U.S. patent application number 15/745915 was filed with the patent office on 2018-11-15 for chiral diaryl macrocycles and uses thereof.
The applicant listed for this patent is TP THERAPEUTICS, INC.. Invention is credited to Jingrong J. CUI, Wei DENG, Zhongdong HUANG, Yishan LI, Evan W. ROGERS, Dayong ZHAI.
Application Number | 20180325901 15/745915 |
Document ID | / |
Family ID | 57834656 |
Filed Date | 2018-11-15 |
United States Patent
Application |
20180325901 |
Kind Code |
A1 |
CUI; Jingrong J. ; et
al. |
November 15, 2018 |
CHIRAL DIARYL MACROCYCLES AND USES THEREOF
Abstract
This disclosure relates to the use of certain diaryl macrocycle
compounds, specifically
(7S13R)-11-fluoro-7,13-di-methyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one in the
treatment of disease in mammals. This disclosure also relates to
compositions including such compounds, and to methods of using such
compositions in the treatment of diseases in mammals, especially in
humans.
Inventors: |
CUI; Jingrong J.; (San
Diego, CA) ; LI; Yishan; (San Diego, CA) ;
ROGERS; Evan W.; (San Diego, CA) ; ZHAI; Dayong;
(San Diego, CA) ; DENG; Wei; (San Diego, CA)
; HUANG; Zhongdong; (San Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
TP THERAPEUTICS, INC. |
San Diego |
CA |
US |
|
|
Family ID: |
57834656 |
Appl. No.: |
15/745915 |
Filed: |
July 20, 2016 |
PCT Filed: |
July 20, 2016 |
PCT NO: |
PCT/US2016/043132 |
371 Date: |
January 18, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62195081 |
Jul 21, 2015 |
|
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|
62302231 |
Mar 2, 2016 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/519 20130101;
A61P 35/00 20180101 |
International
Class: |
A61K 31/519 20060101
A61K031/519; A61P 35/00 20060101 A61P035/00 |
Claims
1. A method of treating disease in a patient comprising,
administering to the patient a therapeutically effective amount of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof.
2. The method of claim 1, wherein the disease is mediated by a
tyrosine kinase selected from the group consisting of ALK, ROS1,
TRKA, TRKB, TRKC, JAK2, SRC, FYN, LYN, YES, FGR, FAK, ARK5, and
combinations thereof.
3.-7. (canceled)
8. The method of claim 1 7, wherein the disease is cancer.
9. (canceled)
10. The method of claim 1, wherein the disease is a cancer mediated
by a genetically altered ALK.
11. The method of claim 1, wherein the disease is a cancer mediated
by a fusion protein comprising a fragment of a protein encoded by
an ALK gene and a fragment of a protein encoded by a gene selected
from the group consisting of NPM, EML4, TPR, TFG, ATIC, CLTC1,
TPM4, MSN AL017 and MYH9.
12. (canceled)
13. The method of claim 10, wherein the genetically altered ALK is
one or more of an EML4-ALK fusion protein, an NPM-ALK fusion
protein, or a TPR-ALK fusion protein.
14. (canceled)
15. The method of claim 13, wherein the one or more of an EML4-ALK
fusion protein, an NPM-ALK fusion protein, or a TPR-ALK fusion
protein comprises at least one resistance mutation.
16. The method of claim 13, wherein the EML4-ALK fusion protein
comprises at least one mutation selected from the group consisting
of L1196M, G1202R, D1203R, L1152P/R, F1174C/L/V, C1156Y, I1171N,
G1123S, S106Y, G1269S/A, and 1151T insertion.
17.-30. (canceled)
31. The method of claim 13, wherein the TPR-ALK fusion protein
comprises a L1196M point mutation.
32. (canceled)
33. The method of claim 1, wherein the disease is a cancer mediated
by ALK having one or more point mutations selected from the group
consisting of R1050H, F1174C/I/L/S/V, F1245C/I/L/V, R1275L/Q,
T1151M, M1166R, I1170N, I1170S, I1171N, I1183T, L1196M, A1200V,
L1204F, L1240V, D1270G, Y1278S, R1192P, G1128A, G1286R, and
T13431.
34.-39. (canceled)
40. The method of claim 1, wherein the disease is a cancer mediated
by a genetically altered ROS1.
41. The method of claim 1, wherein the disease is a cancer mediated
by a fusion protein comprising a fragment of a protein encoded by
an ROS1 gene and a fragment of a protein encoded by a gene selected
from the group consisting of FIG, TPM3, SDC4, SLC34A2, CD74, EZR,
and LRIG3.
42. (canceled)
43. The method of claim 40, wherein the genetically altered ROS1 is
one or more of a CD74-ROS1 fusion protein, a SDC4-ROS1 fusion
protein, or a SLC34A2-ROS1fusion protein.
44. (canceled)
45. The method of claim 43, wherein the one or more of a CD74-ROS1
fusion protein, a SDC4-ROS1 fusion protein, or a SLC34A2-ROS1fusion
protein comprises at least one resistance mutation.
46. The method of claim 43, wherein the CD74-ROS1 fusion protein
comprises a G2032R, L2026M or D2033N point mutation.
47.-49. (canceled)
50. The method of claim 43 4-7, wherein the SDC4-ROS1 fusion
protein comprises a G2032R point mutation.
51.-53. (canceled)
54. The method of claim 43, wherein the SLC34A2-ROS1 fusion protein
comprises a G2032R point mutation.
55.-56. (canceled)
57. The method of claim 1, wherein the disease is a cancer mediated
by TRKA, TRKB or TRKC.
58. The method of claim 1, wherein the disease is a cancer mediated
by a genetically altered TRKA, TRKB or TRKC.
59. (canceled)
60. The method of claim 58, wherein the genetically altered TRKA is
a TPM3-TRKA or LMNA-TRKA fusion protein.
61.-69. (canceled)
70. The method of claim 1, wherein the disease is a cancer mediated
by JAK1, JAK2 or JAK3.
71. The method of claim 1, wherein the disease is a cancer mediated
by a genetically altered JAK2.
72. (canceled)
73. The method of claim 71, wherein the genetically altered JAK2 is
a TEL-JAK2 fusion protein, or a PCM1-JAK2 fusion protein.
74. (canceled)
75. The method of claim 71, wherein the genetically altered JAK2
comprises a V617F point mutation.
76. The method of claim 1, wherein the disease is a cancer mediated
by SRC.
77.-78. (canceled)
79. The method of claim 1, wherein the disease is pain mediated by
TRKA, TRKB or TRKC.
80.-82. (canceled)
83. The method of claim 1 wherein the disease is selected from the
group consisting of psoriasis, rheumatoid arthritis, polycythemia
vera, essential thrombocythemia, ulcerative colitis, and myeloid
metaplasia with myelofibrosis.
84. (canceled)
85. A method of treating cancer in a patient previously shown to
express a genetically altered tyrosine or serine/threonine kinase
comprising, administering to the patient a therapeutically
effective amount of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof.
86. The method of claim 85, wherein the genetically altered
tyrosine kinase is selected from the group consisting of a
genetically altered ALK, genetically altered ROS1, genetically
altered TRK and genetically altered JAK.
87. The method of claim 86, wherein the genetically altered ALK is
a fusion protein comprising a fragment of a protein encoded by an
ALK gene and a fragment of a protein encoded by a gene selected
from the group consisting of NPM, EML4, TPR, TFG, ATIC, CLTC1,
TPM4, MSN ALO17 and MYH9.
88. (canceled)
89. The method of claim 86, wherein the genetically altered ALK is
one or more of an EML4-ALK fusion protein, an NPM-ALK fusion
protein, or a TPR-ALK fusion protein.
90.-100. (canceled)
101. The method of claim 85, wherein the bypass resistance
mechanism is mediated by SRC.
102.-103. (canceled)
104. The method of claim 86, wherein the genetically altered ROS1
is a fusion protein comprising a fragment of a protein encoded by
an ROS1 gene and a fragment of a protein encoded by a gene selected
from the group consisting of FIG, TPM3, SDC4, SLC34A2, CD74, EZR,
and LRIG3.
105. (canceled)
106. The method of claim 104, wherein the genetically altered ROS1
is a CD74-ROS1 fusion protein, a SDC4-ROS1 fusion protein, or a
SLC34A2-ROS1fusion protein.
107.-120. (canceled)
121. The method of claim 86, wherein the genetically altered TRK is
a TPM3-TRKA or LMNA-TRKA fusion protein.
122.-127. (canceled)
128. The method of claim 86, wherein the genetically altered JAK is
a TEL-JAK2 fusion protein or a PCM1-JAK2 fusion protein.
129. (canceled)
130. A method of treating cancer in a patient comprising; i.
identifying a genetically altered tyrosine or serine/threonine
kinase in the patient, and ii. administering to the patient a
therapeutically effective amount of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyraz-
olo[4,3-f][1,4,8,10]benzoxatriaza-cyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof.
131. The method of claim 130, wherein the step of identifying
comprises subjecting a patient sample to a test selected from the
group consisting of FISH, IHC, PCR and gene sequencing.
132. A method of identifying a patient for treatment with
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-j][1,4,8,10]benzoxatriaza-cyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof, comprising diagnosing the
patient with a cancer mediated by a genetically altered tyrosine or
serine/threonin kinase.
133.-143. (canceled)
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority under 35 U.S.C. .sctn.
119(e) to U.S. Provisional Application Ser. No. 62/195,081, filed
Jul. 21, 2015 and U.S. Provisional Application Ser. No. 62/302,231,
filed Mar. 2, 2016, both of which are incorporated herein by
reference in their entirety.
TECHNICAL FIELD
[0002] This disclosure relates to the use of certain diaryl
macrocycle compounds, specifically
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one in the
treatment of disease in mammals. This disclosure also relates to
compositions including such compounds, and to methods of using such
compositions in the treatment of diseases in mammals, especially in
humans.
BACKGROUND
[0003] Protein kinases are key regulators for cell growth,
proliferation and survival. Genetic and epigenetic alterations
accumulate in cancer cells leading to abnormal activation of signal
transduction pathways which drive malignant processes. (Manning,
G.; Whyte, D. B.; Martinez, R.; Hunter, T.; Sudarsanam, S. The
protein kinase complement of the human genome. Science 2002, 298,
1912-1934). Pharmacological inhibition of these signaling pathways
presents promising intervention opportunities for targeted cancer
therapies. (Sawyers, C. Targeted cancer therapy. Nature 2004, 432,
294-297).
[0004] Anaplastic lymphoma kinase (ALK), along with leukocyte
tyrosine kinase (LTK), belongs to the insulin receptor (IR)
superfamily of receptor tyrosine kinases. ALK is mainly expressed
in the central and peripheral nervous systems suggesting a
potential role in normal development and function of the nervous
system. (Pulford K, et al Cell Mol. Life Sci. 2004, 61, 2939). ALK
was first discovered as a fusion protein, NPM (nucleophosmin)-ALK
encoded by a fusion gene arising from the t(2;5)(p23;q35)
chromosomal translocation in anaplastic large cell lymphoma (ALCL)
cell lines in 1994. (Morris S W, et al Science 1994, 263, 1281.)
More than twenty distinct ALK translocation partners have been
discovered in many cancers, including ALCL (60-90% incidence),
inflammatory myofibroblastic tumours (IMT, 50-60%), non-small cell
lung carcinomas (NSCLC, 3-7%), colorectal cancers (CRC, 0-2.4%),
breast cancers (0-2.4%), and other carcinomas with rare incidence.
(Grande E, et al Mol. Cancer Ther. 2011, 10, 569.) Oncogenic point
mutations of ALK have been discovered in both familial and sporadic
cases of neuroblastoma. (Mosse Y P, et al Nature 2008, 455,
930-935.) Both fusion and mutant ALKs are highly oncogenic, which
generate considerable interest and efforts in developing ALK
inhibitors for the treatment of haematopoietic, solid, and
mesenchymal tumors with abnormal ALK gene. (Grande, E, et al Mol.
Cancer Ther. 2011, 10, 569-579.). Crizotinib was approved by the US
Food and Drug Administration for the treatment of ALK-positive
non-small cell lung cancer. Similar with many targeted therapies of
kinase inhibitors, crizotinib drug resistance developed in about 10
months. Mechanisms of drug resistance include target gene
amplification or overexpression, development of secondary missense
mutations, and use of alternative signaling pathway (so-called
"bypass resistance"). As a result, second-generation ALK inhibitors
have been developed to be more potent against wild and many mutant
ALKs. One such mutation is the gatekeeper mutation ALK.sup.L1196M.
Ceritinib was approved by the US Food and Drug Administration for
the treatment of patients with ALK-positive non-small cell lung
cancer showing disease progression or who are intolerant to
crizotinib. Although many second generation ALK inhibitors have
been investigated in clinical trials, new ALK mutations resistant
to the second generation ALK inhibitors have emerged. For example,
the G1202R mutation has been found in tumors resistant to
crizotinib, ceritinib, and alectinib. (Politi K, Clin Cancer Res.
2014, 20, 5576.) Novel isoforms of ALK consisting primarily of the
intracellular tyrosine kinase domain was found to express in
.about.11% of melanomas and sporadically in other human cancer
types, but not in normal tissues (Wiesner T, et al Nature 2015,
526, 453-457). These new ALK isoforms stimulate multiple oncogenic
signalling pathways, and are sensitive to ALK inhibitors,
suggesting potential clinical benefits from ALK inhibition.
[0005] Non-small cell lung cancers harboring ALK gene
rearrangements are sensitive to treatment with the ALK inhibitor
crizotinib. However, the emergence of drug resistance is universal
and rapidly limits clinical applicability. The mechanisms of
resistance include ALK gene amplification, acquired ALK missense
mutations, bypass pathway activation, and epithelial-mesenchymal
transition (EMT) (Katayama R 2012). (Katayama R., et al Sci Transl
Med. 2012, 4(120):120ra17) Bypass and EMT constitute majority of
the acquired resistant population. It is worth to note that 30-40%
of ALK fusion positive patients have intrinsic resistance to ALK
inhibitor treatment. ALK rearranged NSCLCs are typically
adenocarcinoma characterized by a solid signetring cell pattern
that is frequently associated with a metastatic phenotype and
linked to an epithelial-mesenchymal transition (EMT) phenotype.
(Voena C, et al. Oncotarget, 2016, Apr. 23, 8955) The H2228 cell
line with EML4-ALK v3 fusion gene displayed a mesenchymal phenotype
with directly suppressing E-cadherin and up-regulating vimentin
expression, as well as expression of other genes involved in EMT.
H2228 cell line confers intrinsic resistance to crizotinib and
other ALK inhibitors. Therefore, it is necessary to develop a
polypharmacology ALK inhibitor being able to target EMT and
metastasis. Bypass resistance occurs when the original driver
oncogene and a secondary bypass track redundantly maintain
downstream signaling to promote cell survival and proliferation.
For example, ALK inhibition in patient-derived ALK models has been
shown to up-regulate SRC activity. The combination of a Src
tyrosine kinase inhibitor with an ALK inhibitor was shown to
effectively suppress downstream signaling, generated a synergistic
inhibition effect, and re-sensitized the ALK inhibitors in the
patient-derived ALK models in vitro and in vivo. (Crystal A S,
Science. 2014, 346, 1480.) The identification of new ALK inhibitors
that can counter broad secondary ALK mutations including
ALK.sup.G1202R and inhibit Src signaling will be important and
highly desired for effectively overcoming ALK drug resistance and
sustaining the response to ALK inhibitor treatment.
[0006] ROS1 protein is a receptor tyrosine kinase, closely related
to the ALK/LTK and insulin receptor kinase family. Although normal
physiologic functions of human ROS1 kinase have not been fully
understood, the abnormal expression and variable constitutively
activating fusion forms of ROS1 kinase have been reported in a
number of cancers including glioblastoma, non-small cell lung
cancer, cholangiocarcinoma, ovarian cancer, gastric adenocarcinoma,
colorectal cancer, inflammatory myofibroblastic tumor,
angiosarcoma, and epithelioid hemangioendothelioma. (Kurtis D D, et
al Clin Cancer Res 2013, 19 (15), 1.) FIG-ROS1 fusion protein was
the first fusion protein of ROS1 discovered in 2003 in a human
glioblastoma multiforme. (Charest A, et al Genes Chromosomes Cancer
2003, 37, 58) Several fusion proteins with ROS1 kinase including
TPM3, SDC4, SLC34A2, CD74, EZR, and LRIG3 have been reported from
human lung cancers, suggesting the oncogenic role of ROS1 kinase in
lung cancers. (Takeuchi K, et al Nat. Med. 2012, 18, 378) The
survey of activated tyrosine kinases signaling in 23
cholangiocarcinoma patients confirmed the presence of FIG-ROS
kinase fusion in 8.7% of cholangiocarcinoma patients. (Gu T L, et
al PLoS One. 2011, 6, e15640.) More and more ROS1 fusion partners
including KDELR2, CCDC6, MSN, LIMA1, CLTC, NF.kappa.B2, NCOR2,
CEP85L, TMEM106B, HLA-A, MYOSA, PPFIBP1, ERC1, PWWP2A, CLIP1,
ZCCHC8, SHTN1, TFG, and YWHAE have been reported from various human
cancers (Uquen A, et al Future Oncol. 2016, Jun. 3, Epub ahead of
print) Taken together, ROS1 kinase is a promising molecular based
target candidate for cancers with aberrant ROS kinase activities.
The ALK/MET/ROS1 inhibitor crizotinib has demonstrated marked
efficacy in patients with NSCLC whose tumors are positive for ROS1
genetic abnormalities. (Shaw A T, et al N Engl J Med 2015, 372,
683). As expected ROS1 rearrangement-positive patients who
responded to crizotinib eventually experienced disease progression.
The secondary ROS1.sup.G2032R mutation and bypass signaling are
associated with the resistance. (Awad M M, et al N Engl J Med 2013,
368, 2395) It is desired to develop the next generation of ROS1
inhibitor to overcome the resistance.
[0007] The tropomyosin-related receptor tyrosine kinases (Trks) are
high-affinity receptors for neurotrophins (NTs), a nerve growth
factor (NGF) family. Trk was originally cloned as an oncogene fused
with the tropomyosin gene in the extracellular domain. The
activating mutations caused by chromosomal rearrangements or
mutations in TRK family have been reported in many cancers.
(Vaishnavi A, et al Cancer Discov. 2015, 5, 25) Because Trks play
important roles in pain sensation as well as tumour cell growth and
survival signaling, inhibitors of Trk receptor kinases might
provide benefit for pain and cancer treatment.
[0008] The Janus family of kinases (JAKs) include JAK1, JAK2, JAK3
and TYK2, and are cytoplastic non-receptor tyrosine kinases
required for the physiologic signaling of cytokines and growth
factors. (Quintas-Cardama A, et al., Nat. Rev. Drug Discov. 2011,
10(2), 127) Aberrant regulation of JAK/STAT pathways has been
implicated in multiple human pathological diseases, including
cancer (JAK2) and rheumatoid arthritis (JAK1, JAK3). A
gain-of-function mutation of JAK2 (JAK2V617F) has been discovered
with high frequency in MPN patients. (Levine R L, et al. Cancer
Cell 2005, 7, 387) The mutation in the JH2 pseudokinase domain of
JAK2 leads to constitutively kinase activity. Cells containing the
JAK2V617F mutation acquire cytokine-independent growth ability and
often become tumor, providing strong rationale for the development
of JAK inhibitors as a targeted therapy. In addition,
hyperactivation of the JAK2/signal transducers and activators of
transcription 3 (JAK2/STAT3) is responsible for abnormal dendritic
cell differentiation leading to abnormal dendritic cell
differentiation and accumulation of immunosuppressive myeloid cells
in cancer (Nefedova Y, et al. Cancer Res 2005, 65, 9525). In
Pten-null senescent tumors, activation of the JAK2/STAT3 pathway
establishes an immunosuppressive tumor microenvironment that
contributes to tumor growth and chemoresistance (Toso A, et al.
Cell Reports 2014, 9, 75). JAK2 gene fusions with the TEL(ETV6)
(TEL-JAK2) and PCM1 genes have been found in leukemia patients.
(Lacronique V, et al. Science 1997, 278, 5341, 1309-12. Reiter A,
et al. Cancer Res. 2005, 65, 7, 2662-7.) It was reported that
JAK/STAT3 signaling pathway was aberrantly increased in EGFR
inhibitor-resistant EGFR-mutant non-small cell lung cancer (NSCLC)
cells, and JAK2 inhibition overcomes acquired resistance to EGFR
inhibitors that support the use of combination therapy with JAK and
EGFR inhibitors for the treatment of EGFR-dependent NSCLC. (Gao S
P, et al. Sci Signal. 2016, 9 (421):ra33) JAK/STAT3 signaling
promotes cancer hallmarks in the tumor and its environment,
including proliferation, survival, angiogenesis, tumor metabolism
while suppressing antitumor immunity. (Buchert M, et al. Oncogene,
2016, 35, 939-951) Inhibition of cytokine-dependent activation of
the JAK/STAT3 pathway with J AK inhibitors may also afford
orthogonal treatment opportunities for other oncogene-addicted
cancer cells that have gained drug resistance. Focal amplification
of JAK2 gene was observed in postchemotherapy triple-negative
breast cancers (TNBCs) in a group of 9p24-amplified tumors,
suggesting a role in tumorigenicity and chemoresistance. (Balko J
M, et al. Sci Transl Med. 2016, 8(334):ra53) Therefore,
pharmacologic inhibition of the JAK2 signaling pathway can be an
important new therapeutic strategy to enhance antitumor
activity.c-Src is a nonreceptor tyrosine kinase. The Src family
(SFK) comprises of eight members in humans (Src, Fyn, Yes, Lyn,
Lck, Hck, Blk and Fgr) with a molecular weight between 52-62 KDa.
Src and its family members are deregulated in many types of
cancer.Src is a key downstream transducer of many RTKs, including
EGFR, HER2, and c-Met. Activation of Src signaling has been
implicated in conferring therapeutic resistance to targeted
antiendocrine therapies, receptor tyrosine kinase therapies,
traditional chemotherapies, and radiation therapies. (Zhang S, et
al Trends Pharmacol Sci. 2012, 33, 122). SRC can promote signaling
from growth factor receptors in a number of ways including
participation in signaling pathways required for DNA synthesis,
control of receptor turn-over, actin cytoskeleton rearrangement,
migration, adhesion, invasion, motility, and survival. (Bromann PA,
Oncogene 2004, 23, 7957-7968) A prominent role of Src in tumor
progression-related events such as the epithelial mesenchymal
transition (EMT) and the development of metastasis have been
reported through the interaction with the potent metastasis
suppressor, N-myc downstream regulated gene 1 (NDRG1), that
regulates cancer cell migration by inhibiting Src activity. (Liu W,
et al. Oncotarget. 2015, 6: 35522-35541) Although EGFR inhibitors
have achieved a significant success in the majority of NSCLC
patients harbor EGFR-activating mutations, a subset of patients
with EGFR mutations are refractory to EGFR-TKIs. Resistance to EGFR
inhibitors reportedly involves SRC activation and induction of
epithelial-to-mesenchymal transition (EMT). The primary resistance
to EGFR-TKIs is associated with higher levels of CRIPTO1
expression. CRIPTO1 activated SRC and ZEB1 to promote EMT via
microRNA-205 (miR-205) downregulation. Therefore, co-targeting EGFR
and SRC may overcome intrinsic EGFR-inhibitor resistance in
patients with CRIPTO1-positive, EGFR-mutated NSCLC. (Park, K-S, et
al. J Clin Invest. 2014, 124(7):3003-3015) Focal Adhesion Kinase
(FAK) is a 125 kDa non-receptor tyrosine kinase and plays a
significant role in adhesion, survival, motility, metastasis,
angiogenesis, lymphangiogenesis, cancer stem cell functions, tumor
microenvironment and epithelial to mesenchymal transition (EMT).
(Golubovskaya V M, Front Biosci (Landmark Ed).; 19: 687-706)
Nuclear FAK controls chemokine transcription, Tregs, and evasion of
antitumor immunity, and the small-molecule FAK kinase inhibitor
VS-4718 drives depletion of Tregs and promotes a CD8+ T
cell-mediated anti-tumor response. (Serrels A, et al, Cells 2015,
163, 160-173).Therefore, FAK inhibitors may trigger immune-mediated
tumor regression. FAK is hyperactivated in human pancreatic ductal
adenocarcinoma (PDAC) and correlates with immunosuppressive tumor
microenvironment (TME). Targeting focal adhesion kinase renders
pancreatic cancers responsive to checkpoint immunotherapy by
overcoming the fibrotic and immunosuppressive PDAC TME in mouse
models. (Jiang H, et al. Nat Med. 2016, Jul. 4 [Epub ahead of
print]). Recently it was reported that saracatinib, a selective SRC
inhibitor, can re-sensitize ALK inhibitor-resistant cell lines,
demonstrating a therapeutic role of SRC inhibition in overcoming
ALK inhibitor resistance. (Crystal A S, et al. Science 2014, 346,
1480-1486) Therefore, Src/FAK inhibitor may play important roles in
combinatorial regimens in overcoming resistance to current
anticancer therapies and in preventing metastatic recurrence, EMT
and cancer treatment resistance. AMP-activated protein kinase
family member 5 (ARK5), also called NAUK1 is an upstream regulator
of AMPK and limits protein synthesis via inhibition of rapamycin 1
(mTORC1) signalling pathway. ARK5 maintains expression of
mitochondrial respiratory chain complexes and respiratory capacity
for efficient glutamine metabolism. ARK5 is highly expressed in
both primary NSCLC tissues and cell lines, that is functionally
associated with NSCLC metastasis and a predictor of poor progriosi,
for NSCLC patients. ARK5 modulated the migration and invasion of
NSCLC cells and played crucial roles in mTOR pathway. L, et al. Br
J Cancer. 2014, 111(12):2316-27) It was reported that ARK5 confers
doxorubicinresistance in HCC via inducing EMT. (Xu T, et al. Cancer
Lett. 2016, 377(2):140-8) Deregulated expression of the MYC
oncoprotein is associated with many human tumors. MYC promotes cell
growth and proliferation, and alters cellular metabolism.
Inhibition of ARK5 leads to a collapse of cellular ATP levels in
cells expressing deregulated MYC, and prolongs survival in
MYC-driven mouse models of hepatocellular carcinoma. (Liu L, et al.
Nature, 2012, 483, 608-612) Therefore, Targeting cellular energy
homeostasis by ARK5 inhibitor is a valid therapeutic strategy to
eliminate tumor cells with deregulated MYC expression.
[0009] Src is a non-receptor tyrosine kinase that is deregulated in
many types of cancer, and a key downstream transducer of many RTKs,
including EGFR, HER2, and c-Met. Activation of Src signaling has
been implicated in conferring therapeutic resistance to targeted
antiendocrine therapies, receptor tyrosine kinase therapies,
traditional chemotherapies, and radiation therapies. (Zhang S, et
al Trends Pharmacol Sci. 2012, 33, 122). Src inhibitor may play
important roles in combinatorial regimens in overcoming resistance
to current anticancer therapies and in preventing metastatic
recurrence. Cytoplasmic tyrosine kinases (also known as
non-receptor tyrosine kinases) of the Src family (SFKs) play
important roles in signal transduction induced by a large number of
extracellular stimuli including growth factors and integrins.
Elevated SFK activity is found in more than 80% of human colorectal
cancer (CRC) and this has been associated with poor clinical
outcome. (Summy J M, et al. Cancer Metastasis Rev. 2003, 22,
337-358) The SFK member Yes regulates specific oncogenic signalling
pathways important for colon cancer progression that is not shared
with c-Src. (Scancier F. et al. PLoS One. 2011, 6(2): e17237)
WASF2FGR fusion genes were found in lung squamous carcinoma,
ovarian serous cystadenocarcinoma, and skin cutaneous melanoma.
(Stransky N, et al. Nature Communications 2014, 5, 4846) Estrogen
receptorpositive (ER.sup.+) breast cancers adapt to hormone
deprivation and become resistant to antiestrogen therapy. Mutations
in the inhibitory SH2 domain of the SRC family kinase (SFK) LYN
were related to ER.sup.+ tumors that remained highly proliferative
after treatment with the aromatase inhibitor letrozole. LYN was
upregulated in multiple ER.sup.+ breast cancer lines resistant to
long-term estrogen deprivation. (Schwarz L J, et al. J Clin Invest.
2014, 124, 5490-5502) Therefore, targeting LYN will be a rational
strategy overcoming the escape from antiestrogens in a subset of
ER.sup.+ breast cancers. It was reported that LYN was overexpressed
in castrate-resistant prostate cancer (CRPC), enhanced AR
transcriptional activity, and accelerated CRPC progression, and
targeting Lyn kinase induced AR dissociation from the molecular
chaperone Hsp90, leading to its ubiquitination and proteasomal
degradation. (Zardan A., et al. Oncogenesis 2014, 3, e115) The Lyn
tyrosine kinase is a potential therapeutic target for the treatment
of CRPC. The Src family kinase FYN is involved in signal
transduction pathways in the nervous system, as well as the
development and activation of T lymphocytes under normal
physiological conditions. Activation of Fyn is observed in various
cancers, including melanoma, glioblastoma, squamous cell carcinoma,
prostate and breast cancers. (Elias D., et al. Pharmacological
Research 2015, 100, 250-254) Fyn was upregulated in
tamoxifen-resistant breast cancer cell lines and plays a key role
in the resistance mechanism. Peripheral T-cell lymphomas (PTCLs)
are a heterogeneous group of aggressive non Hodgkin lymphomas with
poor prognosis. FYN activating mutations were found in PTCL, and
promoted the growth of cells transformed via expression of
activated FYN mutant alleles. SRC kinase inhibitors may play
important roles in the treatment of PTCLs. (Couronne L, et al.
Blood 2013, 122, 811).
[0010] Discoidin domain receptors (DDRs) are activated by matrix
collagens and have been implicated in numerous cellular functions
such as proliferation, differentiation, adhesion, migration, and
invasion. DDRs play a role in cancer progression by regulating the
interactions of tumor cells with their surrounding collagen matrix.
DDR1 is a direct p53 transcriptional target, and the activation of
DDR1 is associated with p53-dependent DNA damage. DDR1 activated
the MAPK cascade in a Ras-dependent manner. Inhibition of DDR1
function led to increased apoptosis of wild-type p53-containing
cells in response to genotoxic stress through a caspase-dependent
pathway. (Ongusaha P P, et al. EMBO J. 2003, 22, 1289-1301) DDRs
were identified as one of several major activated tyrosine kinases
carrying somatic mutations in lung cancer (Hammerman P S, et al.
Cancer Discov. 2011, 1, 78-89.), serous and clear cell endometrial
cancer (Rudd M L, et al. BMC Cancer 2014, 14, 884), as well as in
acute myeloid leukemia. (Tomasson M H, et al. Blood 2008,
111:4797-4808) Advanced Kirsten rat sarcoma viral oncogene homolog
(KRAS)-mutant lung adenocarcinoma is challenging because of a lack
of effective targeted therapies. The concomitant inhibition of both
DDR1 and Notch signaling induced the regression of KRAS;TP53-mutant
patient-derived lung xenografts (PDX), indicating the combined
inhibition of DDR1 and Notch signaling could be an effective
targeted therapy for patients with KRAS-mutant lung adenocarcinoma.
(Ambrogia C, et al, Nature Medicine, 2016, 22, 270-277).
[0011] It is desirable to prepare compounds that have activity
against disease-driving kinase inhibitors, especially compounds
that have activity against multiple kinases, including against
multiple genetically altered kinases for use as therapeutic agents
in treating diseases. New compounds with polypharmacology profiles
are also desired for targeting the primary oncogene drivers and
their acquired resistance mechanisms including secondary mutations,
bypath signaling, EMT, cancer stemness, and metastasis.
SUMMARY
[0012] Compounds of the formula I
##STR00001##
[0013] wherein X.sup.1, X.sup.2, Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5, Z.sup.6, Z.sup.7, M, R.sup.1, R.sup.2, R.sup.3,
R.sup.4, R.sup.5 and R.sup.6 are defined as described herein have
been shown to have activity against wild-type and mutant ALK
(anaplastic lymphoma kinase), wild-type and mutant ROS1 (ROS1
proto-oncogene receptor tyrosine kinase), the TRK family of kinases
(tropomyosin-related receptor tyrosine kinases, TRKA/B/C), JAK2 of
the Janus family of kinases and SRC (c-Src family of protein
tyrosine kinases (SFKs)).
[0014] One such compound is
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one (also herein
referred to as "Compound 1"), represented by the formula
##STR00002##
[0015] has been shown to be a potent small-molecule multi-target
kinase inhibitor showing activity against wild-type and mutant ALK
(anaplastic lymphoma kinase), wild-type and mutant ROS1 (ROS1
proto-oncogene receptor tyrosine kinase), the TRK family of kinases
(tropomyosin-related receptor tyrosine kinases, TRKA/B/C), JAK2 of
the Janus family of kinases and SRC (c-Src family of protein
tyrosine kinases (SFKs)). Compound 1 has properties, including
anti-tumor properties, which are pharmacologically mediated through
inhibition of receptor and non-receptor tyrosine kinases. Compound
1 is disclosed in International Patent Application No.
PCT/US2015/012597, which is incorporated herein by reference in its
entirety.
[0016] In one aspect, the present disclosure provide a method of
treating disease in a patient comprising, administering to the
patient a therapeutically effective amount of a compound of the
formula I
##STR00003##
[0017] wherein
[0018] M is CR.sup.4a or N;
[0019] X.sup.1 and X.sup.2 are independently S, S(O), S(O).sub.2, O
or N(R.sup.9);
[0020] R.sup.1 is 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.6 cycloalkyl,
C.sub.6-C.sub.10 aryl, --C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8;
wherein each hydrogen atom in 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.6
cycloalkyl and C.sub.6-C.sub.10 aryl is independently optionally
substituted by deuterium, halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)C.sub.1-C.sub.6 alkyl, --NHC(O)NH.sub.2,
--NHC(O)NHC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)NHC.sub.1-C.sub.6
alkyl, --NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)OC.sub.1-C.sub.6 alkyl,
--NHS(O)(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl)S(O)(C.sub.1-C.sub.6 alkyl),
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl),
--NHS(O)NH.sub.2, NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0021] each of R.sup.2 and R.sup.3 is independently H, deuterium,
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.6 cycloalkyl, C.sub.6-C.sub.10 aryl,
--C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8; wherein each hydrogen atom
in 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.6 cycloalkyl and C.sub.6-C.sub.10 aryl is
independently optionally substituted by deuterium, halogen, --OH,
--CN, --OC.sub.1-C.sub.6 alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)C.sub.1-C.sub.6 alkyl,
--NHC(O)NH.sub.2, --NHC(O)NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NHC.sub.1-C.sub.6 alkyl, --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)OC.sub.1-C.sub.6 alkyl, --NHS(O)(C.sub.1-C.sub.6 alkyl),
--NHS(O).sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl), --NHS(O)NH.sub.2,
NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O)NH.sub.2,
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0022] R.sup.4, R.sup.4a and R.sup.5 are each independently H,
fluoro, chloro, bromo, C.sub.1-C.sub.6 alkyl, --OH, --CN,
--OC.sub.1-C.sub.6 alkyl, --NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl).sub.2 or --CF.sub.3;
[0023] R.sup.6 is H, C.sub.1-C.sub.6 alkyl or 3- to 7-membered
heterocycloalkyl, wherein each hydrogen atom in C.sub.1-C.sub.6
alkyl or 3- to 7-membered heterocycloalkyl is independently
optionally substituted by halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --CO.sub.2H, --CO.sub.2C.sub.1-C.sub.6 alkyl,
--CONH.sub.2, --CONH(C.sub.1-C.sub.6 alkyl), --CON(C.sub.1-C.sub.6
alkyl).sub.2, cycloalkyl, or monocyclic heterocycloalkyl;
[0024] each R.sup.7 and R.sup.8 is independently H, deuterium,
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.6 cycloalkyl, 3- to 7-membered
heterocycloalkyl, C.sub.6-C.sub.10 aryl or heteroaryl;
[0025] each R.sup.9 is independently H, deuterium, 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.6 cycloalkyl, 3- to 7-membered heterocycloalkyl,
C.sub.6-C.sub.10 aryl, or mono- or bicyclic heteroaryl; wherein
each hydrogen atom in 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is independently optionally substituted by
deuterium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl or --OR.sup.7;
[0026] each Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6,
or Z.sup.7 is independently N, NH, or C(R.sup.10), wherein each
R.sup.10 is independently H, deuterium, halogen, C.sub.1-C.sub.6
alkyl, --OC.sub.1-C.sub.6 alkyl, --OH, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --NH(phenyl), --NH(heteroaryl), --CN,
or CF.sub.3, and
[0027] provided that at least one of Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4 Z.sup.5, Z.sup.6 or Z.sup.7 is N or NH;
[0028] or a pharmaceutically acceptable salt thereof.
[0029] In another aspect, the present disclosure provides a method
of treating cancer in a patient previously shown to express a
genetically altered tyrosine or serine/throenine kinase comprising,
administering to the patient a therapeutically effective amount of
a compound of the formula I
##STR00004##
[0030] wherein
[0031] M is CR.sup.4a or N;
[0032] X.sup.1 and X.sup.2 are independently S, S(O), S(O).sub.2, O
or N(R.sup.9);
[0033] R.sup.1 is 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.6 cycloalkyl,
C.sub.6-C.sub.10 aryl, --C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8;
wherein each hydrogen atom in 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.6
cycloalkyl and C.sub.6-C.sub.10 aryl is independently optionally
substituted by deuterium, halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)C.sub.1-C.sub.6 alkyl, --NHC(O)NH.sub.2,
--NHC(O)NHC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)NHC.sub.1-C.sub.6
alkyl, --NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)OC.sub.1-C.sub.6 alkyl,
--NHS(O)(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl)S(O)(C.sub.1-C.sub.6 alkyl),
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl),
--NHS(O)NH.sub.2, NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0034] each of R.sup.2 and R.sup.3 is independently H, deuterium,
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.6 cycloalkyl, C.sub.6-C.sub.10 aryl,
--C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8; wherein each hydrogen atom
in 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.6 cycloalkyl and C.sub.6-C.sub.10 aryl is
independently optionally substituted by deuterium, halogen, --OH,
--CN, --OC.sub.1-C.sub.6 alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, NHC(O)C.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)C.sub.1-C.sub.6 alkyl,
--NHC(O)NH.sub.2, --NHC(O)NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NHC.sub.1-C.sub.6 alkyl, --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)OC.sub.1-C.sub.6 alkyl, --NHS(O)(C.sub.1-C.sub.6 alkyl),
--NHS(O).sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl), --NHS(O)NH.sub.2,
NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O)NH.sub.2,
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
.sub.-P(.sub.0)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6
cycloalkyl, or 3- to 7 0173membered heterocycloalkyl;
[0035] R.sup.4, R.sup.4a and R.sup.5 are each independently H,
fluoro, chloro, bromo, C.sub.1-C.sub.6 alkyl, --OH, --CN,
--OC.sub.1-C.sub.6 alkyl, --NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl).sub.2 or --CF.sub.3;
[0036] R.sup.6 is H, C.sub.1-C.sub.6 alkyl or 3- to 7-membered
heterocycloalkyl, wherein each hydrogen atom in C.sub.1-C.sub.6
alkyl or 3- to 7-membered heterocycloalkyl is independently
optionally substituted by halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --CO.sub.2H, --CO.sub.2C.sub.1-C.sub.6 alkyl,
--CONH.sub.2, --CONH(C.sub.1-C.sub.6 alkyl), --CON(C.sub.1-C.sub.6
alkyl).sub.2, cycloalkyl, or monocyclic heterocycloalkyl;
[0037] each R.sup.7 and R.sup.8 is independently H, deuterium,
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.6 cycloalkyl, 3- to 7-membered
heterocycloalkyl, C.sub.6-C.sub.10 aryl or heteroaryl;
[0038] each R.sup.9 is independently H, deuterium, 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.6 cycloalkyl, 3- to 7-membered heterocycloalkyl,
C.sub.6-C.sub.10 aryl, or mono- or bicyclic heteroaryl; wherein
each hydrogen atom in 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is independently optionally substituted by
deuterium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl or --OR.sup.7;
[0039] each Z.sup.1, Z.sup.2, Z.sub.3, Z.sup.4, Z.sup.5, Z.sup.6 or
Z.sup.7 is independently N, NH, or C(R.sup.10), wherein each
R.sup.10 is independently H, deuterium, halogen, C.sub.1-C.sub.6
alkyl, --OC.sub.1-C.sub.6 alkyl, --OH, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --NH(phenyl), --NH(heteroaryl), --CN,
or CF.sub.3, and
[0040] provided that at least one of Z.sup.1, Z.sup.2, or Z.sup.7
is N or NH;
[0041] or a pharmaceutically acceptable salt thereof.
[0042] In another aspect, the present disclosure provides a method
of treating cancer in a patient comprising;
[0043] i. identifying a genetically altered tyrosine or serine
threonine kinase in the patient, and
[0044] ii. administering to the patient a therapeutically effective
amount of a compound of the formula I
##STR00005##
[0045] wherein
[0046] M is CR.sup.4a or N;
[0047] X.sup.1 and X.sup.2 are independently S, S(O), S(O).sub.2, O
or N(R.sup.9);
[0048] R.sup.1 is 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.6 cycloalkyl,
C.sub.6-C.sub.10 aryl, --C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8;
wherein each hydrogen atom in 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.6
cycloalkyl and C.sub.6-C.sub.10 aryl is independently optionally
substituted by deuterium, halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)C.sub.1-C.sub.6 alkyl, --NHC(O)NH.sub.2,
--NHC(O)NHC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)NHC.sub.1-C.sub.6
alkyl, --NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)OC.sub.1-C.sub.6 alkyl,
--NHS(O)(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl)S(O)(C.sub.1-C.sub.6 alkyl),
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl),
--NHS(O)NH.sub.2, NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0049] each of R.sup.2 and R.sup.3 is independently H, deuterium,
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.6 cycloalkyl, C.sub.6-C.sub.10 aryl,
--C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8; wherein each hydrogen atom
in 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.6 cycloalkyl and C.sub.6-C.sub.10 aryl is
independently optionally substituted by deuterium, halogen, --OH,
--CN, --OC.sub.1-C.sub.6 alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)C.sub.1-C.sub.6 alkyl,
--NHC(O)NH.sub.2, --NHC(O)NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NHC.sub.1-C.sub.6 alkyl, --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)OC.sub.1-C.sub.6 alkyl, --NHS(O)(C.sub.1-C.sub.6 alkyl),
--NHS(O).sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl), --NHS(O)NH.sub.2,
NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O)NH.sub.2,
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0050] R.sup.4, R.sup.4a and R.sup.5 are each independently H,
fluoro, chloro, bromo, C.sub.1-C.sub.6 alkyl, --OH, --CN,
--OC.sub.1-C.sub.6 alkyl, --NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl).sub.2 or --CF.sub.3;
[0051] R.sup.6 is H, C.sub.1-C.sub.6 alkyl or 3- to 7-membered
heterocycloalkyl, wherein each hydrogen atom in C.sub.1-C.sub.6
alkyl or 3- to 7-membered heterocycloalkyl is independently
optionally substituted by halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --CO.sub.2H, --CO.sub.2C.sub.1-C.sub.6 alkyl,
--CONH.sub.2, --CONH(C.sub.1-C.sub.6 alkyl), --CON(C.sub.1-C.sub.6
alkyl).sub.2, cycloalkyl, or monocyclic heterocycloalkyl;
[0052] each R.sup.7 and R.sup.8 is independently H, deuterium,
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.6 cycloalkyl, 3- to 7-membered
heterocycloalkyl, C.sub.6-C.sub.10 aryl or heteroaryl;
[0053] each R.sup.9 is independently H, deuterium, 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.6 cycloalkyl, 3- to 7-membered heterocycloalkyl,
C.sub.6-C.sub.10 aryl, or mono- or bicyclic heteroaryl; wherein
each hydrogen atom in 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is independently optionally substituted by
deuterium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl or --OR.sup.7;
[0054] each Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6 or
Z.sup.7 is independently N, NH, or)C(R.sup.10), wherein each
R.sup.10 is independently H, deuterium, halogen, C.sub.1-C.sub.6
alkyl, --OC.sub.1-C.sub.6 alkyl, --OH, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --NH(phenyl), --NH(heteroaryl), --CN,
or CF.sub.3, and
[0055] provided that at least one of Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5, Z.sup.6 or Z.sup.7 is N or NH;
[0056] or a pharmaceutically acceptable salt thereof.
[0057] In another aspect, the present disclosure provides a method
of identifying a patient for treatment with a compound of the
formula I
##STR00006##
[0058] wherein
[0059] M is CR.sup.4a or N;
[0060] X.sup.1 and X.sup.2 are independently S, S(O), S(O).sub.2, O
or N(.sup.9);
[0061] R.sup.1 is 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.6 cycloalkyl,
C.sub.6-C.sub.10 aryl, --C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8;
wherein each hydrogen atom in 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.6
cycloalkyl and C.sub.6-C.sub.10 aryl is independently optionally
substituted by deuterium, halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)C.sub.1-C.sub.6 alkyl, --NHC(O)NH.sub.2,
--NHC(O)NHC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)NHC.sub.1-C.sub.6
alkyl, --NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)OC.sub.1-C.sub.6 alkyl,
--NHS(O)(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl)S(O)(C.sub.1-C.sub.6 alkyl),
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl),
--NHS(O)NH.sub.2, NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0062] each of R.sup.2 and R.sup.3 is independently H, deuterium,
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.6 cycloalkyl, C.sub.6-C.sub.10 aryl,
--C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8; wherein each hydrogen atom
in 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.6 cycloalkyl and C.sub.6-C.sub.10 aryl is
independently optionally substituted by deuterium, halogen, --OH,
--CN, --OC.sub.1-C.sub.6 alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)C.sub.1-C.sub.6 alkyl,
--NHC(O)NH.sub.2, --NHC(O)NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NHC.sub.1-C.sub.6 alkyl, --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)OC.sub.1-C.sub.6 alkyl, --NHS(O)(C.sub.1-C.sub.6 alkyl),
--NHS(O).sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl), --NHS(O)NH.sub.2,
NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O)NH.sub.2,
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0063] R.sup.4, R.sup.4a and R.sup.5 are each independently H,
fluoro, chloro, bromo, C.sub.1-C.sub.6 alkyl, --OH, --CN,
--OC.sub.1-C.sub.6 alkyl, --NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl).sub.2 or --CF.sub.3;
[0064] R.sup.6 is H, C.sub.1-C.sub.6 alkyl or 3- to 7-membered
heterocycloalkyl, wherein each hydrogen atom in C.sub.1-C.sub.6
alkyl or 3- to 7-membered heterocycloalkyl is independently
optionally substituted by halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --CO.sub.2H, --CO.sub.2C.sub.1-C.sub.6 alkyl,
--CONH.sub.2, --CONH(C.sub.1-C.sub.6 alkyl), --CON(C.sub.1-C.sub.6
alkyl).sub.2, cycloalkyl, or monocyclic heterocycloalkyl;
[0065] each R.sup.7 and R.sup.8 is independently H, deuterium,
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.6 cycloalkyl, 3- to 7-membered
heterocycloalkyl, C.sub.6-C.sub.10 aryl or heteroaryl;
[0066] each R.sup.9 is independently H, deuterium, 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.6 cycloalkyl, 3- to 7-membered heterocycloalkyl,
C.sub.6-C.sub.10 aryl, or mono- or bicyclic heteroaryl; wherein
each hydrogen atom in 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is independently optionally substituted by
deuterium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl or --OR.sup.7;
[0067] each Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6 or
Z.sup.7 is independently N, NH, or C(R.sup.10), wherein each
R.sup.10 is independently H, deuterium, halogen, C.sub.1-C.sub.6
alkyl, --OC.sub.1-C.sub.6 alkyl, --OH, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --NH(phenyl), --NH(heteroaryl), --CN,
or CF.sub.3, and
[0068] provided that at least one of Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5, Z.sup.6 or Z.sup.7 is N or NH;
[0069] or a pharmaceutically acceptable salt thereof,
[0070] comprising diagnosing the patient with a cancer mediated by
a genetically altered tyrosine or serine/threonine kinase.
[0071] In another aspect, the present disclosure provides a use of
compound of the formula I
##STR00007##
[0072] wherein
[0073] M is CR.sup.4a or N;
[0074] X.sup.1 and X.sup.2 are independently S, S(O), S(O).sub.2, O
or N(R.sup.9);
[0075] R.sup.1 is 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.6 cycloalkyl,
C.sub.6-C.sub.10 aryl, --C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8;
wherein each hydrogen atom in 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.6
cycloalkyl and C.sub.6-C.sub.10 aryl is independently optionally
substituted by deuterium, halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)C.sub.1-C.sub.6 alkyl, --NHC(O)NH.sub.2,
--NHC(O)NHC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)NHC.sub.1-C.sub.6
alkyl, --NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)OC.sub.1-C.sub.6 alkyl,
--NHS(O)(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl)S(O)(C.sub.1-C.sub.6 alkyl),
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl),
--NHS(O)NH.sub.2, NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0076] each of R.sup.2 and R.sup.3 is independently H, deuterium,
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.6 cycloalkyl, C.sub.6-C.sub.10 aryl,
--C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8; wherein each hydrogen atom
in 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.6 cycloalkyl and C.sub.6-C.sub.10 aryl is
independently optionally substituted by deuterium, halogen, --OH,
--CN, --OC.sub.1-C.sub.6 alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)C.sub.1-C.sub.6 alkyl,
--NHC(O)NH.sub.2, --NHC(O)NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NHC.sub.1-C.sub.6 alkyl, --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)OC.sub.1-C.sub.6 alkyl, --NHS(O)(C.sub.1-C.sub.6 alkyl),
--NHS(O).sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl), --NHS(O)NH.sub.2,
NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O)NH.sub.2,
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
.sub.-P(.sub.0)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6
cycloalkyl, or 3- to 7-membered heterocycloalkyl;
[0077] R.sup.4, R.sup.4a and R.sup.5 are each independently H,
fluoro, chloro, bromo, C.sub.1-C.sub.6 alkyl, --OH, --CN,
--OC.sub.1-C.sub.6 alkyl, --NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl).sub.2 or --CF.sub.3;
[0078] R.sup.6 is H, C.sub.1-C.sub.6 alkyl or 3- to 7-membered
heterocycloalkyl, wherein each hydrogen atom in C.sub.1-C.sub.6
alkyl or 3- to 7-membered heterocycloalkyl is independently
optionally substituted by halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --CO.sub.2H, --CO.sub.2C.sub.1-C.sub.6 alkyl,
--CONH.sub.2, --CONH(C.sub.1-C.sub.6 alkyl), --CON(C.sub.1-C.sub.6
alkyl).sub.2, cycloalkyl, or monocyclic heterocycloalkyl;
[0079] each R.sup.7 and R.sup.8 is independently H, deuterium,
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.6 cycloalkyl, 3- to 7-membered
heterocycloalkyl, C.sub.6-C.sub.10 aryl or heteroaryl;
[0080] each R.sup.9 is independently H, deuterium, 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.6 cycloalkyl, 3- to 7-membered heterocycloalkyl,
C.sub.6-C.sub.10 aryl, or mono- or bicyclic heteroaryl; wherein
each hydrogen atom in 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is independently optionally substituted by
deuterium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl or --OR.sup.7;
[0081] each Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6 or
Z.sup.7 is independently N, NH, or C(R.sup.10), wherein each
R.sup.10 is independently H, deuterium, halogen, C.sub.1-C.sub.6
alkyl, --OC.sub.1-C.sub.6 alkyl, --OH, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --NH(phenyl), --NH(heteroaryl), --CN,
or --CF.sub.3, and
[0082] provided that at least one of Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5, Z.sup.6 or Z.sup.7 is N or NH;
[0083] or a pharmaceutically acceptable salt thereof, in the
preparation of a medicament for the treatment of a disease in a
patient.
[0084] In another aspect, the present disclosure provides a use of
compound of the formula I
##STR00008##
[0085] wherein
[0086] M is CR.sup.4a or N;
[0087] X.sup.1 and X.sup.2 are independently S, S(O), S(O).sub.2, O
or N(R.sup.9);
[0088] R.sup.1 is 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.6 cycloalkyl,
C.sub.6-C.sub.10 aryl, --C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8;
wherein each hydrogen atom in 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.6
cycloalkyl and C.sub.6-C.sub.10 aryl is independently optionally
substituted by deuterium, halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)C.sub.1-C.sub.6 alkyl, --NHC(O)NH.sub.2,
--NHC(O)NHC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)NHC.sub.1-C.sub.6
alkyl, --NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)OC.sub.1-C.sub.6 alkyl,
--NHS(O)(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl)S(O)(C.sub.1-C.sub.6 alkyl),
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl),
--NHS(O)NH.sub.2, NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0089] each of R.sup.2 and R.sup.3 is independently H, deuterium,
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.6 cycloalkyl, C.sub.6-C.sub.10 aryl,
--C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8; wherein each hydrogen atom
in 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.6 cycloalkyl and C.sub.6-C.sub.10 aryl is
independently optionally substituted by deuterium, halogen, --OH,
--CN, --OC.sub.1-C.sub.6 alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, NHC(O)C.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)C.sub.1-C.sub.6 alkyl,
--NHC(O)NH.sub.2, --NHC(O)NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NHC.sub.1-C.sub.6 alkyl, --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)OC.sub.1-C.sub.6 alkyl, --NHS(O)(C.sub.1-C.sub.6 alkyl),
--NHS(O).sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl), --NHS(O)NH.sub.2,
NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O)NH.sub.2,
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
.sub.-P(.sub.0)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6
cycloalkyl, or 3- to 7-membered heterocycloalkyl;
[0090] R.sup.4, R.sup.4a and R.sup.5 are each independently H,
fluoro, chloro, bromo, C.sub.1-C.sub.6 alkyl, --OH, --CN,
--OC.sub.1-C.sub.6 alkyl, --NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl).sub.2 or --CF.sub.3;
[0091] R.sup.6 is H, C.sub.1-C.sub.6 alkyl or 3- to 7-membered
heterocycloalkyl, wherein each hydrogen atom in C.sub.1-C.sub.6
alkyl or 3- to 7-membered heterocycloalkyl is independently
optionally substituted by halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --CO.sub.2H, --CO.sub.2C.sub.1-C.sub.6 alkyl,
--CONH.sub.2, --CONH(C.sub.1-C.sub.6 alkyl), --CON(C.sub.1-C.sub.6
alkyl).sub.2, cycloalkyl, or monocyclic heterocycloalkyl;
[0092] each R.sup.7 and R.sup.8 is independently H, deuterium,
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.6 cycloalkyl, 3- to 7-membered
heterocycloalkyl, C.sub.6-C.sub.10 aryl or heteroaryl;
[0093] each R.sup.9 is independently H, deuterium, 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.6 cycloalkyl, 3- to 7-membered heterocycloalkyl,
C.sub.6-C.sub.10 aryl, or mono- or bicyclic heteroaryl; wherein
each hydrogen atom in 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is independently optionally substituted by
deuterium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl or --OR.sup.7;
[0094] each Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6 or
Z.sup.7 is independently N, NH, or)C(R.sup.10, wherein each
R.sup.10 is independently H, deuterium, halogen, C.sub.1-C.sub.6
alkyl, --O--C.sub.1-C.sub.6 alkyl, --OH, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --NH(phenyl), --NH(heteroaryl), --CN,
or --CF.sub.3, and
[0095] provided that at least one of Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5, Z.sup.6 or Z.sup.7 is N or NH;
[0096] or a pharmaceutically acceptable salt thereof, for treating
cancer in a patient.
[0097] In another aspect, the present disclosure provides the use
of compound of the formula I
##STR00009##
[0098] wherein
[0099] M is CR.sup.4a or N;
[0100] X.sup.1 and X.sup.2 are independently S, S(O), S(O).sub.2, O
or N(.sup.9);
[0101] R.sup.1 is 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.6 cycloalkyl,
C.sub.6-C.sub.10 aryl, --C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8;
wherein each hydrogen atom in 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.6
cycloalkyl and C.sub.6-C.sub.10 aryl is independently optionally
substituted by deuterium, halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)C.sub.1-C.sub.6 alkyl, --NHC(O)NH.sub.2,
--NHC(O)NHC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)NHC.sub.1-C.sub.6
alkyl, --NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)OC.sub.1-C.sub.6 alkyl,
--NHS(O)(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl)S(O)(C.sub.1-C.sub.6 alkyl),
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl),
--NHS(O)NH.sub.2, NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0102] each of R.sup.2 and R.sup.3 is independently H, deuterium,
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.6 cycloalkyl, C.sub.6-C.sub.10 aryl,
--C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8; wherein each hydrogen atom
in 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.6 cycloalkyl and C.sub.6-C.sub.10 aryl is
independently optionally substituted by deuterium, halogen, --OH,
--CN, --OC.sub.1-C.sub.6 alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)C.sub.1-C.sub.6 alkyl,
--NHC(O)NH.sub.2, --NHC(O)NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NHC.sub.1-C.sub.6 alkyl, --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)OC.sub.1-C.sub.6 alkyl, --NHS(O)(C.sub.1-C.sub.6 alkyl),
--NHS(O).sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl), --NHS(O)NH.sub.2,
NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O)NH.sub.2,
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to7- membered heterocycloalkyl;
[0103] R.sup.4, R.sup.4a and R.sup.5 are each independently H,
fluoro, chloro, bromo, C.sub.1-C.sub.6 alkyl, --OH, --CN,
--OC.sub.1-C.sub.6 alkyl, --NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl).sub.2 or --CF.sub.3;
[0104] R.sup.6 is H, C.sub.1-C.sub.6 alkyl or 3- to 7-membered
heterocycloalkyl, wherein each hydrogen atom in C.sub.1-C.sub.6
alkyl or 3- to 7-membered heterocycloalkyl is independently
optionally substituted by halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --CO.sub.2H, --CO.sub.2C.sub.1-C.sub.6 alkyl,
--CONH.sub.2, --CONH(C.sub.1-C.sub.6 alkyl), --CON(C.sub.1-C.sub.6
alkyl).sub.2, cycloalkyl, or monocyclic heterocycloalkyl;
[0105] each R.sup.7 and R.sup.8 is independently H, deuterium,
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.6 cycloalkyl, 3- to 7-membered
heterocycloalkyl, C.sub.6-C.sub.10 aryl or heteroaryl;
[0106] each R.sup.9 is independently H, deuterium, 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.6 cycloalkyl, 3- to 7-membered heterocycloalkyl,
C.sub.6-C.sub.10 aryl, or mono- or bicyclic heteroaryl; wherein
each hydrogen atom in 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is independently optionally substituted by
deuterium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl or --OR.sup.7;
[0107] each Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6 or
Z.sup.7 is independently N, NH, or)C(R.sup.10, wherein each
R.sup.10 is independently H, deuterium, halogen, C.sub.1-C.sub.6
alkyl, --O--C.sub.1-C.sub.6 alkyl, --OH, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --NH(phenyl), --NH(heteroaryl), --CN,
or --CF.sub.3, and
[0108] provided that at least one of Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5, Z.sup.6 or Z.sup.7 is N or NH;
[0109] or a pharmaceutically acceptable salt thereof, for treating
pain in a patient.
[0110] In another aspect, the present disclosure provides use of a
compound of the formula I
##STR00010##
[0111] wherein
[0112] M is CR.sup.4a or N;
[0113] X.sup.1 and X.sup.2 are independently S, S(O), S(O).sub.2, O
or N(.sup.9);
[0114] R.sup.1 is 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.6 cycloalkyl,
C.sub.6-C.sub.10 aryl, --C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8;
wherein each hydrogen atom in 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.6
cycloalkyl and C.sub.6-C.sub.10 aryl is independently optionally
substituted by deuterium, halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)C.sub.1-C.sub.6 alkyl, --NHC(O)NH.sub.2,
--NHC(O)NHC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)NHC.sub.1-C.sub.6
alkyl, --NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)OC.sub.1-C.sub.6 alkyl,
--NHS(O)(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl)S(O)(C.sub.1-C.sub.6 alkyl),
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl),
--NHS(O)NH.sub.2, NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0115] each of R.sup.2 and R.sup.3 is independently H, deuterium,
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.6 cycloalkyl, C.sub.6-C.sub.10 aryl,
--C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8; wherein each hydrogen atom
in 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.6 cycloalkyl and C.sub.6-C.sub.10 aryl is
independently optionally substituted by deuterium, halogen, --OH,
--CN, --OC.sub.1-C.sub.6 alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)C.sub.1-C.sub.6 alkyl,
--NHC(O)NH.sub.2, --NHC(O)NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NHC.sub.1-C.sub.6 alkyl, --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)OC.sub.1-C.sub.6 alkyl, --NHS(O)(C.sub.1-C.sub.6 alkyl),
--NHS(O).sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl), --NHS(O)NH.sub.2,
NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O)NH.sub.2,
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
.sub.-P(.sub.0)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6
cycloalkyl, or 3- to 7-membered heterocycloalkyl;
[0116] R.sup.4, R.sup.4a and R.sup.5 are each independently H,
fluoro, chloro, bromo, C.sub.1-C.sub.6 alkyl, --OH, --CN,
--OC.sub.1-C.sub.6 alkyl, --NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl).sub.2 or --CF.sub.3;
[0117] R.sup.6 is H, C.sub.1-C.sub.6 alkyl or 3- to 7-membered
heterocycloalkyl, wherein each hydrogen atom in C.sub.1-C.sub.6
alkyl or 3- to 7-membered heterocycloalkyl is independently
optionally substituted by halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --CO.sub.2H, --CO.sub.2C.sub.1-C.sub.6 alkyl,
--CONH.sub.2, --CONH(C.sub.1-C.sub.6 alkyl), --CON(C.sub.1-C.sub.6
alkyl).sub.2, cycloalkyl, or monocyclic heterocycloalkyl;
[0118] each R.sup.7 and R.sup.8 is independently H, deuterium,
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.6 cycloalkyl, 3- to 7-membered
heterocycloalkyl, C.sub.6-C.sub.10 aryl or heteroaryl;
[0119] each R.sup.9 is independently H, deuterium, 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.6 cycloalkyl, 3- to 7-membered heterocycloalkyl,
C.sub.6-C.sub.10 aryl, or mono- or bicyclic heteroaryl; wherein
each hydrogen atom in 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is independently optionally substituted by
deuterium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl or --OR.sup.7;
[0120] each Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6 or
Z.sup.7 is independently N, NH, or)C(R.sup.10, wherein each
R.sup.10 is independently H, deuterium, halogen, C.sub.1-C.sub.6
alkyl, --OC.sub.1-C.sub.6 alkyl, --OH, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --NH(phenyl), --NH(heteroaryl), --CN,
or --CF.sub.3, and
[0121] provided that at least one of Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5, Z.sup.6 or Z.sup.7 is N or NH;
[0122] or a pharmaceutically acceptable salt thereof, for treating
cancer in a patient previously shown to express a genetically
altered tyrosine or serine/threonine kinase.
[0123] In another aspect, the present disclosure provide a use a
compound of the formula I
##STR00011##
[0124] wherein
[0125] M is CR.sup.4a or N;
[0126] X.sup.1 and X.sup.2 are independently S, S(O), S(O).sub.2, O
or N(.sup.9);
[0127] R.sup.1 is 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.6 cycloalkyl,
C.sub.6-C.sub.10 aryl, --C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8;
wherein each hydrogen atom in 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.6
cycloalkyl and C.sub.6-C.sub.10 aryl is independently optionally
substituted by deuterium, halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)C.sub.1-C.sub.6 alkyl, --NHC(O)NH.sub.2,
--NHC(O)NHC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)NHC.sub.1-C.sub.6
alkyl, --NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)OC.sub.1-C.sub.6 alkyl,
--NHS(O)(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl)S(O)(C.sub.1-C.sub.6 alkyl),
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl),
--NHS(O)NH.sub.2, NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0128] each of R.sup.2 and R.sup.3 is independently H, deuterium,
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.6 cycloalkyl, C.sub.6-C.sub.10 aryl,
--C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8; wherein each hydrogen atom
in 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.6 cycloalkyl and C.sub.6-C.sub.10 aryl is
independently optionally substituted by deuterium, halogen, --OH,
--CN, --OC.sub.1-C.sub.6 alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)C.sub.1-C.sub.6 alkyl,
--NHC(O)NH.sub.2, --NHC(O)NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NHC.sub.1-C.sub.6 alkyl, --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)OC.sub.1-C.sub.6 alkyl, --NHS(O)(C.sub.1-C.sub.6 alkyl),
--NHS(O).sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl), --NHS(O)NH.sub.2,
NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O)NH.sub.2,
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0129] R.sup.4, R.sup.4a and R.sup.5 are each independently H,
fluoro, chloro, bromo, C.sub.1-C.sub.6 alkyl, --OH, --CN,
--OC.sub.1-C.sub.6 alkyl, --NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl).sub.2 or --CF.sub.3;
[0130] R.sup.6 is H, C.sub.1-C.sub.6 alkyl or 3- to 7-membered
heterocycloalkyl, wherein each hydrogen atom in C.sub.1-C.sub.6
alkyl or 3- to 7-membered heterocycloalkyl is independently
optionally substituted by halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --CO.sub.2H, --CO.sub.2C.sub.1-C.sub.6 alkyl,
--CONH.sub.2, --CONH(C.sub.1-C.sub.6 alkyl), --CON(C.sub.1-C.sub.6
alkyl).sub.2, cycloalkyl, or monocyclic heterocycloalkyl;
[0131] each R.sup.7 and R.sup.8 is independently H, deuterium,
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.6 cycloalkyl, 3- to 7-membered
heterocycloalkyl, C.sub.6-C.sub.10 aryl or heteroaryl;
[0132] each R.sup.9 is independently H, deuterium, 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.6 cycloalkyl, 3- to 7-membered heterocycloalkyl,
C.sub.6-C.sub.10 aryl, or mono- or bicyclic heteroaryl; wherein
each hydrogen atom in 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is independently optionally substituted by
deuterium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl or --OR.sup.7;
[0133] each Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6 or
Z.sup.7 is independently N, NH, or C(R.sup.10), wherein each
R.sup.10 is independently H, deuterium, halogen, C.sub.1-C.sub.6
alkyl, --OC.sub.1-C.sub.6 alkyl, --OH, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --NH(phenyl), --NH(heteroaryl), --CN,
or --CF.sub.3, and
[0134] provided that at least one of Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5, Z.sup.6 or Z.sup.7 is N or NH;
[0135] or a pharmaceutically acceptable salt thereof, for treating
cancer in a patient, wherein the patient has been previously
treated with a cancer therapeutic, and the cancer has developed
resistance to the cancer therapeutic.
[0136] In another aspect, the present disclosure provides the use
of a compound of the formula I
##STR00012##
[0137] wherein
[0138] M is CR.sup.4a or N;
[0139] X.sup.1 and X.sup.2 are independently S, S(O), S(O).sub.2, O
or N(.sup.9);
[0140] R.sup.1 is 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.6 cycloalkyl,
C.sub.6-C.sub.10 aryl, --C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8;
wherein each hydrogen atom in 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.6
cycloalkyl and C.sub.6-C.sub.10 aryl is independently optionally
substituted by deuterium, halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)C.sub.1-C.sub.6 alkyl, --NHC(O)NH.sub.2,
--NHC(O)NHC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)NHC.sub.1-C.sub.6
alkyl, --NHC(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)OC.sub.1-C.sub.6 alkyl,
--NHS(O)(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl)S(O)(C.sub.1-C.sub.6 alkyl),
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl),
--NHS(O)NH.sub.2, NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0141] each of R.sup.2 and R.sup.3 is independently H, deuterium,
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.6 cycloalkyl, C.sub.6-C.sub.10 aryl,
--C(O)OR.sup.7 or --C(O)NR.sup.7R.sup.8; wherein each hydrogen atom
in 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.6 cycloalkyl and C.sub.6-C.sub.10 aryl is
independently optionally substituted by deuterium, halogen, --OH,
--CN, --OC.sub.1-C.sub.6 alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, --NHC(O)C.sub.1-C.sub.6
alkyl, --N(C.sub.1-C.sub.6 alkyl)C(O)C.sub.1-C.sub.6 alkyl,
--NHC(O)NH.sub.2, --NHC(O)NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl)C(O)NH.sub.2, --N(C.sub.1-C.sub.6
alkyl)C(O)NHC.sub.1-C.sub.6 alkyl, --NHC(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --N(C.sub.1-C.sub.6 alkyl)C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --NHC(O)OC.sub.1-C.sub.6 alkyl, --N(C.sub.1-C.sub.6
alkyl)C(O)OC.sub.1-C.sub.6 alkyl, --NHS(O)(C.sub.1-C.sub.6 alkyl),
--NHS(O).sub.2(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2(C.sub.1-C.sub.6 alkyl), --NHS(O)NH.sub.2,
NHS(O).sub.2NH.sub.2, --N(C.sub.1-C.sub.6 alkyl)S(O)NH.sub.2,
--N(C.sub.1-C.sub.6 alkyl)S(O).sub.2NH.sub.2,
--NHS(O)NH(C.sub.1-C.sub.6 alkyl), --NHS(O).sub.2NH(C.sub.1-C.sub.6
alkyl), --NHS(O)N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHS(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O)NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl)S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --N(C.sub.1-C.sub.6
alkyl)S(O).sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --CO.sub.2H,
--C(O)OC.sub.1-C.sub.6 alkyl, --C(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.6 alkyl), --C(O)N(C.sub.1-C.sub.6
alkyl).sub.2, --SC.sub.1-C.sub.6 alkyl, --S(O)C.sub.1-C.sub.6
alkyl, --S(O).sub.2C.sub.1-C.sub.6 alkyl, --S(O)NH(C.sub.1-C.sub.6
alkyl), --S(O).sub.2NH(C.sub.1-C.sub.6 alkyl),
--S(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --S(O).sub.2N(C.sub.1-C.sub.6
alkyl).sub.2, --P(C.sub.1-C.sub.6 alkyl).sub.2,
--P(O)(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.3-C.sub.6 cycloalkyl, or
3- to 7-membered heterocycloalkyl;
[0142] R.sup.4, R.sup.4a and R.sup.5 are each independently H,
fluoro, chloro, bromo, C.sub.1-C.sub.6 alkyl, --OH, --CN,
--OC.sub.1-C.sub.6 alkyl, --NHC.sub.1-C.sub.6 alkyl,
--N(C.sub.1-C.sub.6 alkyl).sub.2 or --CF.sub.3;
[0143] R.sup.6 is H, C.sub.1-C.sub.6 alkyl or 3- to 7-membered
heterocycloalkyl, wherein each hydrogen atom in C.sub.1-C.sub.6
alkyl or 3- to 7-membered heterocycloalkyl is independently
optionally substituted by halogen, --OH, --CN, --OC.sub.1-C.sub.6
alkyl, --NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --CO.sub.2H, --CO.sub.2C.sub.1-C.sub.6 alkyl,
--CONH.sub.2, --CONH(C.sub.1-C.sub.6 alkyl), --CON(C.sub.1-C.sub.6
alkyl).sub.2, cycloalkyl, or monocyclic heterocycloalkyl;
[0144] each R.sup.7 and R.sup.8 is independently H, deuterium,
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.6 cycloalkyl, 3- to 7-membered
heterocycloalkyl, C.sub.6-C.sub.10 aryl or heteroaryl;
[0145] each R.sup.9 is independently H, deuterium, 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.6 cycloalkyl, 3- to 7-membered heterocycloalkyl,
C.sub.6-C.sub.10 aryl, or mono- or bicyclic heteroaryl; wherein
each hydrogen atom in 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is independently optionally substituted by
deuterium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl or --OR.sup.7;
[0146] each Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, Z.sup.5, Z.sup.6 or
Z.sup.7 is independently N, NH, or)C(R.sup.10, wherein each
R.sup.10 is independently H, deuterium, halogen, C.sub.1-C.sub.6
alkyl, --OC.sub.1-C.sub.6 alkyl, --OH, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --NH(phenyl), --NH(heteroaryl), --CN,
or --CF.sub.3, and
[0147] provided that at least one of Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5, Z.sup.6 or Z.sup.7 is N or NH;
[0148] or a pharmaceutically acceptable salt thereof, for treating
cancer in a patient previously shown to express a genetically
altered tyrosine or serine/threonine kinase, wherein the patient
has been previously treated with a cancer therapeutic, and the
cancer has developed resistance to the cancer therapeutic.
[0149] In some embodiments of the aspects described above, the
compound is
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriaza-cyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof.
[0150] In some embodiments, the disease is mediated by a tyrosine
or serine/threonine kinase selected from the group consisting of
ALK, ROS1, TRKA, TRKB, TRKC, JAK2, SRC, FAK, ARK5, and combinations
thereof. In some embodiments the disease is mediated by a receptor
tyrosine kinase. In some embodiments, the receptor tyrosine kinase
is selected from the group consisting of ALK, ROS1, TRKA, TRKB and
TRKC. In some embodiments, the receptor tyrosine kinase is selected
from the group consisting of ALK, ROS1, TRKA, TRKB and TRKC. In
some embodiments, the disease is mediated by a non-receptor kinase.
In some embodiments, the non-receptor kinase is JAK2, FYN, LYN,
YES, FGR, SRC, FAK or ARK5. In some embodiments, the non-receptor
kinase is JAK2, SRC, FAK or ARK5. In some embodiments, the disease
is mediated by a non-receptor tyrosine kinase. In some embodiments,
the non-receptor tyrosine kinase is JAK2, SRC or FAK. In some
embodiments, the disease is mediated by a non-receptor
serine/threonine kinase. In some embodiments, the non-receptor
serine/threonine kinase is ARK5. In some embodiments, the disease
is mediated by a protein tyrosine kinase. In some embodiments, the
protein tyrosine kinase is TXK. In some embodiments, the disease is
mediated by a discoidin domain receptor. In some embodiments, the
discoidin domain receptor is DDR1. In some embodiments, the disease
is selected from the group consisting of cancer, psoriasis,
rheumatoid arthritis, polycythemia vera, essential thrombocythemia,
ulcerative colitis, and myeloid metaplasia with myelofibrosis and
pain.
[0151] In some embodiments, the disease or cancer is a cancer
mediated by ALK. In some embodiments, the disease or cancer is a
cancer mediated by a genetically altered ALK. In some embodiments,
the disease or cancer is a cancer mediated by a fusion protein
comprising a fragment of a protein encoded by an ALK gene and a
fragment of a protein which will form coiled-coil interaction to
facilitate the protein dimerization or oligomerization. In some
embodiments, the disease or cancer is a cancer mediated by a fusion
protein comprising a fragment of a protein encoded by an ALK gene
and a fragment of a protein encoded by a gene selected from the
group consisting of NPM, EML4, TPR, TFG, ATIC, CLTC1, TPM4, MSN
ALO17 and MYH9. In some embodiments, the fusion protein comprises a
fragment of a protein encoded by an ALK gene and a fragment of a
protein encoded by an EML4 gene. In some embodiments, the
genetically altered ALK is an EML4-ALK fusion protein. In some
embodiments, the EML4-ALK fusion protein is a wild-type protein. In
some embodiments, the EML4-ALK fusion protein comprises at least
one resistance mutation. In some embodiments, the EML4-ALK fusion
protein comprises at least one mutation selected from the group
consisting of L1196M, G1202R, D1203R, L1152P/R, F1174C/L/V, C1156Y,
I1171N, G1123S, S1206Y, G1269S/A, and 1151T insertion.
[0152] In some embodiments, the fusion protein comprises a fragment
of a protein encoded by an ALK gene and a fragment of a protein
encoded by a NPM gene. In some embodiments, the genetically altered
ALK is a NPM-ALK fusion protein. In some embodiments, the fusion
protein comprises a fragment of a protein encoded by an ALK gene
and a fragment of a protein encoded by a TPR gene. In some
embodiments, the genetically altered ALK is a TPR-ALK fusion
protein. In some embodiments, the TPR-ALK fusion protein is a
wild-type protein. In some embodiments, the TPR-ALK fusion protein
comprises at least one resistance mutation. In some embodiments,
the TPR-ALK fusion protein comprises a L1196M point mutation.
[0153] In some embodiments, the disease or cancer is a cancer
mediated by ALK. In some embodiments, the disease or cancer is a
cancer mediated by a genetically altered ALK. In some embodiments,
the disease or cancer is a cancer mediated by ALK having one or
more point mutations. In some embodiments, the disease or cancer is
a cancer mediated by ALK having one or more point mutations
selected from the group consisting of R1050H, F1174C/I/L/S/V,
F1245C/I/L/V, R1275L/Q, T1151M, M1166R, 11170N, 11170S, 11171N,
I1183T, L1196M, A1200V, L1204F, L1240V, D1270G, Y1278S, R1192P,
G1128A, G1286R, and T13431. In some embodiments, the point mutation
is a mutation at F1174. In some embodiments, the point mutation is
a mutation of ALK at F1245. In some embodiments, the point mutation
is a mutation of ALK at R1275.
[0154] In some embodiments, the disease or cancer is a cancer
mediated by ROS1. In some embodiments, the disease or cancer is a
cancer mediated by a genetically altered ROS1. In some embodiments,
the disease or cancer is a cancer mediated by a fusion protein
comprising a fragment of a protein encoded by an ROS1 gene and a
fragment of a protein which will form coiled-coil interaction to
facilitate the protein dimerization or oligomerization. In some
embodiments, the disease or cancer is a cancer mediated by a fusion
protein comprising a fragment of a protein encoded by a ROS1 gene
and a fragment of a protein encoded by a gene selected from the
group consisting of FIG, TPM3, SDC4, SLC34A2, CD74, EZR, and LRIG3.
In some embodiments, the fusion protein comprises a fragment of a
protein encoded by an ROS1 gene and a fragment of a protein encoded
by a CD74 gene. In some embodiments, the genetically altered ROS1
is a CD74-ROS1 fusion protein. In some embodiments, the CD74-ROS1
fusion protein is a wild-type protein. In some embodiments, the
CD74-ROS1 fusion protein comprises at least one resistance
mutation. In some embodiments, the CD74-ROS1 fusion protein
comprises a G2032R point mutation. In some embodiments, the
CD74-ROS1 fusion protein comprises a L2026M point mutation. In some
embodiments, the CD74-ROS1 fusion protein comprises a D2033N point
mutation. In some embodiments, the genetically altered ROS1 is a
SDC4-ROS1 fusion protein. In some embodiments, the SDC4-ROS1 fusion
protein is a wild-type protein. In some embodiments, the SDC4-ROS1
fusion protein comprises at least one resistance mutation. In some
embodiments, the SDC4-ROS1 fusion protein comprises a G2032R point
mutation. In some embodiments, the genetically altered ROS1 is a
SLC34A2-ROS1 fusion protein. In some embodiments, the SLC34A2-ROS1
fusion protein is a wild-type protein. In some embodiments, the
SLC34A2-ROS1 fusion protein comprises at least one resistance
mutation. In some embodiments, the SLC34A2-ROS1 fusion protein
comprises a G2032R point mutation.
[0155] In some embodiments, the disease or cancer is a cancer
mediated by TRKA. In some embodiments, the disease or cancer is a
cancer mediated by a genetically altered TRKA. In some embodiments,
the disease or cancer is a cancer mediated by a fusion protein
comprising a fragment of a protein encoded by a TRKA gene and a
fragment of a protein which will form coiled-coil interaction to
facilitate the protein dimerization or oligomerization. In some
embodiments, the disease or cancer is a cancer mediated by a fusion
protein comprising a fragment of a protein encoded by a TRKA gene
and a fragment of a protein encoded by a TPM3 gene. In some
embodiments, the genetically altered TRKA is a TPM3-TRKA fusion
protein. In some embodiments, the TPM3-TRKA fusion protein is a
wild-type protein. In some embodiments, the TPM3-TRKA fusion
protein comprises at least one resistance mutation.
[0156] In some embodiments, the disease or cancer is a cancer
mediated by a fusion protein comprising a fragment of a protein
encoded by a TRKA gene and a fragment of a protein encoded by a
LMNA gene. In some embodiments, the genetically altered TRKA is a
LMNA-TRKA fusion protein. In some embodiments, the LMNA-TRKA fusion
protein is a wild-type protein. In some embodiments, the LMNA-TRKA
fusion protein comprises at least one resistance mutation. In some
embodiments, the LMNA-TRKA fusion protein is a wild-type protein.
In some embodiments, the LMNA-TRKA fusion protein comprises at
least one resistance mutation comprising a G595R point
mutation.
[0157] In some embodiments, the disease or cancer is a cancer
mediated by TRKB. In some embodiments, the disease or cancer is a
cancer mediated by a genetically altered TRKB. In some embodiments,
the disease or cancer is a cancer mediated by a fusion protein
comprising a fragment of a protein encoded by a TRKB gene and a
fragment of a protein which will form coiled-coil interaction to
facilitate the protein dimerization or oligomerization. In some
embodiments, the disease or cancer is a cancer mediated by a fusion
protein comprising a fragment of a protein encoded by a TRKB gene
and a fragment of a protein encoded by a QKI gene or TEL gene. In
some embodiments, the disease or cancer is a cancer mediated by a
fusion protein comprising a fragment of a protein encoded by a TRKB
gene and a fragment of a protein encoded by a QKI gene. In some
embodiments, the disease or cancer is a cancer mediated by a fusion
protein comprising a fragment of a protein encoded by a TRKB gene
and a fragment of a protein encoded by a TEL gene. In some
embodiments, the genetically altered TRKB is a QKI-TRKB or TEL-TRKB
fusion protein. In some embodiments, the genetically altered TRKB
is a TEL-TRKB fusion protein. In some embodiments, the genetically
altered TRKB is a QKI-TRKB fusion protein. In some embodiments, the
QKI-TRKB or TEL-TRKB fusion protein is a wild-type protein. In some
embodiments, the QKI-TRKB fusion protein is a wild-type protein. In
some embodiments, the TEL-TRKB fusion protein is a wild-type
protein. In some embodiments, the QKI-TRKB or TEL-TRKB fusion
protein comprises at least one resistance mutation. In some
embodiments, the QKI-TRKB fusion protein comprises at least one
resistance mutation. In some embodiments, the TEL-TRKB fusion
protein comprises at least one resistance mutation. In some
embodiments, the TEL-TRKB fusion protein comprises a G639R point
mutation.
[0158] In some embodiments, the disease or cancer is a cancer
mediated by TRKC. In some embodiments, the disease or cancer is a
cancer mediated by a genetically altered TRKC. In some embodiments,
the disease or cancer is a cancer mediated by a fusion protein
comprising a fragment of a protein encoded by a TRKC gene and a
fragment of a protein which will form coiled-coil interaction to
facilitate the protein dimerization or oligomerization. In some
embodiments, the disease or cancer is a cancer mediated by a fusion
protein comprising a fragment of a protein encoded by a TRKC gene
and a fragment of a protein encoded by an ETV6 gene. In some
embodiments, the genetically altered TRKC is an ETV6-TRKC fusion
protein. In some embodiments, the ETV6-TRKC fusion protein is a
wild-type protein. In some embodiments, the ETV6-TRKC fusion
protein comprises at least one resistance mutation. In some
embodiments, the ETV6-TRKC fusion protein comprises a G623R point
mutation.
[0159] In some embodiments, the disease or cancer is a cancer
mediated by JAK1, JAK2 or JAK3. In some embodiments, the disease or
cancer is a cancer mediated by a genetically altered JAK2. In some
embodiments, the disease or cancer is a cancer mediated by a fusion
protein comprising a fragment of a protein encoded by a JAK2 gene
and a fragment of a protein which will form coiled-coil interaction
to facilitate the protein dimerization or oligomerization. In some
embodiments, the disease or cancer is a cancer mediated by a fusion
protein comprising a fragment of a protein encoded by a JAK2 gene
and a fragment of a protein encoded by a TEL or PCM1 gene. In some
embodiments, the genetically altered JAK2 is a TEL-JAK2 fusion
protein. In some embodiments, the genetically altered JAK2 is a
PCM1-JAK2 fusion protein. In some embodiments, the disease or
cancer is a cancer mediated by point mutation(s) of JAK2. In some
embodiments, the genetically altered JAK2 has the JAK2V617F
mutation.
[0160] In some embodiments, the disease is pain. In some
embodiments, the disease is pain mediated by TRKA, TRKB or TRKC. In
some embodiments, the pain is mediated by TRKA. In some
embodiments, the pain is mediated by TRKB. In some embodiments, the
pain is mediated by TRKC. In some embodiments, the disease is
selected from the group consisting of psoriasis, rheumatoid
arthritis, polycythemia vera, essential thrombocythemia, ulcerative
colitis, and myeloid metaplasia with myelofibrosis. In some
embodiments, the disease or cancer is a cancer exhibiting bypass
resistance.
[0161] In some embodiments, the disease or cancer is a cancer
mediated by FGR. In some embodiments, the disease or cancer is a
cancer mediated by a genetically altered FGR. In some embodiments,
the fusion protein comprises a fragment of a protein encoded by a
FGR gene and a fragment of a protein encoded by a WASF2 gene. In
some embodiments, the genetically altered FGR is a WASF2-FGR fusion
protein. In some embodiments, the WASF2-FGR fusion protein is a
wild-type protein. In some embodiments, the WASF2-FGR fusion
protein comprises at least one resistance mutation.
[0162] In some embodiments, the cancer is selected from the group
consisting of ALCL, NSCLC, neuroblastoma, inflammatory
myofibroblastic tumor, adult renal cell carcinoma, pediatric renal
cell carcinoma, breast cancer, colonic adenocarcinoma,
glioblastoma, glioblastoma multiforme and anaplastic thyroid
cancer.
[0163] In some embodiments, the cancer is selected from the group
consisting of glioblastoma, glioblastoma multiforme, NSCLC,
cholangiocarcinoma, ovarian cancer, gastric adenocarcinoma,
colorectal cancer, inflammatory myofibroblastic tumor,
angiosarcoma, and epithelioid hemangioendothelioma.
[0164] In some embodiments, the cancer is selected from the group
consisting of glioblastoma, glioblastoma multiforme, NSCLC,
cholangiocarcinoma, intrahepatic cholangiocarcinoma, colorectal
cancer, thyroid papillary cancer, spitzoid neoplasms, sarcoma,
astrocytoma, brain lower grade glioma, secretory breast carcinoma,
mammary analogue carcinoma, breast cancer, acute myeloid leukemia,
congenital mesoblastic nephroma, congenital fibrosarcomas, Ph-like
acute lymphoblastic leukemia, colon adenocarcinoma, thyroid
carcinoma, skin cutaneous melanoma, head and neck squamous cell
carcinoma and pediatric glioma.
[0165] In some embodiments, the cancer is selected from the group
consisting of NSCLC, neuroblastoma, breast cancer, colon cancer and
prostate cancer. In some embodiments, the cancer is NSCLC. In some
embodiments, the cancer is neuroblastoma. In some embodiments, the
cancer is colorectal cancer.
[0166] In some embodiments, the patient has been previously treated
with a cancer therapeutic. In some embodiments, the patient has
been previously treated with a cancer therapeutic, and the cancer
has developed resistance to the cancer therapeutic. In some
embodiments, the resistance is a primary intrinsic resistance. In
some embodiments, the resistance is an acquired resistance from
mutation(s). In some embodiments, the resistance is a bypass
resistance. In some embodiments, the resistance is an EMT-based
resistance.
[0167] Additional embodiments, features, and advantages of the
disclosure will be apparent from the following detailed description
and through practice of the disclosure. The compounds of the
present disclosure can be described as embodiments in any of the
following enumerated clauses. It will be understood that any of the
embodiments described herein can be used in connection with any
other embodiments described herein to the extent that the
embodiments do not contradict one another.
[0168] 1. A method of treating disease in a patient comprising,
administering to the patient a therapeutically effective amount of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof.
[0169] 2. The method of clause 1, wherein the disease is mediated
by a tyrosine or serine/threonine kinase selected from the group
consisting of ALK, ROS1, TRKA, TRKB, TRKC, JAK2, SRC, FYN, LYN,
YES, FGR, FAK, and ARK5, and combinations thereof; or ALK, ROS1,
TRKA, TRKB, TRKC, JAK2, SRC, FAK, and ARK5, and combinations
thereof.
[0170] 3. The method of clause 1, wherein the disease is mediated
by a receptor tyrosine kinase.
[0171] 4. The method of clause 3, wherein the receptor tyrosine
kinase is selected from the group consisting of ALK, ROS1, TRKA,
TRKB and TRKC.
[0172] 5. The method of clause 1, wherein the disease is mediated
by a non-receptor kinase.
[0173] 6. The method of clause 5, wherein the non-receptor kinase
is JAK2, FYN, LYN, YES, FGR, SRC, FAK or ARK5, including the
non-receptor tyrosine kinase JAK2, FYN, LYN, YES, FGR, SRC or FAK,
or the non-receptor serine/threonine kinase ARK5.
[0174] 7. The method of any one of clauses 1 to 6, wherein the
disease is selected from the group consisting of cancer, psoriasis,
rheumatoid arthritis, polycythemia vera, essential thrombocythemia,
ulcerative colitis, and myeloid metaplasia with myelofibrosis and
pain.
[0175] 8. The method of any one of clauses 1 to 6, wherein the
disease is cancer.
[0176] 9. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by ALK.
[0177] 10. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by a genetically altered ALK.
[0178] 11. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by a fusion protein comprising a
fragment of a protein encoded by an ALK gene and a fragment of a
protein encoded by a gene selected from the group consisting of
NPM, EML4, TPR, TFG, ATIC, CLTC1, TPM4, MSN ALO17 and MYH9.
[0179] 12. The method of clause 11, wherein the fusion protein
comprises a fragment of a protein encoded by an ALK gene and a
fragment of a protein encoded by an EML4 gene.
[0180] 13. The method of clause 10, wherein the genetically altered
ALK is an EML4-ALK fusion protein.
[0181] 14. The method of clause 13, wherein the EML4-ALK fusion
protein is a wild-type protein.
[0182] 15. The method of clause 13, wherein the EML4-ALK fusion
protein comprises at least one resistance mutation.
[0183] 16. The method of clause 13, wherein the EML4-ALK fusion
protein comprises at least one mutation selected from the group
consisting of L1196M, G1202R, D1203R, L1152P/R, F1174C/L/V, C1156Y,
I1171N, G1123S, S1206Y, G1269S/A, and 1151T insertion.
[0184] 17. The method of clause 16, wherein the mutation is
L1196M.
[0185] 18. The method of clause 16, wherein the mutation is
G1202R.
[0186] 19. The method of clause 16, wherein the mutation is
L1152P.
[0187] 20. The method of clause 16, wherein the mutation is
F1174C.
[0188] 21. The method of clause 16, wherein the mutation is
C1156Y.
[0189] 22. The method of clause 16, wherein the mutation is
I1171N.
[0190] 23. The method of clause 16, wherein the mutation is
G1269S.
[0191] 24. The method of clause 16, wherein the mutation is 1151T
insertion.
[0192] 25. The method of clause 11, wherein the fusion protein
comprises a fragment of a protein encoded by an ALK gene and a
fragment of a protein encoded by a NPM gene.
[0193] 26. The method of clause 10, wherein the genetically altered
ALK is a NPM-ALK fusion protein.
[0194] 27. The method of clause 11, wherein the fusion protein
comprises a fragment of a protein encoded by an ALK gene and a
fragment of a protein encoded by a TPR gene.
[0195] 28. The method of clause 10, wherein the genetically altered
ALK is a TPR-ALK fusion protein.
[0196] 29. The method of clause 28, wherein the TPR-ALK fusion
protein is a wild-type protein.
[0197] 30. The method of clause 28, wherein the TPR-ALK fusion
protein comprises at least one resistance mutation.
[0198] 31. The method of clause 28, wherein the TPR-ALK fusion
protein comprises a L1196M point mutation.
[0199] 32. The method of any one of clauses 1 to 4, wherein the
disease is a cancer mediated by ALK having one or more point
mutations.
[0200] The method of any one of claim 1 to 4 or 32, wherein the
disease is a cancer mediated by ALK having one or more point
mutations selected from the group consisting of R1050H,
F1174C/I/L/S/V, F1245C/I/L/V, R1275L/Q, T1151M, M1166R, 11170N,
11170S, I1171N, I1183T, L1196M, A1200V, L1204F, L1240V, D1270G,
Y1278S, R1192P, G1128A, G1286R, and T13431.
[0201] 34. The method of any one of clauses 1 to 4, 32 or 33,
wherein the point mutation is a mutation of ALK at F1174.
[0202] 35. The method of any one of clauses 1 to 4, 32 or 33,
wherein the point mutation is a mutation of ALK at F1245.
[0203] 36. The method of any one of clauses 1 to 4, 32 or 33,
wherein the point mutation is a mutation of ALK at R1275.
[0204] 37. The method of any one of clauses 9 to 36, wherein the
cancer is selected from the group consisting of ALCL, NSCLC,
neuroblastoma, inflammatory myofibroblastic tumor, adult renal cell
carcinoma, pediatric renal cell carcinoma, breast cancer, colonic
adenocarcinoma, glioblastoma, glioblastoma multiforme and
anaplastic thyroid cancer.
[0205] 38. The method of any one of clauses 9 to 37, wherein the
cancer is NSCLC.
[0206] 39. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by ROS1.
[0207] 40. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by a genetically altered ROS1.
[0208] 41. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by a fusion protein comprising a
fragment of a protein encoded by an ROS1 gene and a fragment of a
protein encoded by a gene selected from the group consisting of
FIG, TPM3, SDC4, SLC34A2, CD74, EZR, and LRIG3.
[0209] 42. The method of clause 41, wherein the fusion protein
comprises a fragment of a protein encoded by an ROS1 gene and a
fragment of a protein encoded by a CD74 gene.
[0210] 43. The method of clause 40, wherein the genetically altered
ROS1 is a CD74-ROS1 fusion protein.
[0211] 44. The method of clause 43, wherein the CD74-ROS1 fusion
protein is a wild-type protein.
[0212] 45. The method of clause 43, wherein the CD74-ROS1 fusion
protein comprises at least one resistance mutation.
[0213] 46. The method of clause 43, wherein the CD74-ROS1 fusion
protein comprises a G2032R, L2026M or D2033N point mutation.
[0214] 47. The method of clause 40, wherein the genetically altered
ROS1 is a SDC4-ROS1 fusion protein.
[0215] 48. The method of clause 47, wherein the SDC4-ROS1 fusion
protein is a wild-type protein.
[0216] 49. The method of clause 47, wherein the SDC4-ROS1 fusion
protein comprises at least one resistance mutation.
[0217] 50. The method of clause 47, wherein the SDC4-ROS1 fusion
protein comprises a G2032R point mutation.
[0218] 51. The method of clause 40, wherein the genetically altered
ROS1 is a SLC34A2-ROS1 fusion protein.
[0219] 52. The method of clause 51, wherein the SLC34A2-ROS1 fusion
protein is a wild-type protein.
[0220] 53. The method of clause 51, wherein the SLC34A2-ROS1 fusion
protein comprises at least one resistance mutation.
[0221] 54. The method of clause 51, wherein the SLC34A2-ROS1 fusion
protein comprises a G2032R point mutation.
[0222] 55. The method of any one of clauses 1 to 4, 7, 8 or 39 to
54, wherein the cancer is selected from the group consisting of
glioblastoma, glioblastoma multiforme, NSCLC, cholangiocarcinoma,
ovarian cancer, gastric adenocarcinoma, colorectal cancer,
inflammatory myofibroblastic tumor, angiosarcoma, and epithelioid
hemangioendothelioma.
[0223] 56. The method of any one of clauses 1 to 4, 7, 8 or 39 to
55, wherein the cancer is NSCLC.
[0224] 57. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by TRKA.
[0225] 58. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by a genetically altered TRKA.
[0226] 59. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by a fusion protein comprising a
fragment of a protein encoded by a TRKA gene and a fragment of a
protein encoded by a TPM3 gene or LMNA gene.
[0227] 60. The method of clause 58, wherein the genetically altered
TRKA is a TPM3-TRKA or LMNA-TRKA fusion protein.
[0228] 61. The method of clause 60, wherein the TPM3-TRKA or
LMNA-TRKA fusion protein is a wild-type protein.
[0229] 62. The method of clause 60, wherein the TPM3-TRKA or
LMNA-TRKA fusion protein comprises at least one resistance
mutation, including a LMNA-TRKA fusion protein comprising a G595R
point mutation.
[0230] 63. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by TRKB.
[0231] 64. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by a genetically altered TRKB,
including QKI-TRKB or TEL-TRKB, including TEL-TRKB comprising a
G639R point mutation.
[0232] 65. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by TRKC.
[0233] 66. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by a genetically altered TRKC,
including a ETV6-TRKC fusion protein, inclusing a genetically
altered ETV6-TRKC fusion protein, including a ETV6-TRKC fusion
protein comprising a G326R point mutation.
[0234] 67. The method of any one of clauses 1 to 4, 7, 8 or 57 to
66, wherein the cancer is selected from the group consisting of
glioblastoma, glioblastoma multiforme, NSCLC, cholangiocarcinoma,
intrahepatic cholangiocarcinoma, colorectal cancer, thyroid
papillary cancer, spitzoid neoplasms, sarcoma, astrocytoma, brain
lower grade glioma, secretory breast carcinoma, mammary analogue
carcinoma, breast cancer, acute myeloid leukemia, congenital
mesoblastic nephroma, congenital fibrosarcomas, Ph-like acute
lymphoblastic leukemia, colon adenocarcinoma, thyroid carcinoma,
skin cutaneous melanoma, head and neck squamous cell carcinoma and
pediatric glioma.
[0235] 68. The method of any one of clauses 1 to 4, 7, 8 or 57 to
67, wherein the cancer is NSCLC.
[0236] 69. The method of any one of clauses 1 to 4, 7, 8 or 57 to
67, wherein the cancer is colorectal cancer.
[0237] 70. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by JAK1, JAK2 or JAK3.
[0238] 71. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by a genetically altered JAK2.
[0239] 72. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by a fusion protein comprising a
fragment of a protein encoded by a JAK2 gene and a fragment of a
protein encoded by a TEL or PCM1 gene.
[0240] 73. The method of clause 71, wherein the genetically altered
JAK2 is a TEL-JAK2 fusion protein.
[0241] 74. The method of clause 71, wherein the genetically altered
JAK2 is a PCM1-JAK2 fusion protein.
[0242] 75. The method of clause 71, wherein the genetically altered
JAK2 comprises a V617F point mutation.
[0243] 76. The method of any one of clauses 1 to 4, 7 or 8, wherein
the disease is a cancer mediated by SRC.
[0244] 77. The method of clause 76, wherein the cancer is selected
from the group consisting of glioblastoma, glioblastoma multiforme,
NSCLC, cholangiocarcinoma, intrahepatic cholangiocarcinoma,
colorectal cancer, thyroid papillary cancer, spitzoid neoplasms,
sarcoma, astrocytoma, brain lower grade glioma, secretory breast
carcinoma, mammary analogue carcinoma, breast cancer, acute myeloid
leukemia, congenital mesoblastic nephroma, congenital
fibrosarcomas, Ph-like acute lymphoblastic leukemia, colon
adenocarcinoma, thyroid carcinoma, skin cutaneous melanoma, head
and neck squamous cell carcinoma and pediatric glioma.
[0245] 78. The method of any one of clauses 1 to 4, wherein the
disease is pain.
[0246] 79. The method of any one of clauses 1 to 4, wherein the
disease is pain mediated by TRKA, TRKB or TRKC.
[0247] 80. The method of clause 79, wherein the pain is mediated by
TRKA.
[0248] 81. The method of clause 79, wherein the pain is mediated by
TRKB.
[0249] 82. The method of clause 79, wherein the pain is mediated by
TRKC.
[0250] 83. The method of any one of clauses 1 to 3, 5 or 6 wherein
the disease is selected from the group consisting of psoriasis,
rheumatoid arthritis, polycythemia vera, essential thrombocythemia,
ulcerative colitis, and myeloid metaplasia with myelofibrosis.
[0251] 84. The method of any one of clauses 1, 5, 6 or 8, wherein
the disease is a cancer exhibiting bypass resistance.
[0252] 85. A method of treating cancer in a patient previously
shown to express a genetically altered tyrosine or serine/threonine
kinase comprising, administering to the patient a therapeutically
effective amount of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-
1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one,
or a pharmaceutically acceptable salt thereof.
[0253] 86. The method of clause 85, wherein the genetically altered
tyrosine kinase is selected from the group consisting of a
genetically altered ALK, genetically altered ROS1, genetically
altered TRK and genetically altered JAK.
[0254] 87. The method of clause 86, wherein the genetically altered
ALK is a fusion protein comprising a fragment of a protein encoded
by an ALK gene and a fragment of a protein encoded by a gene
selected from the group consisting of NPM, EML4, TPR, TFG, ATIC,
CLTC1, TPM4, MSN ALO17 and MYH9.
[0255] 88. The method of clause 87, wherein the fusion protein
comprises a fragment of a protein encoded by an ALK gene and a
fragment of a protein encoded by an EML4 gene.
[0256] 89. The method of clause 86, wherein the genetically altered
ALK is an EML4-ALK fusion protein.
[0257] 90. The method of clause 89, wherein the EML4-ALK fusion
protein is a wild-type protein.
[0258] 91. The method of clause 89, wherein the EML4-ALK fusion
protein comprises at least one resistance mutation.
[0259] 92. The method of clause 89, wherein the EML4-ALK fusion
protein comprises at least one mutation selected from the group
consisting of L1196M, G1202R, D1203R, L1152P/R, F1174C/L/V, C1156Y,
I1171N, G1123S, S1206Y, G1269S/A, and 1151T insertion.
[0260] 93. The method of clause 87, wherein the fusion protein
comprises a fragment of a protein encoded by an ALK gene and a
fragment of a protein encoded by a NPM gene.
[0261] 94. The method of clause 86, wherein the genetically altered
ALK is a NPM-ALK fusion protein.
[0262] 95. The method of clause 87, wherein the fusion protein
comprises a fragment of a protein encoded by an ALK gene and a
fragment of a protein encoded by a TPR gene.
[0263] 96. The method of clause 86, wherein the genetically altered
ALK is a TPR-ALK fusion protein.
[0264] 97. The method of clause 96, wherein the TPR-ALK fusion
protein is a wild-type protein.
[0265] 98. The method of clause 96, wherein the TPR-ALK fusion
protein comprises at least one resistance mutation.
[0266] 99. The method of clause 98, wherein the TPR-ALK fusion
protein comprises a L1196M point mutation.
[0267] 100. The method of any one of clauses 85 to 99, wherein the
cancer exhibits a bypass resistance mechanism.
[0268] 101. The method of clause 100, wherein the bypass resistance
mechanism is mediated by SRC.
[0269] 102. The method of any one of clauses 85 to 101, wherein the
cancer is selected from the group consisting of ALCL, NSCLC,
neuroblastoma, inflammatory myofibroblastic tumor, adult renal cell
carcinoma, pediatric renal cell carcinoma, breast cancer, colonic
adenocarcinoma, glioblastoma, glioblastoma multiforme and
anaplastic thyroid cancer.
[0270] 103. The method of any one of clauses 85 to 102, wherein the
cancer is NSCLC.
[0271] 104. The method of clause 86, wherein the genetically
altered ROS1 is a fusion protein comprising a fragment of a protein
encoded by an ROS1 gene and a fragment of a protein encoded by a
gene selected from the group consisting of FIG, TPM3, SDC4,
SLC34A2, CD74, EZR, and LRIG3.
[0272] 105. The method of clause 104, wherein the fusion protein
comprises a fragment of a protein encoded by an ROS1 gene and a
fragment of a protein encoded by a CD74 gene.
[0273] 106. The method of clause 104, wherein the genetically
altered ROS1 is a CD74-ROS1 fusion protein.
[0274] 107. The method of clause 106, wherein the CD74-ROS1 fusion
protein is a wild-type protein.
[0275] 108. The method of clause 106, wherein the CD74-ROS1 fusion
protein comprises at least one resistance mutation.
[0276] 109. The method of clause 106, wherein the CD74-ROS1 fusion
protein comprises a G2032R, L2026M or D2033N point mutation.
[0277] 110. The method of clause 105, wherein the genetically
altered ROS1 is a SDC4-ROS1 fusion protein.
[0278] 111. The method of clause 110, wherein the SDC4-ROS1 fusion
protein is a wild-type protein.
[0279] 112. The method of clause 110, wherein the SDC4-ROS1 fusion
protein comprises at least one resistance mutation.
[0280] 113. The method of clause 110, wherein the SDC4-ROS1 fusion
protein comprises a G2032R point mutation.
[0281] 114. The method of clause 105, wherein the genetically
altered ROS1 is a SLC34A2-ROS1 fusion protein.
[0282] 115. The method of clause 114, wherein the SLC34A2-ROS1
fusion protein is a wild-type protein.
[0283] 116. The method of clause 114, wherein the SLC34A2-ROS1
fusion protein comprises at least one resistance mutation.
[0284] 117. The method of clause 114, wherein the SLC34A2-ROS1
fusion protein comprises a G2032R point mutation.
[0285] 118. The method of any one of clauses 85, 86 or 104 to 117,
wherein the cancer is selected from the group consisting of
glioblastoma, glioblastoma multiforme, NSCLC, cholangiocarcinoma,
ovarian cancer, gastric adenocarcinoma, colorectal cancer,
inflammatory myofibroblastic tumor, angiosarcoma, and epithelioid
hemangioendothelioma.
[0286] 119. The method of any one of clauses 85, 86 or 104 to 118,
wherein the cancer is NSCLC.
[0287] 120. The method of clause 86, wherein the genetically
altered TRK is a fusion protein comprising a fragment of a protein
encoded by a TRKA gene and a fragment of a protein encoded by a
TPM3 gene or LMNA gene.
[0288] 121. The method of clause 86, wherein the genetically
altered TRK is a TPM3-TRKA or LMNA-TRKA fusion protein.
[0289] 122. The method of clause 121, wherein the TPM3-TRKA or
LMNA-TRKA fusion protein is a wild-type protein.
[0290] 123. The method of clause 121, wherein the TPM3-TRKA or
LMNA-TRKA fusion protein comprises at least one resistance
mutation.
[0291] 124. The method of any one of clauses 86, 87 or 120 to 123,
wherein the cancer is selected from the group consisting of
glioblastoma, glioblastoma multiforme, NSCLC, cholangiocarcinoma,
intrahepatic cholangiocarcinoma, colorectal cancer, thyroid
papillary cancer, spitzoid neoplasms, sarcoma, astrocytoma, brain
lower grade glioma, secretory breast carcinoma, mammary analogue
carcinoma, breast cancer, acute myeloid leukemia, congenital
mesoblastic nephroma, congenital fibrosarcomas, Ph-like acute
lymphoblastic leukemia, colon adenocarcinoma, thyroid carcinoma,
skin cutaneous melanoma, head and neck squamous cell carcinoma and
pediatric glioma.
[0292] 125. The method of any one of clauses 85, 86 or 120 to 123,
wherein the cancer is NSCLC.
[0293] 126. The method of any one of clauses 79, 80 or 120 to 123,
wherein the cancer is colorectal cancer.
[0294] 127. The method of clause 86, wherein the genetically
altered JAK is fusion protein comprising a fragment of a protein
encoded by a JAK2 gene and a fragment of a protein encoded by a TEL
or PCM1 gene.
[0295] 128. The method of clause 86, wherein the genetically
altered JAK is a TEL-JAK2 fusion protein.
[0296] 129. The method of clause 86, wherein the genetically
altered JAK is a PCM1-JAK2 fusion protein.
[0297] 130. A method of treating cancer in a patient comprising;
[0298] i. identifying a genetically altered tyrosine or
serine/threonine kinase in the patient, and [0299] ii.
administering to the patient a therapeutically effective amount of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriaza- cyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof.
[0300] 131. The method of clause 130, wherein the step of
identifying comprises subjecting a patient sample to a test
selected from the group consisting of FISH, IHC, PCR and gene
sequencing.
[0301] 132. A method of identifying a patient for treatment with
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriaza-cyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof, comprising diagnosing the
patient with a cancer mediated by a genetically altered tyrosine or
serine/threonine kinase.
[0302] 133. The method of clause 132, wherein the diagnosing
comprises subjecting a patient sample to a biological test or
biological assay selected from the group consisting of FISH, IHC,
PCR and gene sequencing.
[0303] 134. The method of any one of the preceding clauses, wherein
the patient has been previously treated with a cancer
therapeutic.
[0304] 135. The method of any one of the preceding clauses, wherein
the patient has been previously treated with a cancer therapeutic,
and the cancer has developed resistance to the cancer
therapeutic.
[0305] 136. The method of clause 129, wherein the resistance is an
acquired resistance.
[0306] 137. The method of clause 129, wherein the resistance is a
bypass resistance.
[0307] 138. Use of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof, in the preparation of a
medicament for the treatment of a disease in a patient.
[0308] 139. Use of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof, for treating cancer in a
patient.
[0309] 140. Use of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof, for treating pain in a
patient.
[0310] 141. Use of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof, for treating cancer in a
patient previously shown to express a genetically altered tyrosine
or serine/threonine kinase.
[0311] 142. Use of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof, for treating cancer in a
patient, wherein the patient has been previously treated with a
cancer therapeutic, and the cancer has developed resistance to the
cancer therapeutic.
[0312] 143. Use of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one, or a
pharmaceutically acceptable salt thereof, for treating cancer in a
patient previously shown to express a genetically altered tyrosine
or serine/threonine kinase, wherein the patient has been previously
treated with a cancer therapeutic, and the cancer has developed
resistance to the cancer therapeutic.
BRIEF DESCRIPTION OF THE DRAWINGS
[0313] FIG. 1 shows the effect of Compound 1 on apoptosis of
Karpas-299 cells. FIG. lA shows Karpas-299 cell apoptosis after
incubation in various concentrations of Compound 1 for 48 hours;
FIG. 1B shows Karpas-299 cells were collected, lysed and analyzed
by SDS-PAGE and immunoblotted with PARP and Actin antibodies.
[0314] FIG. 2 shows the effect of Compound 1 on wound healing in
HCC78 and HT-1080 cells.
[0315] FIG. 3 shows gels demonstrating that Compound 1
down-regulates EGFR expression. FIG. 3A shows results after 4 hour
exposure to Compound 1 at 0 nM, 30 nM, 100 nM, 300 nM and 1000 nM;
FIG. 3A shows results after 24 hour exposure to Compound 1 at 0 nM,
30 nM, 100 nM, 300 nM and 1000 nM.
[0316] FIG. 4 shows gels demonstrating that Compound 1
down-regulates CD44 expression as compared to the same experiment
with crizotinib which does not show down-regulation of CD44
expression. FIG. 4A shows results after exposure to Compound 1 at 0
nM, 30 nM, 100 nM, 300 nM and 1000 nM; FIG. 4A shows results after
exposure to crizotinib at 0 nM, 30 nM, 100 nM, 300 nM and 1000
nM.
[0317] FIG. 5 shows the effect of Compound 1 on tumor growth in the
Karpas-299 in vivo model at various doses, (.smallcircle.) control,
(.box-solid.) 15 mg/kg, (.cndot.) 50 mg/kg.
[0318] FIG. 6 shows the stability of animal body weight in the
Karpas-299 in vivo model following administration of Compound 1 at
various doses, (.smallcircle.) control, (.box-solid.) 15 mg/kg,
(.cndot.) 50 mg/kg.
[0319] FIG. 7 shows the effect of Compound 1 on tumor growth in the
NIH3T3 EML4-ALK WT in vivo model at various doses, (.smallcircle.)
control, (.box-solid.) 15 mg/kg, (.cndot.) 50 mg/kg.
[0320] FIG. 8 shows the stability of animal body weight in the
NIH3T3 EML4-ALK WT in vivo model following administration of
Compound 1 at various doses, (.smallcircle.) control, (.box-solid.)
15 mg/kg, (.cndot.) 50 mg/kg.
[0321] FIG. 9 shows the effect of Compound 1 on tumor growth in the
NIH3T3 SDC4-ROS1 WT in vivo model at various doses, (.smallcircle.)
control, (.box-solid.) 15 mg/kg, (.cndot.) 50 mg/kg.
[0322] FIG. 10 shows the effect of Compound 1 on tumor growth in
the KM12 in vivo model at various doses, (.smallcircle.) control,
(.box-solid.) 15 mg/kg, (.cndot.) 75 mg/kg.
[0323] FIG. 11 shows the stability of animal body weight in the
KM12 in vivo model following administration of Compound 1 at
various doses, (.smallcircle.) control, (.box-solid.) 15 mg/kg,
(.cndot.) 75 mg/kg.
[0324] FIG. 12 shows the inhibition of NPM-ALK phosphorylation by
Compound 1 in the Karpas-299 in vivo model.
[0325] FIG. 13 shows the effect of Compound 1 on tumor growth in
the KM12 in vivo model at various doses, (.smallcircle.) control,
(.box-solid.) 1 mg/kg, (.cndot.) 3 mg/kg, (1) 15 mg/kg.
[0326] FIG. 14 shows the stability of animal body weight in the
KM12 in vivo model following administration of Compound 1 at
various doses, (.smallcircle.) control, (.box-solid.) 1 mg/kg,
(.cndot.) 3 mg/kg, (1) 15 mg/kg.
[0327] FIG. 15 shows the effect of Compound 1 on tumor growth in
the Ba/F3 EML4-ALK WT in vivo model at various doses,
(.smallcircle.) control, (.box-solid.) 15 mg/kg, (.cndot.) 75
mg/kg.
[0328] FIG. 16 shows the stability of animal body weight in the
Ba/F3 EML4-ALK WT in vivo model following administration of
Compound 1 at various doses, (.smallcircle.) control, (.box-solid.)
15 mg/kg, (.cndot.) 75 mg/kg.
[0329] FIG. 17 shows the effect of Compound 1 on tumor growth in
the Ba/F3 EML4-ALK G1202R in vivo model at various doses,
(.smallcircle.) control, (.box-solid.) 15 mg/kg, (.cndot.) 75
mg/kg.
[0330] FIG. 18 shows the stability of animal body weight in the
Ba/F3 EML4-ALK G1202R in vivo model following administration of
Compound 1 at various doses, (.smallcircle.) control, (.box-solid.)
15 mg/kg, (.cndot.) 75 mg/kg.
[0331] FIG. 19 shows the effect of Compound 1 on tumor growth in
the Ba/F3 CD74-ROS1 WT in vivo model at various doses,
(.smallcircle.) control, (.box-solid.) 15 mg/kg, (.cndot.) 75
mg/kg.
[0332] FIG. 20 shows the stability of animal body weight in the
Ba/F3 CD74-ROS1 WT in vivo model following administration of
Compound 1 at various doses, (.smallcircle.) control, (.box-solid.)
15 mg/kg, (.cndot.) 75 mg/kg.
[0333] FIG. 21 shows the effect of Compound 1 on tumor growth in
the Ba/F3 CD74-ROS1 G2032R in vivo model at various doses,
(.smallcircle.) control, (.box-solid.) 15 mg/kg, (.cndot.) 75
mg/kg.
[0334] FIG. 22 shows the stability of animal body weight in the
Ba/F3 CD74-ROS1 G2032R in vivo model following administration of
Compound 1 at various doses, (.smallcircle.) control, (.box-solid.)
15 mg/kg, (.cndot.) 75 mg/kg.
[0335] FIG. 23 shows the effect of Compound 1 on tumor growth in
the NIH3T3 LMNA-TRKA WT in vivo model at various doses,
(.smallcircle.) control, (.box-solid.) 3 mg/kg, (.cndot.) 15
mg/kg.
[0336] FIG. 24 shows the stability of animal body weight in the
NIH3T3 LMNA-TRKA WT in vivo model following administration of
Compound 1 at various doses, (.smallcircle.) control, (.box-solid.)
3 mg/kg, (.cndot.) 15 mg/kg.
[0337] FIG. 25 shows the effect of Compound 1 on tumor growth in
the NIH3T3 LMNA-TRKA G595R in vivo model at various doses,
(.smallcircle.) control, (.box-solid.) 3 mg/kg, (.cndot.) 15 mg/kg,
(1) 60 mg/kg.
[0338] FIG. 26 shows the stability of animal body weight in the
NIH3T3 LMNA-TRKA G595R in vivo model following administration of
Compound 1 at various doses, (.smallcircle.) control, (.box-solid.)
3 mg/kg, (.cndot.) 15 mg/kg, (1) 60 mg/kg.
DETAILED DESCRIPTION
[0339] Before the present disclosure is further described, it is to
be understood that this disclosure is not limited to particular
embodiments described, as such may, of course, vary. It is also to
be understood that the terminology used herein is for the purpose
of describing particular embodiments only, and is not intended to
be limiting, since the scope of the present disclosure will be
limited only by the appended claims.
[0340] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as is commonly understood by one
of ordinary skill in the art to which this disclosure belongs. All
patents, applications, published applications and other
publications referred to herein are incorporated by reference in
their entireties. If a definition set forth in this section is
contrary to or otherwise inconsistent with a definition set forth
in a patent, application, or other publication that is herein
incorporated by reference, the definition set forth in this section
prevails over the definition incorporated herein by reference.
[0341] As used herein and in the appended claims, the singular
forms "a," "an," and "the" include plural referents unless the
context clearly dictates otherwise. It is further noted that the
claims may be drafted to exclude any optional element. As such,
this statement is intended to serve as antecedent basis for use of
such exclusive terminology as "solely," "only" and the like in
connection with the recitation of claim elements, or use of a
"negative" limitation.
[0342] As used herein, the terms "including," "containing," and
"comprising" are used in their open, non-limiting sense.
[0343] To provide a more concise description, some of the
quantitative expressions given herein are not qualified with the
term "about". It is understood that, whether the term "about" is
used explicitly or not, every quantity given herein is meant to
refer to the actual given value, and it is also meant to refer to
the approximation to such given value that would reasonably be
inferred based on the ordinary skill in the art, including
equivalents and approximations due to the experimental and/or
measurement conditions for such given value. Whenever a yield is
given as a percentage, such yield refers to a mass of the entity
for which the yield is given with respect to the maximum amount of
the same entity that could be obtained under the particular
stoichiometric conditions. Concentrations that are given as
percentages refer to mass ratios, unless indicated differently.
[0344] Except as otherwise noted, the methods and techniques of the
present embodiments are generally performed according to
conventional methods well known in the art and as described in
various general and more specific references that are cited and
discussed throughout the present specification. See, e.g., Loudon,
Organic Chemistry, Fourth Edition, New York: Oxford University
Press, 2002, pp. 360-361, 1084-1085; Smith and March, March's
Advanced Organic Chemistry: Reactions, Mechanisms, and Structure,
Fifth Edition, Wiley-Interscience, 2001.
[0345] Definitions
[0346] As used herein, the term "alkyl" includes a chain of carbon
atoms, which is optionally branched and contains from 1 to 20
carbon atoms. It is to be further understood that in certain
embodiments, alkyl may be advantageously of limited length,
including C.sub.1-C.sub.12, C.sub.1-C.sub.10, C.sub.1-C.sub.9,
C.sub.1-C.sub.8, C.sub.1-C.sub.7, C.sub.1-C.sub.6, and
C.sub.1-C.sub.4, Illustratively, such particularly limited length
alkyl groups, including C.sub.1-C.sub.8, C.sub.1-C.sub.7,
C.sub.1-C.sub.6, and C.sub.1-C.sub.4, and the like may be referred
to as "lower alkyl." Illustrative alkyl groups include, but are not
limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,
sec-butyl, tert-butyl, pentyl, 2-pentyl, 3-pentyl, neopentyl,
hexyl, heptyl, octyl, and the like. Alkyl may be substituted or
unsubstituted. Typical substituent groups include cycloalkyl, aryl,
heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, mercapto,
alkylthio, arylthio, cyano, halo, carbonyl, oxo, (.dbd.O),
thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl,
N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy, nitro, and
amino, or as described in the various embodiments provided herein.
It will be understood that "alkyl" may be combined with other
groups, such as those provided above, to form a functionalized
alkyl. By way of example, the combination of an "alkyl" group, as
described herein, with a "carboxy" group may be referred to as a
"carboxyalkyl" group. Other non-limiting examples include
hydroxyalkyl, aminoalkyl, and the like.
[0347] As used herein, the term "alkenyl" includes a chain of
carbon atoms, which is optionally branched, and contains from 2 to
20 carbon atoms, and also includes at least one carbon-carbon
double bond (i.e. C.dbd.C). It will be understood that in certain
embodiments, alkenyl may be advantageously of limited length,
including C.sub.2-C.sub.12, C.sub.2-C.sub.9, C.sub.2-C.sub.8,
C.sub.2-C.sub.7, C.sub.2-C.sub.6, and C.sub.2-C.sub.4.
Illustratively, such particularly limited length alkenyl groups,
including C.sub.2-C.sub.8, C.sub.2-C.sub.7, C.sub.2-C.sub.6, and
C.sub.2-C.sub.4 may be referred to as lower alkenyl. Alkenyl may be
unsubstituted, or substituted as described for alkyl or as
described in the various embodiments provided herein. Illustrative
alkenyl groups include, but are not limited to, ethenyl,
1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl, and the like.
[0348] As used herein, the term "alkynyl" includes a chain of
carbon atoms, which is optionally branched, and contains from 2 to
20 carbon atoms, and also includes at least one carbon-carbon
triple bond (i.e. CC). It will be understood that in certain
embodiments, alkynyl may each be advantageously of limited length,
including C.sub.2-C.sub.12, C.sub.2-C.sub.9, C.sub.2-C.sub.8,
C.sub.2-C.sub.7, C.sub.2-C.sub.6, and C.sub.2-C.sub.4.
Illustratively, such particularly limited length alkynyl groups,
including C.sub.2-C.sub.8, C.sub.2-C.sub.7, C.sub.2-C.sub.6, and
C.sub.2-C.sub.4 may be referred to as lower alkynyl. Alkenyl may be
unsubstituted, or substituted as described for alkyl or as
described in the various embodiments provided herein. Illustrative
alkenyl groups include, but are not limited to, ethynyl,
1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl, and the like.
[0349] As used herein, the term "aryl" refers to an all-carbon
monocyclic or fused-ring polycyclic groups of 6 to 12 carbon atoms
having a completely conjugated pi-electron system. It will be
understood that in certain embodiments, aryl may be advantageously
of limited size such as C.sub.6-C.sub.10 aryl. Illustrative aryl
groups include, but are not limited to, phenyl, naphthalenyl and
anthracenyl. The aryl group may be unsubstituted, or substituted as
described for alkyl or as described in the various embodiments
provided herein.
[0350] As used herein, the term "cycloalkyl" refers to a 3 to 15
member all-carbon monocyclic ring, including an all-carbon
5-member/6-member or 6-member/6-member fused bicyclic ring, or a
multicyclic fused ring (a "fused" ring system means that each ring
in the system shares an adjacent pair of carbon atoms with each
other ring in the system) group, where one or more of the rings may
contain one or more double bonds but the cycloalkyl does not
contain a completely conjugated pi-electron system. It will be
understood that in certain embodiments, cycloalkyl may be
advantageously of limited size such as C.sub.3-C.sub.13,
C.sub.3-C.sub.9, C.sub.3-C.sub.6 and C.sub.4-C.sub.6. Cycloalkyl
may be unsubstituted, or substituted as described for alkyl or as
described in the various embodiments provided herein. Illustrative
cycloalkyl groups include, but are not limited to, cyclopropyl,
cyclobutyl, cyclopentyl, cyclopentenyl, cyclopentadienyl,
cyclohexyl, cyclohexenyl, cycloheptyl, adamantyl, norbornyl,
norbornenyl, 9H-fluoren-9-yl, and the like. Illustrative examples
of cycloalkyl groups shown in graphical representations include the
following entities, in the form of properly bonded moieties:
##STR00013##
[0351] As used herein, the term "heterocycloalkyl" refers to a
monocyclic or fused ring group having in the ring(s) from 3 to 12
ring atoms, in which at least one ring atom is a heteroatom, such
as nitrogen, oxygen or sulfur, the remaining ring atoms being
carbon atoms. Heterocycloalkyl may optionally contain 1, 2, 3 or 4
heteroatoms. Heterocycloalkyl may also have one of more double
bonds, including double bonds to nitrogen (e.g. C.dbd.N or N.dbd.N)
but does not contain a completely conjugated pi-electron system. It
will be understood that in certain embodiments, heterocycloalkyl
may be advantageously of limited size such as 3- to 7-membered
heterocycloalkyl, 5- to 7-membered heterocycloalkyl, and the like.
Heterocycloalkyl may be unsubstituted, or substituted as described
for alkyl or as described in the various embodiments provided
herein. Illustrative heterocycloalkyl groups include, but are not
limited to, oxiranyl, thianaryl, azetidinyl, oxetanyl,
tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl, piperidinyl,
1,4-dioxanyl, morpholinyl, 1,4-dithianyl, piperazinyl, oxepanyl,
3,4-dihydro-2H-pyranyl, 5,6-dihydro-2H-pyranyl, 2H-pyranyl, 1, 2,
3, 4-tetrahydropyridinyl, and the like. Illustrative examples of
heterocycloalkyl groups shown in graphical representations include
the following entities, in the form of properly bonded
moieties:
##STR00014##
[0352] As used herein, the term "heteroaryl" refers to a monocyclic
or fused ring group of 5 to 12 ring atoms containing one, two,
three or four ring heteroatoms selected from nitrogen, oxygen and
sulfur, the remaining ring atoms being carbon atoms, and also
having a completely conjugated pi-electron system. It will be
understood that in certain embodiments, heteroaryl may be
advantageously of limited size such as 3- to 7-membered heteroaryl,
5- to 7-membered heteroaryl, and the like. Heteroaryl may be
unsubstituted, or substituted as described for alkyl or as
described in the various embodiments provided herein. Illustrative
heteroaryl groups include, but are not limited to, pyrrolyl,
furanyl, thiophenyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl,
pyridinyl, pyrimidinyl, quinolinyl, isoquinolinyl, purinyl,
tetrazolyl, triazinyl, pyrazinyl, tetrazinyl, quinazolinyl,
quinoxalinyl, thienyl, isoxazolyl, isothiazolyl, oxadiazolyl,
thiadiazolyl, triazolyl, benzimidazolyl, benzoxazolyl,
benzthiazolyl, benzisoxazolyl, benzisothiazolyl and carbazoloyl,
and the like. Illustrative examples of heteroaryl groups shown in
graphical representations, include the following entities, in the
form of properly bonded moieties:
##STR00015##
[0353] As used herein, "hydroxy" or "hydroxyl" refers to an --OH
group.
[0354] As used herein, "alkoxy" refers to both an --O-(alkyl) or an
--O-(unsubstituted cycloalkyl) group. Representative examples
include, but are not limited to, methoxy, ethoxy, propoxy, butoxy,
cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and
the like.
[0355] As used herein, "aryloxy" refers to an --O-aryl or an
--O-heteroaryl group. Representative examples include, but are not
limited to, phenoxy, pyridinyloxy, furanyloxy, thienyloxy,
pyrimidinyloxy, pyrazinyloxy, and the like, and the like.
[0356] As used herein, "mercapto" refers to an --SH group.
[0357] As used herein, "alkylthio" refers to an --S-(alkyl) or an
--S-(unsubstituted cycloalkyl) group. Representative examples
include, but are not limited to, methylthio, ethylthio, propylthio,
butylthio, cyclopropylthio, cyclobutylthio, cyclopentylthio,
cyclohexylthio, and the like.
[0358] As used herein, "arylthio" refers to an -S-aryl or an
-S-heteroaryl group. Representative examples include, but are not
limited to, phenylthio, pyridinylthio, furanylthio, thienylthio,
pyrimidinylthio, and the like.
[0359] As used herein, "halo" or "halogen" refers to fluorine,
chlorine, bromine or iodine.
[0360] As used herein, "cyano" refers to a --CN group.
[0361] The term "oxo" represents a carbonyl oxygen. For example, a
cyclopentyl substituted with oxo is cyclopentanone.
[0362] The term "substituted" means that the specified group or
moiety bears one or more substituents. The term "unsubstituted"
means that the specified group bears no substituents. Where the
term "substituted" is used to describe a structural system, the
substitution is meant to occur at any valency-allowed position on
the system. In some embodiments, "substituted" means that the
specified group or moiety bears one, two, or three substituents. In
other embodiments, "substituted" means that the specified group or
moiety bears one or two substituents. In still other embodiments,
"substituted" means the specified group or moiety bears one
substituent.
[0363] As used herein, "optional" or "optionally" means that the
subsequently described event or circumstance may but need not
occur, and that the description includes instances where the event
or circumstance occurs and instances in which it does not. For
example, "wherein each hydrogen atom in 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.6
cycloalkyl, 3- to 7-membered heterocycloalkyl, C.sub.6-C.sub.10
aryl, or mono- or bicyclic heteroaryl is independently optionally
substituted by C.sub.1-C.sub.6 alkyl" means that an alkyl may be
but need not be present on any of the 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.6
cycloalkyl, 3- to 7-membered heterocycloalkyl, C.sub.6-C.sub.10
aryl, or mono- or bicyclic heteroaryl by replacement of a hydrogen
atom for each alkyl group, and the description includes situations
where the 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is substituted with an alkyl group and
situations where the 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.6 cycloalkyl, 3- to
7-membered heterocycloalkyl, C.sub.6-C.sub.10 aryl, or mono- or
bicyclic heteroaryl is not substituted with the alkyl group.
[0364] As used herein, "independently" means that the subsequently
described event or circumstance is to be read on its own relative
to other similar events or circumstances. For example, in a
circumstance where several equivalent hydrogen groups are
optionally substituted by another group described in the
circumstance, the use of "independently optionally" means that each
instance of a hydrogen atom on the group may be substituted by
another group, where the groups replacing each of the hydrogen
atoms may be the same or different. Or for example, where multiple
groups exist all of which can be selected from a set of
possibilities, the use of "independently" means that each of the
groups can be selected from the set of possibilities separate from
any other group, and the groups selected in the circumstance may be
the same or different.
[0365] As used herein, the term "pharmaceutically acceptable salt"
refers to those salts which counter ions which may be used in
pharmaceuticals. See, generally, S.M. Berge, et al.,
"Pharmaceutical Salts," J. Pharm. Sci., 1977, 66, 1-19. Preferred
pharmaceutically acceptable salts are those that are
pharmacologically effective and suitable for contact with the
tissues of subjects without undue toxicity, irritation, or allergic
response. A compound described herein may possess a sufficiently
acidic group, a sufficiently basic group, both types of functional
groups, or more than one of each type, and accordingly react with a
number of inorganic or organic bases, and inorganic and organic
acids, to form a pharmaceutically acceptable salt. Such salts
include:
[0366] (1) acid addition salts, which can be obtained by reaction
of the free base of the parent compound with inorganic acids such
as hydrochloric acid, hydrobromic acid, nitric acid, phosphoric
acid, sulfuric acid, and perchloric acid and the like, or with
organic acids such as acetic acid, oxalic acid, (D) or (L) malic
acid, maleic acid, methane sulfonic acid, ethanesulfonic acid,
p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid,
succinic acid or malonic acid and the like; or
[0367] (2) salts formed when an acidic proton present in the parent
compound either is replaced by a metal ion, e.g., an alkali metal
ion, an alkaline earth ion, or an aluminum ion; or coordinates with
an organic base such as ethanolamine, diethanolamine,
triethanolamine, trimethamine, N-methylglucamine, and the like.
[0368] Pharmaceutically acceptable salts are well known to those
skilled in the art, and any such pharmaceutically acceptable salt
may be contemplated in connection with the embodiments described
herein. Examples of pharmaceutically acceptable salts include
sulfates, pyrosulfates, bisulfates, sulfites, bisulfites,
phosphates, monohydrogen-phosphates, dihydrogenphosphates,
metaphosphates, pyrophosphates, chlorides, bromides, iodides,
acetates, propionates, decanoates, caprylates, acrylates, formates,
isobutyrates, caproates, heptanoates, propiolates, oxalates,
malonates, succinates, suberates, sebacates, fumarates, maleates,
butyne-1,4-dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates,
methylbenzoates, dinitrobenzoates, hydroxybenzoates,
methoxybenzoates, phthalates, sulfonates, methylsulfonates,
propylsulfonates, besylates, xylenesulfonates,
naphthalene-1-sulfonates, naphthalene-2-sulfonates, phenylacetates,
phenylpropionates, phenylbutyrates, citrates, lactates,
.gamma.-hydroxybutyrates, glycolates, tartrates, and mandelates.
Lists of other suitable pharmaceutically acceptable salts are found
in Remington's Pharmaceutical Sciences, 17th Edition, Mack
Publishing Company, Easton, Pa., 1985.
[0369] For a compound of Formula I that contains a basic nitrogen,
a pharmaceutically acceptable salt may be prepared by any suitable
method available in the art, for example, treatment of the free
base with an inorganic acid, such as hydrochloric acid, hydrobromic
acid, sulfuric acid, sulfamic acid, nitric acid, boric acid,
phosphoric acid, and the like, or with an organic acid, such as
acetic acid, phenylacetic acid, propionic acid, stearic acid,
lactic acid, ascorbic acid, maleic acid, hydroxymaleic acid,
isethionic acid, succinic acid, valeric acid, fumaric acid, malonic
acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid,
oleic acid, palmitic acid, lauric acid, a pyranosidyl acid, such as
glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such
as mandelic acid, citric acid, or tartaric acid, an amino acid,
such as aspartic acid or glutamic acid, an aromatic acid, such as
benzoic acid, 2-acetoxybenzoic acid, naphthoic acid, or cinnamic
acid, a sulfonic acid, such as laurylsulfonic acid,
p-toluenesulfonic acid, methanesulfonic acid, or ethanesulfonic
acid, or any compatible mixture of acids such as those given as
examples herein, and any other acid and mixture thereof that are
regarded as equivalents or acceptable substitutes in light of the
ordinary level of skill in this technology.
[0370] The disclosure also relates to pharmaceutically acceptable
prodrugs of the compounds of Formula I, and treatment methods
employing such pharmaceutically acceptable prodrugs. The term
"prodrug" means a precursor of a designated compound that,
following administration to a subject, yields the compound in vivo
via a chemical or physiological process such as solvolysis or
enzymatic cleavage, or under physiological conditions (e.g., a
prodrug on being brought to physiological pH is converted to the
compound of Formula I). A "pharmaceutically acceptable prodrug" is
a prodrug that is non-toxic, biologically tolerable, and otherwise
biologically suitable for administration to the subject.
Illustrative procedures for the selection and preparation of
suitable prodrug derivatives are described, for example, in "Design
of Prodrugs", ed. H. Bundgaard, Elsevier, 1985.
[0371] The present disclosure also relates to pharmaceutically
active metabolites of compounds of Formula I, and uses of such
metabolites in the methods of the disclosure. A "pharmaceutically
active metabolite" means a pharmacologically active product of
metabolism in the body of a compound of Formula I or salt thereof.
Prodrugs and active metabolites of a compound may be determined
using routine techniques known or available in the art. See, e.g.,
Bertolini et al., J. Med. Chem. 1997, 40, 2011-2016; Shan et al.,
J. Pharm. Sci. 1997, 86 (7), 765-767; Bagshawe, Drug Dev. Res.
1995, 34, 220-230; Bodor, Adv. Drug Res. 1984, 13, 255-331;
Bundgaard, Design of Prodrugs (Elsevier Press, 1985); and Larsen,
Design and Application of Prodrugs, Drug Design and Development
(Krogsgaard-Larsen et al., eds., Harwood Academic Publishers,
1991).
[0372] Any formula depicted herein is intended to represent a
compound of that structural formula as well as certain variations
or forms. For example, a formula given herein is intended to
include a racemic form, or one or more enantiomeric,
diastereomeric, or geometric isomers, or a mixture thereof.
Additionally, any formula given herein is intended to refer also to
a hydrate, solvate, or polymorph of such a compound, or a mixture
thereof.
[0373] As used herein, the term "genetically altered" refers to a
permanent alteration in the DNA sequence that makes up a gene that
can result in a change in the protein sequence encoded by the gene.
A gene that is "genetically altered" as described herein, can
possess changes in DNA sequence, and/or protein sequence encoded by
the DNA sequence, that range in size; for example, a single
nucleotide (a.k.a. a single nucleotide polymorphism, SNP or point
mutation), a multiple nucleotide polymorphism (MNPs), a large
segment of a chromosome that includes multiple genes, such as a
gene fusion, and the like. Examples of gene fusions include, but
are not limited to, those which are the result of a chromosomal
inversion in which a portion of a chromosomal DNA encoding one or
more genes rearranges to provide a fusion of two genes not
ordinarily in communication in the DNA sequence, such as EML4-ALK
(see for example, Soda, M. et al., Nature, 2007, 448, 561-567),
those which are the result of a deletion in the DNA sequence (an
"interstitial deletion") in which part of a DNA sequence of a
chromosome is deleted to provide a fusion of two genes not
ordinarily in communication in the DNA sequence, such as
TMPRSS2-ERG (see for example, Yu J. et al., Cancer Cell, 2010, 17,
5, 443-54), or those which are the result of a translocation in
which a portion of chromosomal DNA is spliced and inserted into the
same or a different chromosome to provide a fusion of two genes not
ordinarily in communication in the DNA sequence, such as BCR-ABL
(see for example, Advani, A.S. et al. Leukemia Research, 2002, 26,
8, 713-720). One of skill in the art will readily appreciate that
such gene fusions can be found in multiple variants depending on
the individual in which the gene fusion has occurred, and each of
such variants is contemplated by the methods described herein.
[0374] A "genetically altered" gene, or the protein encoded by such
gene, can occur as hereditary mutations which can be inherited from
a parent and are sometimes referred to as germline mutations, or a
"genetically altered" gene, or the protein encoded by such gene,
can occur as an acquired (or somatic) mutation that occurs at some
point during a person's life. In some instances, a "genetically
altered" gene can be described as a de novo (new) mutation, and can
be either hereditary or somatic. It will be further understood that
"genetically altered" can refer to a situation in which more than
one of the changes in DNA sequence described herein can occur in a
patient simultaneously, such as a SNP (or point mutation) and a
translocation. Such situations can arise from, but are not solely
the result of, so-called "acquired resistance" in which a patient
having been treated with a kinase inhibitor can develop a mutation
in the DNA sequence that reduces the effectiveness of the
treatment. Non-limiting examples of such acquired resistance
mutations include the point mutation L1196M, G1202R, L1152P,
F1174C, C1156Y, 11171N, G1269S, and the 1151T insertion that occur
in the EML4-ALK gene fusion.
[0375] As used herein, the term "intrinsic resistance" refers to
the pre-existing resistance of disease cells, especially cancer
cells, to drug treatment, especially chemotherapy treatment. It
will be appreciated that intrinsic resistance can result in
resistance of the cells to a single drug, a small group of
structurally related drugs, or a several drugs of differing
chemical structure (so-called "multidrug resistance" or "MDR").
(Monti, E. 2007. Molecular Determinants of Intrinsic Multidrug
Resistance in Cancer Cells and Tumors In B. Teicher (Ed.), Cancer
Drug Resistance (pp. 241-260). Totowa, N.J.: Humana Press Inc.). It
will be appreciated that intrinsic resistance can be the result of
one or more host-related factors and/or the genetic make-up of the
cells. Such factors include but are not limited to
immunomodulation; pharmacogenetic factors such as failure to
achieve optimal serum drugs levels due to altered ADME or low
tolerance to drug-induced side effects; restricted drug access to
the tumor site; and microenvironmental cues. Such genetic make-up
factors include, but are not limited to altered expression of drug
transporters; qualitative alterations of drug target(s);
quantitative alterations of drug target(s); changes in
intracellular drug handling/metabolism; changes in DNA repair
activities, and alteration in apoptotic pathways. (Gottesman, M.
M., Annu. Rev. Med., 2002, 53, 516-527).
[0376] As used herein, the term "therapeutically effective amount"
refers to that amount of active compound or pharmaceutical agent
that elicits the biological or medicinal response in a patient,
which includes alleviation of the symptoms of the disease or
disorder being treated. In one aspect, the therapeutically
effective amount is that which may treat or alleviate the disease
or symptoms. The specific therapeutically-effective dose level for
any particular patient will depend upon a variety of factors,
including the disorder being treated and the severity of the
disorder; activity of the specific compound employed; the specific
composition employed; the age, body weight, general health, gender
and diet of the patient: the time of administration, route of
administration, and rate of excretion of the specific compound
employed; the duration of the treatment; drugs used in combination
or coincidentally with the specific compound employed; and like
factors. An exemplary dose is in the range of about from about 0.1
mg to 1 g daily, or about 1 mg to 50 mg daily, or about 50 to 250
mg daily, or about 250 mg to 1 g daily. The total dosage may be
given in single or divided dosage units (e.g., BID, TID, QID).
[0377] As used herein, the term "disease" includes, but is not
limited to, cancer, pain, psoriasis, rheumatoid arthritis,
polycythemia vera, essential thrombocythemia, ulcerative colitis,
and myeloid metaplasia with myelofibrosis.
[0378] As used herein, the term "cancer" includes, but is not
limited to, ALCL, NSCLC, neuroblastoma, inflammatory
myofibroblastic tumor, adult renal cell carcinoma, pediatric renal
cell carcinoma, breast cancer, ER.sup.+breast cancer, colonic
adenocarcinoma, glioblastoma, glioblastoma multiforme, anaplastic
thyroid cancer, cholangiocarcinoma, ovarian cancer, gastric
adenocarcinoma, colorectal cancer, inflammatory myofibroblastic
tumor, angiosarcoma, epithelioid hemangioendothelioma, intrahepatic
cholangiocarcinoma, thyroid papillary cancer, spitzoid neoplasms,
sarcoma, astrocytoma, brain lower grade glioma, secretory breast
carcinoma, mammary analogue carcinoma, acute myeloid leukemia,
congenital mesoblastic nephroma, congenital fibrosarcomas, Ph-like
acute lymphoblastic leukemia, thyroid carcinoma, skin cutaneous
melanoma, head and neck squamous cell carcinoma, pediatric glioma
CML, prostate cancer, lung squamous carcinoma, ovarian serous
cystadenocarcinoma, skin cutaneous melanoma, castrate-resistant
prostate cancer, Hodgkin lymphoma, and serous and clear cell
endometrial cancer.
Embodiments
[0379] In some embodiments, the methods described herein relate to
the treatment of disease comprising administering to a patient in
need of treatment a therapeutically effective amount of a compound
having activity against at least one tyrosine kinase selected from
the group consisting of ALK, ROS1, TRK, JAK and SRC. In some
embodiments, the compound has activity against at least one
tyrosine or serine/threonine kinase selected from the group
consisting of ALK, ROS1, TRKA, TRKB, TRKC, JAK2, SRC, FAK and ARK5.
In some embodiments, the compound has activity against at least two
tyrosine or serine/threonine kinases selected from the group
consisting of ALK, ROS1, TRKA, TRKB, TRKC, JAK2, SRC, FAK and ARK5.
In some embodiments, the at least one, or the at least two of the
tyrosine or serine/threonine kinases are genetically altered. In
some embodiments, the compound is of the formula I
##STR00016##
[0380] wherein X.sup.1, X.sup.2, Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4, Z.sup.5, Z.sup.6, Z.sup.7, M, R.sup.1, R.sup.2, R.sup.3,
R.sup.4, R.sup.5 and R.sup.6 are defined as described herein, or a
pharmaceutically acceptable salt thereof.
[0381] It will be appreciated that the disease can be any of a
number of diseases associated with the tyrosine kinases described
herein against which the compounds of the formula I have activity.
For example, the methods described herein can be used for the
treatment of diseases such as cancer, pain, psoriasis, rheumatoid
arthritis, polycythemia vera, essential thrombocythemia, ulcerative
colitis, myeloid metaplasia with myelofibrosis, and the like. It
will be appreciated that the disease can be any disease associated
with the activity of a tyrosine kinase described herein. It will be
further appreciated that the disease can be any disease associated
with a genetically altered tyrosine or serine/threonine kinase
selected from the group consisting of ALK, ROS1, TRKA, TRKB, TRKC,
JAK2, SRC, FAK and ARK5. It will be further appreciated that the
disease can be any disease associated with up-regulation of JAK2,
SRC, FAK or ARK5. In some embodiments, the disease is a cancer
mediated by or associated with a tyrosine kinase. In some
embodiments, the disease is a cancer mediated by or associated with
a serine/threonine kinase. In some embodiments, the disease is a
cancer mediated by or associated with a genetically altered
tyrosine kinase. In some embodiments, the disease is a cancer
mediated by or associated with a genetically altered
serine/threonine kinase. In some embodiments, the disease is a
cancer mediated by or associated with up-regulation of JAK2, SRC,
FAK or ARK5. In some embodiments, the disease is a cancer mediated
by or associated with a genetically altered tyrosine kinase
selected from the group consisting of ALK, ROS1, TRKA, TRKB, TRKC,
JAK2, SRC and FAK. In some embodiments, the disease is a cancer
mediated by or associated with a genetically altered
serine/threonine kinase, such as ARK5.
[0382] It will be appreciated that the cancer can be any cancer
mediated by or associated with a genetically altered tyrosine
kinase selected from the group consisting of ALK, ROS1, TRKA, TRKB,
TRKC and JAK2, or up-regulation of JAK2, SRC, FAK or ARK5,
including but not limited to, ALCL, NSCLC, neuroblastoma,
inflammatory myofibroblastic tumor, adult renal cell carcinoma,
pediatric renal cell carcinoma, breast cancer, colonic
adenocarcinoma, glioblastoma, glioblastoma multiforme, anaplastic
thyroid cancer, cholangiocarcinoma, ovarian cancer, gastric
adenocarcinoma, colorectal cancer, inflammatory myofibroblastic
tumor, angiosarcoma, epithelioid hemangioendothelioma, intrahepatic
cholangiocarcinoma, thyroid papillary cancer, spitzoid neoplasms,
sarcoma, astrocytoma, brain lower grade glioma, secretory breast
carcinoma, mammary analogue carcinoma, acute myeloid leukemia,
congenital mesoblastic nephroma, congenital fibrosarcomas, Ph-like
acute lymphoblastic leukemia, thyroid carcinoma, skin cutaneous
melanoma, head and neck squamous cell carcinoma, pediatric glioma
CML, and prostate cancer. In some embodiments, the cancer is NSCLC.
In some embodiments, the cancer is colorectal cancer. In some
embodiments, the cancer is neuroblastoma.
[0383] In some embodiments, the present disclosure provides methods
of treating disease in a patient that has received a prior
treatment with one or more therapeutic agents. In some embodiments,
the patient has been previously treated with one or more
chemotherapeutic agents. In still other embodiments, the patent has
been previously treated with one or more chemotherapeutic agents
and developed an acquired resistance to the treatment. In still
other embodiments, the patent has been previously treated with one
or more chemotherapeutic agents and developed bypass resistance to
the treatment. In still other embodiments, the patent has been
previously treated with one or more chemotherapeutic agents and
developed bypass resistance to the treatment regulated by SRC or
JAK2.
[0384] Other chemotherapeutic agents which the patient may be been
treated with prior to treatment with one or more of the compounds
described herein include but are not limited to kinase inhibitors,
adrenocorticoids and corticosteroids, alkylating agents, peptide
and peptidomimetic signal transduction inhibitors, antiandrogens,
antiestrogens, androgens, aclamycin and aclamycin derivatives,
estrogens, antimetabolites, platinum compounds, amanitins, plant
alkaloids, mitomycins, discodermolides, microtubule inhibitors,
epothilones, inflammatory and proinflammatory agents, purine
analogs, pyrimidine analogs, camptothecins and dolastatins. In some
embodiments, the chemotherapeutic agent the patient received
previous to treatment with one or more compounds described herein
can be one or more of afatinib, axitinib, alectinib, bosutinib,
brigatini, cabozantinib, ceritinib, crizotinib, dabrefenib,
dasatinib, erlotinib, everolimus, gefitinib, ibrutinib, imatinib,
lapatinib, lenvatinib, nilotinib, nintedanib, palbociclib,
pazopanib, ponatinib, regorafenib, ruxolitinib, sirolimus,
sorafenib, sunitinib, tofacitinib, temsirolimus, trametinib,
vandetanib, vemurafenib, methotrexate, busulfan, carboplatin,
chlorambucil, cisplatin, tamoxiphen, taxol, paclitaxel, docetaxel,
cytosine arabinoside, cyclophosphamide, daunomycin, rhizoxin,
prednisone, hydroxyurea, teniposide, vincristine, vinblastine,
eribulin, camptothecin, irinotecan, geldanamycin, estramustine and
nocodazole. In some embodiments, the methods described herein
provide treatment of a patient previously treated with a kinase
inhibitor selected from the group consisting of afatinib,
alectinib, axitinib, bosutinib, brigatini, cabozantinib, ceritinib,
crizotinib, dabrefenib, dasatinib, erlotinib, everolimus,
gefitinib, ibrutinib, imatinib, lapatinib, lenvatinib, nilotinib,
nintedanib, palbociclib, pazopanib, ponatinib, regorafenib,
ruxolitinib, sirolimus, sorafenib, sunitinib, tofacitinib,
temsirolimus, trametinib, vandetanib and vemurafenib. In some
embodiments, the patient was previously treated with
crizotinib.
[0385] Pharmaceutical Compositions
[0386] For treatment purposes, pharmaceutical compositions
comprising the compounds described herein may further comprise one
or more pharmaceutically-acceptable excipients. A
pharmaceutically-acceptable excipient is a substance that is
non-toxic and otherwise biologically suitable for administration to
a subject. Such excipients facilitate administration of the
compounds described herein and are compatible with the active
ingredient. Examples of pharmaceutically-acceptable excipients
include stabilizers, lubricants, surfactants, diluents,
anti-oxidants, binders, coloring agents, bulking agents,
emulsifiers, or taste-modifying agents. In preferred embodiments,
pharmaceutical compositions according to the invention are sterile
compositions. Pharmaceutical compositions may be prepared using
compounding techniques known or that become available to those
skilled in the art.
[0387] Sterile compositions are also contemplated by the invention,
including compositions that are in accord with national and local
regulations governing such compositions.
[0388] The pharmaceutical compositions and compounds described
herein may be formulated as solutions, emulsions, suspensions, or
dispersions in suitable pharmaceutical solvents or carriers, or as
pills, tablets, lozenges, suppositories, sachets, dragees,
granules, powders, powders for reconstitution, or capsules along
with solid carriers according to conventional methods known in the
art for preparation of various dosage forms. Pharmaceutical
compositions of the invention may be administered by a suitable
route of delivery, such as oral, parenteral, rectal, nasal,
topical, or ocular routes, or by inhalation. Preferably, the
compositions are formulated for intravenous or oral
administration.
[0389] For oral administration, the compounds the invention may be
provided in a solid form, such as a tablet or capsule, or as a
solution, emulsion, or suspension. To prepare the oral
compositions, the compounds of the invention may be formulated to
yield a dosage of, e.g., from about 0.1 mg to 1 g daily, or about 1
mg to 50 mg daily, or about 50 to 250 mg daily, or about 250 mg to
1 g daily. Oral tablets may include the active ingredient(s) mixed
with compatible pharmaceutically acceptable excipients such as
diluents, disintegrating agents, binding agents, lubricating
agents, sweetening agents, flavoring agents, coloring agents and
preservative agents. Suitable inert fillers include sodium and
calcium carbonate, sodium and calcium phosphate, lactose, starch,
sugar, glucose, methyl cellulose, magnesium stearate, mannitol,
sorbitol, and the like. Exemplary liquid oral excipients include
ethanol, glycerol, water, and the like. Starch,
polyvinyl-pyrrolidone (PVP), sodium starch glycolate,
microcrystalline cellulose, and alginic acid are exemplary
disintegrating agents. Binding agents may include starch and
gelatin. The lubricating agent, if present, may be magnesium
stearate, stearic acid, or talc. If desired, the tablets may be
coated with a material such as glyceryl monostearate or glyceryl
distearate to delay absorption in the gastrointestinal tract, or
may be coated with an enteric coating.
[0390] Capsules for oral administration include hard and soft
gelatin capsules. To prepare hard gelatin capsules, active
ingredient(s) may be mixed with a solid, semi-solid, or liquid
diluent. Soft gelatin capsules may be prepared by mixing the active
ingredient with water, an oil, such as peanut oil or olive oil,
liquid paraffin, a mixture of mono and di-glycerides of short chain
fatty acids, polyethylene glycol 400, or propylene glycol.
[0391] Liquids for oral administration may be in the form of
suspensions, solutions, emulsions, or syrups, or may be lyophilized
or presented as a dry product for reconstitution with water or
other suitable vehicle before use. Such liquid compositions may
optionally contain: pharmaceutically-acceptable excipients such as
suspending agents (for example, sorbitol, methyl cellulose, sodium
alginate, gelatin, hydroxyethylcellulose, carboxymethylcellulose,
aluminum stearate gel and the like); non-aqueous vehicles, e.g.,
oil (for example, almond oil or fractionated coconut oil),
propylene glycol, ethyl alcohol, or water; preservatives (for
example, methyl or propyl p-hydroxybenzoate or sorbic acid);
wetting agents such as lecithin; and, if desired, flavoring or
coloring agents.
[0392] For parenteral use, including intravenous, intramuscular,
intraperitoneal, intranasal, or subcutaneous routes, the agents of
the invention may be provided in sterile aqueous solutions or
suspensions, buffered to an appropriate pH and isotonicity or in
parenterally acceptable oil. Suitable aqueous vehicles include
Ringer's solution and isotonic sodium chloride. Such forms may be
presented in unit-dose form such as ampoules or disposable
injection devices, in multi- dose forms such as vials from which
the appropriate dose may be withdrawn, or in a solid form or
pre-concentrate that can be used to prepare an injectable
formulation. Illustrative infusion doses range from about 1 to
1000m/kg/minute of agent admixed with a pharmaceutical carrier over
a period ranging from several minutes to several days.
[0393] For nasal, inhaled, or oral administration, the inventive
pharmaceutical compositions may be administered using, for example,
a spray formulation also containing a suitable carrier. The
inventive compositions may be formulated for rectal administration
as a suppository.
[0394] For topical applications, the compounds of the present
invention are preferably formulated as creams or ointments or a
similar vehicle suitable for topical administration. For topical
administration, the inventive compounds may be mixed with a
pharmaceutical carrier at a concentration of about 0.1% to about
10% of drug to vehicle. Another mode of administering the agents of
the invention may utilize a patch formulation to effect transdermal
delivery.
[0395] Any formula given herein is also intended to represent
unlabeled forms as well as isotopically labeled forms of the
compounds. Isotopically labeled compounds have structures depicted
by the formulas given herein except that one or more atoms are
replaced by an atom having a selected atomic mass or mass number.
Examples of isotopes that can be incorporated into compounds of the
disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen,
phosphorous, fluorine, chlorine, and iodine, such as .sup.2H,
.sup.3H, .sup.11C, .sup.13C, .sup.14C, .sup.15N, .sup.18O,
.sup.17O, .sup.31P, .sup.32P, .sup.35S, .sup.18F, .sup.36Cl, and
.sup.125I, respectively. Such isotopically labelled compounds are
useful in metabolic studies (preferably with .sup.14C), reaction
kinetic studies (with, for example .sup.2H or .sup.3H), detection
or imaging techniques [such as positron emission tomography (PET)
or single-photon emission computed tomography (SPECT)] including
drug or substrate tissue distribution assays, or in radioactive
treatment of patients. Further, substitution with heavier isotopes
such as deuterium (i.e., .sup.2H) may afford certain therapeutic
advantages resulting from greater metabolic stability, for example
increased in vivo half-life or reduced dosage requirements.
Isotopically labeled compounds of this disclosure and prodrugs
thereof can generally be prepared by carrying out the procedures
disclosed in the schemes or in the examples and preparations
described below by substituting a readily available isotopically
labeled reagent for a non-isotopically labeled reagent.
[0396] Drug Combinations
[0397] The compounds described herein may be used in pharmaceutical
compositions or methods in combination with one or more additional
active ingredients in the treatment of the diseases and disorders
described herein. Further additional active ingredients include
other therapeutics or agents that mitigate adverse effects of
therapies for the intended disease targets. Such combinations may
serve to increase efficacy, ameliorate other disease symptoms,
decrease one or more side effects, or decrease the required dose of
an inventive compound. The additional active ingredients may be
administered in a separate pharmaceutical composition from a
compound of the present invention or may be included with a
compound of the present invention in a single pharmaceutical
composition. The additional active ingredients may be administered
simultaneously with, prior to, or after administration of a
compound of the present invention.
[0398] Combination agents include additional active ingredients are
those that are known or discovered to be effective in treating the
diseases and disorders described herein, including those active
against another target associated with the disease. For example,
compositions and formulations of the invention, as well as methods
of treatment, can further comprise other drugs or pharmaceuticals,
e.g., other active agents useful for treating or palliative for the
target diseases or related symptoms or conditions.
[0399] Other chemotherapeutic agents suitable for use in
combination in the methods described herein include but are not
limited to kinase inhibitors, adrenocorticoids and corticosteroids,
alkylating agents, peptide and peptidomimetic signal transduction
inhibitors, antiandrogens, antiestrogens, androgens, aclamycin and
aclamycin derivatives, estrogens, antimetabolites, platinum
compounds, amanitins, plant alkaloids, mitomycins, discodermolides,
microtubule inhibitors, epothilones, inflammatory and
proinflammatory agents, purine analogs, pyrimidine analogs,
camptothecins and dolastatins. In some embodiments,
chemotherapeutic agents suitable for combination treatments in the
methods described herein include but are not limited to one or more
of afatinib, alectinib, axitinib, bosutinib, brigatini,
cabozantinib, ceritinib, crizotinib, dabrefenib, dasatinib,
erlotinib, everolimus, gefitinib, ibrutinib, imatinib, lapatinib,
lenvatinib, nilotinib, nintedanib, palbociclib, pazopanib,
ponatinib, regorafenib, ruxolitinib, sirolimus, sorafenib,
sunitinib, tofacitinib, temsirolimus, trametinib, vandetanib,
vemurafenib, methotrexate, busulfan, carboplatin, chlorambucil,
cisplatin, tamoxiphen, taxol, paclitaxel, docetaxel, cytosine
arabinoside, cyclophosphamide, daunomycin, rhizoxin, prednisone,
hydroxyurea, teniposide, vincristine, vinblastine, eribulin,
camptothecin, irinotecan, geldanamycin, estramustine and
nocodazole. chemotherapeutic agents suitable for combination
treatments in the methods described herein include but are not
limited to one or more kinase inhibitor selected from the group
consisting of afatinib, alectinib, axitinib, bosutinib, brigatini,
cabozantinib, ceritinib, crizotinib, dabrefenib, dasatinib,
erlotinib, everolimus, gefitinib, ibrutinib, imatinib, lapatinib,
lenvatinib, nilotinib, nintedanib, palbociclib, pazopanib,
ponatinib, regorafenib, ruxolitinib, sirolimus, sorafenib,
sunitinib, tofacitinib, temsirolimus, trametinib, vandetanib and
vemurafenib. In some embodiments, the patient was previously
treated with crizotinib. For pain indications, suitable combination
agents include anti-inflammatories such as NSAIDs. The
pharmaceutical compositions of the invention may additional
comprise one or more of such active agents, and methods of
treatment may additionally comprise administering an effective
amount of one or more of such active agents.
[0400] Diagnostic Tests
[0401] In some embodiments, the present disclosure provides methods
for treating disease in a patient previously identified as having a
genetically altered tyrosine or serine/threonine kinase, such as a
gene fusion. In some embodiments, the present disclosure provides
methods for treating cancer in a patient previously identified as
having a genetically altered tyrosine or serine/threonine kinase,
such as a gene fusion. In some embodiments, the present disclosure
provides methods for treating disease in a patient comprising (i)
identifying a genetically altered tyrosine or serine/threonine
kinase in the patient, and (ii) administering to the patient a
therapeutically effective amount of a compound useful in the
treatment of such disease.
[0402] It will be appreciated that the diagnosing or identifying a
patient as having a genetically altered tyrosine or serine
threonine kinase can be accomplished by any number of diagnostic
tests known to one of skill in the art. For example, such
diagnostic tests include, but are not limited to, fluorescence in
situ hybridization (FISH), polymerase chain reaction (PCR),
immunohistochemistry (IHC), whole genome sequencing, next
generation sequencing, and the like. It will also be appreciated
that any of the methods known in the art and applicable to
diagnosing a patient or identifying a patient in connection with
the present disclosure involve the transformation of a biological
sample from one state of matter to another by direct modification,
synthesis or by direct non-covalent connection to provide a
modified sample that can be used to determine whether the subject
has or does not have a genetically altered tyrosine or serine
threonine kinase. In some embodiments, "diagnosing" or
"identifying" with respect to the disease state of a patient means
applying a diagnostic test, such as FISH, PCR or IHC, to a
biological sample obtained from the patient.
[0403] It will be appreciated that FISH is a test that "maps" the
genetic material in a person's cells. This test can be used to
visualize specific genes or portions of genes. FISH is a
cytogenetic technique that uses fluorescent probes that bind to
only those parts of the chromosome with a high degree of sequence
complementarity. Such FISH tests can be used to identify a patient
with a genetically altered tyrosine or serine/threonine kinase by
any method known in the art, and such test can be used in
combination with the methods described herein as either a means of
prior identification of a patient for treatment, or the concomitant
identification of a patient for treatment.
[0404] It will be appreciated that IHC refers to the process of
detecting antigens (e.g., proteins) in cells of a tissue section by
exploiting the principle of antibodies binding specifically to
antigens in biological tissues. Immunohistochemical staining is
widely used in the diagnosis of abnormal cells such as those found
in cancerous tumors. Specific molecular markers are characteristic
of particular cellular events such as proliferation or cell death
(apoptosis). Visualising an antibody-antigen interaction can be
accomplished in a number of ways. In the most common instance, an
antibody is conjugated to an enzyme, such as peroxidase, that can
catalyse a colour-producing reaction. Alternatively, the antibody
can also be tagged to a fluorophore, such as fluorescein or
rhodamine. Such IHC tests can be used to identify a patient with a
genetically altered tyrosine or serine/threonine kinase by any
method known in the art, and such test can be used in combination
with the methods described herein as either a means of prior
identification of a patient for treatment, or the concomitant
identification of a patient for treatment.
[0405] It will be appreciated that PCR refers to a technology in
molecular biology used to amplify a single copy or a few copies of
a piece of DNA across several orders of magnitude, generating
thousands to millions of copies of a particular DNA sequence. Such
PCR tests can be used to identify a patient with a genetically
altered tyrosine or serine/threonine kinase by any method known in
the art, and such test can be used in combination with the methods
described herein as either a means of prior identification of a
patient for treatment, or the concomitant identification of a
patient for treatment.
[0406] It will be appreciated that whole genome sequencing or
next-generation sequencing refers to a process that determines the
complete DNA sequence of an organism's genome at a single time.
This entails sequencing all of an organism's chromosomal DNA as
well as DNA contained in the mitochondria. Such whole genome
sequencing tests can be used to identify a patient with a
genetically altered tyrosine or serine/threonine kinase by any
method known in the art, and such test can be used in combination
with the methods described herein as either a means of prior
identification of a patient for treatment, or the concomitant
identification of a patient for treatment.
EXAMPLES
[0407] The examples and preparations provided below further
illustrate and exemplify particular aspects of embodiments of the
disclosure. It is to be understood that the scope of the present
disclosure is not limited in any way by the scope of the following
examples.
[0408] Abbreviations
[0409] The examples described herein use materials, including but
not limited to, those described by the following abbreviations
known to those skilled in the art:
TABLE-US-00001 g grams eq equivalents mmol millimoles mol moles mL
milliliters L liters EtOAc or EA ethyl acetate MeCN acetonitrile
DCM dichloromethane DEAD diethyl azodicarboxylate MHz megahertz
.delta. chemical shift THF tetrahydrofuran PE petroleum ether
R.sub.f retardation factor DMSO-d.sub.6 deuterated dimethyl
sulfoxide CDCl.sub.3 deuterated chloroform n-BuOH n-butanol DIEA
n,n-diisopropylethylamine TMSCl trimethylsilyl chloride min minutes
hrs, hr or h hours TLC thin layer chromatography M molar MS mass
spectrum m/z mass-to-charge ratio FDPP pentafluorophenyl
diphenylphosphinate DMAP 4-(dimethylamino)pyridine DMF
N,N-dimethylformamide
[0410] Synthesis of Compound 1
[0411] Compound 1 was prepared according to the following synthetic
scheme:
##STR00017## ##STR00018##
Example 1
Preparation of 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate
(1a)
Step 1: Preparation of ethyl
5-oxo-4H-pyrazolo[1,5-a]pyrimidine-3-carboxylate (1-2)
[0412] To a mixture of ethyl 5-amino-1H-pyrazole-4-carboxylate
(Sigma-Aldrich, 150.00 g, 1.08 mmol) and ethyl
(E)-3-ethoxyprop-2-enoate (Sigma-Aldrich, 292.16 g, 2.03 mol) in
DMF (3.2 L) was added Cs.sub.2CO.sub.3 (656.77 g, 2.02 mol) in one
portion at 20.degree. C. under N.sub.2. The mixture was stirred at
110.degree. C. for 6 h. TLC (PE:EtOAc=1:1) showed the reaction was
completed. The mixture was cooled to 20.degree. C. and filtered
through a celite pad. The filter cake was washed with ethyl acetate
(3.times.30 mL). The filtrate was added to H.sub.2O (2 L) and
acidified with HOAc to pH=4. The resultant precipitate was filtered
to afford 1-2 (173.00 g, 834.98 mmol, 86.36% yield) as a white
solid: .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.54 (d, J=7.91
Hz, 1H), 8.12 (s, 1H), 6.13 (d, J=7.91 Hz, 1H), 4.27 (q, J=7.11 Hz,
2H), 1.28 (t, J=7.09 Hz, 3H).
Step 2: Preparation of
5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate (1)
[0413] To a mixture of 1-2 (158.00 g, 762.59 mmol) in MeCN (1.6 L)
was added POCl.sub.3 (584.64 g, 3.81 mol) at 20.degree. C. under
N.sub.2. The mixture was stirred at 100.degree. C. for 2 h. TLC
(PE:EA=1:1) showed the reaction was completed. The mixture was
cooled to 20.degree. C. and poured into ice-water (5000 mL) in
portions at 0.degree. C. and stirred for 20 min. The precipitate
was filtered and dried to afford 1a (110.00 g, 487.52 mmol, 63.93%
yield) as a white solid: .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.33 (d, J=7.28 Hz, 1H), 8.66 (s, 1H), 7.41 (d, J=7.15 Hz,
1H), 4.31 (q, J=7.15 Hz, 2H), 1.32 (t, J=7.09 Hz, 3H).
Example 2
Preparation of (R)-2-(1-aminoethyl)-4-fluorophenol (2).
Step 1: Preparation of
(R)--N--(5-fluoro-2-hydroxybenzylidene)-2-methylpropane-2-sulfinamide
(2-2)
[0414] To a solution of (R)-2-methylpropane-2-sulfinamide
(Sigma-Aldrich, 150.00 g, 1.24 mol, 1.00 eq.) and
5-fluoro-2-hydroxybenzaldehyde (2-1) (Sigma-Aldrich, 173.74 g, 1.24
mol, 1.00 eq.) in DCM (2.00 L) was added Cs.sub.2CO.sub.3 (646.43
g, 1.98 mol, 1.60 eq.). The mixture was stirred at 16.degree. C.
for 16 hours. TLC (PE:EtOAc=5:1) showed the reaction was completed.
The reaction mixture was quenched by addition of H.sub.2O (1000 mL)
at 0.degree. C. and then extracted with EtOAc (500 mL.times.4). The
combined organic layers were washed with brine (1000 mL) and dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to give 2-2 (230.00 g, 945.33 mmol, 76.24% yield).
.sup.1HNMR (CDCl.sub.3, 400 MHz) .delta. 8.64 (s, 1H), 7.22-7.11
(m, 2H), 7.03-6.95 (m, 1H), 1.28 (s, 9H).
Step 2: Preparation of
(R)--N--((R)-1-(5-fluoro-2-hydroxyphenyl)ethyl)-2-methylpropane-2-sulfina-
mide (2-3R)
[0415] To a solution of
(R)--N--(5-fluoro-2-hydroxybenzylidene)-2-methylpropane-2-sulfinamide
(2-2) (200.00 g, 822.03 mmol, 1.00 eq.) in THF (2.5 L) was added
MeMgBr (490.09 g, 4.11 mol, 5.00 eq.) drop-wise at -65.degree. C.
under N.sub.2 over a period of 30 min. The mixture was then warmed
to ambient temperature and stirred for 18 hours. TLC (PE:EtOAc=1:1)
showed the reaction was complete with the production of two
diastereomers. The reaction mixture was quenched by addition of
H.sub.2O (2 L) at 0.degree. C., the mixture was extracted with
EtOAc (500 mL.times.3). The combined organic layers were washed
with brine (500 mL), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to give a residue. The residue
was purified by column chromatography (SiO.sub.2, Petroleum
ether/Ethyl acetate=50/1 to 1:1) to give
(R)--N--((R)-1-(5-fluoro-2-hydroxyphenyl)ethyl)-2-methylpropane-2-sulfina-
mide (2-3R) (125 g, the top, less polar spot with Rf: 0.5,
PE:EA=1:1). .sup.1HNMR (CDCl.sub.3, 400MHz) .delta.: 9.17 (s, 1H),
6.68 (dd, J=3.0, 8.8 Hz, 1H), 6.47 (dt, J=3.0, 8.4 Hz, 1H), 6.31
(dd, J=4.8, 8.8 Hz, 1H), 5.11 (d, J=8.0 Hz, 1H), 4.28 (quin, J=7.2
Hz, 1H), 1.43 (d, J=6.8 Hz, 3H), 1.20 (s, 9H).
Step 3: Preparation of (R)-2-(1-aminoethyl)-4-fluorophenol (2)
[0416] A solution of
(R)--N--((R)-1-(5-fluoro-2-hydroxyphenyl)ethyl)-2-methylpropane-2-sulfina-
mide (2-3R) (125 g, 481.99 mmol, 1.00 eq.) in HCl/dioxane (1.5 L,
4N) was stirred at ambient temperature for 2 hours. TLC
(PE:EtOAc=2:1) showed the reaction was complete. The mixture was
filtered to give (R)-2-(1-aminoethyl)-4-fluorophenol (2) HCl salt
(85 g, 443.56 mmol, 90.03% yield) as a white solid. .sup.1HNMR
(d-DMSO, 400 MHz) .delta. 10.24 (s, 1H), 8.48 (br. s., 3H), 7.31
(dd, J=2.9, 9.7 Hz, 1H), 7.05-6.99 (m, 1H), 6.98-6.93 (m, 1H),
4.59-4.45 (m, 1H), 1.46 (d, J=6.8 Hz, 3H).
Example 3
Preparation of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one (Compound
1)
Step 1: Preparation of ethyl
(R)-5-((1-(5-fluoro-2-hydroxyphenyl)ethyl)amino)pyrazolo[1,5-a]pyrimidine-
-3-carboxylate (3)
[0417] To a solution of (R)-2-(1-aminoethyl)-4-fluorophenol (2) (85
g, 443.56 mmol, 1.00 eq.) and ethyl
5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate (1a) (100.08 g,
443.56 mmol, 1.00 eq.) in n-BuOH (2 L) was added DIEA (343.96 g,
2.66 mol, 6.00 eq.). The mixture was stirred at 120.degree. C. for
2 hrs. TLC (PE:EtOAc=1:1) showed the reaction was completed. The
reaction mixture was diluted with H.sub.2O (500 mL) at 16.degree.
C., and extracted with EtOAc (500 mLx3). The combined organic
layers were washed with brine (500 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to give a residue. The residue was purified by column
chromatography (SiO.sub.2, Petroleum ether/Ethyl acetate=10/1 to
1:3) to give ethyl
(R)-5-((1-(5-fluoro-2-hydroxyphenyl)ethyl)amino)pyrazolo[1,5-a]pyrimidine-
-3-carboxylate (3) (122 g, 349.34 mmol, 78.76% yield, ee>99%
purity) as a white solid. .sup.1HNMR (CDCl.sub.3, 400MHz) .delta.
9.28 (br. s., 1H), 8.26 (s, 1H), 8.14 (d, J=7.5 Hz, 1H), 6.95-6.89
(m, 2H), 6.87-6.80 (m, 1H), 6.18 (d, J=7.5 Hz, 1H), 5.98 (d, J=8.3
Hz, 1H), 5.71-5.54 (m, 1H), 4.50-4.35 (m, 2H), 1.60 (d, J=6.8 Hz,
3H), 1.42 (t, J=7.2 Hz, 3H).
Step 2: Preparation of ethyl
5-(((R)-1-(2-(((S)-1-((tert-butoxycarbonyl)amino)propan-2-yl)oxy)-5-fluor-
ophenyl)ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate (4)
[0418] A mixture of ethyl
(R)-5-((1-(5-fluoro-2-hydroxyphenyl)ethyl)amino)pyrazolo[1,5-a]pyrimidine-
-3-carboxylate (3) (10.00 g, 29.04 mmol) and tert-butyl
(R)-(2-hydroxypropyl)carbamate (Combi-Blocks, 7.63 g, 43.56 mmol)
was azetrope dried from DCM/toluene, and then re-dissolved in DCM
(11.62 mL). To the solution was added PPh.sub.3 (11.43 g, 43.56
mmol), and the mixture was stirred until the starting materials
were completely dissolved. To the solution was added DEAD (8.81 g,
43.56 mmol) over 5 min with mixing. The reaction was stirred for 3
hours. The reaction mixture was diluted with DCM (125 mL), followed
by addition of aqueous NaOH solution (2M, 100 mL). The mixture was
stirred vigorously for 12 hours and the layers were separated. The
aqueous layer was further extracted with DCM (3.times.50 mL). The
combined extracts were washed with brine (50 mL), dried with
Na.sub.2SO.sub.4, and concentrated under reduced pressure. The
residue was purified with flash chromatography (Teledyne ISCO
system, silica (330 g), 0-40% ethyl acetate in hexane to provide
ethyl
5-(((R)-1-(2-(((S)-1-((tert-butoxycarbonyl)amino)propan-2-yl)oxy)-5-
-fluorophenyl)ethyl)amino)-pyrazolo[1,5-a]pyrimidine-3-carboxylate
(4) (8.88 g, 60.9% yield). LC-MS m/z 502.2 (M+H).sup.+. .sup.1HNMR
(400 MHz, CHLOROFORM-d) .delta. 8.24 (s, 1H), 8.21 (d, J=7.6 Hz,
1H), 7.04 (d, J=8.4 Hz, 1H), 6.87 (d, J=6.0 Hz, 2H), 6.13 (d, J=7.2
Hz, 1H), 5.91 (br. s., 1H), 4.58 (d, J=3.6 Hz, 1H), 4.43-4.28 (m,
2H), 3.52-3.34 (m, 2H), 1.54 (d, J=6.8 Hz, 3H), 1.47-1.36 (m, 12H),
1.30 (d, J=6.4 Hz, 3H).
Step 3: Preparation of
5-(((R)-1-(2-(((S)-1-((tert-butoxycarbonyl)amino)propan-2-yl)oxy)-5-fluor-
ophenyl)ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid
(5)
[0419] To a solution of ethyl
5-(((R)-1-(2-(((S)-1-((tert-butoxycarbonyl)amino)propan-2-yl)oxy)-5-fluor-
ophenyl)ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate (4)
(6.98 g, 13.92 mmol, 1 eq.) in methanol (65 mL) and THF (20 mL) was
added LiOH (2M, 47.9 mL, 95.8 mmol). The mixture was heated at
70.degree. C. for 3 hrs, cooled to ambient temperature, and then
quenched with aq. HCl (2M, 95.8 mL) to adjust pH<5. The reaction
mixture was extracted with CH.sub.2Cl.sub.2 (3.times.50 mL), and
dried over Na.sub.2SO.sub.4. After filtration, evaporation, and
high vacuum dry, a white solid of
5-(((R)-1-(2-(((S)-1-((tert-butoxycarbonyl)amino)propan-2-yl)oxy)-5-fluor-
ophenyl)ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (5)
was obtained which was used in the next step without further
purification. LC-MS m/z 474.2 (M+H).sup.+.
Step 4: Preparation of
5-(((R)-1-(2-(((S)-1-aminopropan-2-yl)oxy)-5-fluorophenyl)ethyl)amino)pyr-
azolo[1,5-a]pyrimidine-3-carboxylic acid (6)
[0420] To a solution of
5-(((R)-1-(2-(((S)-1-((tert-butoxycarbonyl)amino)propan-2-yl)oxy)-5-fluor-
ophenyl)ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (5)
(6.59 g, 13.92 mmol) in CH.sub.2Cl.sub.2 (130 mL) was added HCl in
dioxane (4 M, 30.4 mL). Keep stirring at room temperature for 2
hours until the reaction was shown to be completed by LC-MS. The
reaction mixture was concentrated, and high vacuum dried to provide
compound 6 as a white solid which was used in the next step without
further purification. LC-MS m/z 374.2 (M+H).sup.+.
Step 5: Preparation of
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one (Compound
1)
[0421]
5-(((R)-1-(2-(((S)-1-aminopropan-2-yl)oxy)-5-fluorophenyl)ethyl)ami-
no)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (6) (5.20 g, 13.93
mmol) was dissolved in DMF (75 mL) to make Solution A. To a
solution of Hunig's base (DIPEA) (14.40 g, 111.4 mmol) in DMF (150
mL) and DCM (350 mL) was added solution A (25 mL) and one third of
the total FDPP (5.62 g, 14.63 mmol) sequentially. The reaction was
stirred for 1 hour, and LC-MS showed the completion of the coupling
reaction. The same process was repeated for 2 more times. The final
solution was stirred at ambient temperature for 63 hour (or until
the reaction was shown to be completed by LC-MS). The reaction was
quenched by addition of aqueous Na.sub.2CO.sub.3 solution (2M, 150
mL), and the mixture was stirred for 15 min, and extracted with DCM
(3.times.150 mL). The combined extracts were dried with
Na.sub.2SO.sub.4, concentrated under reduced pressure, and purified
on a flash chromatography (Teledyne ISCO system, silica (220 g),
0-7.5% methanol in dichloromethane) to provide
(7S,13R)-11-fluoro-7,13-dimethyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-
[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one (Compound 1)
(4.38 g, 12.33 mmol, 88.5% yield) as a white solid. LC-MS: m/z
[M+H].sup.+ 356.2. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
9.82 (dd, J=8.02, 2.29 Hz, 1H), 8.81 (d, J=6.87 Hz, 1H), 8.58 (d,
J=7.45 Hz, 1H), 8.04 (s, 1H), 7.12 (dd, J=9.45, 3.15 Hz, 1H),
6.99-7.05 (m, 1H), 6.94-6.99 (m, 1H), 6.36 (d, J=7.45 Hz, 1H), 5.53
(m, 1H), 4.45-4.52 (m, 1H), 3.90 (ddd, J=13.46, 8.31, 4.01 Hz, 1H),
3.10-3.17 (m, 1H), 1.46 (d, J=6.30 Hz, 3H), 1.44 (d, J=7.45 Hz,
3H).
[0422] In-Vitro Assays
[0423] Materials and Methods
[0424] Kinase Binding Assay Method
[0425] Kinase binding assays were performed at DiscoveRx using the
general KINOMEscan K.sub.d Protocol (Fabian, M. A. et al., "A small
molecule-kinase interaction map for clinical kinase inhibitors,"
Nat. Biotechnol. 2005, 23(3):329-36). For most assays,
kinase-tagged T7 phage strains were prepared in an E. coli host
derived from the BL21 strain. E. coli were grown to log-phase and
infected with T7 phage and incubated with shaking at 32.degree. C.
until lysis. The lysates were centrifuged and filtered to remove
cell debris. The remaining kinases were produced in HEK-293 cells
and subsequently tagged with DNA for qPCR detection.
Streptavidin-coated magnetic beads were treated with biotinylated
small molecule ligands for 30 minutes at room temperature to
generate affinity resins for kinase assays. The liganded beads were
blocked with excess biotin and washed with blocking buffer
(SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove
unbound ligand and to reduce nonspecific binding. Binding reactions
were assembled by combining kinases, liganded affinity beads, and
test compounds in 1.times. binding buffer (20% SeaBlock,
0.17.times. PBS, 0.05% Tween 20, 6 mM DTT). All reactions were
performed in polystyrene 96-well plates in a final volume of 0.135
mL. The assay plates were incubated at room temperature with
shaking for 1 hour and the affinity beads were washed with wash
buffer (1.times. PBS, 0.05% Tween 20). The beads were then
re-suspended in elution buffer (1.times. PBS, 0.05% Tween 20, 0.5
.mu.M non-biotinylated affinity ligand) and incubated at room
temperature with shaking for 30 minutes. The kinase concentration
in the eluates was measured by qPCR.
[0426] Binding constants (K.sub.ds) were calculated with a standard
dose-response curve using the Hill equation:
Response=Background+(Signal-Background)/(1+(Kd.sup.Hill
Slope/Dose.sup.Hill Slope)) The Hill Slope was set to -1. Curves
were fitted using a non-linear least square fit with the
Levenberg-Marquardt algorithm.
[0427] Biochemical Kinase Assay Method
[0428] The biochemical kinase assay was performed at Reaction
Biology Corporation (www.reactionbiology.com, Malvern, Pa.)
following the procedures described in the reference (Anastassiadis
T, et al Nat Biotechnol. 2011, 29, 1039). Specific kinase/substrate
pairs along with required cofactors were prepared in reaction
buffer; 20 mM Hepes pH 7.5, 10 mM MgCl2, 1 mM EGTA, 0.02% Brij35,
0.02 mg/ml BSA, 0.1 mM Na3VO4, 2 mM DTT, 1% DMSO (for specific
details of individual kinase reaction components see Supplementary
Table 2). Compounds were delivered into the reaction, followed 20
minutes later by addition of a mixture of ATP (Sigma, St. Louis
Mo.) and .sup.33P ATP (Perkin Elmer, Waltham Mass.) to a final
concentration of 10 .mu.M. Reactions were carried out at room
temperature for 120 min, followed by spotting of the reactions onto
P81 ion exchange filter paper (Whatman Inc., Piscataway, N.J.).
Unbound phosphate was removed by extensive washing of filters in
0.75% phosphoric acid. After subtraction of background derived from
control reactions containing inactive enzyme, kinase activity data
was expressed as the percent remaining kinase activity in test
samples compared to vehicle (dimethyl sulfoxide) reactions.
IC.sub.50 values and curve fits were obtained using Prism (GraphPad
Software).
[0429] Cell Lines and Cell Culture:
[0430] Human lung cancer cell line NCI-H2228 was obtained from
ATCC. Cell lines 293T, NIH3T3, Ba/F3, and HCC78 were purchased from
DSMZ. Karpas-299 cell line was purchased from Sigma. KM12 cell line
was obtained from NCI.
[0431] NIH3T3 was maintained in DMEM medium supplemented with 10%
fetal bovine serum and 100 U/mL of penicillin/streptomycin. HCC78,
Karpas-299 and H2228 were maintained in RPMI-1640 supplemented with
10% fetal bovine serum with 100 U/mL of penicillin/streptomycin.
Ba/F3 cells were maintained in RPMI-1640 supplemented with 10%
fetal bovine serum, 10% (Vol/Vol) conditioned media from the
WIHI-3B myelomonocytic IL-3 secreting cells and 100 U/mL of
penicillin/streptomycin. BaF3 stable cell lines were maintained in
RPMI-1640 supplemented with 10% fetal bovine serum, 100 U/mL of
penicillin, and 0.5 .mu.g/mL puromycin solution.
[0432] Cloning and Ba/F3 or NIH3T3 Stable Cell Line Creation
[0433] The EML4-ALK gene (variant 1) was synthesized at GenScript
and cloned into pCDH-CMV-MCS-EF1-Puro plasmid (System Biosciences,
Inc). EML4-ALK point mutations G1202R, L1196M, L1152P, F1174C,
C1156Y, 11171N, G1269S, and 1151T insertion were generated at
GenScript by PCR and confirmed by sequencing. Ba/F3-EML4-ALK wild
type and mutants were generated by transducing Ba/F3 cells with
lentivirus containing EML4-ALK wide type or mutants. Stable cell
lines were selected by puromycin treatment, followed by IL-3
withdrawal. Briefly, 5.times.10.sup.6 Ba/F3 cells were transduced
with lentivirus supernatant in the presence of 8 .mu.g/mL protamine
sulfate. The transduced cells were subsequently selected with 1
.mu.g/mL puromycin in the presence of IL3-containing medium
RPMI1640, plus 10% FBS. After 10-12 days of selection, the
surviving cells were further selected for IL3 independent
growth.
[0434] SDC4-ROS1 wild type, CD74-ROS1 wild type, and their G2032R
mutant genes were synthesized at GenScript and cloned into
pCDH-CMV-MCS-EF1-Puro plasmid (System Biosciences, Inc), and
confirmed by sequencing. Ba/F3 SDC4-ROS1, CD74-ROS1 and
corresponding G2032R mutants were generated by transducing Ba/F3
cells with lentivirus containing the fusion genes. Stable cell
lines were selected by puromycin treatment, followed by IL-3
withdrawal. Briefly, 5.times.10.sup.6 Ba/F3 cells were transduced
with lentivirus supernatant in the presence of 8 .mu.g/mL protamine
sulfate. The transduced cells were subsequently selected with 1
.mu.g/mL puromycin in the presense of IL3-containing medium
RPMI1640, plus 10% FBS. After 10-12 days of selection, the
surviving cells were further selected for IL3 independent
growth.
[0435] Ba/F3 TPR-ALK and Ba/F3 TPR-ALK L1196M cell lines were
created at Advanced Cellular Dynamics, Inc. and cell proliferation
assays were performed there. The ACD panel itself is comprised of
92 unique kinases individually expressed in a common lymphoid cell
line (mouse Ba/F3 cells). Each cell line is dependent upon activity
of the recombinant kinase for survival.
[0436] SDC4-ROS1 L2026M and D2033N mutant genes were synthesized at
GenScript and cloned into pCDH-CMV-MCS-EF1-Puro plasmid (System
Biosciences, Inc), and confirmed by sequencing. Ba/F3 CD74-ROS1
L2026M and D2033N mutants were generated by transducing Ba/F3 cells
with lentivirus containing the fusion genes. Stable cell lines were
selected by puromycin treatment, followed by IL-3 withdrawal.
Briefly, 5.times.10.sup.6 Ba/F3 cells were transduced with
lentivirus supernatant in the presence of 8 .mu.g/mL protamine
sulfate. The transduced cells were subsequently selected with 1
.mu.g/mL puromycin in the presense of IL3-containing medium
RPMI1640, plus 10% FBS. After 10-12 days of selection, the
surviving cells were further selected for IL3 independent
growth.
[0437] LMNA-TRKA wild type, and its G595R mutant genes were
synthesized at GenScript and cloned into pCDH-CMV-MCS-EF1-Puro
plasmid (System Biosciences, Inc), and confirmed by sequencing.
Ba/F3 LMNA-TRKA and corresponding G595R mutant were generated by
transducing Ba/F3 cells with lentivirus containing the fusion
genes. Stable cell lines were selected by puromycin treatment,
followed by IL-3 withdrawal. Briefly, 5.times.10.sup.6 Ba/F3 cells
were transduced with lentivirus supernatant in the presence of 8
.mu.g/mL protamine sulfate. The transduced cells were subsequently
selected with 1 .mu.g/mL puromycin in the presense of
IL3-containing medium RPMI1640, plus 10% FBS. After 10-12 days of
selection, the surviving cells were further selected for IL3
independent growth.
[0438] TEL-TRKB (also named as ETV6-TRKB) wild type and its G639R
mutant genes were synthesized at GenScript and cloned into
pCDH-CMV-MCS-EF1-Puro plasmid (System Biosciences, Inc), and
confirmed by sequencing. Ba/F3 TEL-TRKB and corresponding G639R
mutants were generated by transducing Ba/F3 cells with lentivirus
containing the fusion genes. Stable cell lines were selected by
puromycin treatment, followed by IL-3 withdrawal. Briefly,
5.times.10.sup.6 Ba/F3 cells were transduced with lentivirus
supernatant in the presence of 8 .mu.g/mL protamine sulfate. The
transduced cells were subsequently selected with 1 .mu.g/mL
puromycin in the presense of IL3-containing medium RPMI1640, plus
10% FBS. After 10-12 days of selection, the surviving cells were
further selected for IL3 independent growth.
[0439] TEL-TRKC (also named as ETV6-TRKC) wild type and its G623R
mutant genes were synthesized at GenScript and cloned into
pCDH-CMV-MCS-EF1-Puro plasmid (System Biosciences, Inc), and
confirmed by sequencing. Ba/F3 TEL-TRKC and corresponding G623R
mutant were generated by transducing Ba/F3 cells with lentivirus
containing the fusion genes. Stable cell lines were selected by
puromycin treatment, followed by IL-3 withdrawal. Briefly,
5.times.10.sup.6 Ba/F3 cells were transduced with lentivirus
supernatant in the presence of 8 .mu.g/mL protamine sulfate. The
transduced cells were subsequently selected with 1 .mu.g/mL
puromycin in the presense of IL3-containing medium RPMI1640, plus
10% FBS. After 10-12 days of selection, the surviving cells were
further selected for IL3 independent growth.
[0440] NIH3T3 ALK or ROS1 stable cell lines were generated by
transducing the cells with lentivirus containing EML4-ALK wild type
and G1202R mutant genes, SDC4-ROS1 wild type, CD74-ROS1 wild type
and their G2032R mutant genes. The transduced cells were
subsequently selected with 1 .mu.g/mL puromycin.
[0441] NIH3T3 ROS1, or TRKA , or TRKB, or TRKC stable cell lines
were generated by transducing the cells with lentivirus containing
CD74-ROS1 L2026M and D2033N mutant genes, LMNA-TRKA wild type and
G595R mutant genes, TEL-TRKB wild type and G639R mutant genes,
TEL-TRKB wild type and G623R mutant genes, respectively. The
transduced cells were subsequently selected with 1 .mu.g/mL
puromycin.
[0442] Cell Proliferation Assays:
[0443] Two thousand cells per well were seeded in 384 well white
plate for 24 hrs, and then treated with compounds for 72 hours
(37.degree. C., 5% CO.sub.2). Cell proliferation was measured using
CellTiter-Glo luciferase-based ATP detection assay (Promega)
following the manufactures's protocol. IC.sub.50 determinations
were performed using GraphPad Prism software (GraphPad, Inc., San
Diego, Calif.).
[0444] The cell proliferation assay of Ba/F3 TPR-ALK and B a/F3
TPR-ALK L1196M cell lines were performed at Advanced Cellular
Dynamics, Inc. The inhibition of kinase activity leads to cell
death, which is monitored via ATP concentration using CellTiter-Glo
(Promega). Cell lines were maintained in RPMI-1640 culture media
containing 10% fetal calf serum and antibiotics. Cells in
logarithmic-phase growth were harvested and 5,000 cells were
distributed into each well of a 384-well plate in 50 .mu.L of
growth media. Parental cells (only) were seeded in the presence of
10 ng/mL IL3 to support cell growth and survival. Fifty nanoliters
diluted compound were added to appropriate wells, in duplicate, and
the cells were cultured for 48 hours at 37 C in a humidified 5% CO2
atmosphere. Viability was determined by adding 15 .mu.L
CellTiter-Glo and measuring luminescence, which is reported as
relative light units (RLU) measured in counts per second.
[0445] Immunblotting for Cellular Kinase Phosphorylation Assays
[0446] NSCLC cell line H2228 (harboring endogenous EML4-ALK fusion
gene), HCC78 cells (harboring endogenous SLC34A2-ROS1 fusion gene),
Karpas-299 cells (harboring endogenous NPM-ALK fusion gene) or
SET-2 (harboring endogenous JAK2V617F activating mutation) were
cultured in RPMI medium, and KM12 (harboring endogenous TPM3-TRKA
fusion gene) cell line was cultured in DMEM medium, both
supplemented with 10% fetal bovine serum and 100 U/mL of
penicillin/streptomycin. Ba/F3 and NIH3T3 cells stably expressing
ALK or ROS1 (WT or mutant) were culture as mentioned above. Half a
million cells per well were seeded in 24 well plate for 24 hrs, and
then treated with compounds for 4 hours. Cells were collected after
treatment and lysed in RIPA buffer (50 mM Tris, pH 7.4, 150 mM
NaCl, 1% NP-40, 0.5% Deoxycholate, 0.1% SDS) supplemented with 10
mM EDTA, 1.times. Halt protease and phosphatase inhibitors (Thermo
Scientific). Protein lysates (approximately 20 .mu.g) was resolved
on 4-12% Bolt Bis-Tris precasted gels with MES running buffer (Life
Technologies), transferred to nitrocellulose membranes using
Trans-Blot Turbo Transfer System (Bio-Rad) and detected with
antibodies targeting phosphorylated ALK Y1604 (Cell Signaling
Technology), total ALK (Cell Signaling Technology), phosphorylated
ROS1 and total ROS1 (Cell Signaling Technology), phosphorylated TRK
A/B (Cell Signaling Technology), total TRKA antibody (Santa Cruz
Biotechnogy), phosphorylated STAT3 and STATS, total STAT3 and STATS
(Cell Signaling Technology), phosphorylated AKT (Cell Signaling
Technology), total AKT (Cell Signaling Technology), phosphorylated
ERK (Cell Signaling Technology), total ERK (Cell Signaling
Technology) and Tubulin (Sigma). Antibodies were typically
incubated overnight at 4.degree. C. with gentle shake, followed by
washes and incubation with the appropriate HRP-conjugated secondary
antibodies. Membranes were incubated with chemiluminescent
substrate for 5 min at room temperature (SuperSignal West Femto,
Thermo Scientific). The chemiluminescent images were acquired with
a C-DiGit Imaging System (LI-COR Biosciences). The relative density
of the chemiluminescent bands were quantified via Image Studio
Digits from LICOR. The half inhibitory concentration (IC.sub.50)
value is calculated using non-linear regression analysis through
GraphPad Prism software (GraphPad, Inc., San Diego, Calif.).
[0447] NSCLC cell line H2228 (harboring endogenous EML4-ALK fusion
gene) was cultured in RPMI medium supplemented with 10% fetal
bovine serum and 100 U/mL of penicillin/streptomycin. Ba/F3 and
NIH3T3 cells stably expressing ROS1, or TRKA, or TRKB, or TRKC (WT
or mutant) were cultured as mentioned above. Half a million cells
per well were seeded in 24 well plate for 24 hrs, and then treated
with compounds for 4 hours. Cells were collected after treatment
and lysed in RIPA buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 1%
NP-40, 0.5% Deoxycholate, 0.1% SDS) supplemented with 10 mM EDTA,
1.times. Halt protease and phosphatase inhibitors (Thermo
Scientific). Protein lysates (approximately 20 .mu.g) was resolved
on 4-12% Bolt Bis-Tris precasted gels with MES running buffer (Life
Technologies), transferred to nitrocellulose membranes using
Trans-Blot Turbo Transfer System (Bio-Rad) and detected with
antibodies targeting phosphorylated phosphorylated ROS1 and total
ROS1 (Cell Signaling Technology), phosphorylated TRK A/B (Cell
Signaling Technology), total TRKA antibody (Santa Cruz
Biotechnogy), phosphorylated SRC Y416 (Cell Signaling Technology),
total SRC (Cell Signaling Technology), phosphorylated FAK Y576/577
(Cell Signaling Technology), total FAK (Cell Signaling Technology),
phosphorylated paxillin Y118 (Cell Signaling Technology), total
paxillin (Cell Signaling Technology), phosphorylated EGFR and total
EGFR, CD44, and Tubulin (Sigma). Antibodies were typically
incubated overnight at 4.degree. C. with gentle shake, followed by
washes and incubation with the appropriate HRP-conjugated secondary
antibodies. Membranes were incubated with chemiluminescent
substrate for 5 min at room temperature (SuperSignal West Femto,
Thermo Scientific). The chemiluminescent images were acquired with
a C-DiGit Imaging System (LI-COR Biosciences). The relative density
of the chemiluminescent bands were quantified via Image Studio
Digits from LICOR. The half inhibitory concentration (IC.sub.50)
value is calculated using non-linear regression analysis through
GraphPad Prism software (GraphPad, Inc., San Diego, Calif.).
[0448] Apoptosis/Caspase Activity Assays.
[0449] Karpas-299 cells were maintained in RPMI medium supplemented
with 10% fetal bovine serum and antibiotics. Five hundred thousand
cells per well were seeded in 12-well plate and various
concentration of compounds were introduced and incubated for 48
hrs. The cells were then collected and lysed in a lysis buffer (20
mM HEPES, 150 mM NaCl, 10 mM KCl, 5 mM EDTA, 1% NP40) supplemented
with Halt protease and phosphatase inhibitors (Thermo Scientific).
For caspase assays, approximately 20 .sub.i.t.g of cell lysate were
incubated with 20 .mu.l of caspase-3 glo reagent (Promega), and the
enzyme activity was measured by the release of luminescence after
20 min incubation at 37.degree. C. For western blotting, cell
lysate were boiled and analyzed by SDS-PAGE/immunoblotting using
anti-caspase-3 (Cell Signaling Technology), anti-PARP (Cell
Signaling Technology), or anti-actin (Cell Signaling Technology)
antibodies. Antibodies were typically incubated overnight at
4.degree. C. with gentle shake, followed by washes and incubation
with the appropriate HRP-conjugated secondary antibodies. Membranes
were incubated with chemiluminescent substrate for 5 min at room
temperature (SuperSignal West Femto, Thermo Scientific), and the
chemiluminescent images were obtained with a C-DiGit Imaging System
(LI-COR Biosciences).
[0450] Scratch Wound Healing Assay
[0451] HT1080 or HCC78 cells in RPMI medium supplemented with 10%
fetal bovine serum and antibiotics were seeded in 24-well plate.
After 12-24 hours, confluent cell monolayers were gently scraped
with a sterile pipette tip to form a scratch. The plates were
washed with fresh medium, and the cells were incubated with medium
alone or medium containing various concentration of compounds.
After 12-24 hours, the plates were examined and recorded by an EVOS
FL microscopy (Life Technology) to monitor resealing of the cell
monolayer.
[0452] In-Vivo Methods
[0453] Cell lines
[0454] Cell lines were cultured using standard techniques in DMEM
or RPMI-1640 medium (Corning, Inc) with 10% fetal bovine serum
(Thermo Fisher Scientific, Inc) at 37.degree. C. in a humidified
atmosphere with 5% CO.sub.2. For implantation, cells were harvested
and pelleted by centrifugation at 250g for 2 minutes. Cells were
washed once and resuspended in serum-free medium, with 50% matrigel
(v/v) as needed.
[0455] Subcutaneous Xenograft Models in Immune Compromised Mice
[0456] Female athymic nude mice and SCID/Beige mice (5-8 weeks of
age) were obtained from Charles River Laboratory and were housed in
Innovive IVC disposable cages on HEPA filtered ventilated racks
with ad libitum access to rodent chow and water. Five million cells
in 100 .mu.L serum-free medium were implanted subcutaneously in the
right flank region of the mouse. Karpas299 cells were implanted
into the SCID/Beige mice. KM12 cells, NIH3T3 EML4-ALK wild type
mutant cells, and NIH3T3 SCD4-ROS1 wild type mutant cells were
implanted into the athymic nude mice, respectively. All models
except KM12 were implanted with 50% matrigel (Corning, Inc) in the
medium. Tumor size and body weight were measured on designated
days. Tumor size was measured with an electronic caliper and tumor
volume was calculated as the product of length*width.sup.2*0.5.
Mice were randomized by tumor size into treatment groups when tumor
volume reached about 100-200 mm.sup.3 and Compound 1 were
administered orally (BID) at determined dosage.
[0457] Female athymic nude mice and SCID/Beige mice (5-8 weeks of
age) were obtained from Charles River Laboratory and were housed in
Innovive IVC disposable cages on HEPA filtered ventilated racks
with ad libitum access to rodent chow and water. Five million cells
in 100 .mu.L serum-free medium were implanted subcutaneously in the
right flank region of the mouse. Ba/F3 EML4-ALK wild type and
G1202R mutant cells, Ba/F3 CD74-ROS1 wild type and G2032R mutant
cells were implanted into the SCID/Beige mice. NIH3T3 CD74-ROS1
G2032 mutant cells, NIH3T3 LMNA-TRKA wild type and G595R mutant
cells were implanted into the athymic nude mice, respectively. All
models were implanted with 50% matrigel (Corning, Inc) in the
medium. Tumor size and body weight were measured on designated
days. Tumor size was measured with an electronic caliper and tumor
volume was calculated as the product of length*width.sup.2*0.5.
Mice were randomized by tumor size into treatment groups when tumor
volume reached about 100-200 mm.sup.3 and Compound 1 were
administered orally (BID) at determined dosage.
[0458] Tumor Processing and immunoblotting for In Vivo
Pharmacodynamic Studies
[0459] Mice bearing xenograft tumors were humanely euthanized and
tumors were resected and snap frozen in liquid nitrogen and stored
at -80.degree. C. Frozen tumor samples were processed at 4.degree.
C. in 1.times. Cell Lysis Buffer (Cell Signaling Technologies) to
extract proteins. SDS loading samples were prepared by adding one
volume of 4.times. LDS Sample Buffer (Life Technologies, Inc) to
three volumes of protein lysate. Tumor SDS protein samples were
processed by SDS-PAGE and immunoblotted with rabbit
anti-phosphorylated ALK and mouse anti-actin antibodies (Cell
Signaling Technologies). The signals from immunoblot were detected
by C-DiGit Blot Scanner from LI-COR and the signal intensity were
quantified using the Image Studio Digit software (LI-COR).
[0460] Data and Results:
Example 4a
Kinase Binding Affinities and Enzymatic Kinase Activities of
Compound 1
[0461] The kinase binding affinity of Compound 1 was screened at
DiscoveRx in a panel of 460 kinases followed by the determination
of the K.sub.d for the hits. Compound 1 demonstrated strong binding
affinities with ALK, ROS1, TRKA/B/C, JAK2, SRC, FAK and ARKS (Table
1). Furthermore, the enzymatic kinase inhibition activities of
Compound 1 at 10 .mu.M ATP concentration were determined at
Reaction Biology, and the results of IC.sub.50 were summarized in
Table 1.
TABLE-US-00002 TABLE 1 Target ALK ROS1 TRKA TRKB TRKC JAK2 SRC FAK
ARK5 K.sub.d (nM) 5.7 0.19 0.019 0.054 0.088 0.082 12.0 27 3.7
IC.sub.50 (nM) 1.04 0.0706 0.826 0.0517 0.0956 1.04 5.29 6.96
4.46
Example 4b
Enzymatic Kinase Activities of Compound 1
[0462] The kinase binding affinity of Compound 1 was screened at
DiscoveRx in a panel of 460 kinases. The kinase hits of compound 1
were further confired in the recombined enzymatic kinase inhibition
assay using 10 .mu.M ATP concentration at Reaction Biology, and the
IC.sub.50 results were summarized in Table 1b.
TABLE-US-00003 TABLE 1b IC.sub.50 IC.sub.50 IC.sub.50 (nM) at (nM)
at (nM) at 10 .mu.M 10 .mu.M 10 .mu.M Target ATP Target ATP Target
ATP TRKB 0.05 SNARK 13.0 JAK3 50 ROS1 0.07 HCK 16.4 EPHA8 50.2 TRKC
0.1 IRR 18.1 IGFR 111 TRKA 0.83 LCK 18.6 PLK4 126 ALK 1.04 JAK1 19
AXL 149 JAK2 1.04 TYK2 21.6 MARK3 512 FYN 1.05 LTK 21.8 LYN 1.66
DDR2 23 YES 2.15 BTK 23.7 FGR 3.05 TNK2 24.1 TXK 3.17 EPHA1 25.0
ARKS 4.46 BLK 32.3 SRC 5.3 GRK7 35.2 DDR1 5.7 PYK2 39.9 FAK 6.96
RET 47.1
Example 5
Assessment of Compound 1 Activity Against a Panel of ALK
Mutations
[0463] Compound 1 was evaluated against ALK resistant mutations in
enzymatic kinase assays with 10 .mu.M ATP concentration at Reaction
Biology, Inc. The results were summarized in Table 2. Potent
inhibition by Compound 1 was observed among wild type and mutant
ALKs and ROS1s.
TABLE-US-00004 TABLE 2 NPM- ALK ALK ALK ALK ALK ALK ALK Wild T1151M
1151Tins L1152R C1156Y F1174L IC.sub.50 (nM) 1.23 1.04 0.49 2.16
1.23 0.93 1.46 ALK ALK ALK ALK ALK ALK ALK F1174S L1196M G1202R
S1206R G1269A G1269S R1275Q IC.sub.50 (nM) 1.02 1.08 1.21 0.53 5.5
14.1 2.79 ROS1 ROS1 ROS1- WT G2032R TPM3 IC.sub.50 (nM) 0.0706
0.456 0.113
Example 6
Compound 1 Potently Inhibited Cell Proliferation in Primary Cell
Lines with Oncogenic Fusion or Mutated Genes of ALK, ROS1, TRKA or
JAK2
[0464] ALK fusions are major malignancy drivers in multiple cancer
types and cancer cell lines, including lymphoma cell line
Karpas-299 harboring NPM-ALK fusion gene, non-small cell lung
cancer cell line H2228 harboring EML4-ALK fusion gene, non-small
cell lung cancer cell line HCC78 harboring SLC34A2-ROS1 fusion
gene, colorectal cancer cell line KM12 harboring TPM3-TRKA fusion
gene, and leukemia cell line SET-2 harboring JAK2 V617F mutation.
The anti-proliferating activities of Compound 1 on these cell lines
were evaluated and the results were summarized in Table 3.
Example 7
Evaluation of Compound 1 Activity Against a Panel of ALK Gene
Mutations in Engineered Ba/F3 Cell Lines
[0465] Furthermore, we used Ba/F3 cells engineered to express wild
type EML4-ALK fusion gene and mutant EML4-ALKs. The growth of Ba/F3
cells is dependent on interleukin-s (IL-3). With ectopic expression
of EML4-ALK gene, Ba/F3 cell growth becomes IL-3 independent, and
relies on the kinase activity of the oncogenic fusion ALK. Compound
1 potently inhibited the cell proliferation of various Ba/F3 cell
lines with engineered expression of wild and mutant EML4-ALKs, and
the results were summarized in Table 3.
[0466] The inhibition of cell proliferation of Ba/F3 TPR-ALK and
Ba/F3 TPR-ALK L1196M cell lines were performed at Advanced Cellular
Dynamics. Compound 1 demonstrated potent inhibition in both cell
lines (Table 3).
Example 8
Evaluation of Compound 1 Activity Against ROS1 in Engineered Ba/F3
Cell Lines
[0467] Ba/F3 cells were engineered to express the oncogenic
SDC4-ROS1, SDC4-ROS1.sup.G2032R, CD74-ROS 1, and
CD74-ROS1.sup.G2032R fusion genes, respectively. The engineered
Ba/F3 cells with fusion ROS1 genes were used to examine the
inhibiting activity of Compound 1 on wild and mutant ROS1 fusion
genes. The results of cell growth inhibition were summarized in
Table 3.
[0468] Ba/F3 cells were engineered to express the oncogenic
CD74-ROS1.sup.L2026M and CD74-ROS1.sup.D2033N fusion genes,
respectively. The engineered Ba/F3 cells with fusion ROS1 genes
were used to examine the inhibiting activity of Compound 1. The
results of cell growth inhibition were summarized in Table 3.
[0469] Ba/F3 cells were engineered to express the oncogenic
LMNA-TRKA and LMNA-TRKA.sup.G595R fusion genes, respectively. The
engineered Ba/F3 cells with fusion TRKA genes were used to examine
the inhibiting activity of Compound 1. The results of cell growth
inhibition were summarized in Table 3.
[0470] Ba/F3 cells were engineered to express the oncogenic
TEL-TRKB (also named as ETV6-TRKB) and TEL-TRKB.sup.G639R fusion
genes, respectively. The engineered Ba/F3 cells with fusion TRKB
genes were used to examine the inhibiting activity of Compound ls.
The results of cell growth inhibition were summarized in Table
3.
[0471] Ba/F3 cells were engineered to express the oncogenic
TEL-TRKC (also named as ETV6-TRKC), and TEL-TRKC.sup.G623R fusion
genes, respectively. The engineered Ba/F3 cells with fusion TRKC
genes were used to examine the inhibiting activity of Compound 1.
The results of cell growth inhibition were summarized in Table
3.
TABLE-US-00005 TABLE 3 Assays IC.sub.50 (nM) Cell proliferation of
Karpas-299 cell line 23.7 Cell proliferation of NCI-H2228 cell line
73 Cell proliferation of HCC78 cell line 0.3 Cell proliferation of
KM12 cell line 0.3 Cell proliferation of SET-2 cell line 169 Cell
proliferation of Ba/F3 EML4-ALK 17.8 wild type cell line Cell
proliferation of Ba/F3 EML4-ALK 20.5 G1202R cell line Cell
proliferation of Ba/F3 EML4-ALK 85 L1152P cell line Cell
proliferation of Ba/F3 EML4-ALK 50 L1196M cell line Cell
proliferation of Ba/F3 EML4-ALK 54 F1174C cell line Cell
proliferation of Ba/F3 EML4-ALK 98 C1156Y cell line Cell
proliferation of Ba/F3 TPR-ALK 12 wild type cell line Cell
proliferation of Ba/F3 TPR-ALK 13.4 L1196M cell line Cell
proliferation of Ba/F3 CD74-ROS1 0.2 wild type cell line Cell
proliferation of Ba/F3 CD74-ROS1 8.4 G2032R cell line Cell
proliferation of Ba/F3 SDC4-ROS1 0.2 wild type cell line Cell
proliferation of Ba/F3 SDC4-ROS1 5 G2032R cell line Cell
proliferation of Ba/F3 + IL3 parental 1236 Cell proliferation of
Ba/F3 CD74-ROS1 5 L2026M cell line Cell proliferation of Ba/F3
CD74-ROS1 0.2 D2033N cell line Cell proliferation of Ba/F3
LMNA-TRKA cell line 0.2 Cell proliferation of Ba/F3 LMNA-TRKA 0.4
G595R cell line Cell proliferation of Ba/F3 TEL-TRKB 0.2
(ETV6-TRKB) cell line Cell proliferation of Ba/F3 TEL-TRKB 0.6
(ETV6-TRKB) G639R cell line Cell proliferation of Ba/F3 TEL-TRKC
0.2 (ETV6-TRKC) cell line Cell proliferation of Ba/F3 TEL-TRKC 3
(ETV6-TRKC) G623R cell line
Example 9
Mechanism of action of Compound 1 in Cells
[0472] The pharmacodynamic inhibiting activity of Compound 1 on
ALK, ROS1, TRKA and SRC, and the corresponding downstream signaling
in cells was evaluated, and the results were summarized in Table 4.
Compound 1 caused suppression of ALK autophosphorylation as well as
the downstream STAT3 and AKT phosphorylation at IC.sub.50s of
around 1-3 nM in Karpas-299 cell line, which harbors NPM-ALK fusion
gene. Compound 1 inhibited ROS1 autophosphorylation as well as the
downstream STAT3 and AKT phosphorylation at IC.sub.50s around 1-3
nM in HCC78 cell line, which harbors SLC34A2-ROS1 fusion gene.
Compound 1 inhibited TRKA autophosphorylation as well as the
downstream AKT and ERK phosphorylation at IC.sub.50s around 0.3 nM
in KM12 cell line, which harbors TPM3-TRKA fusion gene. Compound 1
inhibited STATS phosphorylation at IC.sub.50 around 158 nM in SET-2
cell line, which harbors JAK2 V617F mutation. Compound 1 inhibited
autophosphorylation of ALK with IC.sub.50 of 20-30 nM in Ba/F3
engineered stable cell lines encoding wild type or G1202R mutant
EML4-ALK vl fusion genes. Compound 1 inhibited autophosphorylation
of ROS1 in Ba/F3 engineered stable cell lines encoding wild type or
G2032R mutant CD74-ROS1 or SDC4-ROS1 fusion genes. The
pharmacodynamic inhibiting activity of Compound 1 on TRKA, TRKB,
TRKC, and FAK signaling in cells was evaluated, and the results
were summarized in Table 4a. Compound 1 inhibited FAK
phosphorylation as well as the SRC substrate paxillin
phosphorylation at IC.sub.50 around 103 nM in NCI-H2228 cell line,
which harbors EML4-ALK fusion gene with upregulated SRC and FAK
signaling. Compound 1 inhibited autophosphorylation of ROS1 in
Ba/F3 engineered stable cell lines encoding L2026M, or D2033N
mutant CD74-ROS1 fusion genes. Compound 1 inhibited
autophosphorylation of TRKA in NIH3T3 engineered stable cell lines
encoding wild type or G595R mutant LMNA-TRKA fusion genes. Compound
1 inhibited autophosphorylation of TRKB in NIH3T3 engineered stable
cell lines encoding wild type or G639R mutant TEL-TRKB, also named
as ETV6-TRKB fusion genes.
TABLE-US-00006 TABLE 4 Assays IC.sub.50 (nM) ALK phosphorylation of
Karpas-299 cell line 0.9 ALK phosphorylation of NCI-H2228 cell line
5.8 ALK phosphorylation of Ba/F3 29.9 EML4-ALK WT cell line ALK
phosphorylation of Ba/F3 EML4-ALK 18.4 G1202R cell line ROS1
phosphorylation of HCC78 cell line 2 ROS1 phosphorylation of Ba/F3
0.3 CD74-ROS1 WT cell line ROS1 phosphorylation of Ba/F3 CD74-ROS1
3 G2032R cell line ROS1 phosphorylation of Ba/F3 0.5 SDC4-ROS1 WT
cell line ROS1 phosphorylation of Ba/F3 SDC4-ROS1 2 G2032R cell
line TRKA phosphorylation of KM12 cell line 0.35 STAT5
phosphorylation of SET-2 cell line 158 SRC phosphorylation of H2228
cell line 102 ROS1 phosphorylation of Ba/F3 CD74-ROS1 10 L2026M
cell line ROS1 phosphorylation of Ba/F3 CD74-ROS1 <1 D2033N cell
line TRKA phosphorylation of NIH3T3 0.01 LMNA-TRKA cell line TRKA
phosphorylation of NIH3T3 LMNA-TRKA 0.1 G595R cell TRKB
phosphorylation of NIH3T3 <0.1 TEL-TRKB cell line TRKB
phosphorylation of NIH3T3 TEL-TRKB 1 G639R cell FAK phosphorylation
of NCI-H2228 cell line 100
[0473] Compound 1 increased the number of apoptotic Karpas-299
cells. After 48 hours treatment with Compound 1, Karpas-299 cells
were lysed and evaluated with the caspase 3/7 cleavage caspase 3
glo assay or PARP in western blotting assay. The results were
presented in FIG. 1.
[0474] Compound 1 inhibited HCC78 or HT1080 cell migration after 12
hour treatment in the scratch wound healing assays. The results
were presented in FIG. 2
[0475] Compound 1 Down-Regulated EGFR Expression in H2228 Cells
[0476] The activation of SRC kinase activity upregulates RTK
expression level, leading to the bypass signaling pathway
resistance. The inhibition of SRC will result in the down
regulation of RTKs. Compound 1 down-regulated EGFR expression in a
dose- and time-dependent manner as shown in FIG. 3.
[0477] Compound 1 Down-Regulated EGFR Expression in H2228 Cells
[0478] CD44 is a biomarker of cancer stemness. A high expression
level of CD44 was discovered in H2228 cells. Compound 1 suppressed
CD44 expression level in H2228 cells in a concentration-dependent
manner after 48 hours treatment as shown in FIG. 4, indicating that
compound 1 has the potential to inhibit cancer stemness.
[0479] In-Vivo Studies
[0480] Antitumor Efficacy of Compound 1 in Xenograft Tumor
Models
[0481] The antitumor efficacy of Compound 1 was evaluated in
several tumor xenograft models representing cancer populations in
which dysregulation of ALK, ROS1 or TRKA is implicated.
Example 10
Karpas 299 ALCL Model
[0482] The NPM-ALK fusion gene in Karpas 299 cells is proved as the
driver for tumor growth. SCID/Beige mice bearing Karpas 299 tumors
(at the average tumor size of 160 mm.sup.3) were dosed with
Compound 1 orally BID for seven days (FIG. 5). The control group of
mice were given vehicle only. Tumor volume (TMV) was measured by
caliper on the indicated days and is shown at mean.+-.sem in FIG.
5. An * denotes that the mean TMVs are significantly lower in the
treated group compared to that of the control group (p<0.05) as
determined by two-way repeated ANOVA followed by post hoc analysis.
Tumor growth inhibition (TGI) was calculated as
100%*{1-[(TMV.sub.Treated Last Day of Treatment-TMV.sub.Treated
First Day of Treatment)/(TMV.sub.Control on Last Day of
Treatment-TMV.sub.Control on First Day of Treatment)]} when
TMV.sub.Treated Last Day of Treatment.ltoreq.TMV.sub.Treated First
Day of Treatment. In the case of TMV.sub.Treaed Last Day of
Treatment<TMV.sub.Treated First Day of Treatment, tumor
regression (REG) was calculated as 100%*(1-TMV.sub.Treated Last Day
of Treatment/TMV.sub.Treated First Day of Treatment). In this
study, Compound 1 demonstrated the ability to inhibit tumor growth
at 59% and 94% at the dosages of 15 mg/kg and 50 mg/kg BID,
respectively. In addition, 4 of 8 mice in the 50 mg/kg treatment
group exhibited tumor regression. Body weight of the mice were
measured on the designated days of the mice and no body weight loss
was observed in the Compound 1 treatment groups (FIG. 6).
Example 11
NIH3T3 EML4-ALK Wildtype (WT) Model
[0483] Athymic nude mice bearing the NIH3T3 EML4-ALK WT tumors (at
the average tumor size of 120 mm.sup.3) were dosed with Compound 1
orally BID for 12 days (FIG. 7). The control group of mice were
given vehicle only. Tumor volume was measured by caliper on the
indicated days and is shown as mean.+-.sem in FIG. 7. An * denotes
that the mean tumor volumes are significantly lower in the treated
group compared to that of the control group (p<0.05) as
determined by two-way repeated ANOVA followed by post hoc analysis.
In this study, treatment of Compound 1 at 50 mg/kg/BID resulted in
41% tumor regression, with 8 of 8 mice exhibited tumor regression.
Compound 1 at the dosage of 15 mg/kg/BID demonstrated the ability
to inhibit tumor growth at a TGI of 90%, with 2 of 8 mice exhibited
tumor regression. Body weight of the mice were measured on the
designated days of the mice and no body weight loss was observed in
the Compound 1 treatment groups (FIG. 8).
Example 12
NIH3T3 SDC4-ROS1 WT Model
[0484] The SDC4-ROS1 fusion gene is considered as the driver for
tumor progression in NSCLC. Athymic nude mice bearing the NIH3T3
SDC4-ROS1 WT tumors (at the average tumor size of 100 mm.sup.3)
were dosed with Compound 1 orally BID for 26 days (FIG. 9). The
control group of mice were given vehicle only. Tumor volume was
measured by caliper on the indicated days and is shown at
mean.+-.sem in FIG. 9. An * denotes that the mean tumor volumes are
significantly less in the treated group compared to that of the
control group (p<0.05) as determined by two-way repeated ANOVA
followed by post hoc analysis. In this study, treatment of Compound
1 at 50 mg/kg resulted in 69% tumor regression, with 7 of 7 mice
exhibited tumor regression. Mice in the 15 mg/kg Compound 1 group
also demonstrated 39% tumor regression, with 7 of 8 mice in this
group exhibited tumor regression.
Example 13
KM12 Colorectal Cancer Model
[0485] The aberrant activity of the TPM3-TRKA is considered as the
underlying driver for tumor growth in the KM12 model. Athymic nude
mice bearing the KM12 tumors (at the average tumor size of 100
mm.sup.3) were dosed with Compound 1 orally BID for seven days
(FIG. 10). Tumor volume was measured by caliper on the indicated
days and is shown at mean.+-.sem in FIG. 10. An * denotes that the
mean tumor volumes are significantly lower in the treated group
compared to that of the control group (p<0.05) as determined by
two-way repeated ANOVA followed by post hoc analysis. In this
study, Compound 1 at the dosage of 15 mg/kg and 75 mg/kg resulted
in 13% and 11% tumor regression, respectively. In the 15 mg/kg
group, 8 of 8 mice exhibited tumor regression. In the 75 mg/kg
group, 5 of 8 mice exhibited tumor regression. No loss of body
weight were observed in mice treated with Compound 1 compared to
those with vehicle control (FIG. 11).
Example 14
Relationship of ALK Inhibition to Anti-Tumor Efficacy Following
Oral Administration of Compound 1
[0486] To evaluate the effect of Compound 1 on the inhibition of
ALK phosphorylation, Karpas 299 tumors were harvested at either 3
hour or 12 hour after last dose of Compound 1 in a repeated dosing
study (15 mg/kg or 50 mg/kg, BID for seven 7 days). The level of
ALK phosphorylation was determined by immunoblotting combined with
signal quantification by the Image Studio Digit Software. The
ability of Compound 1 to inhibit ALK phosphorylation was
illustrated in FIG. 12. At the dose of 50 mg/kg, ALK
phosphorylation was reduced to <5% of the control level at 3
hours post oral administration of Compound 1 and was maintained at
about 10% of the control level at 12 hours post dosing. This level
of phosphorylation inhibition corresponds to 94% TGI. At the dose
of 15 mg/kg, ALK phosphorylation was reduced to <10% of the
control level at 3 hours post oral administration and was
maintained at about 21% of the control level at 12 hours post
dosing. This level of ALK phosphorylation corresponds to 59% of
TGI. These results support the link between inhibition of ALK, the
target of Compound 1, and the degree of antitumor efficacy in a
NPM-ALK-dependent tumor model.
Example 15
Dose Dependent Studies in KM12 Colorectal Cancer Model
[0487] To investigate the dose-dependent effect of Compound 1 on
tumor inhibition in the KM12 colorectal cancer model, athymic nude
mice bearing the KM12 tumors (at the average tumor size of 125
mm.sup.3) were dosed with Compound 1 orally BID for seven days at
dosages equal or lower than 15 mg/kg for seven days (FIG. 13).
Tumor volume was measured by caliper on the indicated days and is
shown at mean.+-.sem in FIG. 13. An * denotes that the mean tumor
volumes are significantly lower in the treated group compared to
that of the control group (p<0.05) as determined by two-way
repeated ANOVA followed by post hoc analysis. In this study,
Compound 1 inhibited tumor growth in a dose-dependent manner.
Treatment with Compound 1 at the dosage of 15 mg/kg resulted in 1%
tumor regression, with 5 of 10 mice exhibiting tumor regression.
Compound 1 at the dosage of 3 mg/kg demonstrated the ability to
inhibit tumor growth at a TGI of 91%, with 2 of 10 mice exhibiting
tumor regression. Compound 1 at the dosage of 1 mg/kg demonstrated
the ability to inhibit tumor growth at a TGI of 77%. No loss of
body weight were observed in mice treated with Compound 1 compared
to those with vehicle control (FIG. 14).
Example 16
Ba/F3 EML4-ALK WT Model
[0488] SCID/Beige mice bearing the Ba/F3 EML4-ALK WT tumors (at the
average tumor size of 190 mm.sup.3) were dosed with Compound 1
orally BID for 14 days (FIG. 15). The control group of mice were
given vehicle only. Tumor volume was measured by caliper on the
indicated days and is shown as mean.+-.sem in FIG. 15. An * denotes
that the mean tumor volumes are significantly lower in the treated
group compared to that of the control group (p<0.05) as
determined by two-way repeated ANOVA followed by post hoc analysis.
In this study, treatment with Compound 1 at 75 mg/kg resulted in
54% tumor regression, with 6 of 8 mice exhibiting tumor regression.
Compound 1 at the dosage of 15 mg/kg/BID demonstrated the ability
to inhibit tumor growth at a TGI of 74%. Body weight of the mice
were measured on the designated days and no body weight loss was
observed in the Compound 1 treatment groups (FIG. 16).
Example 17
Ba/F3 EML4-ALK G1202R Model
[0489] To investigate the effect of Compound 1 on inhibiting the
growth of tumors containing the drug-resistant solvent front
mutations, SCID/Beige mice bearing the Ba/F3 EML4-ALK G1202R tumors
(at the average tumor size of 210 mm.sup.3) were dosed with
Compound 1 orally BID for 17 days (FIG. 17). The control group of
mice were given vehicle only. Tumor volume was measured by caliper
on the indicated days and is shown as mean.+-.sem in FIG. 17. An *
denotes that the mean tumor volumes are significantly lower in the
treated group compared to that of the control group (p<0.05) as
determined by two-way repeated ANOVA followed by post hoc analysis.
In this study, treatment of Compound 1 at 75 mg/kg resulted in
tumor growth inhibition with a TGI of 99%. 4 of 8 mice exhibited
tumor regression in this group. Compound 1 at the dosage of 15
mg/kg demonstrated the ability to inhibit tumor growth at a TGI of
56%. Body weight of the mice were measured on the designated days
and no body weight loss was observed in the Compound 1 treatment
groups (FIG. 18).
Example 18
Ba/F3 CD74-ROS1 WT Model
[0490] The CD74-ROS1 fusion gene is considered as one of the
drivers for tumor progression in NSCLC. SCID/Beige mice bearing the
Ba/F3 CD74-ROS1 WT tumors (at the average tumor size of 200
mm.sup.3) were dosed with Compound 1 orally BID for 12 days (FIG.
19). The control group of mice were given vehicle only. Tumor
volume was measured by caliper on the indicated days and is shown
at mean.+-.sem in FIG. 19. An * denotes that the mean tumor volumes
are significantly less in the treated group compared to that of the
control group (p<0.05) as determined by two-way repeated ANOVA
followed by post hoc analysis. In this study, treatment with
Compound 1 at 75 mg/kg resulted in 100% tumor regression, with 8 of
8 mice exhibiting complete tumor regression. Mice in the 15 mg/kg
Compound 1 group also demonstrated 97% tumor regression, with 8 of
8 mice in this group exhibiting tumor regression. Body weight of
the mice were measured on the designated days and no body weight
loss was observed in the Compound 1 treatment groups (FIG. 20).
Example 19
Ba/F3 CD74-ROS1 G2032R Model
[0491] To investigate the effect of Compound 1 on inhibiting the
growth of tumors containing the drug-resistant solvent front
mutations, SCID/Beige mice bearing the Ba/F3 CD74-ROS1 G2032R
tumors (at the average tumor size of 210 mm.sup.3) were dosed with
Compound 1 orally BID for 11 days (FIG. 21). The control group of
mice were given vehicle only. Tumor volume was measured by caliper
on the indicated days and is shown at mean.+-.sem in FIG. 21. An *
denotes that the mean tumor volumes are significantly less in the
treated group compared to that of the control group (p<0.05) as
determined by two-way repeated ANOVA followed by post hoc analysis.
In this study, treatment with Compound 1 at 75 mg/kg resulted in
100% tumor regression, with 8 of 8 mice exhibiting complete tumor
regression. Compound 1 at the dosage of 15 mg/kg/BID demonstrated
the ability to inhibit tumor growth at a TGI of 99%, with 2 of 8
mice in this group exhibiting tumor regression. Body weight of the
mice were measured on the designated days and no body weight loss
was observed in the Compound 1 treatment groups (FIG. 22).
Example 20
NIH3T3 LMNA-TRKA WT Model
[0492] The LMNA-TRKA fusion gene is considered as one of the
drivers for tumor progression in colorectal cancer. Athymic nude
mice bearing the NIH3T3 LMNA-TRKA WT tumors (at the average tumor
size of 240 mm.sup.3) were dosed with Compound 1 orally BID for
five days (FIG. 23). Tumor volume was measured by caliper on the
indicated days and is shown at mean.+-.sem in FIG. 23. An * denotes
that the mean tumor volumes are significantly lower in the treated
group compared to that of the control group (p<0.05) as
determined by two-way repeated ANOVA followed by post hoc analysis.
In this study, treatment with Compound 1 at the dosage of 15 mg/kg
resulted in 28% tumor regression, with 7 of 8 mice exhibiting tumor
regression. Compound 1 at the dosage of 3 mg/kg demonstrated the
ability to inhibit tumor growth at a TGI of 100%, with 3 of 8 mice
exhibiting tumor regression. No loss of body weight were observed
in mice treated with Compound 1 compared to those with vehicle
control (FIG. 24).
Example 21
NIH3T3 LMNA-TRKA G595R Model
[0493] To investigate the effect of Compound 1 on inhibiting the
growth of tumors containing the drug-resistant solvent front
mutations, athymic nude mice bearing the NIH3T3 LMNA-TRKA G595R
tumors (at the average tumor size of 230 mm.sup.3) were dosed with
Compound 1 orally BID for five days (FIG. 25). The control group of
mice were given vehicle only. Tumor volume was measured by caliper
on the indicated days and is shown at mean.+-.sem in FIG. 25. An *
denotes that the mean tumor volumes are significantly less in the
treated group compared to that of the control group (p<0.05) as
determined by two-way repeated ANOVA followed by post hoc analysis.
In this study, treatment with Compound 1 at 60 mg/kg resulted in
23% tumor regression, with 10 of 10 mice exhibiting tumor
regression. Compound 1 at the dosage of 15 mg/kg demonstrated the
ability to inhibit tumor growth at a TGI of 97%, with 3 of 8 mice
in this group exhibiting tumor regression. Compound 1 at the dosage
of 3 mg/kg demonstrated the ability to inhibit tumor growth at a
TGI of 56%. Body weight of the mice were measured on the designated
days and no body weight loss was observed in the Compound 1
treatment groups (FIG. 26).
* * * * *