all possible
variations thereof is encompassed by the invention unless otherwise
indicated herein or otherwise clearly contradicted by context.
* * * * *