U.S. patent application number 14/687992 was filed with the patent office on 2015-10-22 for bicyclic pyrazolone compounds and methods of use.
This patent application is currently assigned to Calitor Sciences, LLC. The applicant listed for this patent is Yanjun Wu, Ning Xi. Invention is credited to Yanjun Wu, Ning Xi.
Application Number | 20150299219 14/687992 |
Document ID | / |
Family ID | 54321425 |
Filed Date | 2015-10-22 |
United States Patent
Application |
20150299219 |
Kind Code |
A1 |
Xi; Ning ; et al. |
October 22, 2015 |
BICYCLIC PYRAZOLONE COMPOUNDS AND METHODS OF USE
Abstract
The present invention provides substituted bicyclic pyrazolone
compounds, which are used to inhibit or modulate the activity of
receptor tyrosine kinases, especially Axl, Mer, c-Met and Ron. The
invention also provides pharmaceutical compositions comprising the
compound disclosed herein, and a method of preventing, treating or
lessening the severity of a proliferative disorder in a patient
with the compounds or the pharmaceutical compositions disclosed
herein.
Inventors: |
Xi; Ning; (Newbury Park,
CA) ; Wu; Yanjun; (Dongguan, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Xi; Ning
Wu; Yanjun |
Newbury Park
Dongguan |
CA |
US
CN |
|
|
Assignee: |
Calitor Sciences, LLC
Newbury Park
CA
Sunshine Lake Pharma Co., Ltd.
Dongguan
|
Family ID: |
54321425 |
Appl. No.: |
14/687992 |
Filed: |
April 16, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61982729 |
Apr 22, 2014 |
|
|
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Current U.S.
Class: |
424/85.7 ;
514/230.5; 514/300; 514/333; 514/338; 544/105; 546/121; 546/256;
546/275.7 |
Current CPC
Class: |
A61K 31/5383 20130101;
C07D 471/04 20130101; A61P 43/00 20180101; A61P 35/00 20180101;
C07D 498/04 20130101; A61K 31/4439 20130101; C12Q 1/485 20130101;
A61K 9/0095 20130101; A61P 35/04 20180101; A61K 9/0019 20130101;
A61K 31/444 20130101; C07D 487/04 20130101; A61K 31/444 20130101;
A61K 47/38 20130101; A61K 31/437 20130101; A61K 31/5395 20130101;
A61P 11/00 20180101; C12N 9/12 20130101; A61K 31/5383 20130101;
A61P 35/02 20180101; A61K 47/26 20130101; A61K 45/06 20130101; A61K
31/5395 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101;
A61K 2300/00 20130101; A61K 2300/00 20130101; A61K 31/4439
20130101; A61P 9/10 20180101; C12Y 207/10001 20130101 |
International
Class: |
C07D 498/04 20060101
C07D498/04; A61K 31/4439 20060101 A61K031/4439; A61K 31/5383
20060101 A61K031/5383; C07D 471/04 20060101 C07D471/04; A61K 31/444
20060101 A61K031/444; C07D 487/04 20060101 C07D487/04; A61K 45/06
20060101 A61K045/06 |
Claims
1. A compound having Formula (I): ##STR00040## or a stereoisomer, a
tautomer, an N-oxide, a solvate, a metabolite, a pharmaceutically
acceptable salt or a prodrug thereof, wherein: Q is H, OR.sup.a,
NR.sup.aR.sup.b, --C(.dbd.O)NR.sup.aR.sup.b,
--N(R.sup.c)C(.dbd.O)R.sup.d, --N(R.sup.c)C(.dbd.O)OR.sup.a or
--N(R.sup.c)C(.dbd.O)NR.sup.aR.sup.b; U is CR.sup.7 or N, provided
that when U is N, the compound is not
2-oxo-1-phenyl-N-(5-((2-(pyrrolidine-1-carboxamido)pyridin-4-yl)oxy)pyrid-
in-2-yl)-2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
or
1-(buta-1,3-dien-2-yl)-2-oxo-N-(5-((2-(pyrrolidine-1-carboxamido)pyrid-
in-4-yl)oxy)pyridin-2-yl)-1,2,4,5,6,7-hexahydropyrazolo[1,5-.alpha.]pyrazi-
ne-3-carboxamide; X is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
alkenyl, C.sub.3-C.sub.8 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl or
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), wherein
each of the C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 alkenyl,
C.sub.3-C.sub.8 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl and
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl) is
optionally substituted with 1, 2, 3, 4 or 5 substituents
independently selected from F, Cl, Br, CN, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, OR.sup.a,
NR.sup.aR.sup.b, --(C.sub.1-C.sub.4 alkylene)-OR.sup.a and
--(C.sub.1-C.sub.4 alkylene)-NR.sup.aR.sup.b; each Y is
independently 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.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl,
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), OR.sup.a,
NR.sup.aR.sup.b, --(C.sub.1-C.sub.4 alkylene)-OR.sup.a or
--(C.sub.1-C.sub.4 alkylene)-NR.sup.aR.sup.b; m is 0, 1, 2, 3, 4; W
is --(CH.sub.2).sub.n--, --(CH.sub.2).sub.nO--,
--(CH.sub.2).sub.nNH-- or --(CH.sub.2).sub.nS--, wherein n is 0, 1,
2, 3 or 4; each of R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5,
R.sup.6 and R.sup.7 is independently H, F, Cl, Br, CN, N.sub.3,
OR.sup.a, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.2-C.sub.6 alkenyl or C.sub.2-C.sub.6 alkynyl; each of
R.sup.a, R.sup.b and R.sup.c is independently H, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.3-C.sub.6 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-6 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl or
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), or R.sup.a
and R.sup.b taken together with the nitrogen atom to which they are
attached form a 3-8 membered heterocyclic ring, wherein each of the
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.3-C.sub.6
cycloalkyl, --(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.6
cycloalkyl), 3-6 membered heterocyclyl, --(C.sub.1-C.sub.4
alkylene)-(3-6 membered heterocyclyl), C.sub.6-C.sub.10 aryl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered
heteroaryl, --(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl)
and 3-8 membered heterocyclic ring is optionally substituted with
1, 2, 3 or 4 substituents independently selected from F, Cl, CN,
N.sub.3, OH, NH.sub.2, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
alkoxy and C.sub.1-C.sub.6 alkylamino; and R.sup.d is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.5 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl or
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), wherein
each of the C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl and
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl) is
optionally substituted with 1, 2, 3 or 4 substituents independently
selected from F, Cl, Br, CN, OR.sup.a, NR.sup.aR.sup.b,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, --(C.sub.1-C.sub.4 alkylene)-ORB and --(C.sub.1-C.sub.4
alkylene)-NR.sup.aR.sup.b.
2. The compound of claim 1, wherein Q is NR.sup.aR.sup.b,
--C(.dbd.O)NR.sup.aR.sup.b, --N(R.sup.c)C(.dbd.O)R.sup.d or
--N(R.sup.c)C(.dbd.O)NR.sup.aR.sup.b.
3. The compound of claim 1, wherein X is C.sub.1-C.sub.4 alkyl,
C.sub.3-C.sub.6 alkenyl, C.sub.3-C.sub.6 alkynyl, C.sub.3-C.sub.6
cycloalkyl, --(C.sub.1-C.sub.2 alkylene)-(C.sub.3-C.sub.6
cycloalkyl), phenyl or --(C.sub.1-C.sub.2 alkylene)-phenyl, wherein
each of the C.sub.1-C.sub.4 alkyl, C.sub.3-C.sub.6 alkenyl,
C.sub.3-C.sub.6 alkynyl, C.sub.3-C.sub.6 cycloalkyl,
--(C.sub.1-C.sub.2 alkylene)-(C.sub.3-C.sub.6 cycloalkyl), phenyl
and --(C.sub.1-C.sub.2 alkylene)-phenyl is optionally substituted
with 1, 2, 3, 4 or 5 substituents independently selected from F,
Cl, Br, CN, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 alkynyl, OR.sup.a, NR.sup.aR.sup.b,
--(C.sub.1-C.sub.2 alkylene)-OR.sup.a and --(C.sub.1-C.sub.2
alkylene)-NR.sup.aR.sup.b.
4. The compound of claim 1, wherein each Y is independently
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.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl,
--(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl), phenyl,
--(C.sub.1-C.sub.2 alkylene)-phenyl, 5-6 membered heteroaryl,
--(C.sub.1-C.sub.2 alkylene)-(5-6 membered heteroaryl), OR.sup.a,
NR.sup.aR.sup.b, --(C.sub.1-C.sub.2 alkylene)-OR.sup.a or
--(C.sub.1-C.sub.2 alkylene)-NR.sup.aR.sup.b; m is 0, 1 or 2.
5. The compound of claim 1, wherein W is --(CH.sub.2).sub.n--,
--(CH.sub.2).sub.nO-- or --(CH.sub.2).sub.nNH--, wherein n is 0, 1
or 2.
6. The compound of claim 1, wherein each of R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6 and R.sup.7 is independently H,
F, Cl, Me or OMe.
7. The compound of claim 1, wherein each of R.sup.a, R.sup.b and
R.sup.c is independently H, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4
haloalkyl, C.sub.3-C.sub.6 cycloalkyl, --(C.sub.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl
or --(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl), or
R.sup.a and R.sup.b taken together with the nitrogen atom to which
they are attached form a 3-8 membered heterocyclic ring, wherein
each of the C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 haloalkyl,
C.sub.3-C.sub.6 cycloalkyl, --(C.sub.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl,
--(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl) and 3-8
membered heterocyclic ring is optionally substituted with 1, 2, 3
or 4 substituents independently selected from F, Cl, CN, N.sub.3,
OH, NH.sub.2, C.sub.1-C.sub.3 haloalkyl, C.sub.1-C.sub.3 alkoxy and
C.sub.1-C.sub.3 alkylamino.
8. The compound of claim 1, wherein R.sup.d is H, C.sub.1-C.sub.4
alkyl, C.sub.3-C.sub.6 cycloalkyl, --(C.sub.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl
or --(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl),
wherein each of the C.sub.1-C.sub.4 alkyl, C.sub.3-C.sub.6
cycloalkyl, --(C.sub.1-C.sub.2 alkylene)-(C.sub.3-C.sub.6
cycloalkyl), 3-6 membered heterocyclyl and --(C.sub.1-C.sub.2
alkylene)-(3-6 membered heterocyclyl) is optionally substituted
with 1, 2, 3 or 4 substituents independently selected from F, CN,
OR.sup.a, NR.sup.aR.sup.b, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4
alkenyl, C.sub.2-C.sub.4 alkynyl, --(C.sub.1-C.sub.2
alkylene)-OR.sup.a and --(C.sub.1-C.sub.2
alkylene)-NR.sup.aR.sup.b.
9. The compound of claim 1, wherein Q is: ##STR00041##
##STR00042##
10. The compound of claim 1 having one of the following structures:
##STR00043## ##STR00044## ##STR00045## ##STR00046## ##STR00047##
##STR00048##
11. A pharmaceutical composition comprising the compound of claim
1, and a pharmaceutically acceptable excipient, carrier, adjuvant,
vehicle or a combination thereof.
12. The pharmaceutical composition of claim 11 further comprising a
therapeutic agent selected from the group consisting of
chemotherapeutic agents, anti-proliferative agents, agents for
treating atherosclerosis, agents for treating lung fibrosis and
combinations thereof.
13. The pharmaceutical composition of claim 12, wherein the
therapeutic agent is chlorambucil, melphalan, cyclophosphamide,
ifosfamide, busulfan, carmustine, lomustine, streptozocin,
cisplatin, carboplatin, oxaliplatin, dacarbazine, temozolomide,
procarbazine, methotrexate, fluorouracil, cytarabine, gemcitabine,
mercaptopurine, fludarabine, vinblastine, vincristine, vinorelbine,
paclitaxel, docetaxel, topotecan, irinotecan, etoposide,
trabectedin, dactinomycin, doxorubicin, epirubicin, daunorubicin,
mitoxantrone, bleomycin, mitomycin, ixabepilone, tamoxifen,
flutamide, gonadorelin analogues, megestrol, prednisone,
dexamethasone, methylprednisolone, thalidomide, interferon alfa,
leucovorin, sirolimus, temsirolimus, everolimus, afatinib,
alisertib, amuvatinib, apatinib, axitinib, bortezomib, bosutinib,
brivanib, cabozantinib, cediranib, crenolanib, crizotinib,
dabrafenib, dacomitinib, danusertib, dasatinib, dovitinib,
erlotinib, foretinib, ganetespib, gefitinib, ibrutinib, icotinib,
imatinib, iniparib, lapatinib, lenvatinib, linifanib, linsitinib,
masitinib, momelotinib, motesanib, neratinib, nilotinib, niraparib,
oprozomib, olaparib, pazopanib, pictilisib, ponatinib, quizartinib,
regorafenib, rigosertib, rucaparib, ruxolitinib, saracatinib,
saridegib, sorafenib, sunitinib, tasocitinib, telatinib,
tivantinib, tivozanib, tofacitinib, trametinib, vandetanib,
veliparib, vemurafenib, vismodegib, volasertib, alemtuzumab,
bevacizumab, brentuximabvedotin, catumaxomab, cetuximab, denosumab,
gemtuzumab, ipilimumab, nimotuzumab, ofatumumab, panitumumab,
ramucirumab, rituximab, tositumomab, trastuzumab or a combination
thereof.
14. A method of preventing, treating or lessening the severity of a
proliferative disease in a patient by administering to the patient
with a therapeutically effective amount of the compound of claim
1.
15. The method of claim 14, wherein the proliferative disease is
colon cancer, rectal cancer, gastric cancer, gastric
adenocarcinoma, pancreatic cancer, bladder cancer, gallbladder
cancer, breast cancer, kidney cancer, renal cell carcinoma, liver
cancer, hepatocellular carcinoma, lung cancer, skin cancer,
melanoma, thyroid cancer, osteosarcomas, soft tissue sarcoma, a
cancer of the head and neck, a cancer of the central nervous
system, glioma, glioblastomas, ovarian cancer, uterine cancer,
endometrial carcinoma, prostate cancer, acute myeloid leukemia or
acute lymphoblastic leukemia, or a metastasis thereof.
16. The method of claim 14, wherein the proliferative disease is
atherosclerosis or lung fibrosis.
17. A method of preventing, treating or lessening the severity of a
proliferative disease in a patient by administering to the patient
with a therapeutically effective amount of the pharmaceutical
composition of claim 11.
18. The method of claim 17, wherein the proliferative disease is
colon cancer, rectal cancer, gastric cancer, gastric
adenocarcinoma, pancreatic cancer, bladder cancer, gallbladder
cancer, breast cancer, kidney cancer, renal cell carcinoma, liver
cancer, hepatocellular carcinoma, lung cancer, skin cancer,
melanoma, thyroid cancer, osteosarcomas, soft tissue sarcoma, a
cancer of the head and neck, a cancer of the central nervous
system, glioma, glioblastomas, ovarian cancer, uterine cancer,
endometrial carcinoma, prostate cancer, acute myeloid leukemia or
acute lymphoblastic leukemia, or a metastasis thereof.
19. The method of claim 17, wherein the proliferative disease is
atherosclerosis or lung fibrosis.
20. A method of inhibiting or modulating the activity of a protein
kinase in a biological sample comprising contacting the biological
sample with the compound of claim 1.
21. The method of claim 20, wherein the protein kinase is a
receptor tyrosine kinase.
22. The method of claim 21, wherein the receptor tyrosine kinase is
Axl, Mer, c-Met, Ron or a combination thereof.
23. A method of inhibiting or modulating the activity of a protein
kinase in a biological sample comprising contacting the biological
sample with the pharmaceutical composition of claim 11.
24. The method of claim 23, wherein the protein kinase is a
receptor tyrosine kinase.
25. The method of claim 24, wherein the receptor tyrosine kinase is
Axl, Mer, c-Met, Ron or a combination thereof.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application claims the benefit of U.S. Provisional
Application Ser. No. 61/982,729, filed on Apr. 22, 2014, which is
hereby incorporated by reference in its entirety.
FIELD OF THE INVENTION
[0002] This invention relates to novel substituted bicyclic
pyrazolone compounds and salts thereof, which are useful in the
treatment of proliferative diseases, such as cancers, in mammals.
In particular, the invention relates to compounds that inhibit the
protein tyrosine kinase activity, resulting in the inhibition of
inter- and/or intra-cellular signaling. This invention also relates
to a method of using such compounds in the treatment of
hyperproliferative diseases in mammals, especially humans, and to
pharmaceutical compositions containing such compounds.
BACKGROUND OF THE INVENTION
[0003] Protein kinases represent a large family of proteins that
play a central role in the regulation of a wide variety of cellular
processes. Through regulating an array of signaling pathways,
protein kinases control cell metabolism, cell cycle progression,
cell proliferation and cell death, differentiation and survival.
There are over 500 kinases in the human kinome, and over 150 of
these have been shown or are proposed to be involved in the onset
and/or progression of various human diseases including inflammatory
diseases, cardiovascular diseases, metabolic diseases,
neurodegenerative diseases and cancer.
[0004] A partial list of such kinases include abl, AATK, ALK, Akt,
Axl, bmx, bcr-abl, Blk, Brk, Btk, csk, c-kit, c-Met, c-src, c-fins,
CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, cRaf1,
CSF1R, CSK, DDR1, DDR2, EPHA, EPHB, EGFR, ErbB2, ErbB3, ErbB4, Erk,
Fak, fes, FER, FGFR1, FGFR2, FGFR3, FGFR3, FGFR4, FGFR5, Fgr,
flt-1, Fps, Frk, Fyn, GSG2, GSK, Hck, LLK, INSRR, IRAK4, ITK,
IGF-1R, INS-R, Jak, KSR1, KDR, LMTK2, LMTK3, LTK, Lck, Lyn, MATK,
MERTK (Mer), MLTK, MST1R (Ron), MUSK, NPR1, NTRK, MEK, MET, PLK4,
PTK, p38, PDGFR, PIK, PKC, PYK2, RET, ROR1, ROR2, RYK, ros, SGK493,
SRC, SRMS, STYK1, SYK, TEC, TEK, TEX14, TNK1, TNK2, TNNI3K, TXK,
TYK2, Tyro-3, tie, tie2, TRK, Yes, and Zap 70.
[0005] Protein tyrosine kinases are a subclass of protein kinase.
They also may be classified as growth factor receptor (e.g., Axl,
Mer, c-Met (HGFR), Ron, EGFR, PDGFR and FGFR) or non-receptor
(e.g., c-src and bcr-abl) kinases. Receptor tyrosine kinases are
transmembrane proteins that possess an extracellular binding domain
for growth factors, a transmembrane domain, and an intracellular
portion that functions as a kinase to phosphorylate a specific
tyrosine residue in proteins. Abnormal expression or activity of
protein kinases has been directly implicated in the pathogenesis of
myriad human cancers.
[0006] Axl and Mer are both members of the TAM receptor family,
which also includes Tyro3. All three are activated by a common
ligand, Growth Arrest-specific protein 6 (Gas6), and they
ordinarily play an embryonic developmental role in cell survival,
migration and differentiation. The TAM receptors are characterized
by a combination of two immunoglobulin-like domains and dual
fibronectin type III repeats in the extracellular region and a
cytoplasmic kinase domain (Trevor et al., "The anticoagulation
factor protein S and its relative, Gas6, are ligands for the
Tyro3/Axl family of receptor tyrosine kinases" Cell, 1995, 80,
661-670; Varnum et al., "Axl receptor tyrosine kinase stimulated by
the vitamin K-dependent protein encoded by growth-arrest-specific
gene 6" Nature, 1995, 373, 623-626).
[0007] Axl signaling is required to maintain EMT-associated
features including invasiveness and metastasis (Linger et al., "TAM
receptor tyrosine kinases: biologic functions, signaling, and
potential therapeutic targeting in human cancer" Adv. Cancer Res.,
2008, 100, 35-83). Axl overexpression and signaling has been
implicated in several human malignancies, such as colon, breast,
glioma, thyroid, gastric, melanoma, lung cancer, and in renal cell
carcinoma (RCC). A more detailed role of Axl biology has been
proven in glioma, where loss of Axl signaling diminished glioma
tumor growth, and in breast cancer, where Axl drive cell migration,
tube formation, neovascularization and tumor growth. Axl has been
shown to play multiple roles in tumorigenesis and that therapeutic
antibodies against Axl may block Axl functions not only in
malignant tumor cells but also in the tumor stroma. The additive
effect of Axl inhibition with anti-VEGF suggests that blocking Axl
function could be an effective approach for enhancing
antiangiogenic therapy (Li et al., "Axl as a potential therapeutic
target in cancer: role of Axl in tumor growth, metastasis and
angiogenesis" Oncogene, 2009, 28, 3442-3455; and Linger et al.,
"TAM receptor tyrosine kinases: biologic functions, signaling, and
potential therapeutic targeting in human cancer" Adv. Cancer Res.,
2008, 100, 35-83).
[0008] High levels of Axl expression have been correlated with poor
survival in many types of cancer, including breast cancer
(Christine et al., "Axl is an essential epithelial-to-mesenchymal
transition-induced regulator of breast cancer metastasis and
patient survival" Proc. Natl. Acad. Sci. USA, 2010, 107(3),
1124-1129), acute myeloid leukemia (Amer. Soc. Hematol. Annual
Meeting, San Diego 2011), glioblastoma multiforme (Markus et al.,
"Axl and growth arrest-specific gene 6 are frequently overexpressed
in human gliomas and predict poor prognosis in patients with
glioblastoma multiforme" Clin. Cancer Res., 2008, 14, 130-138) and
osteosarcoma (Han et al., "Gas6/Axl mediates tumor cell apoptosis,
migration and invasion and predicts the clinical outcome of
osteosarcoma patients" Biochem. Biophys. Res. Commun., 2013,
435(3), 493-500). In addition, activation of Axl kinase has been
identified as one mechanism by which lung cancers can develop
resistance to therapies targeting EGFR, such as Tarceva (erlotinib)
(Zhang et al., "Activation of the Axl kinase causes resistance to
EGFR-targeted therapy in lung cancer" Nat. Genet., 2012, 44(8),
852-860).
[0009] Mer expression correlates with disease progression. It has
been found that Mer expression was high in metastatic melanomas
(Jennifer et al., "MERTK receptor tyrosine kinase is a therapeutic
target in melanoma" J. Clin. Invest., 2013, 123(5), 2257-2267), and
activation of Mer promotes invasion and survival in glioblastoma
multiforme (Wang et al., "Mer receptor tyrosine kinase promotes
invasion and survival in glioblastoma multiforme" Oncogene, 2013,
32, 872-882). Studies also indicated a role for Mer in acute
lymphoblastic leukemia (ALL). Mer is ectopically expressed in at
least 50% of pediatric T-cell ALL samples as well as in pre-B ALL
samples (Graham et al., "Ectopic expression of the proto-oncogene
Mer in pediatric T-cell acute lymphoblastic leukemia" Clin. Cancer
Res., 2006, 12(9), 2662-2669). Thus, Mer receptor tyrosine kinase
is proposed to be a therapeutic target for various solid or
hematological malignancies.
[0010] Recently a study showed that Mer and Axl were frequently
overexpressed and activated in NSCLC cell lines. Ligand--dependent
Mer or Axl activation stimulated MAPK, AKT and FAK signaling
pathways indicating roles for these RTKs in multiple oncogenic
processes. Abnormal expression and activation of Axl knockdown also
improved in vitro NSCLC sensitivity to chemotherapeutic agents by
promoting apoptosis. When comparing the effects of Mer and Axl
knockdown, Mer inhibition exhibited more complete blockade of tumor
growth while Axl knockdown more robustly improved chemosensitivity.
These results indicated that Mer and Axl play complementary and
overlapping roles in NSCLC and suggest that treatment strategies
targeting both RTKs may be more effective than singly-targeted
agents. Therefore, inhibition of both Axl and Mer is potentially a
therapeutic strategy to target cancer cells (Rachel et al., "Mer or
Axl Receptor Tyrosine Kinase inhibition promotes apoptosis, blocks
growth, and enhances chemosensitivity of human non-small cell lung
cancer" Oncogene, 2013, 32(29), 3420-3431).
[0011] c-Met, also referred to as hepatocyte growth factor receptor
(HGFR), is expressed predominantly in epithelial cells but has also
been identified in endothelial cells, myoblasts, hematopoietic
cells and motor neurons. The natural ligand for c-Met is hepatocyte
growth factor (HGF), also known as scatter factor (SF). In both
embryos and adults, activated c-Met promotes a morphogenetic
program, known as invasive growth, which induces cell spreading,
the disruption of intercellular contacts, and the migration of
cells towards their surroundings (Peschard et al., "From Tpr-Met to
Met, tumorigenesis and tubes" Oncogene, 2007, 26, 1276-1285; and
Christine et al., "MET receptor tyrosine kinase as a therapeutic
anticancer target" Cancer Letters, 2009, 280(1), 1-14).
[0012] A wide variety of human malignancies exhibit sustained c-Met
stimulation, overexpression or mutation, including carcinomas of
the breast, liver, lung, ovary, kidney, thyroid, colon,
glioblastomas and prostate, etc. c-Met is also implicated in
atherosclerosis and lung fibrosis. Invasive growth of certain
cancer cells is drastically enhanced by tumor-stromal interactions
involving the HGF/c-Met pathway. Thus, extensive evidence that
c-Met signaling is involved in the progression and spread of
several cancers and an enhanced understanding of its role in
disease have generated considerable interest in c-Met as major
targets in cancer drug development (Cristina et al., "Molecular
cancer therapy: Can our expectation be MET" Eur. J. Cancer, 2008,
44(5) 641-651; and Peruzzi et al., "Targeting the c-Met signaling
pathway in cancer" Clin. Cancer Res., 2006, 12(12), 3657-3660).
[0013] Ron (MST1R, recepteur d'origine nantais), the other member
of the MET family, is a receptor tyrosine kinase for the ligand
macrophage-stimulating protein (MSP, also known as MST1, and
hepatocyte growth factor-like (HGFL) protein), which is associated
with in vitro and in vivo cell dissociation, motility and matrix
invasion--all of which are surrogate markers of an aggressive
cancer phenotype with metastatic potential. Ron mediates oncogenic
phenotypes in lung, thyroid, pancreas, prostate, colon and breast
cancer cells and predicts a poor prognosis in human breast cancer.
Co-expression of Ron with Met and the induction of Ron expression
by HGF-Met signaling have both been described in hepatocellular
carcinoma. Furthermore, co-expression of Met and Ron portends a
worse prognosis in ovary, breast and bladder cancers. Given Ron and
Met signaling redundancy, it is possible that resistance to Met
inhibition is mediated by Ron signaling (Catenacci et al., "RON
(MST1R) is a novel prognostic marker and therapeutic target for
gastroesophageal adenocarcinoma" Cancer Biol. Ther., 2011, 12(1),
9-46).
[0014] The roles of MSP-Ron signaling axis in cancer pathogenesis
has also been extensively studied in various model systems. Both in
vitro and in vivo evidence has revealed that MSP-Ron signaling is
important for the invasive growth of different types of cancers.
Aberrant Ron activation, which is induced by overexpression of
protein and the generation of oncogenic isoforms and is indicated
by the persistent activation of multi-intracellular signaling
cascades, occurs in various types of cancers. Ron signaling is also
necessary for cancer cell growth and survival. These features
render Ron as a drug target for cancer therapy (Yao et al.,
"MSP-RON signalling in cancer: pathogenesis and therapeutic
potential" Nat. Rev. Cancer, 2013, 13(7), 466-481).
[0015] It is widely known that cancer cells employ multiple
mechanisms to evade tightly regulated cellular processes such as
proliferation, apoptosis and senescence. Thus, most tumors can
escape from the inhibition of any single kinase. System-wide
analysis of tumors identified receptor tyrosine kinase (RTK)
coactivation as an important mechanism by which cancer cells
achieve chemoresistance. One of the strategies to overcome RTK
coactivation may involve therapeutically targeting multiple RTKs
simultaneously in order to shut down oncogenic RTK signaling and
overcome compensatory mechanisms (Alexander et al., "Receptor
tyrosine kinase coactivation metworks in cancer" Cancer Res., 2010,
70, 3857-3860). Anti-tumor approaches in targeting Axl, Mer, c-Met
and/or Ron signaling may circumvent the ability of tumor cells to
overcome Axl, Mer (MERTK), c-Met (HGFR) and/or Ron (MST1R)
inhibition alone and thus may represent improved cancer
therapeutics.
SUMMARY OF THE INVENTION
[0016] The present invention provides substituted bicyclic
pyrazolone compounds which are useful as multiple function
inhibitors, capable of inhibiting, for example, Axl, Mer (MERTK),
c-Met (HGFR) and/or Ron (MST1R). The invention also provides
methods of making the compounds, methods of using such compounds in
the treatment of diseases and conditions proliferative diseases in
mammals and pharmaceutical compositions comprising such
compounds.
[0017] Specifically, in one aspect, provided herein is a compound
having Formula (I):
##STR00001##
or a stereoisomer, a tautomer, an N-oxide, a solvate, a metabolite,
a pharmaceutically acceptable salt or a prodrug thereof, wherein: Q
is H, OR.sup.a, NR.sup.aR.sup.b, --C(.dbd.O)NR.sup.aR.sup.b,
--N(R.sup.c)C(.dbd.O)R.sup.d, --N(R.sup.c)C(.dbd.O)OR.sup.a or
--N(R.sup.c)C(.dbd.O)NR.sup.aR.sup.b; U is CR.sup.7 or N, provided
that when U is N, the compound is not
2-oxo-1-phenyl-N-(5-((2-(pyrrolidine-1-carboxamido)pyridin-4-yl)oxy)pyrid-
in-2-yl)-2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
or
1-(buta-1,3-dien-2-yl)-2-oxo-N-(5-((2-(pyrrolidine-1-carboxamido)pyrid-
in-4-yl)oxy)pyridin-2-yl)-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyrazine-3-c-
arboxamide; X is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 alkenyl,
C.sub.3-C.sub.8 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl or
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), wherein
each of the C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 alkenyl,
C.sub.3-C.sub.8 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl and
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl) is
optionally substituted with 1, 2, 3, 4 or 5 substituents
independently selected from F, Cl, Br, CN, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, OR.sup.a,
NR.sup.aR.sup.b, --(C.sub.1-C.sub.4 alkylene)-OR.sup.a and
--(C.sub.1-C.sub.4 alkylene)-NR.sup.aR.sup.b; each Y is
independently 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.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl,
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), OR.sup.a,
NR.sup.aR.sup.b, --(C.sub.1-C.sub.4 alkylene)-OR.sup.a or
--(C.sub.1-C.sub.4 alkylene)-NR.sup.aR.sup.b; m is 0, 1, 2, 3, 4; W
is --(CH.sub.2).sub.n--, --(CH.sub.2).sub.nO--,
--(CH.sub.2).sub.nNH-- or --(CH.sub.2).sub.nS--, wherein n is 0, 1,
2, 3 or 4; each of R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5,
R.sup.6 and R.sup.7 is independently H, F, Cl, Br, CN, N.sub.3,
OR.sup.a, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.2-C.sub.6 alkenyl or C.sub.2-C.sub.6 alkynyl; each of
R.sup.a, R.sup.b and R.sup.c is independently H, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.3-C.sub.6 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-6 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl or
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), or R.sup.a
and R.sup.b taken together with the nitrogen atom to which they are
attached form a 3-8 membered heterocyclic ring, wherein each of the
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.3-C.sub.6
cycloalkyl, --(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.6
cycloalkyl), 3-6 membered heterocyclyl, --(C.sub.1-C.sub.4
alkylene)-(3-6 membered heterocyclyl), C.sub.6-C.sub.10 aryl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered
heteroaryl, --(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl)
and 3-8 membered heterocyclic ring is optionally substituted with
1, 2, 3 or 4 substituents independently selected from F, Cl, CN,
N.sub.3, OH, NH.sub.2, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
alkoxy and C.sub.1-C.sub.6 alkylamino; and R.sup.d is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl or
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), wherein
each of the C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl and
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl) is
optionally substituted with 1, 2, 3 or 4 substituents independently
selected from F, Cl, Br, CN, OR.sup.a, NR.sup.aR.sup.b,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, --(C.sub.1-C.sub.4 alkylene)-OR.sup.a and
--(C.sub.1-C.sub.4 alkylene)-NR.sup.aR.sup.b.
[0018] In one embodiment, Q is NR.sup.aR.sup.b,
--C(.dbd.O)NR.sup.aR.sup.b, --N(R.sup.c)C(.dbd.O)R.sup.d or
--N(R.sup.c)C(.dbd.O)NR.sup.aR.sup.b.
[0019] In another embodiment, X is C.sub.1-C.sub.4 alkyl,
C.sub.3-C.sub.6 alkenyl, C.sub.3-C.sub.6 alkynyl, C.sub.3-C.sub.6
cycloalkyl, --(C.sub.1-C.sub.2 alkylene)-(C.sub.3-C.sub.6
cycloalkyl), phenyl or --(C.sub.1-C.sub.2 alkylene)-phenyl, wherein
each of the C.sub.1-C.sub.4 alkyl, C.sub.3-C.sub.6 alkenyl,
C.sub.3-C.sub.6 alkynyl, C.sub.3-C.sub.6 cycloalkyl,
--(C.sub.1-C.sub.2 alkylene)-(C.sub.3-C.sub.6 cycloalkyl), phenyl
and --(C.sub.1-C.sub.2 alkylene)-phenyl is optionally substituted
with 1, 2, 3, 4 or 5 substituents independently selected from F,
Cl, Br, CN, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 alkynyl, OR.sup.a, NR.sup.aR.sup.b,
--(C.sub.1-C.sub.2 alkylene)-OR.sup.a and --(C.sub.1-C.sub.2
alkylene)-NR.sup.aR.sup.b.
[0020] In another embodiment, each Y is independently
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.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl,
--(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl), phenyl,
--(C.sub.1-C.sub.2 alkylene)-phenyl, 5-6 membered heteroaryl,
--(C.sub.1-C.sub.2 alkylene)-(5-6 membered heteroaryl), OR.sup.a,
NR.sup.aR.sup.b, --(C.sub.1-C.sub.2 alkylene)-OR.sup.a or
--(C.sub.1-C.sub.2 alkylene)-NR.sup.aR.sup.b; m is 0, 1 or 2.
[0021] In another embodiment, W is --(CH.sub.2).sub.n--,
--(CH.sub.2).sub.nO-- or --(CH.sub.2).sub.nNH--, wherein n is 0, 1
or 2.
[0022] In another embodiment, each of R.sup.1, R.sup.2, R.sup.3,
R.sup.4, R.sup.5, R.sup.6 and R.sup.7 is independently H, F, Cl, Me
or OMe.
[0023] In another embodiment, each of R.sup.a, R.sup.b and R.sup.c
is independently H, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4
haloalkyl, C.sub.3-C.sub.6 cycloalkyl, --(C.sub.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl
or --(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl), or
R.sup.a and R.sup.b taken together with the nitrogen atom to which
they are attached form a 3-8 membered heterocyclic ring, wherein
each of the C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 haloalkyl,
C.sub.3-C.sub.6 cycloalkyl, --(C.sub.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl,
--(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl) and 3-8
membered heterocyclic ring is optionally substituted with 1, 2, 3
or 4 substituents independently selected from F, Cl, CN, N.sub.3,
OH, NH.sub.2, C.sub.1-C.sub.3 haloalkyl, C.sub.1-C.sub.3 alkoxy and
C.sub.1-C.sub.3 alkylamino.
[0024] In one embodiment, R.sup.d is H, C.sub.1-C.sub.4 alkyl,
C.sub.3-C.sub.6 cycloalkyl, --(C.sub.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl
or --(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl),
wherein each of the C.sub.1-C.sub.4 alkyl, C.sub.3-C.sub.6
cycloalkyl, --(C.sub.1-C.sub.2 alkylene)-(C.sub.3-C.sub.6
cycloalkyl), 3-6 membered heterocyclyl and --(C.sub.1-C.sub.2
alkylene)-(3-6 membered heterocyclyl) is optionally substituted
with 1, 2, 3 or 4 substituents independently selected from F, CN,
OR.sup.a, NR.sup.aR.sup.b, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4
alkenyl, C.sub.2-C.sub.4 alkynyl, --(C.sub.1-C.sub.2
alkylene)-OR.sup.a and --(C.sub.1-C.sub.2
alkylene)-NR.sup.aR.sup.b.
[0025] In another embodiment, Q is:
##STR00002## ##STR00003##
[0026] In another aspect, provided herein is a pharmaceutical
composition comprising the compound disclosed herein, and a
pharmaceutically acceptable excipient, carrier, adjuvant, vehicle
or a combination thereof.
[0027] In one embodiment, the pharmaceutical composition disclosed
herein further comprising therapeutic selected from the group
consisting of chemotherapeutic agents, anti-proliferative agents,
agents for treating atherosclerosis, agents for treating lung
fibrosis and combinations thereof.
[0028] In one embodiment, the pharmaceutical composition disclosed
herein is chlorambucil, melphalan, cyclophosphamide, ifosfamide,
busulfan, carmustine, lomustine, streptozocin, cisplatin,
carboplatin, oxaliplatin, dacarbazine, temozolomide, procarbazine,
methotrexate, fluorouracil, cytarabine, gemcitabine,
mercaptopurine, fludarabine, vinblastine, vincristine, vinorelbine,
paclitaxel, docetaxel, topotecan, irinotecan, etoposide,
trabectedin, dactinomycin, doxorubicin, epirubicin, daunorubicin,
mitoxantrone, bleomycin, mitomycin, ixabepilone, tamoxifen,
flutamide, gonadorelin analogues, megestrol, prednisone,
dexamethasone, methylprednisolone, thalidomide, interferon alfa,
leucovorin, sirolimus, temsirolimus, everolimus, afatinib,
alisertib, amuvatinib, apatinib, axitinib, bortezomib, bosutinib,
brivanib, cabozantinib, cediranib, crenolanib, crizotinib,
dabrafenib, dacomitinib, danusertib, dasatinib, dovitinib,
erlotinib, foretinib, ganetespib, gefitinib, ibrutinib, icotinib,
imatinib, iniparib, lapatinib, lenvatinib, linifanib, linsitinib,
masitinib, momelotinib, motesanib, neratinib, nilotinib, niraparib,
oprozomib, olaparib, pazopanib, pictilisib, ponatinib, quizartinib,
regorafenib, rigosertib, rucaparib, ruxolitinib, saracatinib,
saridegib, sorafenib, sunitinib, tasocitinib, telatinib,
tivantinib, tivozanib, tofacitinib, trametinib, vandetanib,
veliparib, vemurafenib, vismodegib, volasertib, alemtuzumab,
bevacizumab, brentuximabvedotin, catumaxomab, cetuximab, denosumab,
gemtuzumab, ipilimumab, nimotuzumab, ofatumumab, panitumumab,
ramucirumab, rituximab, tositumomab, trastuzumab or a combination
thereof.
[0029] In another aspect, provided herein is a method of
preventing, treating or lessening the severity of a proliferative
disease in a patient with the compound or the pharmaceutical
composition disclosed herein.
[0030] In one embodiment, the proliferative disease is colon
cancer, rectal cancer, gastric cancer, gastric adenocarcinoma,
pancreatic cancer, bladder cancer, gallbladder cancer, breast
cancer, kidney cancer, renal cell carcinoma, liver cancer,
hepatocellular carcinoma, lung cancer, skin cancer, melanoma,
thyroid cancer, osteosarcomas, soft tissue sarcoma, a cancer of the
head and neck, a cancer of the central nervous system, glioma,
glioblastomas, ovarian cancer, uterine cancer, endometrial
carcinoma, prostate cancer, acute myeloid leukemia or acute
lymphoblastic leukemia, or metastases thereof.
[0031] In another embodiment, the proliferative disease is
atherosclerosis or lung fibrosis.
[0032] In another aspect, provided herein is the compound or the
pharmaceutical composition disclosed herein for use in preventing,
treating or lessening the severity of a proliferative disease in a
patient.
[0033] In another aspect, provided herein is the use of the
compound or the pharmaceutical composition disclosed herein in the
manufacture of a medicament for preventing, treating or lessening
the severity of a proliferative disease in a patient.
[0034] In another aspect, provided herein is a method of inhibiting
or modulating the activity of a protein kinase in a biological
sample comprising contacting a biological sample with the compound
and the pharmaceutical composition disclosed herein.
[0035] In one embodiment, the protein kinase is a receptor tyrosine
kinase.
[0036] In one embodiment, the receptor tyrosine kinase is Axl, Mer,
c-Met, Ron or a combination thereof.
[0037] In another aspect, provided herein are methods for
preparation, separation and purification of the compounds
represented by Formula (I).
[0038] Biological test results indicate that the compounds provided
herein can be used as preferable inhibitors of Axl, Mer, c-Met and
Ron.
[0039] Any embodiment disclosed herein can be combined other
embodiments as long as they are not contradictory to one another,
even though the embodiments are described under different aspects
of the invention. In addition, any technical feature in one
embodiment can be applied to the corresponding technical feature in
other embodiment as long as they are not contradictory to one
another, even though the embodiments are described under different
aspects of the invention.
[0040] The foregoing merely summarizes certain aspects of the
invention and is not intended to be limiting in nature. These
aspects and other aspects and embodiments are described more fully
below.
DETAILED DESCRIPTION OF THE INVENTION
Definitions and General Terminology
[0041] Reference will now be made in detail to certain embodiments
of the invention, examples of which are illustrated in the
accompanying structures and formulas. The invention is intended to
cover all alternatives, modifications, and equivalents which may be
included within the scope of the present invention as defined by
the claims. One skilled in the art will recognize many methods and
materials similar or equivalent to those described herein, which
could be used in the practice of the present invention. The present
invention is in no way limited to the methods and materials
described herein. In the event that one or more of the incorporated
literature, patents, and similar materials differs from or
contradicts this application, including but not limited to defined
terms, term usage, described techniques, or the like, this
application controls.
[0042] It is further appreciated that certain features of the
invention, which are, for clarity, described in the context of
separate embodiments, can also be provided in combination in a
single embodiment. Conversely, various features of the invention
which are, for brevity, described in the context of a single
embodiment, can also be provided separately or in any suitable
subcombination.
[0043] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as is commonly understood by one
skilled in the art to which this invention belongs. All patents and
publications referred to herein are incorporated by reference in
their entirety.
[0044] As used herein, the following definitions shall apply unless
otherwise indicated. For purposes of this invention, the chemical
elements are identified in accordance with the Periodic Table of
the Elements, CAS version, and the Handbook of Chemistry and
Physics, 75.sup.th Ed. 1994. Additionally, general principles of
organic chemistry are described in Sorrell et al., "Organic
Chemistry", University Science Books, Sausalito: 1999, and Smith et
al., "March's Advanced Organic Chemistry", John Wiley & Sons,
New York: 2007, all of which are incorporated by reference in their
entireties.
[0045] The grammatical articles "a", "an" and "the", as used
herein, are intended to include "at least one" or "one or more"
unless otherwise indicated herein or clearly contradicted by the
context. Thus, the articles are used herein to refer to one or more
than one (i.e. at least one) of the grammatical objects of the
article. By way of example, "a component" means one or more
components, and thus, possibly, more than one component is
contemplated and may be employed or used in an implementation of
the described embodiments.
[0046] A weight percent (wt. %) of a component, unless specifically
stated to the contrary, is based on the total weight of the
formulation or composition in which the component is included.
[0047] As used herein, the term "subject" refers to an animal.
Typically the animal is a mammal. A subject also refers to for
example, primates (e.g., humans, male or female), cows, sheep,
goats, horses, dogs, cats, rabbits, rats, mice, fish, birds and the
like. In certain embodiments, the subject is a primate. In yet
other embodiments, the subject is a human.
[0048] As used herein, "patient" refers to a human (including
adults and children) or other animal. In one embodiment, "patient"
refers to a human.
[0049] The term "comprising" is meant to be open ended, including
the indicated component but not excluding other elements.
[0050] The term "stereoisomers" refers to compounds which have
identical chemical constitution, but differ with regard to the
arrangement of the atoms or groups in space. Stereoisomers include
enantiomer, diastereomers, conformer (rotamer), geometric
(cis/trans) isomer, atropisomer, etc.
[0051] The term "chiral" refers to molecules which have the
property of non-superimposability of the mirror image partner,
while the term "achiral" refers to molecules which are
superimposable on their mirror image partner.
[0052] The term "enantiomers" refers to two stereoisomers of a
compound which are non-superimposable mirror images of one
another.
[0053] The term "diastereomer" refers to a stereoisomer with two or
more centers of chirality and whose molecules are not mirror images
of one another. Diastereomers have different physical properties,
e.g. melting points, boiling points, spectral properties or
biological activities. Mixture of diastereomers may separate under
high resolution analytical procedures such as electrophoresis and
chromatography such as HPLC.
[0054] Stereochemical definitions and conventions used herein
generally follow Parker et al., McGraw-Hill Dictionary of Chemical
Terms (1984) McGraw-Hill Book Company, New York and Eliel et al.,
"Stereochemistry of Organic Compounds", John Wiley & Sons,
Inc., New York, 1994.
[0055] Many organic compounds exist in optically active forms,
i.e., they have the ability to rotate the plane of plane-polarized
light. In describing an optically active compound, the prefixes D
and L, or R and S, are used to denote the absolute configuration of
the molecule about its chiral center(s). The prefixes d and l or
(+) and (-) are employed to designate the sign of rotation of
plane-polarized light by the compound, with (-) or l meaning that
the compound is levorotatory. A compound prefixed with (+) or d is
dextrorotatory. A specific stereoisomer may be referred to as an
enantiomer, and a mixture of such stereoisomers is called an
enantiomeric mixture. A 50:50 mixture of enantiomers is referred to
as a racemic mixture or a racemate, which may occur where there has
been no stereoselection or stereospecificity in a chemical reaction
or process.
[0056] Any asymmetric atom (e.g., carbon or the like) of the
compound(s) disclosed herein can be present in racemic or
enantiomerically enriched, for example the (R)-, (S)- or
(R,S)-configuration. In certain embodiments, each asymmetric atom
has at least 50% enantiomeric excess, at least 60% enantiomeric
excess, at least 70% enantiomeric excess, at least 80% enantiomeric
excess, at least 90% enantiomeric excess, at least 95% enantiomeric
excess, or at least 99% enantiomeric excess in the (R)- or
(S)-configuration.
[0057] Depending on the choice of the starting materials and
procedures, the compounds can be present in the form of one of the
possible stereoisomers or as mixtures thereof, such as racemates
and diastereoisomer mixtures, depending on the number of asymmetric
carbon atoms. Optically active (R)- and (S)-isomers may be prepared
using chiral synthons or chiral reagents, or resolved using
conventional techniques. If the compound contains a double bond,
the substituent may be E or Z configuration. If the compound
contains a disubstituted cycloalkyl, the cycloalkyl substituent may
have a cis- or trans-configuration.
[0058] Any resulting mixtures of stereoisomers can be separated on
the basis of the physicochemical differences of the constituents,
into the pure or substantially pure geometric isomers, enantiomers,
diastereomers, for example, by chromatography and/or fractional
crystallization.
[0059] Any resulting racemates of final products or intermediates
can be resolved into the optical antipodes by methods known to
those skilled in the art, e.g., by separation of the diastereomeric
salts thereof. Racemic products can also be resolved by chiral
chromatography, e.g., high performance liquid chromatography (HPLC)
using a chiral adsorbent. Preferred enantiomers can also be
prepared by asymmetric syntheses. See, for example, Jacques, et
al., Enantiomers, Racemates and Resolutions (Wiley Interscience,
New York, 1981); Principles of Asymmetric Synthesis (2.sup.nd Ed.
Robert et al., Elsevier, Oxford, UK, 2012); Eliel et al.,
Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and
Wilen et al., Tables of Resolving Agents and Optical Resolutions p.
268 (E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind.,
1972). Chiral Separation Techniques: A Practical Approach
(Subramanian, G. Ed., Wiley-VCH Verlag GmbH & Co. KGaA,
Weinheim, Germany, 2007).
[0060] The term "tautomer" or "tautomeric form" refers to
structural isomers of different energies which are interconvertible
via a low energy barrier. Where tautomerization is possible (e.g.
in solution), a chemical equilibrium of tautomers can be reached.
For example, proton tautomers (also known as prototropic tautomers)
includes interconversions via migration of a proton, such as
keto-enol and imine-enamine isomerizations. Valence tautomers
include interconversions by reorganization of some of the bonding
electrons. A specific example of keto-enol tautomerization is the
interconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-one
tautomers. Another example of tautomerization is phenol-keto
tautomerization. A specific example of phenol-keto tautomerization
is the interconversion of pyridin-4-ol and pridin-4(1H)-one
tautomers. Unless otherwise state, all tautomeric forms of the
compounds disclosed herein are within the scope of the
invention.
[0061] The term "prodrug" as used herein, represents a compound
that is transformed in vivo into a compound of Formula (I). Such a
transformation can be affected, for example, by hydrolysis in blood
or enzymatic transformation of the prodrug form to the parent form
in blood or tissue. Prodrugs of the compounds disclosed herein may
be, for example, esters. Esters that may be utilized as prodrugs in
the present invention are phenyl esters, aliphatic
(C.sub.1-C.sub.24) esters, acyloxymethyl esters, carbonates,
carbamates, and amino acid esters. For example, a compound
disclosed herein that contains an OH group may be acylated at this
position in its prodrug form. Other prodrug forms include
phosphates, such as, for example those phosphates resulting from
the phosphonation of an OH group on the parent compound. A thorough
discussion of prodrugs is provided in Higuchi et al., Pro-drugs as
Novel Delivery Systems, Vol. 14, A.C.S. Symposium Series; Roche et
al., Bioreversible Carriers in Drug Design, American Pharmaceutical
Association and Pergamon Press, 1987; Rautio et al., Prodrugs:
Design and Clinical Applications, Nat. Rev. Drug Discovery, 2008,
7, 255-270, and Hecker et al., Prodrugs of Phosphates and
Phosphonates, J. Med. Chem., 2008, 51, 2328-2345, all of which are
incorporated herein by reference.
[0062] A "metabolite" refers to a product produced through
metabolism in the body of a specified compound or salt thereof. The
metabolites of a compound may be identified using routine
techniques known in the art and their activities determined using
tests such as those described herein. Such products may result for
example from the oxidation, reduction, hydrolysis, amidation,
deamidation, esterification, deesterification, enzymatic cleavage,
and the like, of the administered compound. Accordingly, the
invention includes metabolites of compounds disclosed herein,
including compounds produced by a process comprising contacting a
compound disclosed herein with a mammal for a period of time
sufficient to yield a metabolic product thereof.
[0063] A "pharmaceutically acceptable salt" refers to organic or
inorganic salts of a compound disclosed herein. The
pharmaceutically acceptable salts are well known in the art. For
example, Berge et al., describe pharmaceutically acceptable salts
in detail in J. Pharm. Sci., 1977, 66, 1-19, which is incorporated
herein by reference. Some non-limiting examples of the
pharmaceutically acceptable salt include salts of an amino group
formed with inorganic acids such as hydrochloric acid, hydrobromic
acid, phosphoric acid, sulfuric acid and perchloric acid or with
organic acids such as acetic acid, oxalic acid, maleic acid,
tartaric acid, citric acid, succinic acid or malonic acid.
[0064] Other examples of the pharmaceutically acceptable salt
include adipate, alginate, ascorbate, aspartate, benzenesulfonate,
benzoate, bisulfate, borate, butyrate, camphorate,
camphorsulfonate, citrate, cyclopentanepropionate, digluconate,
dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate,
glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate,
hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate,
laurate, lauryl sulfate, malate, maleate, malonate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,
oleate, oxalate, palmitate, pamoate, pectinate, persulfate,
3-phenylpropionate, phosphate, picrate, pivalate, propionate,
stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like.
[0065] Pharmaceutically acceptable salts derived from appropriate
bases include alkali metal, alkaline earth metal, ammonium and
N.sup.+(C.sub.1-C.sub.4 alkyl).sub.4 salts. This invention also
envisions the quaternization of any basic nitrogen-containing
groups of the compounds disclosed herein. Water or oil-soluble or
dispersible products may be obtained by such quaternization.
Representative alkali or alkaline earth metal salts include sodium,
lithium, potassium, calcium, magnesium, and the like. Further
examples of the pharmaceutically acceptable salt include, when
appropriate, nontoxic ammonium, quaternary ammonium, and amine
cations formed using counterions such as halide, hydroxide,
carboxylate, sulfate, phosphate, nitrate, C.sub.1-C.sub.8 sulfonate
and aryl sulfonate.
[0066] A "solvate" refers to an association or complex of one or
more solvent molecules and a compound disclosed herein. Examples of
solvents that form solvates include, but are not limited to, water,
isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid,
and ethanolamine. The term "hydrate" refers to the complex where
the solvent molecule is water.
[0067] As described herein, compounds disclosed herein may
optionally be substituted with one or more substituents, such as
those illustrated below, or as exemplified by particular classes,
subclasses, and species of the invention. It will be appreciated
that the phrase "optionally substituted" is used interchangeably
with the phrase "substituted or unsubstituted". In general, the
term "substituted" refers to the replacement of one or more
hydrogen radicals in a given structure with the radical of a
specified substituent. Unless otherwise indicated, an optionally
substituted group may have a substituent at each substitutable
position of the group. When more than one position in a given
structure can be substituted with more than one substituent
selected from a specified group, the substituent may be either the
same or different at each position.
[0068] At various places in the present specification, substituents
of compounds disclosed herein are disclosed in groups or in ranges.
It is specifically intended that the invention include each and
every individual subcombination of the members of such groups and
ranges. For example, the term "C.sub.1-C.sub.6 alkyl" is
specifically intended to individually disclose methyl, ethyl,
C.sub.3 alkyl, C.sub.4 alkyl, C.sub.5 alkyl, and C.sub.6 alkyl.
[0069] At various places in the present specification, linking
substituents are described. Where the structure clearly requires a
linking group, the Markush variables listed for that group are
understood to be linking groups. For example, if the structure
requires a linking group and the Markush group definition for that
variable lists "alkyl" or "aryl" then it is understood that the
"alkyl" or "aryl" represents a linking alkylene group or arylene
group, respectively.
[0070] The term "alkyl" or "alkyl group" refers to a saturated
linear or branched-chain monovalent hydrocarbon radical of 1 to 20
carbon atoms, wherein the alkyl radical may be optionally
substituted independently with one or more substituents described
below. Unless otherwise specified, the alkyl group contains 1-20
carbon atoms. In one embodiment, the alkyl group contains 1-12
carbon atoms. In another embodiment, the alkyl group contains 1-6
carbon atoms. In still another embodiment, the alkyl group contains
1-4 carbon atoms. In yet another embodiment, the alkyl group
contains 1-3 carbon atoms.
[0071] Some non-limiting examples of the alkyl group include methyl
(Me, --CH.sub.3), ethyl (Et, --CH.sub.2CH.sub.3), 1-propyl (n-Pr,
n-propyl, --CH.sub.2CH.sub.2CH.sub.3), 2-propyl (i-Pr, i-propyl,
--CH(CH.sub.3).sub.2), 1-butyl (n-Bu, n-butyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-methyl-1-propyl (i-Bu,
i-butyl, --CH.sub.2CH(CH.sub.3).sub.2), 2-butyl (s-Bu, s-butyl,
--CH(CH.sub.3)CH.sub.2CH.sub.3), 2-methyl-2-propyl (t-Bu, t-butyl,
--C(CH.sub.3).sub.3), 1-pentyl (n-pentyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-pentyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.3), 3-pentyl
(--CH(CH.sub.2CH.sub.3).sub.2), 2-methyl-2-butyl
(--C(CH.sub.3).sub.2CH.sub.2CH.sub.3), 3-methyl-2-butyl
(--CH(CH.sub.3)CH(CH.sub.3).sub.2), 3-methyl-1-butyl
(--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2), 2-methyl-1-butyl
(--CH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.3), 1-hexyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-hexyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 3-hexyl
(--CH(CH.sub.2CH.sub.3)(CH.sub.2CH.sub.2CH.sub.3)),
2-methyl-2-pentyl (--C(CH.sub.3).sub.2CH.sub.2CH.sub.2CH.sub.3),
3-methyl-2-pentyl (--CH(CH.sub.3)CH(CH.sub.3)CH.sub.2CH.sub.3),
4-methyl-2-pentyl (--CH(CH.sub.3)CH.sub.2CH(CH.sub.3).sub.2),
3-methyl-3-pentyl (--C(CH.sub.3)(CH.sub.2CH.sub.3).sub.2),
2-methyl-3-pentyl (--CH(CH.sub.2CH.sub.3)CH(CH.sub.3).sub.2),
2,3-dimethyl-2-butyl (--C(CH.sub.3).sub.2CH(CH.sub.3).sub.2),
3,3-dimethyl-2-butyl (--CH(CH.sub.3)C(CH.sub.3).sub.3, 1-heptyl,
1-octyl, and the like.
[0072] The term "alkylene" refers to a saturated divalent
hydrocarbon group derived from a straight or branched chain
saturated hydrocarbon by the removal of two hydrogen atoms. Unless
otherwise specified, the alkylene group contains 1-12 carbon atoms.
In one embodiment, the alkylene group contains 1-6 carbon atoms. In
another embodiment, the alkylene group contains 1-4 carbon atoms.
In still another embodiment, the alkylene group contains 1-3 carbon
atoms. In yet another embodiment, the alkylene group contains 1-2
carbon atoms. The alkylene group is exemplified by methylene
(--CH.sub.2--), ethylene (--CH.sub.2CH.sub.2--), isopropylene
(--CH(CH.sub.3)CH.sub.2--), and the like.
[0073] The term "alkenyl" refers to a linear or branched-chain
monovalent hydrocarbon radical of 2 to 12 carbon atoms with at
least one site of unsaturation, i.e., a carbon-carbon, sp2 double
bond, wherein the alkenyl radical may be optionally substituted
independently with one or more substituents described herein, and
includes radicals having "cis" and "trans" orientations, or
alternatively, "E" and "Z" orientations. In one embodiment, the
alkenyl group contains 2-8 carbon atoms. In another embodiment, the
alkenyl group contains 2-6 carbon atoms. In still another
embodiment, the alkenyl group contains 2-4 carbon atoms. Some
non-limiting examples of the alkenyl group include ethylenyl or
vinyl (--CH.dbd.CH.sub.2), allyl (--CH.sub.2CH.dbd.CH.sub.2), and
the like.
[0074] The term "alkynyl" refers to a linear or branched-chain
monovalent hydrocarbon radical of 2 to 12 carbon atoms with at
least one site of unsaturation, i.e., a carbon-carbon, sp triple
bond, wherein the alkynyl radical may be optionally substituted
independently with one or more substituents described herein. In
one embodiment, the alkynyl group contains 2-8 carbon atoms. In
another embodiment, the alkynyl group contains 2-6 carbon atoms. In
still another embodiment, the alkynyl group contains 2-4 carbon
atoms. Some non-limiting examples of the alkynyl group include
ethynyl (--C.ident.CH), propynyl (propargyl, --CH.sub.2C.ident.CH),
--C.ident.C--CH.sub.3, and the like.
[0075] The term "alkoxy" refers to an alkyl group, as previously
defined, attached to the principal carbon atom through an oxygen
atom. Unless otherwise specified, the alkoxy group contains 1-12
carbon atoms. In one embodiment, the alkoxy group contains 1-6
carbon atoms. In another embodiment, the alkoxy group contains 1-4
carbon atoms. In still another embodiment, the alkoxy group
contains 1-3 carbon atoms. The alkoxy radical may be optionally
substituted independently with one or more substituents described
herein.
[0076] Some non-limiting examples of alkoxy groups include methoxy
(MeO, --OCH.sub.3), ethoxy (EtO, --OCH.sub.2CH.sub.3), 1-propoxy
(n-PrO, n-propoxy, --OCH.sub.2CH.sub.2CH.sub.3), 2-propoxy (i-PrO,
i-propoxy, --OCH(CH.sub.3).sub.2), 1-butoxy (n-BuO, n-butoxy,
--OCH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-methyl-1-propoxy (i-BuO,
i-butoxy, --OCH.sub.2CH(CH.sub.3).sub.2), 2-butoxy (s-BuO,
s-butoxy, --OCH(CH.sub.3)CH.sub.2CH.sub.3), 2-methyl-2-propoxy
(t-BuO, t-butoxy, --OC(CH.sub.3).sub.3), 1-pentoxy (n-pentoxy,
--OCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-pentoxy
(--OCH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.3), 3-pentoxy
(--OCH(CH.sub.2CH.sub.3).sub.2), 2-methyl-2-butoxy
(--OC(CH.sub.3).sub.2CH.sub.2CH.sub.3), 3-methyl-2-butoxy
(--OCH(CH.sub.3)CH(CH.sub.3).sub.2), 3-methyl-1-butoxy
(--OCH.sub.2CH.sub.2CH(CH.sub.3).sub.2), 2-methyl-1-butoxy
(--OCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.3), and the like.
[0077] The term "haloalkyl", "haloalkenyl" or "haloalkoxy" refers
to alkyl, alkenyl, or alkoxy, as the case may be, substituted with
one or more halogen atoms.
[0078] The term "carbocycle", "carbocyclyl" or "carbocyclic ring"
refers to a monovalent or multivalent non-aromatic, saturated or
partially unsaturated ring having 3 to 12 carbon atoms as a
monocyclic, bicyclic or tricyclic ring system. A carbobicyclyl
system includes a spiro carbobicyclyl or a fused carbobicyclyl.
Suitable carbocyclyl groups include, but are not limited to,
cycloalkyl, cycloalkenyl, and cycloalkynyl. Further non-limiting
examples of carbocyclyl group include cyclopropyl, cyclobutyl,
cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl,
1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl,
1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, and the
like.
[0079] The term "cycloalkyl" refers to a monovalent or multivalent
saturated ring having 3 to 12 carbon atoms as a monocyclic,
bicyclic, or tricyclic ring system. In one embodiment, the
cycloalkyl contains 3-12 carbon atoms. In another embodiment, the
cycloalkyl contains 3-8 carbon atoms. In still another embodiment,
the cycloalkyl contains 3-6 carbon atoms. The cycloalkyl radical
may be optionally substituted independently with one or more
substituents disclosed herein.
[0080] The term "heterocycle", "heterocyclyl", or "heterocyclic
ring" as used interchangeably herein refers to a saturated or
partially unsaturated monocyclic, bicyclic or tricyclic ring
containing 3-12 ring atoms of which at least one ring atom is
selected from nitrogen, sulfur and oxygen, and which may, unless
otherwise specified, be carbon or nitrogen linked, and of which a
--CH.sub.2-- group can optionally be replaced by a --C(.dbd.O)--
group. Ring sulfur atoms may be optionally oxidized to form
S-oxides. Ring nitrogen atoms maybe optionally oxidized to form
N-oxides. In one embodiment, heterocyclyl may be 3-8 membered
heterocyclyl, which refers to a saturated or partially unsaturated
monocyclic ring containing 3-8 ring atoms. In another embodiment,
heterocyclyl may be 3-6 membered heterocyclyl, which refers to a
saturated or partially unsaturated monocyclic ring containing 3-6
ring atoms. In still another embodiment, heterocyclyl refers to a
7-12 membered heterocyclyl, which refers to a saturated or
partially unsaturated spiro or fused bicyclyl ring containing 7-12
ring atoms.
[0081] Examples of heterocyclyl include, but are not limited to,
oxiranyl, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
2-pyrrolinyl, 3-pyrrolinyl, pyrazolinyl, pyrazolidinyl,
imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl,
tetrahydrothienyl, dihydrothienyl, 1,3-dioxolanyl, dithiolanyl,
tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl,
tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl,
piperazinyl, dioxanyl, thioxanyl, dithianyl, homopiperazinyl,
homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl,
thiazepinyl, indolinyl, 1,2,3,4-tetrahydroisoquinolinyl,
1,3-benzodioxolyl, 2-oxa-5-azabicyclo[2.2.1]hept-5-yl. Some
non-limited examples of heterocyclyl wherein --CH.sub.2-- group is
replaced by --C(.dbd.O)-- moiety are 2-oxopyrrolidinyl,
oxo-1,3-thiazolidinyl, 2-piperidinonyl, 3,5-dioxopiperidinyl and
pyrimidinedionyl. Some non-limited examples of heterocyclyl wherein
the ring sulfur atom is oxidized are sulfolanyl,
1,1-dioxo-thiomorpholinyl. The heterocyclyl group may be optionally
substituted with one or more substituents disclosed herein.
[0082] The terms "fused bicyclic ring", "fused cyclic", "fused
bicyclyl" and "fused cyclyl" are used interchangeably and refer to
a monovalent or multivalent, saturated or partially unsaturated
bridged ring system, which refers to a bicyclic ring system that is
not aromatic. Such a system may contain isolated or conjugated
unsaturation, but not aromatic or heteroaromatic rings in its core
structure (but may have aromatic substitution thereon).
[0083] The terms "spirocyclyl", "spirocyclic", "spiro bicyclyl" and
"spiro bicyclic" are used interchangeably and refer to a monovalent
or multivalent, saturated or partially unsaturated ring system
wherein a ring originating from a particular annular carbon of
another ring. For example, as depicted below in Figure a, a
saturated bridged ring system (ring B and B') is termed as "fused
bicyclyl", whereas ring A and ring B share an atom between the two
saturated ring system, which terms as a "spirocyclyl" or "spiro
bicyclyl". Each ring in the fused bicyclyl or the spiro bicyclyl
can be either a carbocyclyl or a heterocyclyl, and each ring is
optionally substituted independently with one or more substituents
disclosed herein.
##STR00004##
[0084] The term "heterocycloalkyl" refers to a monovalent or
multivalent saturated ring having 3 to 12 ring atoms as a
monocyclic, bicyclic, or tricyclic ring system in which at least
one ring atom is selected from nitrogen, sulfur and oxygen.
[0085] The term "n membered" where n is an integer typically
describes the number of ring-forming atoms in a moiety where the
number of ring-forming atoms is n. For example, piperidinyl is an
example of a 6 membered heterocycloalkyl and
1,2,3,4-tetrahydronaphthyl is an example of a 10 membered
cycloalkyl group.
[0086] The term "unsaturated" refers to a moiety having one or more
units of unsaturation.
[0087] The term "heteroatom" refers to one or more of oxygen,
sulfur, nitrogen, phosphorus, or silicon, including any oxidized
form of nitrogen, sulfur, or phosphorus; the quaternized form of
any basic nitrogen; or a substitutable nitrogen of a heterocyclic
ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in
pyrrolidinyl) or NR (as in N-substituted pyrrolidinyl).
[0088] The term "halogen" refers to Fluoro (F), Chloro (CO, Bromo
(Br), or Iodo (I).
[0089] The term "azido" or "N.sub.3" refers to an azide moiety.
This radical may be attached, for example, to a methyl group to
form azidomethane (methyl azide, MeN); or attached to a phenyl
group to form phenyl azide (PhN.sub.3).
[0090] The term "aryl" refers to monocyclic, bicyclic, and
tricyclic carbocyclic ring systems having a total of 6 to 14 ring
members, preferably, 6 to 12 ring members, and more preferably 6 to
10 ring members, wherein at least one ring in the system is
aromatic and each ring in the system contains 3 to 7 ring members.
An aryl radical is commonly, but not necessarily, attached to the
rest of the molecule via an aromatic ring of the aryl radical. The
term "aryl" may be used interchangeably with the term "aryl ring"
or "aromatic ring". Some non-limiting examples of the aryl group
would include phenyl, naphthyl, and anthryl. The aryl radical is
optionally substituted independently with one or more substituents
described herein.
[0091] The term "heteroaryl" or "heteroaromatic ring" refers to
monocyclic, bicyclic, and tricyclic ring systems having a total of
5 to 14 ring members, preferably 5 to 10 ring members, and more
preferably 5 to 6 ring members, wherein at least one ring in the
system is aromatic, at least one ring in the system contains one or
more heteroatoms, and each ring in the system contains 5 to 7 ring
members. A heteroaryl radical is commonly, but not necessarily,
attached to the rest of the molecule via an aromatic ring of the
heteroaryl radical. The term "heteroaryl" may be used
interchangeably with the term "heteroaryl ring" or the term
"heteroaromatic ring". The heteroaryl radicals are optionally
substituted independently with one or more substituents described
herein. In one embodiment, a 5-10 membered heteroaryl comprises 1,
2, 3 or 4 heteroatoms independently selected from O, S and N. The
heteroaryl radical is optionally substituted independently with one
or more substituents described herein.
[0092] Some non-limiting examples of the heteroaryl group include
2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl,
5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl,
4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl,
2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl,
5-pyrimidinyl, pyridazinyl (e.g., 3-pyridazinyl), 2-thiazolyl,
4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g., 5-tetrazolyl),
triazolyl (e.g., 2-triazolyl and 5-triazolyl), 2-thienyl,
3-thienyl, pyrazolyl (e.g., 2-pyrazolyl), isothiazolyl,
1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl,
1,2,3-triazolyl, 1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl,
1,2,5-thiadiazolyl, pyrazinyl, 1,3,5-triazinyl, and the following
bicycles: benzimidazolyl, benzofuryl, benzothiophenyl, indolyl
(e.g., 2-indolyl), purinyl, quinolinyl (e.g., 2-quinolinyl,
3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g.,
1-isoquinolinyl, 3-isoquinolinyl or 4-isoquinolinyl),
imidazo[1,2-a]pyridyl, pyrazolo[1,5-a]pyridyl,
pyrazolo[1,5-a]pyrimidyl, imidazo[1,2-b]pyridazinyl,
[1,2,4]triazolo[4,3-b]pyridazinyl,
[1,2,4]triazolo[1,5-a]pyrimidinyl and
[1,2,4]triazolo[1,5-a]pyridyl.
[0093] The term "carboxy" or "carboxyl", whether used alone or with
other terms, such as "carboxyalkyl", refers to --CO.sub.2H. The
term "carbonyl", whether used alone or with other terms, such as
"aminocarbonyl", denotes --(C.dbd.O)--.
[0094] The term "alkylamino" embraces "N-alkylamino" and
"N,N-dialkylamino" where amino groups are independently substituted
with one alkyl radical or with two alkyl radicals, respectively.
Some non-limiting examples of alkylamino radicals are "lower
alkylamino" radicals having one or two alkyl radicals of one to six
carbon atoms, attached to a nitrogen atom. Suitable alkylamino
radicals may be mono or dialkylamino such as N-methylamino,
N-ethylamino, N,N-dimethylamino, N,N-diethylamino and the like.
[0095] The term "arylamino" refers to amino groups, which have been
substituted with one or two aryl radicals, such as N-phenylamino.
The arylamino radicals may be further substituted on the aryl ring
portion of the radical.
[0096] The term "aminoalkyl" refers to linear or branched alkyl
radicals having one to about ten carbon atoms any one of which may
be substituted with one or more amino radicals. More preferred
aminoalkyl radicals are "lower aminoalkyl" radicals having 1-6
carbon atoms and one or more amino radicals. Examples of such
radicals include aminomethyl, aminoethyl, aminopropyl, aminobutyl
and aminohexyl.
[0097] The term "protecting group" or "PG" refers to a substituent
that is commonly employed to block or protect a particular
functionality while reacting other functional groups on the
compound. For example, an "amino-protecting group" is a substituent
attached to an amino group that blocks or protects the amino
functionality in the compound. Suitable amino-protecting groups
include acetyl, trifluoroacetyl, t-butoxy-carbonyl (BOC, Boc),
benzyloxycarbonyl (CBZ, Cbz) and 9-fluorenylmethylenoxy-carbonyl
(Fmoc). Similarly, a "hydroxy-protecting group" refers to a
substituent of a hydroxy group that blocks or protects the hydroxy
functionality. Suitable protecting groups include acetyl and silyl.
A "carboxy-protecting group" refers to a substituent of the carboxy
group that blocks or protects the carboxy functionality. Common
carboxy-protecting groups include --CH.sub.2CH.sub.2SO.sub.2Ph,
cyanoethyl, 2-(trimethylsilyl)ethyl,
2-(trimethylsilyl)ethoxy-methyl, 2-(p-toluenesulfonyl)-ethyl,
2-(p-nitrophenylsulfenyl)-ethyl, 2-(diphenylphosphino)-ethyl,
nitroethyl and the like. For a general description of protecting
groups and their use, see T. W. Greene, Protective Groups in
Organic Synthesis, John Wiley & Sons, New York, 1991; and P. J.
Kocienski, Protecting Groups, Thieme, Stuttgart, 2005.
[0098] As used herein, the term "treat", "treating" or "treatment"
of any disease or disorder refers in one embodiment, to
ameliorating the disease or disorder (i.e., slowing or arresting or
reducing the development of the disease or at least one of the
clinical symptoms thereof). In another embodiment "treat",
"treating" or "treatment" refers to alleviating or ameliorating at
least one physical parameter including those which may not be
discernible by the patient. In yet another embodiment, "treat",
"treating" or "treatment" refers to modulating the disease or
disorder, either physically, (e.g., stabilization of a discernible
symptom), physiologically, (e.g., stabilization of a physical
parameter), or both. In yet another embodiment, "treat", "treating"
or "treatment" refers to preventing or delaying the onset or
development or progression of the disease or disorder.
[0099] The terms "cancer" and "cancerous" refer to or describe the
physiological condition in mammals that is typically characterized
by unregulated cell growth. A "tumor" comprises one or more
cancerous cells. Examples of cancer include, but are not limited
to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or
lymphoid malignancies. More particular examples of such cancers
include squamous cell cancer (e.g., epithelial squamous cell
cancer), lung cancer including small-cell lung cancer, non-small
cell lung cancer ("NSCLC"), adenocarcinoma of the lung and squamous
carcinoma of the lung, cancer of the peritoneum, hepatocellular
cancer, gastric or stomach cancer including gastrointestinal
cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian
cancer, liver cancer, bladder cancer, hepatoma, breast cancer,
colon cancer, rectal cancer, colorectal cancer, endometrial or
uterine carcinoma, salivary gland carcinoma, kidney or renal
cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic
carcinoma, anal carcinoma, penile carcinoma, as well as head and
neck cancer.
Description of Compounds of the Invention
[0100] The present invention provides substituted bicyclic
pyrazolone compounds which are potentially useful in the treatment
of diseases associated with inappropriate kinase activity, in
particular inappropriate Axl, Mer (MERTK), c-Met (HGFR) and/or Ron
(MST1R) kinase activity.
[0101] Specifically, in one aspect, provided herein is a compound
having Formula (I):
##STR00005##
or a stereoisomer, a tautomer, an N-oxide, a solvate, a metabolite,
a pharmaceutically acceptable salt or a prodrug thereof, wherein: Q
is H, OR.sup.a, NR.sup.aR.sup.b, --C(.dbd.O)NR.sup.aR.sup.b,
--N(R.sup.c)C(.dbd.O)R.sup.d, --N(R.sup.c)C(.dbd.O)OR.sup.a or
--N(R.sup.c)C(.dbd.O)NR.sup.aR.sup.b; U is CR.sup.7 or N, provided
that when U is N, the compound is not
2-oxo-1-phenyl-N-(5-((2-(pyrrolidine-1-carboxamido)pyridin-4-yl)oxy)pyrid-
in-2-yl)-2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
or
1-(buta-1,3-dien-2-yl)-2-oxo-N-(5-((2-(pyrrolidine-1-carboxamido)pyrid-
in-4-yl)oxy)pyridin-2-yl)-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyrazine-3-c-
arboxamide; X is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 alkenyl,
C.sub.3-C.sub.8 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl or
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), wherein
each of the C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 alkenyl,
C.sub.3-C.sub.8 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl and
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl) is
optionally substituted with 1, 2, 3, 4 or 5 substituents
independently selected from F, Cl, Br, CN, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, OR.sup.a,
NR.sup.aR.sup.b, --(C.sub.1-C.sub.4 alkylene)-OR.sup.a and
--(C.sub.1-C.sub.4 alkylene)-NR.sup.aR.sup.b; each Y is
independently 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.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl,
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), OR.sup.a,
NR.sup.aR.sup.b, --(C.sub.1-C.sub.4 alkylene)-OR.sup.a or
--(C.sub.1-C.sub.4 alkylene)-NR.sup.aR.sup.b; m is 0, 1, 2, 3, 4; W
is --(CH.sub.2).sub.n--, --(CH.sub.2).sub.nO--,
--(CH.sub.2).sub.nNH-- or --(CH.sub.2).sub.nS--, wherein n is 0, 1,
2, 3 or 4; each of R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5,
R.sup.6 and R.sup.7 is independently H, F, Cl, Br, CN, N.sub.3,
OR.sup.a, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.2-C.sub.6 alkenyl or C.sub.2-C.sub.6 alkynyl; each of
R.sup.a, R.sup.b and R.sup.c is independently H, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.3-C.sub.6 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-6 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl or
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), or R.sup.a
and R.sup.b taken together with the nitrogen atom to which they are
attached form a 3-8 membered heterocyclic ring, wherein each of the
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.3-C.sub.6
cycloalkyl, --(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.6
cycloalkyl), 3-6 membered heterocyclyl, --(C.sub.1-C.sub.4
alkylene)-(3-6 membered heterocyclyl), C.sub.6-C.sub.10 aryl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered
heteroaryl, --(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl)
and 3-8 membered heterocyclic ring is optionally substituted with
1, 2, 3 or 4 substituents independently selected from F, Cl, CN,
N.sub.3, OH, NH.sub.2, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
alkoxy and C.sub.1-C.sub.6 alkylamino; and R.sup.d is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl or
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl), wherein
each of the C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl,
--(C.sub.1-C.sub.4 alkylene)-(C.sub.3-C.sub.8 cycloalkyl), 3-8
membered heterocyclyl, --(C.sub.1-C.sub.4 alkylene)-(3-8 membered
heterocyclyl), C.sub.6-C.sub.10 aryl, --(C.sub.1-C.sub.4
alkylene)-(C.sub.6-C.sub.10 aryl), 5-10 membered heteroaryl and
--(C.sub.1-C.sub.4 alkylene)-(5-10 membered heteroaryl) is
optionally substituted with 1, 2, 3 or 4 substituents independently
selected from F, Cl, Br, CN, OR.sup.a, NR.sup.aR.sup.b,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, --(C.sub.1-C.sub.4 alkylene)-OR.sup.a and
--(C.sub.1-C.sub.4 alkylene)-NR.sup.aR.sup.b.
[0102] In one embodiment, Q is NR.sup.aR.sup.b,
--C(.dbd.O)NR.sup.aR.sup.b, --N(R.sup.c)C(.dbd.O)R.sup.d or
--N(R.sup.c)C(.dbd.O)NR.sup.aR.sup.b.
[0103] In another embodiment, X is C.sub.1-C.sub.4 alkyl,
C.sub.3-C.sub.6 alkenyl, C.sub.3-C.sub.6 alkynyl, C.sub.3-C.sub.6
cycloalkyl, --(C.sub.1-C.sub.2 alkylene)-(C.sub.3-C.sub.6
cycloalkyl), phenyl or --(C.sub.1-C.sub.2 alkylene)-phenyl, wherein
each of the C.sub.1-C.sub.4 alkyl, C.sub.3-C.sub.6 alkenyl,
C.sub.3-C.sub.6 alkynyl, C.sub.3-C.sub.6 cycloalkyl,
--(C.sub.1-C.sub.2 alkylene)-(C.sub.3-C.sub.6 cycloalkyl), phenyl
and --(C.sub.1-C.sub.2 alkylene)-phenyl is optionally substituted
with 1, 2, 3, 4 or 5 substituents independently selected from F,
Cl, Br, CN, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 alkynyl, OR.sup.a, NR.sup.aR.sup.b,
--(C.sub.1-C.sub.2 alkylene)-OR.sup.a and --(C.sub.1-C.sub.2
alkylene)-NR.sup.aR.sup.b.
[0104] In another embodiment, each Y is independently
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.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl,
--(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl), phenyl,
--(C.sub.1-C.sub.2 alkylene)-phenyl, 5-6 membered heteroaryl,
--(C.sub.1-C.sub.2 alkylene)-(5-6 membered heteroaryl), OR.sup.a,
NR.sup.aR.sup.b, --(C.sub.1-C.sub.2 alkylene)-OR.sup.a or
--(C.sub.1-C.sub.2 alkylene)-NR.sup.aR.sup.b; m is 0, 1 or 2.
[0105] In another embodiment, W is --(CH.sub.2).sub.n--,
--(CH.sub.2).sub.nO-- or --(CH.sub.2).sub.nNH--, wherein n is 0, 1
or 2.
[0106] In another embodiment, each of R.sup.1, R.sup.2, R.sup.3,
R.sup.4, R.sup.5, R.sup.6 and R.sup.7 is independently H, F,
C.sub.1, Me or OMe.
[0107] In another embodiment, each of R.sup.a, R.sup.b and R.sup.c
is independently H, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4
haloalkyl, C.sub.3-C.sub.6 cycloalkyl, --(C.sub.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl
or --(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl), or
R.sup.a and R.sup.b taken together with the nitrogen atom to which
they are attached form a 3-8 membered heterocyclic ring, wherein
each of the C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 haloalkyl,
C.sub.3-C.sub.6 cycloalkyl, --(C.sub.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl,
--(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl) and 3-8
membered heterocyclic ring is optionally substituted with 1, 2, 3
or 4 substituents independently selected from F, Cl, CN, N.sub.3,
OH, NH.sub.2, C.sub.1-C.sub.3 haloalkyl, C.sub.1-C.sub.3 alkoxy and
C.sub.1-C.sub.3 alkylamino.
[0108] In one embodiment, R.sup.d is H, C.sub.1-C.sub.4 alkyl,
C.sub.3-C.sub.6 cycloalkyl, --(C.sub.1-C.sub.2
alkylene)-(C.sub.3-C.sub.6 cycloalkyl), 3-6 membered heterocyclyl
or --(C.sub.1-C.sub.2 alkylene)-(3-6 membered heterocyclyl),
wherein each of the C.sub.1-C.sub.4 alkyl, C.sub.3-C.sub.6
cycloalkyl, --(C.sub.1-C.sub.2 alkylene)-(C.sub.3-C.sub.6
cycloalkyl), 3-6 membered heterocyclyl and --(C.sub.1-C.sub.2
alkylene)-(3-6 membered heterocyclyl) is optionally substituted
with 1, 2, 3 or 4 substituents independently selected from F, CN,
OR.sup.a, NR.sup.aR.sup.b, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4
alkenyl, C.sub.2-C.sub.4 alkynyl, --(C.sub.1-C.sub.2
alkylene)-OR.sup.a and --(C.sub.1-C.sub.2
alkylene)-NR.sup.aR.sup.b.
[0109] In another embodiment, Q is:
##STR00006## ##STR00007## ##STR00008##
[0110] In another embodiment, the compound disclosed herein having
one of the following structure:
##STR00009## ##STR00010## ##STR00011## ##STR00012## ##STR00013##
##STR00014##
[0111] Unless otherwise stated, all stereoisomers, tautomers,
solvates, metabolites, salts, and pharmaceutically acceptable
prodrugs of the compounds of Formula (I) are within the scope of
the invention.
[0112] The compounds disclosed herein may contain asymmetric or
chiral centers, and therefore exist in different stereoisomeric
forms. It is intended that all stereoisomeric forms of compounds of
Formula (I), including but not limited to, diastereomers,
enantiomers, atropisomers, conformers (rotamers) and geometric
(cis/trans) isomers as well as mixtures thereof such as racemic
mixtures, form part of the present invention.
[0113] In the structures shown herein, where the stereochemistry of
any particular chiral atom is not specified, then all stereoisomers
are contemplated and included as the compounds of the invention.
Where stereochemistry is specified by a solid wedge or dashed line
representing a particular configuration, then that stereoisomer is
so specified and defined.
[0114] The compounds of Formula (I) may exist in different
tautomeric forms, and all such forms are embraced within the scope
of the invention, as defined by the claims.
[0115] The compounds of Formula (I) can be in the form of salts. In
one embodiment, the salts are pharmaceutically acceptable salts.
The phrase "pharmaceutically acceptable" indicates that the
substance or composition must be compatible chemically and/or
toxicologically, with the other ingredients comprising a
formulation, and/or the mammal being treated therewith. In another
embodiment, the salts are not necessarily pharmaceutically
acceptable salts, and which may be useful as intermediates for
preparing and/or purifying compounds of Formula (I) and/or for
separating enantiomers of compounds of Formula (I).
[0116] Pharmaceutically acceptable acid addition salts can be
formed with inorganic acids and organic acids, e.g., acetate,
aspartate, benzoate, besylate, bromide/hydrobromide,
bicarbonate/carbonate, bisulfate/sulfate, camphorsulfonate,
chloride/hydrochloride, chlortheophyllonate, citrate,
ethandisulfonate, fumarate, gluceptate, gluconate, glucuronate,
hippurate, hydroiodide/iodide, isethionate, lactate, lactobionate,
laurylsulfate, malate, maleate, malonate, mandelate, mesylate,
methylsulphate, naphthoate, napsylate, nicotinate, nitrate,
octadecanoate, oleate, oxalate, palmitate, pamoate,
phosphate/hydrogen phosphate/dihydrogen phosphate,
polygalacturonate, propionate, stearate, succinate, subsalicylate,
tartrate, tosylate and trifluoroacetate salts.
[0117] Inorganic acids from which salts can be derived include, for
example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric
acid, phosphoric acid, and the like.
[0118] Organic acids from which salts can be derived include, for
example, acetic acid, propionic acid, glycolic acid, oxalic acid,
maleic acid, malonic acid, succinic acid, fumaric acid, tartaric
acid, citric acid, benzoic acid, mandelic acid, methanesulfonic
acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic
acid, and the like.
[0119] Pharmaceutically acceptable base addition salts can be
formed with inorganic and organic bases.
[0120] Inorganic bases from which salts can be derived include, for
example, ammonium salts and metals from columns I to XII of the
periodic table. In certain embodiments, the salts are derived from
sodium, potassium, ammonium, calcium, magnesium, iron, silver,
zinc, and copper; particularly suitable salts include ammonium,
potassium, sodium, calcium and magnesium salts.
[0121] Organic bases from which salts can be derived include, for
example, primary, secondary, and tertiary amines, substituted
amines including naturally occurring substituted amines, cyclic
amines, basic ion exchange resins, and the like. Certain organic
amines include isopropylamine, benzathine, cholinate,
diethanolamine, diethylamine, lysine, meglumine, piperazine and
tromethamine.
[0122] The pharmaceutically acceptable salts of the present
invention can be synthesized from a basic or acidic moiety, by
conventional chemical methods. Generally, such salts can be
prepared by reacting free acid forms of these compounds with a
stoichiometric amount of the appropriate base (such as Na, Ca, Mg,
or K hydroxide, carbonate, bicarbonate or the like), or by reacting
free base forms of these compounds with a stoichiometric amount of
the appropriate acid. Such reactions are typically carried out in
water or in an organic solvent, or in a mixture of the two.
Generally, use of non-aqueous media like ether, ethyl acetate,
ethanol, isopropanol, or acetonitrile is desirable, where
practicable. Lists of additional suitable salts can be found, e.g.,
in "Remington's Pharmaceutical Sciences", 20th ed., Mack Publishing
Company, Easton, Pa., (1985); and in "Handbook of Pharmaceutical
Salts: Properties, Selection, and Use" by Stahl and Wermuth
(Wiley-VCH, Weinheim, Germany, 2002).
[0123] Furthermore, the compounds disclosed herein, including their
salts, can also be obtained in the form of their hydrates, or
include other solvents such as ethanol, DMSO, and the like, used
for their crystallization. The compounds of the present invention
may inherently or by design form solvates with pharmaceutically
acceptable solvents (including water); therefore, it is intended
that the invention embrace both solvated and unsolvated forms.
[0124] Any formula given herein is also intended to represent
isotopically unenriched forms as well as isotopically enriched
forms of the compounds. Isotopically enriched 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 invention include isotopes of hydrogen, carbon,
nitrogen, oxygen, phosphorous, fluorine, and chlorine, such as
.sup.2H (deuterium, D), .sup.3H, .sup.11C, .sup.13C, .sup.14C,
.sup.15N, .sup.17O, .sup.18O, .sup.18F, .sup.31P, .sup.32P,
.sup.35S, .sup.36Cl, .sup.125I, respectively.
[0125] In another aspect, the compounds of the invention include
isotopically enriched compounds as defined herein, for example
those into which radioactive isotopes, such as .sup.3H, .sup.14C
and .sup.18F, or those into which non-radioactive isotopes, such as
.sup.2H and .sup.13C are present. Such isotopically enriched
compounds are useful in metabolic studies (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. In particular, an .sup.18F-enriched compound
may be particularly desirable for PET or SPECT studies.
Isotopically-enriched compounds of Formula (I) can generally be
prepared by conventional techniques known to those skilled in the
art or by processes analogous to those described in the
accompanying Examples and Preparations using an appropriate
isotopically-labeled reagent in place of the non-labeled reagent
previously employed.
[0126] Further, substitution with heavier isotopes, particularly
deuterium (i.e., .sup.2H or D) may afford certain therapeutic
advantages resulting from greater metabolic stability, for example
increased in vivo half-life or reduced dosage requirements or an
improvement in therapeutic index. It is understood that deuterium
in this context is regarded as a substituent of a compound of
Formula (I). The concentration of such a heavier isotope,
specifically deuterium, may be defined by the isotopic enrichment
factor. The term "isotopic enrichment factor" as used herein means
the ratio between the isotopic abundance and the natural abundance
of a specified isotope. If a substituent in a compound of this
invention is denoted deuterium, such compound has an isotopic
enrichment factor for each designated deuterium atom of at least
3500 (52.5% deuterium incorporation at each designated deuterium
atom), at least 4000 (60% deuterium incorporation), at least 4500
(67.5% deuterium incorporation), at least 5000 (75% deuterium
incorporation), at least 5500 (82.5% deuterium incorporation), at
least 6000 (90% deuterium incorporation), at least 6333.3 (95%
deuterium incorporation), at least 6466.7 (97% deuterium
incorporation), at least 6600 (99% deuterium incorporation), or at
least 6633.3 (99.5% deuterium incorporation). Pharmaceutically
acceptable solvates in accordance with the invention include those
wherein the solvent of crystallization may be isotopically
substituted, e.g. D.sub.2O, acetone-d.sub.6, DMSO-d.sub.6.
[0127] In another aspect, provided herein are intermediates for
preparing the compounds disclosed herein.
[0128] In another aspect, provided herein are methods of preparing,
methods of separating, and methods of purifying the compounds
disclosed herein.
Pharmaceutical Composition, Formulations and Administration of the
Compounds of the Invention
[0129] The present invention provides a pharmaceutical composition
that include a compound of disclosed herein, or a compound listed
in Table 1; and a pharmaceutically acceptable excipient, carrier,
adjuvant, vehicle or a combination thereof. The amount of compound
in the pharmaceutical composition disclosed herein is such that is
effective to detectably inhibit a protein kinase in a biological
sample or in a patient.
[0130] It will also be appreciated that certain compounds disclosed
herein can exist in free form for treatment, or where appropriate,
as a pharmaceutically acceptable derivative thereof. Some
non-limiting examples of pharmaceutically acceptable derivative
include pharmaceutically acceptable prodrugs, salts, esters, salts
of such esters, or any other adduct or derivative which upon
administration to a patient in need is capable of providing,
directly or indirectly, a compound as otherwise described herein,
or a metabolite or residue thereof
[0131] The pharmaceutical compositions disclosed herein may be
prepared and packaged in bulk form wherein a safe and effective
amount of the compound disclosed herein can be extracted and then
given to the patient such as with powders or syrups. Alternatively,
the pharmaceutical compositions disclosed herein may be prepared
and packaged in unit dosage form wherein each physically discrete
unit contains the compound disclosed herein. When prepared in unit
dosage form, the pharmaceutical compositions of the invention
typically may contain, for example, from 0.5 mg to 1 g, or from 1
mg to 700 mg, or from 5 mg to 100 mg of the compound disclosed
herein.
[0132] As used herein, "pharmaceutically acceptable excipient"
means a pharmaceutically acceptable material, composition or
vehicle involved in giving form or consistency to the
pharmaceutical composition. Each excipient must be compatible with
the other ingredients of the pharmaceutical composition when
commingled such that interactions which would substantially reduce
the efficacy of the compound disclosed herein when administered to
a patient and interactions which would result in pharmaceutical
compositions that are not pharmaceutically acceptable are avoided.
In addition, each excipient must be pharmaceutically-acceptable,
e.g., of sufficiently high purity.
[0133] Suitable pharmaceutically acceptable excipients will vary
depending upon the particular dosage form chosen. In addition,
suitable pharmaceutically acceptable excipients may be chosen for a
particular function that they may serve in the composition. For
example, certain pharmaceutically acceptable excipients may be
chosen for their ability to facilitate the production of uniform
dosage forms. Certain pharmaceutically acceptable excipients may be
chosen for their ability to facilitate the production of stable
dosage forms. Certain pharmaceutically acceptable excipients may be
chosen for their ability to facilitate the carrying or transporting
of the compound or compounds disclosed herein once administered to
the patient from one organ, or portion of the body, to another
organ, or portion of the body. Certain pharmaceutically acceptable
excipients may be chosen for their ability to enhance patient
compliance.
[0134] Suitable pharmaceutically acceptable excipients comprise the
following types of excipients: diluents, fillers, binders,
disintegrants, lubricants, glidants, granulating agents, coating
agents, wetting agents, solvents, co-solvents, suspending agents,
emulsifiers, sweetners, flavoring agents, flavor masking agents,
coloring agents, anticaking agents, hemectants, chelating agents,
plasticizers, viscosity increasing agents, antioxidants,
preservatives, stabilizers, surfactants, and buffering agents. The
skilled artisan will appreciate that certain pharmaceutically
acceptable excipients may serve more than one function and may
serve alternative functions depending on how much of the excipient
is present in the formulation and what other excipients are present
in the formulation.
[0135] Skilled artisans possess the knowledge and skill in the art
to enable them to select suitable pharmaceutically-acceptable
excipients in appropriate amounts for use in the invention. In
addition, there are a number of resources that are available to the
skilled artisan which describe pharmaceutically acceptable
excipients and may be useful in selecting suitable pharmaceutically
acceptable excipients. Examples include Remington's Pharmaceutical
Sciences (Mack Publishing Company), The Handbook of Pharmaceutical
Additives (Gower Publishing Limited), and The Handbook of
Pharmaceutical Excipients (the American Pharmaceutical Association
and the Pharmaceutical Press).
[0136] In Remington: The Science and Practice of Pharmacy, 21st
edition, 2005, ed. D. B. Troy, Lippincott Williams & Wilkins,
Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds.
J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York,
the contents of each of which is incorporated by reference herein,
are disclosed various carriers used in formulating pharmaceutically
acceptable compositions and known techniques for the preparation
thereof. Except insofar as any conventional carrier medium is
incompatible with the compounds disclosed herein, such as by
producing any undesirable biological effect or otherwise
interacting in a deleterious manner with any other component(s) of
the pharmaceutically acceptable composition, its use is
contemplated to be within the scope of this invention.
[0137] The pharmaceutical compositions disclosed herein are
prepared using techniques and methods known to those skilled in the
art. Some of the methods commonly used in the art are described in
Remington's Pharmaceutical Sciences (Mack Publishing Company).
[0138] Accordingly, in another aspect the invention is directed to
process for the preparation of a pharmaceutical composition
comprising the compound disclosed herein and a pharmaceutically
acceptable excipient, carrier, adjuvant, vehicle or a combination
thereof, which comprises mixing the ingredients. A pharmaceutical
composition comprising the compound disclosed herein may be
prepared by, for example, admixture at ambient temperature and
atmospheric pressure.
[0139] The compounds disclosed herein will typically be formulated
into a dosage form adapted for administration to the patient by the
desired route of administration. For example, dosage forms include
those adapted for (1) oral administration such as tablets,
capsules, caplets, pills, troches, powders, syrups, elixers,
suspensions, solutions, emulsions, sachets, and cachets; (2)
parenteral administration such as sterile solutions, suspensions,
and powders for reconstitution; (3) transdermal administration such
as transdermal patches; (4) rectal administration such as
suppositories; (5) inhalation such as aerosols, solutions, and dry
powders; and (6) topical administration such as creams, ointments,
lotions, solutions, pastes, sprays, foams, and gels.
[0140] In one embodiment, the compounds disclosed herein will be
formulated for oral administration. In another embodiment, the
compounds disclosed herein will be formulated for inhaled
administration. In a further embodiment, the compounds disclosed
herein will be formulated for intranasal administration. In another
embodiment, the compounds disclosed herein will be formulated for
transdermal administration. In a further embodiment, the compounds
disclosed herein will be formulated for topical administration.
[0141] The pharmaceutical compositions provided herein can be
provided as compressed tablets, tablet triturates, chewable
lozenges, rapidly dissolving tablets, multiple compressed tablets,
or enteric-coating tablets, sugar-coated, or film-coated tablets.
Enteric-coated tablets are compressed tablets coated with
substances that resist the action of stomach acid but dissolve or
disintegrate in the intestine, thus protecting the active
ingredients from the acidic environment of the stomach.
Enteric-coatings include, but are not limited to, fatty acids,
fats, phenyl salicylate, waxes, shellac, ammoniated shellac, and
cellulose acetate phthalates. Sugar-coated tablets are compressed
tablets surrounded by a sugar coating, which may be beneficial in
covering up objectionable tastes or odors and in protecting the
tablets from oxidation. Film-coated tablets are compressed tablets
that are covered with a thin layer or film of a water-soluble
material. Film coatings include, but are not limited to,
hydroxyethylcellulose, sodium carboxymethylcellulose, polyethylene
glycol 4000, and cellulose acetate phthalate. Film coating imparts
the same general characteristics as sugar coating. Multiple
compressed tablets are compressed tablets made by more than one
compression cycle, including layered tablets, and press-coated or
dry-coated tablets.
[0142] The tablet dosage forms can be prepared from the active
ingredient in powdered, crystalline, or granular forms, alone or in
combination with one or more carriers or excipients described
herein, including binders, disintegrants, controlled-release
polymers, lubricants, diluents, and/or colorants. Flavoring and
sweetening agents are especially useful in the formation of
chewable tablets and lozenges.
[0143] The pharmaceutical compositions provided herein can be
provided as soft or hard capsules, which can be made from gelatin,
methylcellulose, starch, or calcium alginate. The hard gelatin
capsule, also known as the dry-filled capsule (DFC), consists of
two sections, one slipping over the other, thus completely
enclosing the active ingredient. The soft elastic capsule (SEC) is
a soft, globular shell, such as a gelatin shell, which is
plasticized by the addition of glycerin, sorbitol, or a similar
polyol. The soft gelatin shells may contain a preservative to
prevent the growth of microorganisms. Suitable preservatives are
those as described herein, including methyl- and propyl-parabens,
and sorbic acid. The liquid, semisolid, and solid dosage forms
provided herein may be encapsulated in a capsule. Suitable liquid
and semisolid dosage forms include solutions and suspensions in
propylene carbonate, vegetable oils, or triglycerides. Capsules
containing such solutions can be prepared as described in U.S. Pat.
Nos. 4,328,245; 4,409,239; and 4,410,545. The capsules may also be
coated as known by those of skill in the art in order to modify or
sustain dissolution of the active ingredient.
[0144] The pharmaceutical compositions provided herein can be
provided in liquid and semisolid dosage forms, including emulsions,
solutions, suspensions, elixirs, and syrups. An emulsion is a
two-phase system, in which one liquid is dispersed in the form of
small globules throughout another liquid, which can be oil-in-water
or water-in-oil. Emulsions may include a pharmaceutically
acceptable nonaqueous liquid or solvent, emulsifying agent, and
preservative. Suspensions may include a pharmaceutically acceptable
suspending agent and preservative. Aqueous alcoholic solutions may
include a pharmaceutically acceptable acetal, such as a di(lower
alkyl) acetal of a lower alkyl aldehyde, e.g., acetaldehyde diethyl
acetal; and a water-miscible solvent having one or more hydroxyl
groups, such as propylene glycol and ethanol. Elixirs are clear,
sweetened, and hydroalcoholic solutions. Syrups are concentrated
aqueous solutions of a sugar, for example, sucrose, and may also
contain a preservative. For a liquid dosage form, for example, a
solution in a polyethylene glycol may be diluted with a sufficient
quantity of a pharmaceutically acceptable liquid carrier, e.g.,
water, to be measured conveniently for administration.
[0145] Other useful liquid and semisolid dosage forms include, but
are not limited to, those containing the active ingredient(s)
provided herein, and a dialkylated mono- or poly-alkylene glycol,
including, 1,2-dimethoxymethane, diglyme, triglyme, tetraglyme,
polyethylene glycol-350-dimethyl ether, polyethylene
glycol-550-dimethyl ether, polyethylene glycol-750-dimethyl ether,
wherein 350, 550, and 750 refer to the approximate average
molecular weight of the polyethylene glycol. These formulations can
further comprise one or more antioxidants, such as butylated
hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl
gallate, vitamin E, hydroquinone, hydroxycoumarins, ethanolamine,
lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoric
acid, bisulfite, sodium metabisulfite, thiodipropionic acid and its
esters, and dithiocarbamates.
[0146] Where appropriate, dosage unit formulations for oral
administration can be microencapsulated. The composition can also
be prepared to prolong or sustain the release as for example by
coating or embedding particulate material in polymers, wax or the
like.
[0147] The pharmaceutical compositions provided herein for oral
administration can be also provided in the forms of liposomes,
micelles, microspheres, or nanosystems. Micellar dosage forms can
be prepared as described in U.S. Pat. No. 6,350,458.
[0148] The pharmaceutical compositions provided herein can be
provided as non-effervescent or effervescent, granules and powders,
to be reconstituted into a liquid dosage form. Pharmaceutically
acceptable carriers and excipients used in the non-effervescent
granules or powders may include diluents, sweeteners, and wetting
agents. Pharmaceutically acceptable carriers and excipients used in
the effervescent granules or powders may include organic acids and
a source of carbon dioxide.
[0149] Coloring and flavoring agents can be used in all of the
above dosage forms.
[0150] The compounds disclosed herein may also be coupled with
soluble polymers as targetable drug carriers. Such polymers can
include polyvinylpyrrolidone, pyran copolymer,
polyhydroxypropylmethacrylamidephenol,
polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysine
substituted with palmitoyl residues. Furthermore, the compounds
disclosed herein may be coupled to a class of biodegradable
polymers useful in achieving controlled release of a drug, for
example, polylactic acid, polyepsilon caprolactone, polyhydroxy
butyric acid, polyorthoesters, polyacetals, polydihydropyrans,
polycyanoacrylates and cross-linked or amphipathic block copolymers
of hydrogels.
[0151] The pharmaceutical compositions provided herein can be
formulated as immediate or modified release dosage forms, including
delayed-, sustained-, pulsed-, controlled-, targeted-, and
programmed-release forms.
[0152] The pharmaceutical compositions provided herein can be
co-formulated with other active ingredients which do not impair the
desired therapeutic action, or with substances that supplement the
desired action.
[0153] The pharmaceutical compositions provided herein can be
administered parenterally by injection, infusion, or implantation,
for local or systemic administration. Parenteral administration, as
used herein, include intravenous, intraarterial, intraperitoneal,
intrathecal, intraventricular, intraurethral, intrasternal,
intracranial, intramuscular, intrasynovial, intravesical, and
subcutaneous administration.
[0154] The pharmaceutical compositions provided herein can be
formulated in any dosage forms that are suitable for parenteral
administration, including solutions, suspensions, emulsions,
micelles, liposomes, microspheres, nanosystems, and solid forms
suitable for solutions or suspensions in liquid prior to injection.
Such dosage forms can be prepared according to conventional methods
known to those skilled in the art of pharmaceutical science (see,
Remington: The Science and Practice of Pharmacy, supra).
[0155] The pharmaceutical compositions intended for parenteral
administration can include one or more pharmaceutically acceptable
carriers and excipients, including, but not limited to, aqueous
vehicles, water-miscible vehicles, non-aqueous vehicles,
antimicrobial agents or preservatives against the growth of
microorganisms, stabilizers, solubility enhancers, isotonic agents,
buffering agents, antioxidants, local anesthetics, suspending and
dispersing agents, wetting or emulsifying agents, complexing
agents, sequestering or chelating agents, cryoprotectants,
lyoprotectants, thickening agents, pH adjusting agents, and inert
gases.
[0156] Suitable aqueous vehicles include, but are not limited to,
water, saline, physiological saline or phosphate buffered saline
(PBS), sodium chloride injection, Ringers injection, isotonic
dextrose injection, sterile water injection, dextrose and lactated
Ringers injection. Non-aqueous vehicles include, but are not
limited to, fixed oils of vegetable origin, castor oil, corn oil,
cottonseed oil, olive oil, peanut oil, peppermint oil, safflower
oil, sesame oil, soybean oil, hydrogenated vegetable oils,
hydrogenated soybean oil, and medium-chain triglycerides of coconut
oil, and palm seed oil. Water-miscible vehicles include, but are
not limited to, ethanol, 1,3-butanediol, liquid polyethylene glycol
(e.g., polyethylene glycol 300 and polyethylene glycol 400),
propylene glycol, glycerin, N-methyl-2-pyrrolidone,
N,N-dimethylacetamide, and dimethyl sulfoxide.
[0157] Suitable antimicrobial agents or preservatives include, but
are not limited to, phenols, cresols, mercurials, benzyl alcohol,
chlorobutanol, methyl and propyl p-hydroxybenzoates, thimerosal,
benzalkonium chloride (e.g., benzethonium chloride), methyl- and
propyl-parabens, and sorbic acid. Suitable isotonic agents include,
but are not limited to, sodium chloride, glycerin, and dextrose.
Suitable buffering agents include, but are not limited to,
phosphate and citrate. Suitable antioxidants are those as described
herein, including bisulfite and sodium metabisulfite. Suitable
local anesthetics include, but are not limited to, procaine
hydrochloride. Suitable suspending and dispersing agents are those
as described herein, including sodium carboxymethylcelluose,
hydroxypropyl methylcellulose, and polyvinylpyrrolidone. Suitable
emulsifying agents include those described herein, including
polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan
monooleate 80, and triethanolamine oleate. Suitable sequestering or
chelating agents include, but are not limited to EDTA. Suitable pH
adjusting agents include, but are not limited to, sodium hydroxide,
hydrochloric acid, citric acid, and lactic acid. Suitable
complexing agents include, but are not limited to, cyclodextrins,
including a-cyclodextrin, .beta.-cyclodextrin,
hydroxypropyl-P-cyclodextrin, sulfobutylether-.beta.-cyclodextrin,
and sulfobutylether 7-P-cyclodextrin (CAPTISOL.RTM., CyDex, Lenexa,
Kans.).
[0158] The pharmaceutical compositions provided herein can be
formulated for single or multiple dosage administration. The single
dosage formulations are packaged in an ampoule, a vial, or a
syringe. The multiple dosage parenteral formulations must contain
an antimicrobial agent at bacteriostatic or fungistatic
concentrations. All parenteral formulations must be sterile, as
known and practiced in the art.
[0159] In one embodiment, the pharmaceutical compositions are
provided as ready-to-use sterile solutions. In another embodiment,
the pharmaceutical compositions are provided as sterile dry soluble
products, including lyophilized powders and hypodermic tablets, to
be reconstituted with a vehicle prior to use. In yet another
embodiment, the pharmaceutical compositions are provided as
ready-to-use sterile suspensions. In yet another embodiment, the
pharmaceutical compositions are provided as sterile dry insoluble
products to be reconstituted with a vehicle prior to use. In still
another embodiment, the pharmaceutical compositions are provided as
ready-to-use sterile emulsions.
[0160] The pharmaceutical compositions provided herein can be
formulated as immediate or modified release dosage forms, including
delayed-, sustained-, pulsed-, controlled-, targeted-, and
programmed-release forms.
[0161] The pharmaceutical compositions can be formulated as a
suspension, solid, semi-solid, or thixotropic liquid, for
administration as an implanted depot. In one embodiment, the
pharmaceutical compositions provided herein are dispersed in a
solid inner matrix, which is surrounded by an outer polymeric
membrane that is insoluble in body fluids but allows the active
ingredient in the pharmaceutical compositions diffuse through.
[0162] Suitable inner matrixes include polymethylmethacrylate,
polybutyl-methacrylate, plasticized or unplasticized
polyvinylchloride, plasticized nylon, plasticized polyethylene
terephthalate, natural rubber, polyisoprene, polyisobutylene,
polybutadiene, polyethylene, ethylene-vinyl acetate copolymers,
silicone rubbers, polydimethylsiloxanes, silicone carbonate
copolymers, hydrophilic polymers, such as hydrogels of esters of
acrylic and methacrylic acid, collagen, cross-linked polyvinyl
alcohol, and cross-linked partially hydrolyzed polyvinyl
acetate.
[0163] Suitable outer polymeric membranes include polyethylene,
polypropylene, ethylene/propylene copolymers, ethylene/ethyl
acrylate copolymers, ethylene/vinyl acetate copolymers, silicone
rubbers, polydimethyl siloxanes, neoprene rubber, chlorinated
polyethylene, polyvinylchloride, vinyl chloride copolymers with
vinyl acetate, vinylidene chloride, ethylene and propylene, ionomer
polyethylene terephthalate, butyl rubber epichlorohydrin rubbers,
ethylene/vinyl alcohol copolymer, ethylene/vinyl acetate/vinyl
alcohol terpolymer, and ethylene/vinyloxyethanol copolymer.
[0164] In another aspect, The pharmaceutical compositions disclosed
herein can be formulated in any dosage forms that are adapted for
administration to a patient by inhalation, for example as a dry
powder, an aerosol, a suspension, or a solution composition. In one
embodiment, the pharmaceutical compositions disclosed herein can be
formulated in a dosage form adapted for administration to a patient
by inhalation as a dry powder. In a further embodiment, the
pharmaceutical compositions disclosed herein can be formulated in a
dosage form adapted for administration to a patient by inhalation
via a nebulizer. Dry powder compositions for delivery to the lung
by inhalation typically comprise the compounds disclosed herein as
a finely divided powder together with one or more
pharmaceutically-acceptable excipients as finely divided powders.
Pharmaceutically-acceptable excipients particularly suited for use
in dry powders are known to those skilled in the art and include
lactose, starch, mannitol, and mono-, di-, and polysaccharides. The
finely divided powder may be prepared by, for example,
micronisation and milling. Generally, the size-reduced (eg
micronised) compound can be defined by a D.sub.50 value of about 1
to about 10 microns (for example as measured using laser
diffraction).
[0165] Aerosols may be formed by suspending or dissolving the
compound disclosed herein in a liquified propellant. Suitable
propellants include halocarbons, hydrocarbons, and other liquified
gases. Representative propellants include: trichlorofluoromethane
(propellant 11), dichlorofluoromethane (propellant 12),
dichlorotetrafluoroethane (propellant 114), tetrafluoroethane
(HFA-134a), 1,1-difluoroethane (HFA-152a), difluoromethane
(HFA-32), pentafluoroethane (HFA-12), heptafluoropropane
(HFA-227a), perfluoropropane, perfluorobutane, perfluoropentane,
butane, isobutane, and pentane. Aerosols comprising the compound
disclosed herein will typically be administered to a patient via a
metered dose inhaler (MDI). Such devices are known to those skilled
in the art.
[0166] The aerosol may contain additional
pharmaceutically-acceptable excipients typically used with MDIs
such as surfactants, lubricants, cosolvents and other excipients to
improve the physical stability of the formulation, to improve valve
performance, to improve solubility, or to improve taste.
[0167] Pharmaceutical compositions adapted for transdermal
administration may be presented as discrete patches intended to
remain in intimate contact with the epidermis of the patient for a
prolonged period of time. For example, the active ingredient may be
delivered from the patch by iontophoresis as generally described in
Pharmaceutical Research, 3(6), 318 (1986).
[0168] Pharmaceutical compositions adapted for topical
administration may be formulated as ointments, creams, suspensions,
lotions, powders, solutions, pastes, gels, sprays, aerosols or
oils. Ointments, creams and gels, may, for example, be formulated
with an aqueous or oily base with the addition of suitable
thickening and/or gelling agent and/or solvents. Such bases may
thus, for example, include water and/or oil such as liquid paraffin
or a vegetable oil such as arachis oil or castor oil, or a solvent
such as polyethylene glycol. Thickening agents and gelling agents
which may be used according to the nature of the base include soft
paraffin, aluminium stearate, cetostearyl alcohol, polyethylene
glycols, woolfat, beeswax, carboxypolymethylene and cellulose
derivatives, and/or glyceryl monostearate and/or non-ionic
emulsifying agents.
[0169] Lotions may be formulated with an aqueous or oily base and
will in general also contain one or more emulsifying agents,
stabilising agents, dispersing agents, suspending agents or
thickening agents.
[0170] Powders for external application may be formed with the aid
of any suitable powder base, for example, talc, lactose or starch.
Drops may be formulated with an aqueous or nonaqueous base also
comprising one or more dispersing agents, solubilizing agents,
suspending agents or preservatives.
[0171] Topical preparations may be administered by one or more
applications per day to the affected area; over skin areas
occlusive dressings may advantageously be used. Continuous or
prolonged delivery may be achieved by an adhesive reservoir
system.
[0172] For treatments of the eye or other external tissues, for
example mouth and skin, the compositions may be applied as a
topical ointment or cream. When formulated in an ointment, the
compound disclosed herein may be employed with either a paraffinic
or a water-miscible ointment base. Alternatively, the compound
disclosed herein may be formulated in a cream with an oil-in-water
cream base or a water-in-oil base.
Use of the Compounds and Compositions of the Invention
[0173] The present invention provides a method of using the
compound disclosed herein, or the pharmaceutical composition
comprising the compound disclosed herein for the treatment,
prevention, or amelioration of a disease that are mediated or
otherwise affected via receptor tyrosine kinase, especially Axl,
Mer, c-Met and/or Ron kinase activity.
[0174] In one embodiment, provided herein is a method of using the
compound or the pharmaceutical composition disclosed herein for the
treatment, prevention or amelioration of a disease that is mediated
or otherwise affected via inappropriate Axl kinase activity. In
another embodiment, a disease is related to the inappropriate
activity of Mer kinase. In still another embodiment, a disease is
related to the inappropriate activity of c-Met kinase. In yet
another embodiment, a disease is related to the inappropriate
activity of Ron kinase.
[0175] The compound disclosed herein would be useful for, but not
limited to, the prevention or treatment of proliferative diseases,
condition, or disorder in a patient by administering to the patient
a compound or a composition disclosed herein in an effective
amount. Such diseases, conditions, or disorders include cancer,
particularly metastatic cancer, atherosclerosis, and lung
fibrosis.
[0176] The compounds disclosed herein would be useful for the
treatment of neoplasia including cancer and metastasis, including,
but not limited to: carcinoma such as cancer of the bladder,
breast, colon, kidney, liver, lung (including small cell lung
cancer), esophagus, gall-bladder, ovary, pancreas, stomach, cervix,
thyroid, prostate, and skin (including squamous cell carcinoma);
hematopoietic tumors of lymphoid lineage (including leukemia, acute
lymphocitic leukemia, acute lymphoblastic leukemia, B-cell
lymphoma, T-cell-lymphoma, Hodgkin's lymphoma, non-Hodgkin's
lymphoma, hairy cell lymphoma and Burkett's lymphoma);
hematopoietic tumors of myeloid lineage (including acute and
chronic myelogenous leukemias, myelodysplastic syndrome and
promyelocytic leukemia); tumors of mesenchymal origin (including
fibrosarcoma and rhabdomyosarcoma, and other sarcomas, e.g. soft
tissue and bone); tumors of the central and peripheral nervous
system (including astrocytoma, neuroblastoma, glioma and
schwannomas); and other tumors (including melanoma, seminoma,
teratocarcinoma, osteosarcoma, xeroderoma pigmentosum,
keratoacanthoma, thyroid follicular cancer and Kaposi's
sarcoma).
[0177] The compounds disclosed herein also would be useful for
treatment of ophthalmological conditions such as corneal graft
rejection, ocular neovascularization, retinal neovascularization
including neovascularization following injury or infection,
diabetic retinopathy, retrolental fibroplasia and neovascular
glaucoma; retinal ischemia; vitreous hemorrhage; ulcerative
diseases such as gastric ulcer; pathological, but non-malignant,
conditions such as hemangiomas, including infantile hemaginomas,
angiofibroma of the nasopharynx and avascular necrosis of bone; and
disorders of the female reproductive system such as endometriosis.
The compounds are also useful for the treatment of edema, and
conditions of vascular hyperpermeability.
[0178] The compounds disclosed herein are also useful in the
treatment of diabetic conditions such as diabetic retinopathy and
microangiopathy. The compounds disclosed herein are also useful in
the reduction of blood flow in a tumor in a subject. The compounds
disclosed herein are also useful in the reduction of metastasis of
a tumor in a subject.
[0179] Besides being useful for human treatment, the compounds
disclosed herein are also useful for veterinary treatment of
companion animals, exotic animals and farm animals, including
mammals, rodents, and the like. More preferred animals include
horses, dogs, and cats. As used herein, the compounds disclosed
herein include the pharmaceutically acceptable derivatives
thereof.
Combination Therapy
[0180] A compound disclosed herein can be administered as the sole
active agent or it can be administered in combination with other
therapeutic agents, including other compounds that demonstrate the
same or a similar therapeutic activity and that are determined to
be safe and efficacious for such combined administration.
[0181] In one aspect, provided herein is a method of treating,
preventing, or ameliorating a disease or disorder comprising
administering a safe and effective amount of a combination
comprising the compound disclosed herein together with one or more
other therapeutically active agents. In one embodiment, the
combinations comprising one or two other therapeutic agents.
[0182] In another aspect, provided herein is a product comprising a
compound disclosed herein and at least one other therapeutic agent
as a combined preparation for simultaneous, separate or sequential
use in therapy. In one embodiment, the therapy is the treatment of
a disease or disorder mediated by the activity of the Axl, Mer,
c-Met and/or Ron kinase. Products provided as a combined
preparation include a composition comprising the compound disclosed
herein and the other therapeutic agent(s) together in the same
pharmaceutical composition, or the compound disclosed herein and
the other therapeutic agent(s) in separate form, e.g. in the form
of a kit.
[0183] In another aspect, provided herein is a pharmaceutical
composition comprising a compound disclosed herein and another
therapeutic agent(s). In one embodiment, the pharmaceutical
composition may comprise a pharmaceutically acceptable excipient,
carrier, adjuvant or vehicle as described above.
[0184] In another aspect, provided herein is a kit comprising two
or more separate pharmaceutical compositions, at least one of which
contains a compound disclosed herein. In one embodiment, the kit
comprises means for separately retaining said compositions, such as
a container, divided bottle, or divided foil packet. An example of
such a kit is a blister pack, as typically used for the packaging
of tablets, capsules and the like.
[0185] The invention also provides the use of a compound disclosed
herein for treating a disease or condition mediated by the activity
of the Axl, Mer, c-Met and/or Ron kinase, wherein the patient has
previously (e.g. within 24 hours) been treated with another
therapeutic agent. The invention also provides the use of another
therapeutic agent for treating a disease or condition mediated by
the activity of the Axl, Mer, c-Met and/or Ron kinase, wherein the
patient has previously (e.g. within 24 hours) been treated with a
compound disclosed herein.
[0186] The therapeutic agents which can be used in combination with
the compound disclosed herein include, but not limited to,
chemotherapeutic agents or other anti-proliferative agents, agents
for treating atherosclerosis, agents for treating lung fibrosis.
Where the compound disclosed herein is administered in conjunction
with other therapeutic agent will of course vary depending on the
type of co-drug employed, on the specific drug employed, on the
condition being treated and so forth.
[0187] The chemotherapeutic agents or other anti-proliferative
agents may refer to aromatase inhibitors; antiestrogens;
topoisomerase I inhibitors; topoisomerase II inhibitors;
microtubule active agents; alkylating agents; histone deacetylase
inhibitors; compounds that induce cell differentiation processes;
cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors;
antineoplastic antimetabolites; platin compounds; compounds
targeting/decreasing a protein or lipid kinase activity and further
anti-angiogenic compounds; compounds which target, decrease or
inhibit the activity of a protein or lipid phosphatase; gonadorelin
agonists; anti-androgens; methionine aminopeptidase inhibitors;
bisphosphonates; biological response modifiers; antiproliferative
antibodies; heparanase inhibitors; inhibitors of Ras oncogenic
isoforms; telomerase inhibitors; proteasome inhibitors; agents used
in the treatment of hematologic malignancies; compounds which
target, decrease or inhibit the activity of Flt-3; Hsp90
inhibitors; temozolomide (TEMODAL.RTM.); and leucovorin.
[0188] The term "aromatase inhibitor", as used herein, relates to a
compound which inhibits the estrogen production, i.e., the
conversion of the substrates androstenedione and testosterone to
estrone and estradiol, respectively. The term includes, but is not
limited to, steroids, especially atamestane, exemestane and
formestane; and, in particular, nonsteroids, especially
aminoglutethimide, roglethimide, pyridoglutethimide, trilostane,
testolactone, ketoconazole, vorozole, fadrozole, anastrozole and
letrozole. Exemestane can be administered, e.g., in the form as it
is marketed, e.g., under the trademark AROMASIN.RTM.. Formestane
can be administered, e.g., in the form as it is marketed, e.g.,
under the trademark LENTARON.RTM.. Fadrozole can be administered,
e.g., in the form as it is marketed, e.g., under the trademark
AFEMA.RTM.. Anastrozole can be administered, e.g., in the form as
it is marketed, e.g., under the trademark ARIMIDEX.RTM.. Letrozole
can be administered, e.g., in the form as it is marketed, e.g.,
under the trademark FEMARA.RTM. or FEMAR.RTM.. Aminoglutethimide
can be administered, e.g., in the form as it is marketed, e.g.,
under the trademark ORIMETEN.RTM.. A combination of the invention
comprising a chemotherapeutic agent which is an aromatase inhibitor
is particularly useful for the treatment of hormone receptor
positive tumors, e.g., breast tumors.
[0189] The term "aromatase inhibitor", as used herein, relates to a
compound which inhibits the estrogen production, i.e., the
conversion of the substrates androstenedione and testosterone to
estrone and estradiol, respectively. The term includes, but is not
limited to, steroids, especially atamestane, exemestane and
formestane; and, in particular, nonsteroids, especially
aminoglutethimide, roglethimide, pyridoglutethimide, trilostane,
testolactone, ketoconazole, vorozole, fadrozole, anastrozole and
letrozole. Exemestane can be administered, e.g., in the form as it
is marketed, e.g., under the trademark AROMASIN.RTM.. Formestane
can be administered, e.g., in the form as it is marketed, e.g.,
under the trademark LENTARON.RTM.. Fadrozole can be administered,
e.g., in the form as it is marketed, e.g., under the trademark
AFEMA.RTM.. Anastrozole can be administered, e.g., in the form as
it is marketed, e.g., under the trademark ARIMIDEX.RTM.. Letrozole
can be administered, e.g., in the form as it is marketed, e.g.,
under the trademark FEMARA.RTM. or FEMAR.RTM.. Aminoglutethimide
can be administered, e.g., in the form as it is marketed, e.g.,
under the trademark ORIMETEN.RTM.. A combination of the invention
comprising a chemotherapeutic agent which is an aromatase inhibitor
is particularly useful for the treatment of hormone receptor
positive tumors, e.g., breast tumors.
[0190] The term "anti-estrogen", as used herein, relates to a
compound which antagonizes the effect of estrogens at the estrogen
receptor level. The term includes, but is not limited to,
tamoxifen, fulvestrant, raloxifene and raloxifene hydrochloride.
Tamoxifen can be administered, e.g., in the form as it is marketed,
e.g., under the trademark NOLVADEX.RTM.. Raloxifene hydrochloride
can be administered, e.g., in the form as it is marketed, e.g.,
under the trademark EVISTA.RTM.. Fulvestrant can be formulated as
disclosed in U.S. Pat. No. 4,659,516 or it can be administered,
e.g., in the form as it is marketed, e.g., under the trademark
FASLODEX.RTM.. A combination of the invention comprising a
chemotherapeutic agent which is an antiestrogen is particularly
useful for the treatment of estrogen receptor positive tumors,
e.g., breast tumors.
[0191] The term "anti-androgen", as used herein, relates to any
substance which is capable of inhibiting the biological effects of
androgenic hormones and includes, but is not limited to,
bicalutamide (CASODEX), which can be formulated, e.g., as disclosed
in U.S. Pat. No. 4,636,505.
[0192] The term "gonadorelin agonist", as used herein, includes,
but is not limited to, abarelix, goserelin and goserelin acetate.
Goserelin is disclosed in U.S. Pat. No. 4,100,274 and can be
administered, e.g., in the form as it is marketed, e.g., under the
trademark ZOLADEX.RTM.. Abarelix can be formulated, e.g., as
disclosed in U.S. Pat. No. 5,843,901. The term "topoisomerase I
inhibitor", as used herein, includes, but is not limited to,
topotecan, gimatecan, irinotecan, camptothecian and its analogues,
9-nitrocamptothecin and the macromolecular camptothecin conjugate
PNU-166148 (compound A1 in WO 99/17804). Irinotecan can be
administered, e.g., in the form as it is marketed, e.g., under the
trademark CAMPTOSAR.RTM.. Topotecan can be administered, e.g., in
the form as it is marketed, e.g., under the trademark
HYCAMTIN.RTM..
[0193] The term "topoisomerase II inhibitor", as used herein,
includes, but is not limited to, the anthracyclines, such as
doxorubicin, including liposomal formulation, e.g., CAELYX.RTM.;
daunorubicin; epirubicin; idarubicin; nemorubicin; the
anthraquinones mitoxantrone and losoxantrone; and the
podophillotoxines etoposide and teniposide. Etoposide can be
administered, e.g., in the form as it is marketed, e.g., under the
trademark ETOPOPHOS.RTM.. Teniposide can be administered, e.g., in
the form as it is marketed, e.g., under the trademark VM
26-BRISTOL.RTM.. Doxorubicin can be administered, e.g., in the form
as it is marketed, e.g., under the trademark ADRIBLASTIN.RTM. or
ADRIAMYCIN.RTM..
[0194] Epirubicin can be administered, e.g., in the form as it is
marketed, e.g., under the trademark FARMORUBICIN.RTM.. Idarubicin
can be administered, e.g., in the form as it is marketed, e.g.,
under the trademark ZAVEDOS.RTM.. Mitoxantrone can be administered,
e.g., in the form as it is marketed, e.g., under the trademark
NOVANTRON.RTM..
[0195] The term "microtubule active agent" relates to microtubule
stabilizing, microtubule destabilizing agents and microtublin
polymerization inhibitors including, but not limited to, taxanes,
e.g., paclitaxel and docetaxel; vinca alkaloids, e.g., vinblastine,
especially vinblastine sulfate; vincristine, especially vincristine
sulfate and vinorelbine; discodermolides; cochicine; and
epothilones and derivatives thereof, e.g., epothilone B or D or
derivatives thereof. Paclitaxel may be administered, e.g., in the
form as it is marketed, e.g., TAXOL.RTM.. Docetaxel can be
administered, e.g., in the form as it is marketed, e.g., under the
trademark TAXOTERE.RTM.. Vinblastine sulfate can be administered,
e.g., in the form as it is marketed, e.g., under the trademark
VINBLASTIN R.P.RTM.. Vincristine sulfate can be administered, e.g.,
in the form as it is marketed, e.g., under the trademark
FARMISTIN.RTM.. Discodermolide can be obtained, e.g., as disclosed
in U.S. Pat. No. 5,010,099. Also included are epothilone
derivatives which are disclosed in WO 98/10121, U.S. Pat. No.
6,194,181, WO 98/25929, WO 98/08849, WO 99/43653, WO 98/22461 and
WO 00/31247. Especially preferred are epothilone A and/or B.
[0196] The term "alkylating agent", as used herein, includes, but
is not limited to, cyclophosphamide, ifosfamide, melphalan or
nitrosourea (BCNU or Gliadel). Cyclophosphamide can be
administered, e.g., in the form as it is marketed, e.g., under the
trademark CYCLOSTIN.RTM.. Ifosfamide can be administered, e.g., in
the form as it is marketed, e.g., under the trademark
HOLOXAN.RTM..
[0197] The term "histone deacetylase inhibitors" or "HDAC
inhibitors" relates to compounds which inhibit the histone
deacetylase and which possess antiproliferative activity. This
includes compounds disclosed in WO 02/22577, especially
N-hydroxy-3-[4-[[(2-hydroxyethyl)[2-(1H-indol-3-yl)ethyl]-amino]methyl]ph-
enyl]-2E-2-propenamide,
N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]--
2E-2-propenamide and pharmaceutically acceptable salts thereof. It
further especially includes suberoylanilide hydroxamic acid
(SAHA).
[0198] The term "antineoplastic antimetabolite" includes, but is
not limited to, 5-fluorouracil or 5-FU; capecitabine; gemcitabine;
DNA demethylating agents, such as 5-azacytidine and decitabine;
methotrexate and edatrexate; and folic acid antagonists, such as
pemetrexed. Capecitabine can be administered, e.g., in the form as
it is marketed, e.g., under the trademark XELODA.RTM.. Gemcitabine
can be administered, e.g., in the form as it is marketed, e.g.,
under the trademark GEMZAR.RTM.. Also included is the monoclonal
antibody trastuzumab which can be administered, e.g., in the form
as it is marketed, e.g., under the trademark HERCEPTIN.RTM..
[0199] The term "platin compound", as used herein, includes, but is
not limited to, carboplatin, cis-platin, cisplatinum and
oxaliplatin. Carboplatin can be administered, e.g., in the form as
it is marketed, e.g., under the trademark CARBOPLAT.RTM..
Oxaliplatin can be administered, e.g., in the form as it is
marketed, e.g., under the trademark ELOXATIN.RTM..
[0200] The term "compounds targeting/decreasing a protein or lipid
kinase activity; or a protein or lipid phosphatase activity; or
further anti-angiogenic compounds", as used herein, includes, but
is not limited to, protein tyrosine kinase and/or serine and/or
threonine kinase inhibitors or lipid kinase inhibitors, e.g.,
a) compounds targeting, decreasing or inhibiting the activity of
the platelet-derived growth factor-receptors (PDGFR), such as
compounds which target, decrease or inhibit the activity of PDGFR,
especially compounds which inhibit the PDGF receptor, e.g., a
N-phenyl-2-pyrimidine-amine derivative, e.g., imatinib, SU101,
SU6668 and GFB-111; b) compounds targeting, decreasing or
inhibiting the activity of the fibroblast growth factor-receptors
(FGFR); c) compounds targeting, decreasing or inhibiting the
activity of the insulin-like growth factor receptor I (IGF-IR),
such as compounds which target, decrease or inhibit the activity of
IGF-IR, especially compounds which inhibit the IGF-IR receptor,
such as those compounds disclosed in WO 02/092599; d) compounds
targeting, decreasing or inhibiting the activity of the Trk
receptor tyrosine kinase family; e) compounds targeting, decreasing
or inhibiting the activity of the Axl receptor tyrosine kinase
family; f) compounds targeting, decreasing or inhibiting the
activity of the c-Met receptor; g) compounds targeting, decreasing
or inhibiting the activity of the Kit/SCFR receptor tyrosine
kinase; h) compounds targeting, decreasing or inhibiting the
activity of the c-kit receptor tyrosine kinases--(part of the PDGFR
family), such as compounds which target, decrease or inhibit the
activity of the c-Kit receptor tyrosine kinase family, especially
compounds which inhibit the c-Kit receptor, e.g., imatinib; i)
compounds targeting, decreasing or inhibiting the activity of
members of the c-Abl family and their gene-fusion products, e.g.,
BCR-Abl kinase, such as compounds which target decrease or inhibit
the activity of c-Abl family members and their gene fusion
products, e.g., a N-phenyl-2-pyrimidine-amine derivative, e.g.,
imatinib, PD180970, AG957, NSC 680410 or PD173955 from ParkeDavis;
j) compounds targeting, decreasing or inhibiting the activity of
members of the protein kinase C (PKC) and Raf family of
serine/threonine kinases, members of the MEK, SRC, JAK, FAK, PDK
and Ras/MAPK family members, or PI(3) kinase family, or of the
PI(3)-kinase-related kinase family, and/or members of the
cyclin-dependent kinase family (CDK) and are especially those
staurosporine derivatives disclosed in U.S. Pat. No. 5,093,330,
e.g., midostaurin; examples of further compounds include, e.g.,
UCN-01; safingol; BAY 43-9006; Bryostatin 1; Perifosine;
Ilmofosine; RO 318220 and RO 320432; GO 6976; Isis 3521;
LY333531/LY379196; isochinoline compounds, such as those disclosed
in WO 00/09495; FTIs; PD184352; or QAN697 (a PI3K inhibitor); k)
compounds targeting, decreasing or inhibiting the activity of
protein-tyrosine kinase inhibitors, such as compounds which target,
decrease or inhibit the activity of protein-tyrosine kinase
inhibitors include imatinib mesylate (GLEEVEC.RTM.) or tyrphostin.
A tyrphostin is preferably a low molecular weight (Mr<1500)
compound, or a pharmaceutically acceptable salt thereof, especially
a compound selected from the benzylidenemalonitrile class or the
S-arylbenzenemalonirile or bisubstrate quinoline class of
compounds, more especially any compound selected from the group
consisting of Tyrphostin A23/RG-50810, AG 99, Tyrphostin AG 213,
Tyrphostin AG 1748, Tyrphostin AG 490, Tyrphostin B44, Tyrphostin
B44 (+) enantiomer, Tyrphostin AG 555, AG 494, Tyrphostin AG 556,
AG957 and adaphostin
(4-{[(2,5-dihydroxyphenyl)methyl]amino}-benzoic acid adamantyl
ester, NSC 680410, adaphostin; and l) compounds targeting,
decreasing or inhibiting the activity of the epidermal growth
factor family of receptor tyrosine kinases (EGFR, ErbB2, ErbB3,
ErbB4 as homo- or hetero-dimers), such as compounds which target,
decrease or inhibit the activity of the epidermal growth factor
receptor family are especially compounds, proteins or antibodies
which inhibit members of the EGF receptor tyrosine kinase family,
e.g., EGF receptor, ErbB2, ErbB3 and ErbB4 or bind to EGF or EGF
related ligands, and are in particular those compounds, proteins or
monoclonal antibodies generically and specifically disclosed in WO
97/02266, e.g., the compound of Example 39, or in EP 0564409; WO
99/03854; EP 0520722; EP 0566226; EP 0787722; EP 0837063; U.S. Pat.
No. 5,747,498; WO 98/10767; WO 97/30034; WO 97/49688; WO 97/38983
and, especially, WO 96/30347, e.g., compound known as CP 358774; WO
96/33980, e.g., compound ZD 1839; and WO 95/03283, e.g., compound
ZM105180, e.g., trastuzumab (HERCEPTIN), cetuximab, Iressa,
Tarceva, OS-774, CI-1033, EKB-569, GW-2016, E1.1, E2.4, E2.5, E6.2,
E6.4, E2.11, E6.3 or E7.6.3; and 7H-pyrrolo-[2,3-d]pyrimidine
derivatives which are disclosed in WO 03/013541.
[0201] Further anti-angiogenic compounds include compounds having
another mechanism for their activity, e.g., unrelated to protein or
lipid kinase inhibition, e.g., thalidomide (THALOMID.RTM.) and
TNP-470.
[0202] Compounds which target, decrease or inhibit the activity of
a protein or lipid phosphatase are, e.g., inhibitors of phosphatase
1, phosphatase 2A, PTEN or CDC25, e.g., okadaic acid or a
derivative thereof.
[0203] Compounds that induce cell differentiation processes are
e.g. retinoic acid, .alpha.-, .gamma.- or .delta.-tocopherol or
.alpha.-, .gamma.- or .delta.-tocotrienol.
[0204] The term cyclooxygenase inhibitor, as used herein, includes,
but is not limited to, e.g., Cox-2 inhibitors, 5-alkyl substituted
2-arylaminophenylacetic acid and derivatives, such as celecoxib
(CELEBREX.RTM.), rofecoxib (VIOXX.RTM.), etoricoxib, valdecoxib or
a 5-alkyl-2-arylaminophenylacetic acid, e.g.,
5-methyl-2-(2'-chloro-6'-fluoroanilino)phenyl acetic acid or
lumiracoxib.
[0205] The term "bisphosphonates", as used herein, includes, but is
not limited to, etridonic, clodronic, tiludronic, pamidronic,
alendronic, ibandronic, risedronic and zoledronic acid. "Etridonic
acid" can be administered, e.g., in the form as it is marketed,
e.g., under the trademark DIDRONEL.RTM.. "Clodronic acid" can be
administered, e.g., in the form as it is marketed, e.g., under the
trademark BONEFOS.RTM.. "Tiludronic acid" can be administered,
e.g., in the form as it is marketed, e.g., under the trademark
SKELID.RTM.. "Pamidronic acid" can be administered, e.g., in the
form as it is marketed, e.g., under the trademark AREDIA.TM..
"Alendronic acid" can be administered, e.g., in the form as it is
marketed, e.g., under the trademark FOSAMAX.RTM.. "Ibandronic acid"
can be administered, e.g., in the form as it is marketed, e.g.,
under the trademark BONDRANAT.RTM.. "Risedronic acid" can be
administered, e.g., in the form as it is marketed, e.g., under the
trademark ACTONEL.RTM.. "Zoledronic acid" can be administered,
e.g., in the form as it is marketed, e.g., under the trademark
ZOMETA.RTM..
[0206] The term "mTOR inhibitors" relates to compounds which
inhibit the mammalian target of rapamycin (mTOR) and which possess
antiproliferative activity, such as sirolimus (Rapamune),
everolimus (Certican.TM.), CCI-779 and ABT578.
[0207] The term "heparanase inhibitor", as used herein, refers to
compounds which target, decrease or inhibit heparin sulphate
degradation. The term includes, but is not limited to, PI-88.
[0208] The term "biological response modifier", as used herein,
refers to a lymphokine or interferons, e.g., interferon
.gamma..
[0209] The term "inhibitor of Ras oncogenic isoforms", e.g., H-Ras,
K-Ras or N-Ras, as used herein, refers to compounds which target,
decrease or inhibit the oncogenic activity of Ras, e.g., a
"farnesyl transferase inhibitor", e.g., L-744832, DK8G557 or R1
15777 (Zarnestra).
[0210] The term "telomerase inhibitor", as used herein, refers to
compounds which target, decrease or inhibit the activity of
telomerase. Compounds which target, decrease or inhibit the
activity of telomerase are especially compounds which inhibit the
telomerase receptor, e.g., telomestatin.
[0211] The term "methionine aminopeptidase inhibitor", as used
herein, refers to compounds which target, decrease or inhibit the
activity of methionine aminopeptidase. Compounds which target,
decrease or inhibit the activity of methionine aminopeptidase are,
e.g., bengamide or a derivative thereof.
[0212] The term "proteasome inhibitor", as used herein, refers to
compounds which target, decrease or inhibit the activity of the
proteasome. Compounds which target, decrease or inhibit the
activity of the proteasome include, e.g., PS-341 and MLN 341.
[0213] The term "matrix metalloproteinase inhibitor" or "MMP
inhibitor", as used herein, includes, but is not limited to,
collagen peptidomimetic and nonpeptidomimetic inhibitors,
tetracycline derivatives, e.g., hydroxamate peptidomimetic
inhibitor batimastat and its orally bioavailable analogue
marimastat (BB-2516), prinomastat (AG3340), metastat (NSC 683551)
BMS-279251, BAY 12-9566, TAA211, MMI270B or AAJ996.
[0214] The term "agents used in the treatment of hematologic
malignancies", as used herein, includes, but is not limited to,
FMS-like tyrosine kinase inhibitors, e.g., compounds targeting,
decreasing or inhibiting the activity of FMS-like tyrosine kinase
receptors (Flt-3R); interferon, 1-b-D-arabinofuransylcytosine
(ara-c) and bisulfan; and ALK inhibitors, e.g., compounds which
target, decrease or inhibit anaplastic lymphoma kinase.
[0215] Compounds which target, decrease or inhibit the activity of
FMS-like tyrosine kinase receptors (Flt-3R) are especially
compounds, proteins or antibodies which inhibit members of the
Flt-3R receptor kinase family, e.g., PKC412, midostaurin, a
staurosporine derivative, SU1 1248 and MLN518.
[0216] The term "HSP90 inhibitors", as used herein, includes, but
is not limited to, compounds targeting, decreasing or inhibiting
the intrinsic ATPase activity of HSP90; degrading, targeting,
decreasing or inhibiting the HSP90 client proteins via the
ubiquitin proteasome pathway. Compounds targeting, decreasing or
inhibiting the intrinsic ATPase activity of HSP90 are especially
compounds, proteins or antibodies which inhibit the ATPase activity
of HSP90, e.g., 17-allylamino, 17-demethoxygeldanamycin (17AAG), a
geldanamycin derivative, other geldanamycin related compounds,
radicicol and HDAC inhibitors.
[0217] The term "antiproliferative antibodies", as used herein,
includes, but is not limited to, trastuzumab (HERCEPTIN.TM.),
Trastuzumab-DM1, erlotinib (TARCEVA.TM.), bevacizumab
(AVASTIN.TM.), rituximab (RITUXAN.RTM.), PR064553 (anti-CD40) and
2C4 antibody. By antibodies is meant, e.g., intact monoclonal
antibodies, polyclonal antibodies, multispecific antibodies formed
from at least two intact antibodies, and antibodies fragments so
long as they exhibit the desired biological activity. For the
treatment of acute myeloid leukemia (AML), compounds disclosed
herein can be used in combination with standard leukemia therapies,
especially in combination with therapies used for the treatment of
AML. In particular, compounds disclosed herein can be administered
in combination with, e.g., farnesyl transferase inhibitors and/or
other drugs useful for the treatment of AML, such as Daunorubicin,
Adriamycin, Ara-C, VP-16, Teniposide, Mitoxantrone, Idarubicin,
Carboplatinum and PKC412.
[0218] The structure of the active agents identified by code nos.,
generic or trade names may be taken from the actual edition of the
standard compendium "The Merck Index" or from databases, e.g.,
Patents International, e.g., IMS World Publications.
[0219] The above-mentioned compounds, which can be used in
combination with a compound disclosed herein, can be prepared and
administered as described in the art, such as in the documents
cited above.
[0220] A compound disclosed herein may also be used to advantage in
combination with known therapeutic processes, e.g., surgery, the
administration of hormones or especially radiation (in but a few
examples, gamma radiation, neutron beam radiotherapy, electron beam
radiotherapy, proton therapy, brachytherapy, and systemic
radioactive isotopes, to name a few). A compound disclosed herein
may in particular be used as a radiosensitizer, especially for the
treatment of tumors which exhibit poor sensitivity to
radiotherapy.
[0221] By "combination", there is meant either a fixed combination
in one dosage unit form, or a kit of parts for the combined
administration where a compound disclosed herein and a combination
partner may be administered independently at the same time or
separately within time intervals that especially allow that the
combination partners show a cooperative, e.g., synergistic, effect
or any combination thereof. The terms "coadministration" or
"combined administration" or the like as utilized herein are meant
to encompass administration of the selected combination partner to
a single subject in need thereof (e.g. a patient), and are intended
to include treatment regimens in which the agents are not
necessarily administered by the same route of administration or at
the same time. The term "pharmaceutical combination" as used herein
means a product that results from the mixing or combining of more
than one active ingredient and includes both fixed and non-fixed
combinations of the active ingredients. The term "fixed
combination" means that the active ingredients, e.g. a compound
disclosed herein and a combination partner, are both administered
to a patient simultaneously in the form of a single entity or
dosage. The term "non-fixed combination" means that the active
ingredients, e.g. a compound disclosed herein and a combination
partner, are both administered to a patient as separate entities
either simultaneously, concurrently or sequentially with no
specific time limits, wherein such administration provides
therapeutically effective levels of the two compounds in the body
of the patient. The latter also applies to cocktail therapy, e.g.
the administration of three or more active ingredients.
Methods of Treatment
[0222] In one embodiment, the methods of treatment disclosed herein
comprise administering a safe and effective amount of a compound or
a pharmaceutically composition disclosed herein to a patient in
need thereof. Individual embodiments disclosed herein include
methods of treating any one of the above-mentioned disorders by
administering a safe and effective amount of a compound disclosed
herein or a pharmaceutical composition containing a compound
disclosed herein to a patient in need thereof.
[0223] In one embodiment, the compounds disclosed herein or
pharmaceutically compositions containing the compounds disclosed
herein may be administered by any suitable route of administration,
including both systemic administration and topical administration.
Systemic administration includes oral administration, parenteral
administration, transdermal administration and rectal
administration. Parenteral administration is typically by injection
or infusion, including intravenous, intramuscular, and subcutaneous
injection or infusion. Topical administration includes application
to the skin as well as intraocular, otic, intravaginal, inhaled and
intranasal administration. In one embodiment, the compounds
disclosed herein or pharmaceutical compositions containing the
compounds disclosed herein may be administered orally. In another
embodiment, the compounds disclosed herein or pharmaceutically
compositions containing the compounds disclosed herein may be
administered by inhalation. In a further embodiment, the compounds
disclosed herein or pharmaceutical compositions containing the
compounds disclosed herein may be administered intranasally.
[0224] In another embodiment, the compounds disclosed herein or
pharmaceutically compositions containing the compounds disclosed
herein may be administered once or according to a dosing regimen
wherein a number of doses are administered at varying intervals of
time for a given period of time. For example, doses may be
administered one, two, three, or four times per day. In one
embodiment, a dose is administered once per day. In a further
embodiment, a dose is administered twice per day. Doses may be
administered until the desired therapeutic effect is achieved or
indefinitely to maintain the desired therapeutic effect. Suitable
dosing regimens for a compound disclosed herein or a pharmaceutical
composition containing a compound disclosed herein depend on the
pharmacokinetic properties of that compound, such as absorption,
distribution, and half-life, which can be determined by the skilled
artisan. In addition, suitable dosing regimens, including the
duration such regimens are administered, for a compound disclosed
herein or a pharmaceutical composition containing a compound
disclosed herein depend on the disorder being treated, the severity
of the disorder being treated, the age and physical condition of
the patient being treated, the medical history of the patient to be
treated, the nature of concurrent therapy, the desired therapeutic
effect, and like factors within the knowledge and expertise of the
skilled artisan. It will be further understood by such skilled
artisans that suitable dosing regimens may require adjustment given
an individual patient's response to the dosing regimen or over time
as individual patient needs change.
[0225] The compound of the present invention may be administered
either simultaneously with, or before or after, one or more other
therapeutic agent. The compound of the present invention may be
administered separately, by the same or different route of
administration, or together in the same pharmaceutical composition
as the other agents.
[0226] The pharmaceutical composition or combination of the present
invention can be in unit dosage of about 1-1000 mg of active
ingredient(s) for a subject of about 50-70 kg, or about 1-500 mg or
about 1-250 mg or about 1-150 mg or about 0.5-100 mg, or about 1-50
mg of active ingredients. The therapeutically effective dosage of a
compound, the pharmaceutical composition, or the combinations
thereof, is dependent on the species of the subject, the body
weight, age and individual condition, the disorder or disease or
the severity thereof being treated. A physician, clinician or
veterinarian of ordinary skill can readily determine the effective
amount of each of the active ingredients necessary to prevent,
treat or inhibit the progress of the disorder or disease. The
above-cited dosage properties are demonstrable in vitro and in vivo
tests using advantageously mammals, e.g., mice, rats, dogs, monkeys
or isolated organs, tissues and preparations thereof. The compounds
of the present invention can be applied in vitro in the form of
solutions, e.g., aqueous solutions, and in vivo either enterally,
parenterally, advantageously intravenously, e.g., as a suspension
or in aqueous solution.
[0227] In one embodiment, the therapeutically effective dose is
from about 0.1 mg to about 2,000 mg per day of a compound provided
herein. The pharmaceutical compositions therefore should provide a
dosage of from about 0.1 mg to about 2000 mg of the compound. In
certain embodiments, pharmaceutical dosage unit forms are prepared
to provide from about 1 mg to about 2000 mg, from about 10 mg to
about 1000 mg, from about 20 mg to about 500 mg or from about 25 mg
to about 250 mg of the essential active ingredient or a combination
of essential ingredients per dosage unit form. In certain
embodiments, the pharmaceutical dosage unit forms are prepared to
provide about 10 mg, 20 mg, 25 mg, 50 mg, 100 mg, 250 mg, 500 mg,
1000 mg or 2000 mg of the essential active ingredient.
[0228] Additionally, the compounds disclosed herein may be
administered as prodrugs. As used herein, a "prodrug" of a compound
disclosed herein is a functional derivative of the compound which,
upon administration to a patient, eventually liberates the compound
disclosed herein in vivo. Administration of a compound disclosed
herein as a prodrug may enable the skilled artisan to do one or
more of the following: (a) modify the onset of the activity of the
compound in vivo; (b) modify the duration of action of the compound
in vivo; (c) modify the transportation or distribution of the
compound in vivo; (d) modify the solubility of the compound in
vivo; and (e) overcome a side effect or other difficulty
encountered with the compound. Typical functional derivatives used
to prepare prodrugs include modifications of the compound that are
chemically or enzymatically cleavable in vivo. Such modifications,
which include the preparation of phosphates, amides, esters,
thioesters, carbonates, and carbamates, are well known to those
skilled in the art.
General Synthetic Procedures
[0229] In order to illustrate the invention, the following examples
are included. However, it is to be understood that these examples
do not limit the invention and are only meant to suggest a method
of practicing the invention.
[0230] Generally, the compounds in this invention may be prepared
by methods described herein, wherein the substituents are as
defined for Formula (I), above, except where further noted. The
following non-limiting schemes and examples are presented to
further exemplify the invention. Persons skilled in the art will
recognize that the chemical reactions described herein may be
readily adapted to prepare a number of other compounds of the
invention, and alternative methods for preparing the compounds of
this invention are deemed to be within the scope of this invention.
For example, the synthesis of non-exemplified compounds according
to the invention may be successfully performed by modifications
apparent to those skilled in the art, e.g., by appropriately
protecting interfering groups, by utilizing other suitable reagents
known in the art other than those described, and/or by making
routine modifications of reaction conditions. Alternatively, other
reactions disclosed herein or known in the art will be recognized
as having applicability for preparing other compounds of the
invention.
[0231] In the examples described below, unless otherwise indicated
all temperatures are set forth in degrees Celsius. Reagents were
purchased from commercial suppliers such as Aldrich Chemical
Company, Arco Chemical Company and Alfa Chemical Company, Shanghai
Medpep. Co Ltd, Aladdin-Shanghai Jinchun Reagents, Ltd, and were
used without further purification unless otherwise indicated.
Common solvents were purchased from commercial suppliers such as
Shantou XiLong Chemical Factory, Guangdong Guanghua Reagent
Chemical Factory Co. Ltd., Guangzhou Reagent Chemical Factory,
Tainjin YuYu Fine Chemical Ltd., Qingdao Tenglong Reagent Chemical
Ltd., and Qingdao Ocean Chemical Factory.
[0232] Anhydrous THF, dioxane, toluene, and ether were obtained by
refluxing the solvent with sodium. Anhydrous CH.sub.2Cl.sub.2 and
CHCl.sub.3 were obtained by refluxing the solvent with CaH.sub.2.
EtOAc, PE, hexanes, DMA and DMF were treated with anhydrous
Na.sub.2SO.sub.4 prior use.
[0233] The reactions set forth below were done generally under a
positive pressure of nitrogen or argon or with a drying tube
(unless otherwise stated) in anhydrous solvents, and the reaction
flasks were typically fitted with rubber septa for the introduction
of substrates and reagents via syringe. Glassware was oven dried
and/or heat dried.
[0234] Column chromatography was conducted using a silica gel
column. Silica gel (300-400 mesh) was purchased from Qingdao Ocean
Chemical Factory.
[0235] .sup.1H NMR spectra were recorded with a Bruker 400 MHz or
600 MHz spectrometer at ambient temperature. .sup.1H NMR spectra
were obtained as CDCl.sub.3, DMSO-d.sub.6, CD.sub.3OD or
acetone-d.sub.6 solutions (reported in ppm), using TMS (0 ppm) or
chloroform (7.26 ppm) as the reference standard. When peak
multiplicities are reported, the following abbreviations are used:
s (singlet), d (doublet), t (triplet), m (multiplet), br
(broadened), dd (doublet of doublets), dt (doublet of triplets), td
(triplet of doublets). Coupling constants, when given, are reported
in Hertz (Hz).
[0236] Low-resolution mass spectral (MS) data were generally
determined on an Agilent 6120 quadrupole HPLC-MS (Zorbax SB-C18,
2.1.times.30 mm, 3.5 micron, 6 minutes run, 0.6 mL/min flow rate,
5% to 95% (0.1% formic acid in CH.sub.3CN) in (0.1% formic acid in
H.sub.2O)) with UV detection at 210/254 nm and electrospray
ionization (ESI).
[0237] Purities of compounds were assessed by Agilent 1260 pre-HPLC
or Calesep pump 250 pre-HPLC (column: NOVASEP 50/80 mm DAC) with UV
detection at 210 nm and 254 nm.
[0238] The following abbreviations are used throughout the
specification:
AcOH, HAc, CH.sub.3COOH acetic acid Ac.sub.2O acetic anhydride BOC,
Boc butyloxycarbonyl n-BuOH butyl alcohol Cbz-Cl benzyl
chloroformate CDCl.sub.3 chloroform deuterated DCM,
CH.sub.2Cl.sub.2 methylene chloride DIEA, DIPEA, i-Pr.sub.2NEt
N,N-diisopropylethylamine DMF dimethylformamide DMAP
4-dimethylaminopyridine DMSO dimethylsulfoxide EDC, EDCI
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride EDTA
ethylenediaminetetraacetic acid Et.sub.3N, TEA triethylamine EtOAc,
EA ethyl acetate EtOH, CH.sub.3CH.sub.2OH ethanol g gram h hour hex
hexane HCl hydrochloric acid HOAT 1-hydroxy-7-azabenzotriazole
MeCN, CH.sub.3CN acetonitrile MeOH, CH.sub.3OH methanol mL, ml
milliliter NaClO sodium hypochloride Na.sub.2CO.sub.3 sodium
carbonate NaH sodium hydride NaH.sub.2PO.sub.4 sodium dihydrogen
phosphate NaOH sodium hydroxide Na.sub.2SO.sub.4 sodium sulfate
Pd/C palladium on carbon PE petroleum ether (60-90.degree. C.) RT,
rt, r.t. room temperature Rt retention time SOCl.sub.2 thionyl
chloride TFA trifluoroacetic acid
[0239] Representative synthetic procedure for the preparation of
the compound disclosed herein is outlined below in following Scheme
1. Unless otherwise indicated, each of W, R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6 and U carries the definition
disclosed herein. X is C.sub.1, Br, OH or
--OC(.dbd.O)C.sub.1-C.sub.4alkyl.
##STR00015## ##STR00016##
[0240] The compound disclosed herein can be prepared according to
the general synthetic methods illustrated in Scheme 1 and described
in details in the examples. As showing in Scheme 1, phenylhydrazine
(1) is acylated with compound (2) under basic conditions to give
compound (3). Compound (3) is then treated with ethyl
3-chloro-3-oxopropanoate to furnish compound (4). Subsequent
self-cyclization of compound (4) can be accomplished with the aid
of a base, such as NaH, to afford compound (5), which is then
transformed to compound (6) via intramolecular condensation under
basic conditions. The ester group in (6) is converted to carboxyl
through basic hydrolysis. Coupling of carboxylic acid (7) with
compound (8) in the presence of coupling reagent such as EDCI/HOAT
furnishes carbamoyl compound (9). The amide group in compound (9)
is transformed to amine (10) with the aid of NaClO in aq. NaOH
solution. Finally, condensation of compound (10) with compound (11)
affords compound (12).
Examples
Example 1
N-(4-((2-aminopyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,5,6-tetrahydro-1-
H-pyrrolo[1,2-b]pyrazole-3-carboxamide
##STR00017##
[0241] Step 1) 4-chloro-N'-phenylbutanehydrazide
[0242] To a mixture of phenylhydrazine (16.0 g, 148.0 mmol) and 10%
Na.sub.2CO.sub.3 (aq) (250 mL) in DCM (250 mL) was added
4-chlorobutanoyl chloride (20.9 g, 148.0 mmol) via a syringe at
0.degree. C. The reaction was warmed up to r.t. and stirred
overnight, then diluted with DCM (150 mL). The separated organic
phase was washed with 2 M HCl (aq) (300 mL.times.3) followed with
brine (150 mL), then dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The resulted residue was
recrystallized in EtOAc/n-hexane (50 mL/100 mL) to afford the title
compound as a pale solid (14.7 g, 47%).
[0243] LC-MS (ESI, pos, ion) m/z: 213.1 [M+H].sup.+;
[0244] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 2.19-2.20
(m, 2H), 2.49 (t, J=7.1 Hz, 14.2 Hz, 2H), 3.54-3.67 (m, 2H), 5.79
(s, 1H), 6.87-6.80 (m, 2H), 6.92-6.95 (m, 1H), 7.24-7.32 (m, 2H),
7.38 (s, 1H).
Step 2) ethyl
3-(2-(4-chlorobutanoyl)-1-phenylhydrazinyl)-3-oxopropanoate
[0245] To a solution of 3-ethoxy-3-oxopropanoic acid (14.9 g, 112.8
mmol) in toluene (226 mL) was added SOCl.sub.2 (26.6 g, 225.6
mmol). The reaction was heated at 110.degree. C. for 4 h, and then
concentrated in vacuo to give ethyl 3-chloro-3-oxopropanoate as
brown oil, which was used for the next step immediately without
further purification.
[0246] To a suspension of 4-chloro-N'-phenylbutanehydrazide (12 g,
56.4 mmol) and Na.sub.2CO.sub.3 (26.9 g, 253.8 mmol) in DCM (226
mL) was added a solution of ethyl 3-chloro-3-oxopropanoate in DCM
(30 mL) slowly. The reaction was stirred at r.t. overnight, and
then filtered through a pad of Celite.RTM.. The filtrate was
diluted with DCM/water (100 mL/50 mL). The organic phase was
separated and the aqueous phase was extracted with EtOAc (200 mL).
The combined organic phases were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The resulted
residue was purified by a silica gel column chromatography
(n-hexane/EtOAc (v/v)=3/2) to give the title compound as brown oil
(6.2 g, 34%).
[0247] LC-MS (ESI, pos, ion) m/z: 327.2 [M+H].sup.+;
[0248] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 8.41 (s,
1H), 7.44 (m, 5H), 4.13 (t, J=6.7 Hz, 2H), 3.59 (m, 2H), 3.36 (s,
2H), 2.44 (d, J=13.3 Hz, 2H), 2.12 (d, J=12.9 Hz, 2H), 1.27 (t,
J=14.3 Hz, 3H).
Step 3) ethyl
3-oxo-3((2-oxopyrrolidin-1-yl)(phenyl)amino)propanoate
[0249] To a solution of ethyl
3-(2-(4-chlorobutanoyl)-1-phenylhydrazinyl)-3-oxopropanoate (6.2 g,
19 mmol) in DMF (48 mL) was added NaH (60% dispersion in mineral
oil, 2.3 g, 57 mmol) in portions at 0.degree. C. The reaction was
warmed up to r.t. and stirred for 4 h. The mixture was adjusted to
pH=7 with saturated NaH.sub.2PO.sub.4 (aq), and then filtered. The
organic phase was separated and the aqueous phase was extracted
with EtOAc (200 mL.times.3). The combined organic phases were
washed with brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The resulted residue was
purified by a silica gel column chromatography (EtOAc/PE (v/v)=4/5)
to give the title compound as brown oil (5.0 g, 91%).
[0250] LC-MS (ESI, pos, ion) m/z: 290.1 [M+H].sup.+;
[0251] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 7.38-7.52
(m, 5H), 4.17-4.25 (m, 2H), 3.48-3.55 (m, 2H), 3.31 (s, 2H),
2.37-2.54 (m, 2H), 2.01-2.18 (m, 2H), 1.25-1.33 (m, 3H).
Step 4) ethyl
2-oxo-1-phenyl-2,4,5,6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxylate
[0252] A solution of ethyl
3-oxo-3-((2-oxopyrrolidin-1-yl)(phenyl)amino)propanoate (3.3 g,
11.4 mmol) in DBU (10.0 mL) was stirred at 50.degree. C. for 5 h,
then cooled to r.t. and diluted with water (20 mL). The mixture was
adjust to pH 7 with saturated NaH.sub.2PO.sub.4 (aq), and then
extracted with DCM (20 mL.times.3). The combined organic phases
were washed with brine (20 mL), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The resulted
residue was purified by a silica gel column chromatography
(EtOAc/n-hexane (v/v)=4/5) to give the title compound as a white
solid (2.45 g, 79%).
[0253] LC-MS (ESI, pos, ion) m/z: 273.2 [M+H].sup.+;
[0254] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 7.46 (t,
J=7.8 Hz, 2H), 7.41-7.37 (m, 2H), 7.32 (d, J=7.3 Hz, 1H), 4.33 (q,
J=7.1 Hz, 2H), 3.69 (t, J=6.9 Hz, 2H), 3.21 (t, J=7.4 Hz, 2H),
2.51-2.44 (m, 2H), 1.38 (d, J=7.1 Hz, 3H).
Step 5)
2-oxo-1-phenyl-2,4,5,6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carb-
oxylic acid
[0255] To a solution of ethyl
2-oxo-1-phenyl-2,4,5,6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxylate
(2.0 g, 7.3 mmol) in EtOH (10 mL) was added 1.6 M NaOH (aq) (10
mL). The reaction was stirred at r.t. overnight, then concentrated
in vacuo to remove EtOH. The resulted residue was washed with DCM
(10 mL.times.2), and then adjust to pH 2 with 2 M HCl (aq). The
precipitate was collected by filtration to give the title compound
as a white solid (1.5 g, 84%).
[0256] LC-MS (ESI, pos, ion) m/z: 245.1 [M+H].sup.+;
[0257] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 7.55 (t,
J=7.7 Hz, 2H), 7.43 (t, J=9.1 Hz, 3H), 3.82 (t, J=6.9 Hz, 2H), 3.29
(t, J=7.4 Hz, 2H), 2.57 (m, 2H).
Step 6)
N-(4-((2-carbamoylpyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,5,6--
tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
[0258] To a solution of 4-(4-aminophenoxy)picolinamide (450 mg,
1.96 mmol),
2-oxo-1-phenyl-2,4,5,6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carb-
oxylic acid (503 mg, 2.06 mmol) and HOAT (53.4 mg, 0.39 mmol) in
DCM (10 mL) was added EDCI (564 mg, 2.94 mmol) in portions. The
reaction was heated at 45.degree. C. overnight, then cooled to r.t.
and diluted with EtOAc/water (6 mL/6 mL). The mixture was continued
to stir at r.t. for 1 h, and then filtered. The solid was collected
and dried in vacuo at 60.degree. C. for 5 h to afford the title
compound as a pale-yellow solid (667 mg, 75%).
[0259] LC-MS (ESI, pos, ion) m/z: 456.2 [M+H].sup.+;
[0260] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 10.26 (s,
1H), 8.41 (d, J=5.6 Hz, 1H), 7.85 (s, 1H), 7.79-7.74 (m, 3H), 7.55
(t, J=7.8 Hz, 2H), 7.47-7.38 (m, 3H), 7.07 (d, J=8.9 Hz, 2H), 6.98
(dd, J=5.6 Hz, 2.6 Hz, 1H), 5.54 (s, 1H), 3.74 (t, J=6.9 Hz, 2H),
3.35 (t, J=7.4 Hz, 2H), 2.56 (m, 2H).
Step 7)
N-(4-((2-aminopyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,5,6-tetr-
ahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
[0261] To a solution of
N-(4-((2-carbamoylpyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,5,6-tetrahy-
dro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide (667 mg, 1.94 mmol) in
dioxane/MeOH (16 mL, v/v=1/1) was added a mixed solution of NaOCl
(5.5% free Cl ion, 5 mL) and aq. NaOH (2.5 M, 5 mL) dropwise at
0.degree. C. The reaction was stirred at 0.degree. C. for 2 h, then
heated to 80.degree. C. and stirred for 3 h. The mixture was cooled
down to 0.degree. C., followed by an addition of water (15 mL). The
precipitate was collected by filtration to give the title compound
as a pale solid (300 mg, 48%).
[0262] LC-MS (ESI, pos, ion) m/z: 428.2 [M+H].sup.+;
[0263] Q-TOF (ESI, pos, ion) m/z: 428.1717 [M+H].sup.+;
[0264] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 10.22 (s,
1H), 7.93 (d, J=5.7 Hz, 1H), 7.72 (d, J=8.8 Hz, 2H), 7.55 (t, J=7.8
Hz, 2H), 7.48-7.43 (m, 2H), 7.40 (t, J=7.4 Hz, 1H), 7.06 (d, J=8.8
Hz, 2H), 6.32 (dd, J=5.8, 1.9 Hz, 1H), 5.96 (s, 1H), 4.47 (s, 2H),
3.74 (t, J=6.9 Hz, 2H), 3.34 (t, J=7.4 Hz, 2H), 2.56 (m, 2H);
[0265] .sup.13C NMR (150 MHz, CDCl.sub.3) .delta. (ppm): 166.9,
166.3, 162.4, 160.8, 160.0, 149.7, 149.3, 135.8, 134.6, 129.5,
127.5, 123.1, 121.5, 121.2, 104.2, 98.8, 95.0, 49.6, 29.7, 25.6,
22.2.
Example 2
N-(4-((2-(cyclopropanecarboxamido)pyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl--
2,4,5,6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
##STR00018##
[0267] To a solution of
N-(4-((2-aminopyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,5,6-tetrahydro--
1H-pyrrolo[1,2-b]pyrazole-3-carboxamide (150 mg, 0.35 mmol) and
pyridine (1.4 mL) in MeCN (2 mL) was added a solution of
cyclopropanecarbonyl chloride (110 mg, 1.05 mmol) and DMAP (128.3
mg, 1.05 mmol) in MeCN (1.4 mL). The reaction was heated at
60.degree. C. overnight, then cooled to r.t. and diluted with
DCM/water (10 mL/15 mL). The separated organic phase was washed
with water (6 mL.times.3), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The resulted residue was
recrystallized in EtOAc/PE (8 mL, v/v=1/3) to give the title
compound as a pale solid (109 mg, 60%).
[0268] LC-MS (ESI, pos, ion) m/z: 496.3 [M+H].sup.+;
[0269] Q-TOF (ESI, pos, ion) m/z: 496.1981 [M+H].sup.+;
[0270] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 10.21 (s,
1H), 8.44 (s, 1H), 8.09 (d, J=5.8 Hz, 1H), 7.83 (s, 1H), 7.74 (d,
J=8.8 Hz, 2H), 7.54 (t, J=7.8 Hz, 2H), 7.45 (d, J=7.6 Hz, 2H), 7.40
(t, J=7.4 Hz, 1H), 7.06 (d, J=8.8 Hz, 2H), 6.56 (dd, J=5.7 Hz, 2.2
Hz, 1H), 3.73 (t, J=6.9 Hz, 2H), 3.34 (t, J=7.4 Hz, 2H), 2.55 (p,
J=7.1 Hz, 2H), 1.53 (dt, J=11.9 Hz, 3.9 Hz, 1H), 1.12-1.07 (m, 2H),
0.90-0.84 (m, 2H);
[0271] .sup.13C NMR (150 MHz, CDCl.sub.3) .delta. (ppm): 172.1,
166.9, 166.3, 162.4, 160.7, 153.1, 149.6, 148.9, 136.0, 134.7,
129.5, 127.5, 123.1, 121.2, 108.2, 101.9, 98.9, 49.6, 25.6, 22.2,
15.8, 8.3.
Example 3
N-(4-((2-acetamidopyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,5,6-tetrahyd-
ro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
##STR00019##
[0273] To a solution of
N-(4-((2-aminopyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,5,6-tetrahydro--
1H-pyrrolo[1,2-b]pyrazole-3-carboxamide (100 mg, 0.23 mmol) in
acetic anhydride (2 mL) was added Et.sub.3N (142 mg, 1.4 mmol). The
reaction was stirred at 30.degree. C. overnight, and then
concentrated in vacuo. The resulted residue was purified by a
column chromatography on silica gel (100% EtOAc) to afford the
title compound as a pale solid (38 mg, 35%).
[0274] LC-MS (ESI, pos, ion) m/z: 470.2 [M+H].sup.+;
[0275] Q-TOF (ESI, pos, ion) m/z: 470.1827 [M+H].sup.+;
[0276] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 10.22 (s,
1H), 8.27 (s, 1H), 8.08 (d, J=5.8 Hz, 1H), 7.86 (s, 1H), 7.75 (d,
J=8.9 Hz, 2H), 7.54 (t, J=7.9 Hz, 2H), 7.44 (d, J=7.5 Hz, 2H), 7.40
(t, J=7.4 Hz, 1H), 7.07 (d, J=8.9 Hz, 2H), 6.56 (dd, J=5.8 Hz, 2.2
Hz, 1H), 3.73 (t, J=6.9 Hz, 2H), 3.34 (t, J=7.4 Hz, 2H), 2.58-2.50
(m, 2H), 2.18 (s, 3H);
[0277] .sup.13C NMR (150 MHz, CDCl.sub.3) .delta. (ppm): 167.0,
166.3, 162.4, 160.8, 153.0, 149.5, 148.9, 136.1, 134.7, 129.5,
127.5, 123.1, 121.3, 121.2, 108.2, 102.1, 98.9, 49.6, 25.6, 24.7,
22.2.
Example 4
N-(4-((2-aminopyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-2,4,5,6-tet-
rahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
##STR00020##
[0278] Step 1)
N-(4-((2-carbamoylpyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-2,4,5,-
6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
[0279] To a solution of 4-(4-amino-2-fluorophenoxy)picolinamide
(322 mg, 1.30 mmol),
2-oxo-1-phenyl-2,4,5,6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxylic
acid (325 mg, 1.33 mmol) and HOAT (35.4 mg, 0.26 mmol) in DCM (10
mL) was added EDCI (382 mg, 1.99 mmol) in portions. The reaction
was heated at 50.degree. C. overnight, then cooled to r.t. and
concentrated in vacuo. The residue was diluted with 1 M HCl and
stirred for 30 min. The precipitate was collected by filtration to
give the title compound as a pale solid (260 mg, 42%).
[0280] LC-MS (ESI, pos, ion) m/z: 474.3 [M+H].sup.+;
[0281] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.40 (s,
1H), 8.55 (d, J=5.6 Hz, 1H), 8.15 (s, 1H), 8.01 (d, J=11.9 Hz, 1H),
7.74 (s, 1H), 7.60-7.51 (m, 4H), 7.45-7.39 (m, 4H), 7.23 (dd, J=5.6
Hz, 2.6 Hz, 1H), 3.83 (t, J=7.0 Hz, 2H), 3.18 (t, J=7.4 Hz, 2H),
2.46-2.42 (m, 2H).
Step 2)
N-(4-((2-aminopyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-2,4-
,5,6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
[0282] To a solution of
N-(4-((2-carbamoylpyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-2,4,5,-
6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide (261 mg, 0.551
mmol) in dioxane/MeOH (10 mL, v/v=1/1) was added a mixed solution
of NaOCl (5.5% free C.sub.1 ion, 1.7 mL) and 2.5 M NaOH (aq) (0.3
mL) dropwise at 0.degree. C. The reaction was stirred at 0.degree.
C. for 4 h, then heated to 80.degree. C. and stirred further for 1
h. The mixture was cooled down to 0.degree. C., followed by an
addition of water (15 mL). The precipitate was collected by
filtration to give the title compound as a brown solid (60 mg,
24%).
[0283] LC-MS (ESI, pos, ion) m/z: 446.2 [M+H].sup.+;
[0284] Q-TOF (ESI, pos, ion) m/z: 446.1627 [M+H].sup.+;
[0285] .sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. (ppm): 10.34 (s,
1H), 7.94 (d, J=12.9 Hz, 1H), 7.80 (d, J=5.5 Hz, 1H), 7.59-7.50 (m,
4H), 7.43 (t, J=6.9 Hz, 1H), 7.35 (d, J=8.3 Hz, 1H), 7.28 (t, J=8.8
Hz, 1H), 6.17 (d, J=3.6 Hz, 1H), 5.96 (s, 2H), 5.80 (s, 1H), 3.82
(s, 2H), 3.17 (d, J=6.9 Hz, 2H), 2.47-2.40 (m, 2H);
[0286] .sup.13C NMR (150 MHz, DMSO-d.sub.6) .delta. (ppm): 165.6,
165.6, 162.1, 161.6, 160.9, 153.3, 150.0, 135.9, 134.7, 129.8,
127.8, 124.6, 123.9, 116.0, 108.3, 108.2, 101.5, 96.5, 92.8, 49.9,
26.0, 22.2.
Example 5
N-(4-((2-(cyclopropanecarboxamido)pyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo--
1-phenyl-2,4,5,6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
##STR00021##
[0288] To a solution of
N-(4-((2-aminopyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-2,4,5,6-te-
trahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide (22 mg, 0.05 mmol)
and pyridine (0.2 mL) in MeCN (0.3 mL) was added a solution of
cyclopropanecarbonyl chloride (15.5 mg, 0.15 mmol) and DMAP (18.3
mg, 0.15 mmol) in MeCN (0.2 mL). The reaction was heated at
60.degree. C. overnight, then cooled to r.t. and diluted with
DCM/water (6 mL/15 mL). The separated organic phase was washed with
water (6 mL.times.3), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The resulted residue was
purified by a column chromatography on silica gel (EtOAc/PE
(v/v)=5/1) to afford the title compound as a pale-yellow solid (20
mg, 67%).
[0289] LC-MS (ESI, pos, ion) m/z: 514.3 [M+H].sup.+;
[0290] Q-TOF (ESI, pos, ion) m/z: 514.1883 [M+H].sup.+;
[0291] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 10.31 (s,
1H), 8.38 (s, 1H), 8.11 (d, J=5.8 Hz, 1H), 7.90 (dd, J=12.4 Hz, 2.2
Hz, 1H), 7.82 (s, 1H), 7.55 (t, J=7.8 Hz, 2H), 7.46-7.39 (m, 3H),
7.12 (t, J=8.7 Hz, 1H), 6.58 (dd, J=5.7 Hz, 2.2 Hz, 1H), 4.14 (q,
J=7.1 Hz, 1H), 3.75 (t, J=6.9 Hz, 2H), 3.34 (t, J=7.4 Hz, 2H),
2.59-2.52 (m, 2H), 1.56-1.50 (m, 1H), 1.09 (dt, J=7.9 Hz, 4.0 Hz,
2H), 0.90-0.86 (m, 2H).
Example 6
N-(4-((2-acetamidopyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-2,4,5,6-
-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
##STR00022##
[0293] To a solution of
N-(4-((2-aminopyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-2,4,5,6-te-
trahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide (22 mg, 0.05 mmol)
in acetic anhydride (1 mL) was added Et.sub.3N (30 mg, 0.3 mmol).
The reaction was stirred at 30.degree. C. for 4 h, and then
concentrated in vacuo. The resulted residue was purified by a
column chromatography on silica gel (100% EtOAc) to afford the
title compound as a pale-yellow solid (20 mg, 74%).
[0294] LC-MS (ESI, pos, ion) m/z: 488.3 [M+H].sup.+;
[0295] Q-TOF (ESI, pos, ion) m/z: 488.1733 [M+H].sup.+;
[0296] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 10.32 (s,
1H), 8.32 (s, 1H), 8.10 (d, J=5.6 Hz, 1H), 7.96-7.83 (m, 2H), 7.54
(t, J=7.4 Hz, 2H), 7.43 (dd, J=14.1 Hz, 7.7 Hz, 4H), 7.13 (t, J=8.6
Hz, 1H), 6.57 (d, J=3.8 Hz, 1H), 3.75 (t, J=6.6 Hz, 2H), 3.34 (t,
J=7.0 Hz, 2H), 2.62-2.50 (m, 2H), 2.18 (s, 3H).
Example 7
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-1,2,4,5,6,7-he-
xahydropyrazolo[1,5-a]pyridine-3-carboxamide
##STR00023##
[0297] Step 1) 5-chloro-N'-phenylpentanehydrazide
[0298] To a mixture of phenylhydrazine (21.6 g, 200.0 mmol) and 10%
Na.sub.2CO.sub.3 (aq) (340 mL) in DCM (340 mL) was added
5-chloropentanoyl chloride (31.0 g, 200.0 mmol) via a syringe at
0.degree. C. The reaction was warmed up to r.t., stirred overnight,
and then diluted with DCM (150 mL). The separated organic phase was
washed with 2 M HCl (aq) (250 mL.times.5) followed with brine (250
mL), then dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The resulted residue was recrystallized in
EtOAc/n-hexane (80 mL/240 mL) to afford the title compound as a
pale solid (19.6 g, 44%).
[0299] LC-MS (ESI, pos, ion) m/z: 227.2 [M+H].sup.+;
[0300] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 7.44 (s,
1H), 7.27-7.22 (m, 2H), 6.95 (m, 2H), 6.79 (d, J=7.8 Hz, 1H), 5.78
(s, 1H), 3.58 (m, 2H), 2.32 (m, 2H), 1.87 (m, 2H), 1.81 (m,
2H).
Step 2) ethyl
3-(2-(5-chloropentanoyl)-1-phenylhydrazinyl)-3-oxopropanoate
[0301] To a solution of 3-ethoxy-3-oxopropanoic acid (38.5 g, 291
mmol) in toluene (290 mL) was added SOCl.sub.2 (68.6 g, 582 mmol).
The reaction was heated at 110.degree. C. for 6 h, and then
concentrated in vacuo to give ethyl 3-chloro-3-oxopropanoate as
brown oil, which was used for the next step immediately without
further purification.
[0302] To a suspension of 5-chloro-N'-phenylpentanehydrazide (22 g,
97 mmol) and Na.sub.2CO.sub.3 (31 g, 291 mmol) in DCM (582 mL) was
added a solution of ethyl 3-chloro-3-oxopropanoate in DCM (50 mL)
slowly. The reaction was stirred at r.t. overnight, and then
filtered through a pad of Celite.RTM.. The filtrate was diluted
with DCM/water (100 mL/50 mL). The separated organic phase was
washed with water (150 mL.times.2) followed by brine (150 mL), then
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The resulted residue was purified by a silica gel column
chromatography (n-hexane/EtOAc (v/v)=3/2) to give the title
compound as brown oil (25 g, 76%).
[0303] LC-MS (ESI, pos, ion) m/z: 341.2 [M+H].sup.+;
[0304] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 8.28 (s,
1H), 7.46-7.38 (m, 5H), 4.31-4.17 (m, 2H), 4.14 (m, 2H), 3.52 (m,
2H), 3.36 (s, 2H), 2.29 (m, 2H), 1.33 (m, 2H), 1.29-1.22 (m,
3H).
Step 3) ethyl
3-oxo-3-((2-oxopiperidin-1-yl)(phenyl)amino)propanoate
[0305] To a solution of ethyl
3-(2-(5-chloropentanoyl)-1-phenylhydrazinyl)-3-oxopropanoate (25 g,
73 mmol) in DMF (219 mL) was added NaH (60% dispersion in mineral
oil, 8.8 g, 219 mmol) in portions at 0.degree. C. The reaction was
warmed up to r.t. and stirred overnight. The mixture was adjusted
to pH 7 with saturated NaH.sub.2PO.sub.4 (aq), and then filtered.
The organic phase was separated and the aqueous phase was extracted
with EtOAc (200 mL.times.4). The combined organic phases were
washed with brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo to give the title compound as
brown oil (20 g, 90%).
[0306] LC-MS (ESI, pos, ion) m/z: 305.2 [M+H].sup.+.
Step 4) ethyl
2-oxo-1-phenyl-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxylate
[0307] A solution of ethyl
3-oxo-3-((2-oxopiperidin-1-yl)(phenyl)amino)propanoate (20 g, 66
mmol) in DBU (20.0 mL) was stirred at 50.degree. C. overnight, then
cooled to r.t. and diluted with water (20 mL). The mixture was
adjusted to pH 7 with saturated NaH.sub.2PO.sub.4 (aq), and then
extracted with DCM (100 mL.times.6). The combined organic phases
were washed with brine (100 mL), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the
title compound as a yellow solid (17 g, 90%).
[0308] LC-MS (ESI, pos, ion) m/z: 287.1 [M+H].sup.+;
[0309] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 7.50-7.47
(m, 2H), 7.40-7.36 (m, 1H), 7.34-7.32 (m, 2H), 4.33 (q, J=7.1 Hz,
2H), 3.53 (t, J=5.9 Hz, 2H), 3.22 (t, J=6.5 Hz, 2H), 2.07-2.04 (m,
2H), 1.93-1.89 (m, 2H), 1.39 (t, J=7.1 Hz, 3H).
Step 5)
2-oxo-1-phenyl-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carb-
oxylic acid
[0310] To a solution of ethyl
2-oxo-1-phenyl-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxylate
(18 g, 62.8 mmol) in EtOH (100 mL) was added 1.6 M NaOH (aq) (100
mL). The reaction was stirred at r.t. overnight, and then
concentrated in vacuo to remove EtOH. The resulted residue was
washed with DCM (20 mL.times.2), and adjusted to pH 2 with 2 M HCl
(aq). The precipitate was collected by filtration to give the title
compound as a pale yellow solid (10.7 g, 66%).
[0311] LC-MS (ESI, pos, ion) m/z: 259.1 [M+H].sup.+;
[0312] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 7.58-7.55
(m, 2H), 7.52-7.49 (m, 1H), 7.39-7.36 (m, 2H), 3.60 (t, J=6.0 Hz,
2H), 3.28 (t, J=6.5 Hz, 2H), 2.13-2.08 (m, 2H), 1.96-1.92 (m,
2H).
Step 6)
N-(5-hydroxypyridin-2-yl)-2-oxo-1-phenyl-1,2,4,5,6,7-hexahydropyra-
zolo[1,5-a]pyridine-3-carboxamide
[0313] To a solution of 6-aminopyridin-3-ol hydrochloride (2.55 g,
17.4 mmol) in DMF (23 mL) was added KOH (1.624 g, 29 mmol). The
mixture was stirred at r.t. for 30 min, followed by an addition of
2-oxo-1-phenyl-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxylic
acid (3.0 g, 11.6 mmol), HOAT (316 mg, 2.32 mmol) and EDCI (4.48 g,
23.2 mmol). The reaction was heated at 60.degree. C. overnight,
then cooled to r.t., diluted with water (230 mL) and extracted with
DCM (100 mL.times.3). The combined organic phases were washed with
brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The resulted residue was recrystallized in
DCM/EtOAc/PE (30 mL/30 mL/30 mL) to give the title compound as a
yellow solid (2.8 g, 69%).
[0314] LC-MS (ESI, pos, ion) m/z: 351.1 [M+H].sup.+;
[0315] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 10.72 (s,
1H), 8.05 (d, J=8.9 Hz, 1H), 7.82 (d, J=2.8 Hz, 1H), 7.57 (d, J=7.7
Hz, 2H), 7.50 (dd, J=8.5 Hz, 6.2 Hz, 1H), 7.47-7.43 (m, 2H), 7.19
(dd, J=8.9 Hz, 2.9 Hz, 1H), 3.55 (t, J=5.8 Hz, 2H), 3.19 (d, J=6.3
Hz, 2H), 1.97 (d, J=5.7 Hz, 2H), 1.81 (d, J=5.8 Hz, 2H).
Step 7)
N-(5-((2-carbamoylpyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-1,-
2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
[0316] To a solution of 4-chloropicolinamide (1.44 g, 9.2 mmol) and
N-(5-hydroxypyridin-2-yl)-2-oxo-1-phenyl-1,2,4,5,6,7-hexahydropyrazolo[1,-
5-a]pyridine-3-carboxamide (2.80 g, 8.0 mmol) in DMF (16 mL) was
added t-BuOK (1.80 g, 16.0 mmol). The reaction was heated at
130.degree. C. overnight, then cooled to r.t. and diluted with
water (160 mL). The mixture was continued to stir at r.t.
overnight, then filtered. The solid was dried in vacuo to give the
title compound as a brown solid (2.9 g, 78%).
[0317] LC-MS (ESI, pos, ion) m/z: 471.1 [M+H].sup.+;
[0318] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 11.15 (s,
1H), 8.44 (d, J=5.6 Hz, 1H), 8.38 (d, J=9.1 Hz, 1H), 8.17 (d, J=2.7
Hz, 1H), 7.84 (s, 1H), 7.76 (d, J=2.5 Hz, 1H), 7.55 (t, J=7.6 Hz,
2H), 7.48-7.42 (m, 3H), 7.41-7.38 (m, 2H), 6.98 (dd, J=5.6 Hz, 2.6
Hz, 1H), 5.60 (s, 1H), 3.60 (t, J=5.9 Hz, 2H), 3.41 (t, J=6.4 Hz,
2H), 2.12-2.08 (m, 2H), 1.99-1.94 (m, 2H).
Step 8)
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-1,2,4,-
5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
[0319] To a solution of
N-(5-((2-carbamoylpyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-1,2,4,5,6-
,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide (2.9 g, 6.2 mmol)
in dioxane/MeOH (62 mL, v/v=1/1) was added a mixed solution of
NaOCl (5.5% free C.sub.1 ion, 30 mL) and 2.5 M NaOH (aq) (3.1 mL)
at 0.degree. C. The reaction was stirred at 0.degree. C. for 4 h,
then heated to 60.degree. C. and stirred overnight. The mixture was
cooled down to 0.degree. C., followed by an addition of water (360
mL). The precipitate was collected by filtration to give the title
compound as a pale-yellow solid (1.02 g, 37%).
[0320] LC-MS (ESI, pos, ion) m/z: 443.1 [M+H].sup.+;
[0321] Q-TOF (ESI, pos, ion) m/z: 443.1840 [M+H].sup.+;
[0322] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 11.05 (s,
1H), 8.29 (d, J=9.0 Hz, 1H), 8.16 (d, J=2.8 Hz, 1H), 7.80 (d, J=5.8
Hz, 1H), 7.64 (dd, J=9.0 Hz, 2.8 Hz, 1H), 7.59 (t, J=7.7 Hz, 2H),
7.51 (t, J=7.4 Hz, 1H), 7.46 (d, J=7.5 Hz, 2H), 6.17 (dd, J=5.8 Hz,
2.2 Hz, 1H), 5.95 (s, 2H), 5.83 (d, J=2.0 Hz, 1H), 3.57 (t, J=5.8
Hz, 2H), 3.21 (t, J=6.3 Hz, 2H), 2.01-1.95 (m, 2H), 1.84-1.79 (m,
2H);
[0323] .sup.13C NMR (150 MHz, CDCl.sub.3) .delta. (ppm): 165.8,
162.9, 162.1, 161.6, 153.8, 150.1, 149.4, 146.6, 141.5, 133.0,
131.5, 129.9, 129.4, 128.0, 114.4, 102.2, 95.9, 93.8, 46.6, 23.9,
22.2, 18.8.
Example 8
N-(5-((2-(cyclopropanecarboxamido)pyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-p-
henyl-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
##STR00024##
[0325] To a solution of
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-1,2,4,5,6,7-h-
exahydropyrazolo[1,5-a]pyridine-3-carboxamide (100 mg, 0.226 mmol)
in pyridine (1 mL) was added cyclopropanecarbonyl chloride (142 mg,
1.36 mmol) dropwise at 0.degree. C. The reaction was warmed up to
r.t. and stirred overnight. The mixture was diluted with water (10
mL) and extracted with DCM (6 mL.times.3). The combined organic
phases were washed with brine (10 mL), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The resulted
residue was purified by a column chromatography on silica gel
(DCM/MeOH (v/v)=40/1) to afford the title compound as a pale-yellow
solid (37 mg, 32%).
[0326] LC-MS (ESI, pos, ion) m/z: 511.1 [M+H].sup.+;
[0327] Q-TOF (ESI, pos, ion) m/z: 511.2125 [M+H].sup.+;
[0328] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 11.10 (s,
1H), 8.60 (s, 1H), 8.34 (d, J=9.0 Hz, 1H), 8.15 (d, J=2.7 Hz, 1H),
8.11 (d, J=5.8 Hz, 1H), 7.86 (s, 1H), 7.54 (t, J=7.7 Hz, 2H),
7.47-7.41 (m, 2H), 7.39 (d, J=7.8 Hz, 2H), 6.56 (dd, J=5.7 Hz, 2.1
Hz, 1H), 3.59 (t, J=5.9 Hz, 2H), 3.40 (t, J=6.4 Hz, 2H), 2.13-2.07
(m, 2H), 1.97-1.91 (m, 2H), 1.59-1.52 (m, 1H), 1.11-1.07 (m, 2H),
0.90-0.89 (m, 2H).
Example 9
N-(5-((2-acetamidopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-1,2,4,5,6,-
7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
##STR00025##
[0330] To a solution of
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-1,2,4,5,6,7-h-
exahydropyrazolo[1,5-a]pyridine-3-carboxamide (100 mg, 0.226 mmol)
in acetic anhydride (2 mL) was added Et.sub.3N (137 mg, 1.36 mmol).
The reaction was stirred at 30.degree. C. overnight, and then
concentrated in vacuo. The residue was purified by a column
chromatography on silica gel (DCM/MeOH (v/v)=40/1) to afford the
title compound as a pale-yellow solid (55 mg, 50%).
[0331] LC-MS (ESI, pos, ion) m/z: 485.3 [M+H].sup.+;
[0332] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 11.09 (s,
1H), 10.57 (s, 1H), 8.32 (d, J=9.0 Hz, 1H), 8.21 (dd, J=14.2 Hz,
4.3 Hz, 2H), 7.72-7.66 (m, 2H), 7.60 (t, J=7.7 Hz, 2H), 7.52 (t,
J=7.4 Hz, 1H), 7.48 (d, J=7.8 Hz, 2H), 6.70 (dd, J=5.7 Hz, 2.3 Hz,
1H), 3.58 (t, J=5.8 Hz, 2H), 3.22 (t, J=6.3 Hz, 2H), 2.01-1.96 (m,
2H), 1.85-1.80 (m, 2H), 1.24 (s, 3H).
Example 10
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,6,7-tetrah-
ydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
##STR00026##
[0333] Step 1)
2-(2-(2-(tert-butoxycarbonyl)-2-phenylhydrazinyl)-2-oxoethoxy)acetic
acid
[0334] To a solution of tert-butyl 1-phenylhydrazinecarboxylate
(25.2 g, 121 mmol) and diglycolic anhydride (16.8 g, 145.2 mmol) in
190 mL DMF was added sodium carbonate (12.8 g, 121 mmol) at room
temperature. The reaction was stirred at r.t. overnight, and then
filtered through a pad of Celite.RTM., the solid mass was washed
with EtOAc (50 mL). The filtrate was concentrated in vacuo. The
residue was added H.sub.2O (180 mL) and extracted by EtOAc (50
mL.times.5), the water phase was adjusted to pH 6-7 with
NaH.sub.2PO.sub.4 (aq), and then was extracted with EtOAc (50 mL).
The organic phase was dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo to give the title compound as
yellow oil (28.3 g, 73%).
[0335] LC-MS (ESI, pos, ion): 225.2 [M-100+H].sup.+;
[0336] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.67 (s,
1H), 7.39-7.30 (m, 4H), 7.20-7.10 (m, 1H), 4.15 (s, 4H), 1.42 (s,
9H).
Step 2) tert-butyl
2-(2-(2-hydroxyethoxyl)acetyl)-1-phenylhydrazinecarboxylate
[0337] To a solution of
2-(2-(2-(tert-butoxycarbonyl)-2-phenylhydrazinyl)-2-oxoethoxy)acetic
acid (10.7 g, 33 mmol) in 140 mL THF at 0.degree. C. was added
triethylamine (9.24 mL, 66 mmol) followed by isobutyl
chlorocarbonate (5.2 mL, 39.6 mmol) via a syringe. The reaction
became cloudy due to the formation of Et.sub.3N--HCl salt. The
reaction was stirred at 0.degree. C. for 2 h and then filtered. The
filtrate was cooled to 0.degree. C. and NaBH.sub.4 (5 g, 132 mmol)
in 60 mL of H.sub.2O was added. The reaction was stirred at
0.degree. C. for 1 h, then the mixture was added H.sub.2O (100 mL)
and extracted with EtOAc (200 mL.times.2). The combined organic
phase was washed with 1 M NaH.sub.2PO.sub.4 (100 mL) aqueous
solution, followed with brine (100 mL), then dried over anhydrous
Na.sub.2SO.sub.4, filtered and the filtrate was concentrated in
vacuo to afford title compound as yellow oil (8.7 g, 85%).
[0338] LC-MS (ESI, neg, ion): 309.2 [M-H].sup.-;
[0339] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.54 (s,
1H), 7.60-7.23 (m, 4H), 7.25-7.04 (m, 1H), 4.07 (s, 2H), 3.61-3.50
(m, 4H), 1.42 (s, 9H).
Step 3) tert-butyl(3-oxomorpholino)(phenyl)carbamate
[0340] To a solution of tert-butyl
2-(2-(2-hydroxyethoxyl)acetyl)-1-phenylhydrazine carboxylate (27.6
g, 89 mmol) and PPh.sub.3 (35 g, 133.5 mmol) in dry THF (180 mL) at
0.degree. C. under N.sub.2 atmosphere was added a solution of DIAD
in THF (120 mL) slowly via a syringe. The reaction was warmed up to
r.t. and stirred overnight. The reaction mixture was filtered, the
filtrate was added EtOAc (800 mL) and washed with H.sub.2O (300
mL.times.2), followed with brine (300 mL). The organic phase was
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The resulted residue was purified by a silica gel column
chromatography (EtOAc/hex (v/v)=1/10-1/5) to afford the title
compound as yellow oil (29.7 g, 100%).
[0341] LC-MS (ESI, pos, ion): 193.1 [M-100+H].sup.+;
[0342] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 7.37 (dd,
J=9.8, 3.8 Hz, 4H), 7.25-7.18 (m, 1H), 4.96-4.73 (m, 2H), 4.27 (q,
J=16.5 Hz, 2H), 4.01-3.86 (m, 2H), 1.45 (s, 9H).
Step 4) 4-(phenylamino)morpholin-3-one
[0343] To a solution of
tert-butyl(3-oxomorpholino)(phenyl)carbamate (29.7 g, 102 mmol) in
EtOAc (50 mL) was added a saturated solution of HCl in EtOAc (350
mL). The mixture was stirred at r.t. for 2 h. The mixture was
cooled to 0.degree. C., and adjusted to pH=7-8 with a solution of
NaOH (3 M aq), then the mixture was extracted with EtOAc (100
mL.times.3), the combined organic phase was dried over anhydrous
Na.sub.2SO.sub.4 and concentrated in vacuo to afford the title
compound as a yellow solid (12 g, 61%).
[0344] LC-MS (ESI, pos, ion): 193.1 [M+H].sup.+;
[0345] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 8.24 (s,
1H), 7.19 (dd, J=8.4, 7.4 Hz, 2H), 6.77 (t, J=7.3 Hz, 1H), 6.69 (d,
J=7.6 Hz, 2H), 4.20 (s, 2H), 4.01 (dd, J=14.3, 9.3 Hz, 2H),
3.60-3.48 (m, 2H).
Step 5) ethyl
3-oxo-3-((3-oxomorpholino)(phenyl)amino)propanoate
[0346] To a solution of 4-(phenylamino)morpholin-3-one (9 g, 46.8
mmol) and Na.sub.2CO.sub.3 (32.3 g, 305 mmol) in DCM (400 mL) was
added ethyl malonoyl chloride (42.2 g, 281 mmol). The reaction was
stirred at r.t. for 4 h. The reaction was filtered, the filtrate
was added H.sub.2O (500 mL). The water phase was extracted by EtOAc
(200 mL.times.2), and the combined organic phase was dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The resulted
residue was purified by a silica gel column chromatography
(EtOAc/hex (v/v)=1/10-1/2) to give the title compound as yellow oil
(15 g, 100%).
[0347] LC-MS (ESI, pos, ion): 307.2 [M+H].sup.+;
[0348] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 7.54-7.53
(m, 2H), 7.45 (m, 3H), 4.38-4.32 (m, 2H), 4.18-4.10 (m, 2H),
3.96-3.84 (m, 2H), 3.72-3.61 (m, 2H), 3.30 (s, 2H), 1.27-1.23 (m,
3H).
Step 6) ethyl
2-oxo-1-phenyl-2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carbox-
ylate
[0349] A solution of ethyl
3-oxo-3-((3-oxomorpholino)(phenyl)amino)propanoate (15 g, 49 mmol)
in DBU (16 mL) was warmed to 50.degree. C. and stirred overnight.
The mixture was cooled to r.t. and adjusted to pH=7 with saturated
NaH.sub.2PO.sub.4 aqueous solution. Filtered, and the filtrate was
extracted with EtOAc (50 mL.times.6). The combined organic phase
was washed with brine (50 mL), dried over anhydrous
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by a silica gel column chromatography (EtOAc/hex
(v/v)=7/10-8/10) to give the title compound as a white solid (13 g,
93%).
[0350] LC-MS (ESI, pos, ion): 289.1 [M+H].sup.+;
[0351] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 7.55 (t,
J=7.6 Hz, 2H), 7.46 (t, J=7.4 Hz, 1H), 7.42-7.36 (m, 2H), 4.98 (s,
2H), 4.15 (q, J=7.1 Hz, 2H), 4.07-4.02 (m, 2H), 3.61 (t, J=5.0 Hz,
2H), 1.23 (t, J=7.1 Hz, 3H).
Step 7)
2-oxo-1-phenyl-2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-
-carboxylic acid
[0352] To a solution of ethyl
2-oxo-1-phenyl-2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carbox-
ylate (13 g, 45 mmol) in EtOH (70 mL) was added 1.6 N NaOH aqueous
solution (70 mL). The reaction mixture was stirred at r.t.
overnight. The mixture was concentrated in vacuo to remove EtOH,
the water phase was washed with DCM (30 mL.times.3), and then the
water phase was acidified with 2 N HCl aqueous solution to pH=2 and
stirred at r.t. for 2 h, filtered, the collected solid mass was the
title compound as a pale yellow solid (9.2 g, 79%).
[0353] LC-MS (ESI, pos, ion): 261.1 [M+H].sup.+;
[0354] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 11.55 (s,
1H), 7.62-7.56 (m, 2H), 7.56-7.50 (m, 1H), 7.40 (dd, J=5.3, 3.3 Hz,
2H), 5.17 (s, 2H), 4.19-4.14 (m, 2H), 3.72-3.68 (m, 2H).
Step 8)
N-(5-hydroxypyridin-2-yl)-2-oxo-1-phenyl-2,4,6,7-tetrahydro-1H-pyr-
azolo[5,1-c][1,4]oxazine-3-carboxamide
[0355] To a solution of 6-aminopyridin-3-ol hydrochloride (3.2 g,
21.5 mmol) in DMF (25 mL) was added KOH (1.9 g, 33.8 mmol) at r.t.,
the reaction was stirred at r.t. for 30 min. and then to the
solution was added
2-oxo-1-phenyl-2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3--
carboxylic acid (3.5 g, 13.5 mmol), HOAT (367 mg, 2.7 mmol) and
EDCI (5.2 g, 27.0 mmol) in portions. The reaction was allowed to
warm up to 60.degree. C. and stirred overnight. The reaction was
cooled to r.t. and was added water (250 mL) and stirred for 2 h,
filtered, the collected solid was dried in vacuo at 60.degree. C.
for 5 h to afford the title compound as a brown solid (2.65 g,
56%).
[0356] LC-MS (ESI, pos, ion): 353.2 [M+H].sup.+;
[0357] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.48 (s,
1H), 9.63 (s, 1H), 8.03 (d, J=8.9 Hz, 1H), 7.84 (d, J=2.7 Hz, 1H),
7.60 (t, J=7.4 Hz, 3H), 7.52 (t, J=7.7 Hz, 4H), 7.20 (dd, J=8.9,
2.8 Hz, 1H), 5.11 (s, 2H), 4.09 (t, J=4.7 Hz, 2H), 3.68 (d, J=4.6
Hz, 2H).
Step 9)
N-(5-(2-carbamoylpyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4-
,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
[0358] To a solution of 4-chloropicolinamide (1.07 g, 6.9 mmol) and
N-(5-hydroxypyridin-2-yl)-2-oxo-1-phenyl-2,4,6,7-tetrahydro-1H-pyrazolo[5-
,1-c][1,4]oxazine-3-carboxamide (2.10 g, 6.0 mmol) in DMF (12 mL)
was added t-BuOK (1.35 g, 12.0 mmol). The reaction was warmed up to
130.degree. C. and stirred overnight. The reaction was cooled to
r.t. and diluted with 120 mL of water, continued to stir at r.t.
overnight, filtered, the collected mass was the title compound as a
brown solid (2.2 g, 78%).
[0359] LC-MS (ESI, pos, ion): 473.3 [M+H].sup.+;
[0360] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.86 (s,
1H), 8.54 (d, J=5.6 Hz, 1H), 8.33 (d, J=9.0 Hz, 1H), 8.29 (d, J=2.8
Hz, 1H), 8.14 (s, 1H), 7.78 (dd, J=9.0, 2.9 Hz, 1H), 7.72 (s, 1H),
7.60 (d, J=7.3 Hz, 2H), 7.54 (t, J=6.0 Hz, 3H), 7.44 (d, J=2.6 Hz,
1H), 7.22 (dd, J=5.6, 2.6 Hz, 1H), 5.13 (s, 2H), 4.10 (t, J=5.0 Hz,
2H), 3.70 (t, J=4.9 Hz, 2H).
Step 10)
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,6-
,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
[0361] To a solution of
N-(5-((2-carbamoylpyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,6,7-t-
etrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide (2.1 g, 4.4
mmol) in a mixture solvent of EtOH (35 mL), MeCN (35 mL) and water
(17 mL) at 0.degree. C. was added PhI(OAc).sub.2 (1.8 g, 5.6 mmol).
The mixture was stirred at 0.degree. C. for 30 min, and then
allowed to warm up to r.t. and stirred overnight. The reaction was
filtered, the solid was washed with EtOAc (15 mL.times.2), the
filtrate was concentrated in vacuo, and the residue was purified by
a column chromatography on silica gel (100% DCM to 1% MeOH/DCM) to
afford the title compound as a yellow solid (900 mg, 46%).
[0362] LC-MS (ESI, pos, ion): 445.1 [M+H].sup.+;
[0363] Q-TOF (ESI, pos, ion): 445.1621 [M+H].sup.+;
[0364] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 10.82 (s,
1H), 8.29 (d, J=9.0 Hz, 1H), 8.16 (d, J=2.8 Hz, 1H), 7.95 (d, J=5.7
Hz, 1H), 7.57 (t, J=7.8 Hz, 2H), 7.49 (t, J=7.5 Hz, 1H), 7.46-7.44
(m, 1H), 7.42 (d, J=9.0 Hz, 2H), 6.32-6.30 (m, 1H), 5.96 (s, 1H),
5.28 (s, 2H), 4.53 (s, 2H), 4.19-4.16 (m, 2H), 3.69 (t, J=5.0 Hz,
2H).
Example 11
N-(5-((2-(cyclopropanecarboxamido)pyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-p-
henyl-2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
##STR00027##
[0366] To a solution of
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,6,7-tetra-
hydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide (150 mg, 0.34
mmol) in a mixture solvent of MeCN (2 mL) and pyridine (1.2 mL) at
r.t. was added a solution of cyclopropanecarbonyl chloride (106 mg,
1.01 mmol) and DMAP (123 mg, 1.01 mmol) in MeCN (1.2 mL). The
reaction mixture was allowed to warm up to 60.degree. C. and
stirred overnight. The reaction was cooled to r.t., then diluted
with DCM (15 mL), washed with water (10 mL.times.3), followed with
brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4 and filtered,
the filtrate was concentrated in vacuo, and the residue was
recrystallized in a mixture solution of DCM/EtOAc at the ratio of
1/1 (6 mL) to afford the title compound as a pale solid (125 mg,
72%).
[0367] LC-MS (ESI, pos, ion): 513.0 [M+H].sup.+;
[0368] Q-TOF (ESI, pos, ion): 513.1873 [M+H].sup.+;
[0369] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 10.81 (s,
1H), 8.31 (d, J=8.9 Hz, 1H), 8.17 (d, J=2.8 Hz, 2H), 8.13 (d, J=5.8
Hz, 1H), 7.84 (d, J=2.2 Hz, 1H), 7.57 (t, J=7.6 Hz, 2H), 7.49 (d,
J=7.4 Hz, 1H), 7.46-7.41 (m, 3H), 6.57 (dd, J=5.7, 2.3 Hz, 1H),
5.28 (s, 2H), 4.19-4.14 (m, 2H), 3.69 (t, J=5.0 Hz, 2H), 1.56-1.50
(m, 1H), 1.28 (t, J=7.1 Hz, 2H), 1.12-1.07 (m, 2H).
Example 12
N-(5-((2-acetamidopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,6,7-te-
trahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
##STR00028##
[0371] To a solution of
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,6,7-tetra-
hydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide (100 mg, 0.23
mmol) in Ac.sub.2O (1 mL) was added Et.sub.3N (139 mg, 1.38 mmol).
The suspension was warmed up to 30.degree. C. and stirred
overnight. The reaction was cooled down to r.t. and concentrated in
vacuo, the residue was recrystallized in a mixture solution of
DCM/EtOAc/hex at the ratio of 2/2/1 (5 mL) to afford the title
compound as a pale solid (55 mg, 50%).
[0372] LC-MS (ESI, pos, ion): 487.0 [M+H].sup.+;
[0373] Q-TOF (ESI, pos, ion): 487.1730 [M+H].sup.+;
[0374] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 10.82 (s,
1H), 8.32 (d, J=9.0 Hz, 1H), 8.17 (d, J=2.6 Hz, 1H), 8.12 (s, 1H),
8.03 (s, 1H), 7.88 (s, 1H), 7.57 (t, J=7.8 Hz, 2H), 7.50-7.45 (m,
2H), 7.42 (d, J=7.5 Hz, 2H), 6.56 (d, J=3.8 Hz, 1H), 5.28 (s, 2H),
4.19-4.15 (m, 2H), 3.69 (t, J=4.9 Hz, 2H), 2.20 (s, 3H).
Example 13
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,5,6-tetrah-
ydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
##STR00029##
[0375] Step 1)
N-(5-hydroxypyridin-2-yl)-2-oxo-1-phenyl-2,4,5,6-tetrahydro-1H-pyrrolo[1,-
2-b]pyrazole-3-carboxamide
[0376] To a solution of 6-aminopyridin-3-ol hydrochloride (2.02 g,
13.8 mmol) in DMF (19 mL) was added TEA (2.18 g, 21.5 mmol) at
r.t., the reaction was stirred at r.t. for 30 min. and then to the
solution was added
2-oxo-1-phenyl-2,4,5,6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carbo-
xylic acid (2.1 g, 8.6 mmol), HOAT (293 mg, 2.15 mmol) and EDCI
(2.06 g, 10.75 mmol) in portions. The reaction was allowed to warm
up to 60.degree. C. and stirred overnight. The reaction was cooled
down to r.t. and diluted with water (190 mL) and stirred for 4 h,
filtered and the solid was washed with water (20 mL.times.2), dried
in vacuo at 60.degree. C. for 4 h to afford the title compound as a
brown solid (1.61 g, 56%).
[0377] LC-MS (ESI, pos, ion): 337.1 [M+H].sup.+;
[0378] .sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. (ppm): 10.34 (s,
1H), 9.65 (s, 1H), 8.05 (d, J=8.9 Hz, 1H), 7.84 (d, J=2.8 Hz, 1H),
7.56 (t, J=7.9 Hz, 2H), 7.52-7.49 (m, 2H), 7.41 (t, J=7.3 Hz, 1H),
7.21 (dd, J=8.9, 2.9 Hz, 1H), 3.79 (t, J=6.9 Hz, 2H), 3.16 (t,
J=7.4 Hz, 2H), 2.44-2.39 (m, 2H).
Step 2)
N-(5-((2-carbamoylpyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,-
4,5,6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
[0379] To a solution of 4-chloropicolinamide (862 mg, 5.5 mmol) and
N-(5-hydroxypyridin-2-yl)-2-oxo-1-phenyl-2,4,5,6-tetrahydro-1H-pyrrolo[1,-
2-b]pyrazole-3-carboxamide (1.61 g, 4.8 mmol) in DMF (10 mL) was
added t-BuOK (1.08 g, 9.6 mmol). The reaction was allowed to warm
up to 130.degree. C. and stirred overnight. The reaction was cooled
to r.t. and diluted with 100 mL of water, continued to stir at r.t.
overnight, filtered, the collected mass was dried in vacuo at
60.degree. C. for 4 h to afford the title compound as a brown solid
(1.7 g, 79%).
[0380] LC-MS (ESI, pos, ion): 457.3 [M+H].sup.+;
[0381] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 10.76 (s,
1H), 8.45 (d, J=5.6 Hz, 1H), 8.41 (d, J=9.0 Hz, 1H), 8.19 (d, J=2.8
Hz, 1H), 7.84 (s, 1H), 7.77 (d, J=2.5 Hz, 1H), 7.52 (d, J=7.4 Hz,
2H), 7.46 (d, J=6.8 Hz, 3H), 7.39 (t, J=7.3 Hz, 1H), 7.00 (dd,
J=5.6, 2.6 Hz, 1H), 5.57 (s, 1H), 3.76 (t, J=6.9 Hz, 2H), 3.35 (t,
J=7.4 Hz, 2H), 2.56 (p, J=7.0 Hz, 2H).
Step 3)
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,5,-
6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
[0382] To a solution of
N-(5-((2-carbamoylpyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,5,6-t-
etrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide (1.72 g, 3.8
mmol) in a mixture solvent of EtOAc (28 mL), CH.sub.3CN (28 mL) and
water (14 mL) at 0.degree. C. was added PhI(OAc).sub.2 (1.52 g, 4.7
mmol). The reaction mixture was stirred at 0.degree. C. for 30 min
and then removed to r.t. and stirred overnight. The reaction was
filtered, the solid mass was washed with EtOAc (20 mL) twice, the
filtrate was concentrated in vacuo, and the residue was purified by
a column chromatography on a silica gel (DCM to DCM/MeOH=100/1) to
afford the title compound as a pale yellow solid (450 mg, 28%).
[0383] LC-MS (ESI, pos, ion): 429.3 [M+H].sup.+;
[0384] Q-TOF (ESI, pos, ion): 429.1678 [M+H].sup.+;
[0385] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 10.71 (s,
1H), 8.34 (d, J=9.0 Hz, 1H), 8.16 (d, J=2.8 Hz, 1H), 7.95 (d, J=5.9
Hz, 1H), 7.52 (t, J=7.9 Hz, 2H), 7.46-7.41 (m, 3H), 7.38 (t, J=7.4
Hz, 1H), 6.30 (dd, J=5.9, 2.2 Hz, 1H), 5.96 (d, J=2.1 Hz, 1H), 4.53
(s, 2H), 3.75 (t, J=6.9 Hz, 2H), 3.33 (t, J=7.4 Hz, 2H), 2.55 (p,
J=7.1 Hz, 2H).
Example 14
N-(5-((2-(cyclopropanecarboxamido)pyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-p-
henyl-2,4,5,6-tetrahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
##STR00030##
[0387] To a solution of
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,5,6-tetra-
hydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide (150 mg, 0.35 mmol)
and pyridine (1.3 mL) in CH.sub.3CN (2 mL) was added a solution of
DMAP (128 mg, 1.05 mmol) and cyclopropanecarbonyl chloride (110 mg,
1.05 mmol) in CH.sub.3CN (1.3 mL) at r.t. The reaction mixture was
allowed to warm up to 60.degree. C. and stirred overnight. The
reaction was cooled down to r.t., and was diluted with 20 mL of DCM
and 5 mL of water, the water phase was extracted with DCM (10 mL),
the combined organic phase was washed with brine (10 mL), dried
over Na.sub.2SO.sub.4 and filtered, the filtrate was concentrated
in vacuo, and the residue was added 5 mL of EtOAc and 5 mL of water
then stirred for 4 h, filtered, the collected solid mass was dried
in vacuo at 60.degree. C. for 4 h to afford the title compound as a
pale yellow solid (120 mg, 61%).
[0388] LC-MS (ESI, pos, ion): 497.0 [M+H].sup.+;
[0389] Q-TOF (ESI, pos, ion): 497.1915 [M+H].sup.+;
[0390] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 10.69 (s,
1H), 8.35 (d, J=8.8 Hz, 2H), 8.15 (d, J=2.7 Hz, 1H), 8.11 (d, J=5.8
Hz, 1H), 7.83 (d, J=1.6 Hz, 1H), 7.50 (t, J=7.8 Hz, 2H), 7.45-7.41
(m, 3H), 7.36 (t, J=7.4 Hz, 1H), 6.55 (dd, J=5.8, 2.3 Hz, 1H), 3.73
(t, J=6.9 Hz, 2H), 3.31 (t, J=7.4 Hz, 2H), 2.53 (p, J=7.1 Hz, 2H),
1.50-1.54 (m, 1H), 1.06-1.09 (m, 2H), 0.86-0.88 (m, 2H). .sup.13C
NMR (150 MHz, CDCl.sub.3) .delta. (ppm): 172.20, 166.50, 165.94,
162.63, 160.97, 153.22, 149.18, 146.61, 141.07, 134.80, 130.25,
129.46, 127.35, 123.06, 114.79, 108.05, 101.81, 98.64, 49.77,
25.72, 22.14, 15.92, 8.39.
Example 15
N-(5-((2-acetamidopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,5,6-te-
trahydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide
##STR00031##
[0392] To a solution of
N-(5-((2-aminopyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-2,4,5,6-tetra-
hydro-1H-pyrrolo[1,2-b]pyrazole-3-carboxamide (100 mg, 0.23 mmol)
in Ac.sub.2O (1 mL) was added Et.sub.3N (141 mg, 1.40 mmol). The
suspension was warmed up to 30.degree. C. and stirred overnight.
The reaction was cooled down to r.t. and 4 mL of water and 4 mL of
EtOAc was added, the mixture was continued to stir at r.t. for 2 h,
filtered, the collected solid was washed with EtOAc (2 mL), then
dried in vacuo at 60.degree. C. for 4 h to afford the title
compound as a pale solid (60 mg, 55%).
[0393] LC-MS (ESI, pos, ion): 471.3 [M+H].sup.+;
[0394] Q-TOF (ESI, pos, ion): 471.1776 [M+H].sup.+;
[0395] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 10.70 (s,
1H), 8.36 (d, J=9.0 Hz, 1H), 8.16 (d, J=2.8 Hz, 1H), 8.10 (d, J=5.8
Hz, 1H), 8.05 (s, 1H), 7.86 (s, 1H), 7.51 (t, J=7.9 Hz, 2H),
7.46-7.42 (m, 3H), 7.36 (t, J=7.4 Hz, 1H), 6.54 (dd, J=5.8, 2.3 Hz,
1H), 3.73 (t, J=6.9 Hz, 2H), 3.32 (t, J=7.4 Hz, 2H), 2.53 (p, J=7.1
Hz, 2H), 2.18 (s, 3H).
Example 16
N-(4-((2-aminopyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-2,4,6,7-tet-
rahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
##STR00032##
[0396] Step 1)
N-(4-((2-carbamoylpyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-2,4,6,-
7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
[0397] To a solution of
2-oxo-1-phenyl-2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carbox-
ylic acid (500 mg, 1.92 mmol),
4-(4-amino-2-fluorophenoxy)picolinamide (465.6 mg, 1.89 mmol) and
HOAT (51.4 mg, 0.38 mmol) in DCM (15 mL) was added EDCI (555 mg,
2.89 mmol). The reaction was allowed to warm up to 50.degree. C.
and stirred overnight. The reaction was cooled down to r.t. and
concentrated in vacuo, the residue was purified by a column
chromatography on silica gel (DCM/MeOH (v/v)=100/1-50/1) to afford
the title compound as a white solid (450 mg, 48.7%).
[0398] LC-MS (ESI, pos, ion): 490.3 [M+H].sup.+;
[0399] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.55 (s,
1H), 8.54 (d, J=5.6 Hz, 1H), 8.13 (s, 1H), 8.01-7.94 (m, 1H), 7.73
(s, 1H), 7.65-7.57 (m, 2H), 7.54 (dd, J=7.1, 4.9 Hz, 3H), 7.44-7.35
(m, 3H), 7.22 (dd, J=5.6, 2.7 Hz, 1H), 5.12 (s, 2H), 4.10 (q, J=5.3
Hz, 2H), 3.71 (t, J=4.9 Hz, 2H).
Step 2)
N-(4-((2-aminopyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-2,4-
,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
[0400]
N-(4-((2-carbamoylpyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl--
2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
(450 mg, 0.92 mmol) in a mixture of EtOAc (7 mL), MeCN (7 mL) and
H.sub.2O (3.5 mL) at 0.degree. C. was added PhI(OAc).sub.2 (370.4
mg, 1.15 mmol). The mixture was stirred at 0.degree. C. for 0.5 h.
Then the reaction was warmed up to r.t. and stirred overnight. The
mixture was concentrated in vacuo and the residue was purified by a
column chromatography on silica gel (DCM/MeOH (v/v)=100/1-50/1) to
afford the title compound as a yellow solid (280 mg, 66%).
[0401] LC-MS (ESI, pos, ion): 462.0 [M+H].sup.+;
[0402] Q-TOF (ESI, pos, ion): 462.1578 [M+H].sup.+;
[0403] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.55 (s,
1H), 8.54 (d, J=5.6 Hz, 1H), 8.13 (s, 1H), 8.01-7.94 (m, 1H), 7.73
(s, 1H), 7.65-7.57 (m, 2H), 7.54 (dd, J=7.1, 4.9 Hz, 3H), 7.44-7.35
(m, 3H), 7.22 (dd, J=5.6, 2.7 Hz, 1H), 5.12 (s, 2H), 4.10 (q, J=5.3
Hz, 2H), 3.71 (t, J=4.9 Hz, 2H).
Example 17
N-(4-((2-(cyclopropanecarboxamido)pyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo--
1-phenyl-2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
##STR00033##
[0405] To a solution of
N-(4-((2-aminopyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-2,4,6,7-te-
trahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide (170 mg, 0.37
mmol) and pyridine (1.2 mL) in MeCN (2 mL) was added a solution of
cyclopropanecarbonyl chloride (116 mg, 1.1 mmol) and DMAP (134 mg,
1.1 mmol) in MeCN (1.2 mL). The reaction mixture was allowed to
warmed up to 60.degree. C. and stirred overnight. The reaction was
cooled down to r.t. and concentrated in vacuo, the residue was
purified by a column chromatography on silica gel (EtOAc/PE
(v/v)=5/2-1/1.5) to afford the title compound as a white solid (100
mg, 51.3%).
[0406] LC-MS (ESI, pos, ion): 530.0 [M+H].sup.+;
[0407] Q-TOF (ESI, pos, ion): 530.1837 [M+H].sup.+;
[0408] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.86 (s,
1H), 10.52 (s, 1H), 8.20 (d, J=5.7 Hz, 1H), 8.01-7.87 (m, 1H),
7.67-7.56 (m, 3H), 7.54 (t, J=6.3 Hz, 3H), 7.38-7.27 (m, 2H), 6.72
(dd, J=5.7, 2.4 Hz, 1H), 5.12 (s, 2H), 4.09 (dt, J=17.8, 8.8 Hz,
2H), 3.71 (t, J=4.9 Hz, 2H), 2.04-1.90 (m, 1H), 1.32-1.14 (m, 2H),
0.81-0.72 (m, 2H).
Example 18
N-(4-((2-aminopyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,6,7-tetrahydro-1-
H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
##STR00034##
[0409] Step 1)
N-(4-((2-carbamoylpyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,6,7-tetrahy-
dro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
[0410] To a solution of
2-oxo-1-phenyl-2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carbox-
ylic acid (500 mg, 1.92 mmol), 4-(4-aminophenoxy)picolinamide (431
mg, 1.89 mmol) and HOAT (51.4 mg, 0.38 mmol) in DCM (15 mL) was
added EDCI (555 mg, 2.89 mmol). The reaction was allowed to warmed
up to 50.degree. C. and stirred overnight. The reaction was cooled
down to r.t. and concentrated in vacuo, the residue was purified by
a column chromatography on silica gel (DCM/MeOH (v/v)=100/1-50/1)
to afford the title compound as a white solid (680 mg, 74.15%).
[0411] LC-MS (ESI, pos, ion): 472.3 [M+H].sup.+;
[0412] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.43 (s,
1H), 8.51 (d, J=5.6 Hz, 1H), 8.11 (s, 1H), 7.72 (t, J=11.1 Hz, 3H),
7.64-7.57 (m, 2H), 7.56-7.48 (m, 3H), 7.40 (d, J=2.5 Hz, 1H),
7.23-7.14 (m, 3H), 5.13 (s, 2H), 4.09 (dd, J=10.9, 5.5 Hz, 2H),
3.70 (t, J=4.8 Hz, 2H).
Step 2)
N-(4-((2-aminopyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,6,7-tetr-
ahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
[0413] To a solution of
N-(4-((2-carbamoylpyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,6,7-tetrahy-
dro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide (650 mg, 1.38
mmol) in a mixture of EtOAc (16 mL), MeCN (16 mL) and H.sub.2O (8
mL) at 0.degree. C. was added PhI(OAc).sub.2 (666.76 mg, 2.07
mmol). The mixture was stirred at 0.degree. C. for 0.5 h. Then the
reaction was warmed up to r.t. and stirred overnight. The mixture
was concentrated in vacuo and the residue was purified by a column
chromatography on silica gel (DCM/MeOH (v/v)=100/1-50/1) to afford
the title compound as a yellow solid (561 mg, 91.8%).
[0414] LC-MS (ESI, pos, ion): 444.3 [M+H].sup.+;
[0415] Q-TOF (ESI, pos, ion): 444.1861 [M+H].sup.+;
[0416] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.37 (s,
1H), 7.79 (d, J=5.8 Hz, 1H), 7.67 (d, J=8.8 Hz, 2H), 7.64-7.57 (m,
2H), 7.53 (t, J=6.3 Hz, 3H), 7.10 (d, J=8.8 Hz, 2H), 6.14 (dd,
J=5.8, 2.1 Hz, 1H), 5.92 (s, 2H), 5.82 (d, J=1.9 Hz, 1H), 5.12 (s,
2H), 4.10 (t, J=4.8 Hz, 2H), 3.69 (t, J=4.8 Hz, 2H).
Example 19
N-(4-((2-(cyclopropanecarboxamido)pyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl--
2,4,6,7-tetrahydro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
##STR00035##
[0418] To a solution of
N-(4-((2-aminopyridin-4-yl)oxy)-3-phenyl)-2-oxo-1-phenyl-2,4,6,7-tetrahyd-
ro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide (222 mg, 0.5 mmol)
and pyridine (1.6 mL) in MeCN (2.7 mL) was added a solution of
cyclopropanecarbonyl chloride (157 mg, 1.5 mmol) and DMAP (183 mg,
1.5 mmol) in MeCN (1.6 mL). The reaction mixture was allowed to
warmed up to 60.degree. C. and stirred overnight. The reaction was
cooled down to r.t. and concentrated in vacuo, the residue was
purified by a column chromatography on silica gel (DCM/MeOH
(v/v)=100/0-100/1) to afford the title compound as a white solid
(100 mg, 39.1%).
[0419] LC-MS (ESI, pos, ion): 512.3 [M+H].sup.+;
[0420] Q-TOF (ESI, pos, ion): 512.1884 [M+H].sup.+;
[0421] .sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. (ppm): 10.81 (s,
1H), 10.40 (s, 1H), 8.18 (d, J=5.7 Hz, 1H), 7.72-7.67 (m, 2H), 7.64
(d, J=2.3 Hz, 1H), 7.60 (dd, J=9.6, 5.9 Hz, 2H), 7.55-7.50 (m, 3H),
7.16-7.10 (m, 2H), 6.67 (dd, J=5.7, 2.4 Hz, 1H), 5.12 (s, 2H), 4.10
(t, J=5.0 Hz, 2H), 3.70 (t, J=5.0 Hz, 2H), 1.98 (m, 1H), 1.27-1.23
(m, 2H), 0.77 (m, 2H).
Example 20
N-(4-((2-acetamidopyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-2,4,6,7-tetrahyd-
ro-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide
##STR00036##
[0423] To a solution of
N-(4-((2-aminopyridin-4-yl)oxy)-phenyl)-2-oxo-1-phenyl-2,4,6,7-tetrahydro-
-1H-pyrazolo[5,1-c][1,4]oxazine-3-carboxamide (1.2 g, 2.7 mmol) in
acetyl acetate (20 mL) was added Et.sub.3N (1.63 g, 16.1 mmol). The
reaction mixture was allowed to warm up to 50.degree. C. and
stirred overnight. The reaction was cooled down to r.t. and
concentrated in vacuo, the residue was added a solution of
DCM/EtOAc/PE=2/2/1 (30 mL) and stirred for 2 h. The mixture was
filtered to afford the title compound as a white solid (800 mg,
60%).
[0424] LC-MS (ESI, pos, ion): 486.3 [M+H].sup.+;
[0425] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.51 (s,
1H), 10.40 (s, 1H), 8.17 (d, J=5.7 Hz, 1H), 7.69 (d, J=8.9 Hz, 2H),
7.65 (d, J=1.7 Hz, 1H), 7.63-7.57 (m, 2H), 7.56-7.49 (m, 3H), 7.14
(d, J=8.9 Hz, 2H), 6.64 (dd, J=5.7, 2.3 Hz, 1H), 5.12 (s, 2H), 4.10
(t, J=4.9 Hz, 2H), 3.69 (t, J=4.8 Hz, 2H), 2.04 (s, 3H).
Example 21
N-(5-((2-amino-3-chloropyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl-1,2,4-
,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
##STR00037##
[0426] Step 1)
N-(5-((2-carbamoyl-3-chloropyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl--
1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
[0427] To a solution of 3,4-dichloropicolinamide (263 mg, 1.38
mmol) and
N-(5-hydroxypyridin-2-yl)-2-oxo-1-phenyl-1,2,4,5,6,7-hexahydropyrazolo[1,-
5-a]pyridine-3-carboxamide (420 mg, 1.20 mmol) in DMF (2.4 mL) was
added t-BuOK (269 mg, 2.40 mmol). The reaction was allowed to warm
up to 130.degree. C. and stirred overnight. The reaction was cooled
down to r.t. and diluted with 24 mL of water, continued to stir at
r.t. overnight, filtered, and the collected mass was dried in vacuo
at 60.degree. C. for 5 h to afford the title compound as a brown
solid (489 mg, 81%).
[0428] LC-MS (ESI, pos, ion): 505.2 [M+H].sup.+;
[0429] .sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. (ppm): 11.13 (s,
1H), 8.35 (dd, J=7.5, 3.0 Hz, 2H), 8.27 (d, J=2.9 Hz, 1H), 8.04 (s,
1H), 7.75 (dd, J=9.0, 2.8 Hz, 2H), 7.60 (t, J=7.7 Hz, 2H), 7.53 (t,
J=7.4 Hz, 1H), 7.49-7.46 (m, 2H), 6.93 (d, J=5.6 Hz, 1H), 3.58 (t,
J=5.9 Hz, 2H), 3.22 (t, J=6.3 Hz, 2H), 2.00-1.97 (m, 2H), 1.85-1.80
(m, 2H).
Step 2)
N-(5-((2-amino-3-chloropyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phen-
yl-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
[0430] To a solution of
N-(5-((2-carbamoyl-3-chloropyridin-4-yl)oxy)pyridin-2-yl)-2-oxo-1-phenyl--
1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide (489 mg,
0.97 mmol) in a mixture solvent of EtOAc (7.2 mL), CH.sub.3CN (7.2
mL) and H.sub.2O (3.6 mL) at 0.degree. C. was added PhI(OAc).sub.2
(390 mg, 1.21 mmol). The mixture was continued to stir at 0.degree.
C. for 30 min, then was allowed to warm up to r.t. and stirred
overnight. The reaction was filtered, and the filtrate was
concentrated in vacuo, the residue was purified by a column
chromatography on silica gel (DCM to MeOH/DCM=1/100) to afford the
title compound as a pale yellow solid (260 mg, 56%).
[0431] LC-MS (ESI, pos, ion): 476.9 [M+H].sup.+;
[0432] Q-TOF (ESI, pos, ion): 477.1391 [M+H].sup.+;
[0433] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. (ppm): 11.12 (s,
1H), 8.34 (d, J=9.0 Hz, 1H), 8.17 (d, J=2.7 Hz, 1H), 7.82 (d, J=5.7
Hz, 1H), 7.54 (t, J=7.7 Hz, 2H), 7.48-7.41 (m, 2H), 7.38 (d, J=7.5
Hz, 2H), 6.08 (d, J=5.7 Hz, 1H), 5.00 (s, 2H), 3.59 (t, J=5.9 Hz,
2H), 3.40 (t, J=6.4 Hz, 2H), 2.13-2.07 (m, 2H), 1.98-1.91 (m,
2H).
[0434] .sup.13C NMR (150 MHz, CDCl.sub.3) .delta. (ppm): 163.53,
161.81, 160.77, 156.67, 155.51, 149.52, 146.87, 146.60, 140.85,
133.00, 129.98, 129.64, 128.80, 126.56, 114.68, 103.09, 102.85,
98.21, 47.42, 23.77, 22.60, 19.72.
Example 22
N-(4-((2-amino-3-chloropyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-1,2,4,5,6,7-
-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
##STR00038##
[0435] Step 1) 3,4-dichloropicolinamide
[0436] To a solution of 2,2,6,6-tetramethylpiperidine (12.4 mL,
74.4 mmol) in diethylether (100 mL) at 0.degree. C. was added
n-BuLi in hexane (2.5 M, 46 mL, 115 mmol) via a syringe over 15
min. The resulting solution was stirred at 0.degree. C. for 0.5 h
and at -78.degree. C. for 0.5 h. To this mixture was added a
mixture of 3,4-dichloropyridine (10.00 g, 67.6 mmol) in
diethylether (40 mL) via a syringe over 15 min. The resulting
mixture was stirred at -78.degree. C. for 2 h before the addition
of isocyanatotrimethylsilane (94% pure, 13.4 mL, 101.4 mmol). After
the addition, the cooling bath was removed and the reaction mixture
was allowed to warm to room temperature for 2 h. The reaction
mixture was quenched with acetic acid (13.52 g, 225.2 mmol) and 70
mL of water. The mixture was allowed to stir overnight, and the
white solid that formed was collected through filtration and washed
with water. The filtrate was extracted with EtOAc (50 mL.times.3).
The combined organic phase was washed with brine and dried over
anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo. The residue
was beaten by PE/EtOAc=2/1 (100 mL) to give a white solid. The two
parts of solid was got together to give the title compound (6.95 g,
53.9%).
[0437] LC-MS (ESI, pos, ion): 191 [M+H].sup.+;
[0438] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. (ppm): 8.38 (d,
J=5.0 Hz, 1H), 7.59 (d, J=5.0 Hz, 1H).
Step 2) 4-(4-aminophenoxy)-3-chloropicolinamide
[0439] To a solution of 4-aminophenol hydrochloride (436.8 mg, 3
mmol) in DMF (6 mL) was added t-BuOK (841 mg, 7.5 mmol) at
0.degree. C. The mixture was stirred at 0.degree. C. for 15 min
before 3,4-dichloropicolinamide (570 mg, 3 mmol) was added. The
reaction mixture was allowed to warm up to 80.degree. C. and
stirred overnight. The reaction mixture was concentrated in vacuo.
The residue was added H.sub.2O (10 mL) and filtered. The filter
cake was washed with 2 mL of H.sub.2O and dried in vacuo. The
filtrate was extracted by EtOAc (10 mL.times.2). The organic phase
was washed with brine dried over anhydrous Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was beten by PE/EtOAc (15 mL/5
mL) to give a yellow solid. Two parts of solid was collected
together to give the title compound (510 mg, 64.3%).
[0440] LC-MS (ESI, pos, ion): 264.2 [M+H].sup.+;
[0441] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 8.28 (d,
J=5.6 Hz, 1H), 7.97 (d, J=12.5 Hz, 1H), 7.70 (s, 1H), 6.87 (d,
J=8.7 Hz, 2H), 6.67 (dd, J=12.5, 7.2 Hz, 3H), 5.21 (s, 2H).
Step 3)
N-(4-((2-carbamoyl-3-chloropyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-
-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
[0442] To a solution of
2-oxo-1-phenyl-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxylic
acid (510 mg, 1.98 mmol), 4-(4-aminophenoxy)-3-chloropicolinamide
(510 mg, 1.94 mmol) and HOAT (52 mg, 0.39 mmol) in DCM (15 mL) was
added EDCI (558 mg, 2.91 mmol). The reaction was allowed to warm up
to 50.degree. C. and stirred overnight. The reaction was cooled
down to r.t. and concentrated in vacuo. the residue was beaten by
DCM/EtOAc/H.sub.2O (20 mL/10 mL/10 mL) for 1 h. The mixture was
filtered and the filter cake was dried in vacuo to afford the title
compound as a white solid (539 mg, 55.3%).
[0443] LC-MS (ESI, pos, ion): 504.3 [M+H].sup.+;
[0444] .sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. (ppm): 10.66 (s,
1H), 7.76 (d, J=5.8 Hz, 1H), 7.66 (t, J=10.3 Hz, 2H), 7.58 (t,
J=7.7 Hz, 2H), 7.51 (t, J=7.4 Hz, 1H), 7.46 (d, J=7.4 Hz, 2H), 7.11
(d, J=8.9 Hz, 2H), 6.48 (s, 2H), 5.98 (d, J=5.7 Hz, 1H), 3.57 (t,
J=5.8 Hz, 2H), 3.21 (t, J=6.3 Hz, 2H), 1.98 (dd, J=7.6, 4.3 Hz,
2H), 1.85-1.77 (m, 2H).
Step 4)
N-(4-((2-amino-3-chloropyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl-1,2-
,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
[0445]
N-(4-((2-carbamoyl-3-chloropyridin-4-yl)oxy)phenyl)-2-oxo-1-phenyl--
1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide (539 mg,
1.03 mmol) in a mixture solvent of EtOAc (8 mL), MeCN (8 mL) and
H.sub.2O (4 mL) at 0.degree. C. was added PhI(OAc).sub.2 (500 mg,
1.55 mmol). The mixture was stirred at 0.degree. C. for 0.5 h. Then
the reaction was warmed up to r.t. and stirred overnight. The
mixture was filtered. The filtrate was concentrated in vacuo. The
residue was beaten by PE/EtOAc (4 mL/4 mL) for 1 h. The mixture was
filtered and the filter cake was dried in vacuo to afford the title
compound as a yellow solid (254 mg, 51.9%).
[0446] LC-MS (ESI, pos, ion): 476.0 [M+H].sup.+;
[0447] Q-TOF (ESI, pos, ion): 476.1447 [M+H].sup.+;
[0448] .sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. (ppm): 10.66 (s,
1H), 7.76 (d, J=5.8 Hz, 1H), 7.66 (t, J=10.3 Hz, 2H), 7.58 (t,
J=7.7 Hz, 2H), 7.51 (t, J=7.4 Hz, 1H), 7.46 (d, J=7.4 Hz, 2H), 7.11
(d, J=8.9 Hz, 2H), 6.48 (s, 2H), 5.98 (d, J=5.7 Hz, 1H), 3.57 (t,
J=5.8 Hz, 2H), 3.21 (t, J=6.3 Hz, 2H), 1.98 (dd, J=7.6, 4.3 Hz,
2H), 1.85-1.77 (m, 2H).
Example 23
N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-phenyl-1,-
2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
##STR00039##
[0449] Step 1) 4-amino-2-fluorophenol
[0450] To a solution of 2-fluoro-4-nitrophenol (1.57 g, 10 mmol) in
EtOH (50 mL) and H.sub.2O (18 mL) was added Fe (2.24 g, 40 mmol)
and NH.sub.4C.sub.1 (4.24 g, 80 mmol). The reaction mixture was
stirred at r.t. overnight. The mixture was filtered through a pad
of celite. The filtrate was concentrated in vacuo. The residue was
added H.sub.2O (50 mL) and extracted by EtOAc (60 mL.times.2). The
combined organic phase was washed with brine, dried over anhydrous
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was beaten
by PE/EtOAc (v/v, 10 mL/15 mL), filtered to give the title compound
as a brown solid (700 mg, 55.1%).
[0451] LC-MS (ESI, pos, ion): 128.3 [M+H].sup.+;
[0452] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 8.56 (s,
1H), 6.62 (dd, J=10.0, 8.6 Hz, 1H), 6.34 (dd, J=13.4, 2.6 Hz, 1H),
6.24-6.15 (m, 1H), 4.66 (s, 2H).
Step 2) 4-(4-amino-2-fluorophenoxy)-3-chloropicolinamide
[0453] To a solution of 4-amino-2-fluorophenol (381 mg, 3 mmol) and
3,4-dichloropicolinamide (570 mg, 3 mmol) in DMF (4 mL) was added
t-BuOK (404 mg, 3.6 mmol). The reaction mixture was allowed to warm
up to 80.degree. C. and stirred overnight. The mixture was added
H.sub.2O (15 mL) and filtered. The filter cake was washed with 2 mL
of H.sub.2O and dried in vacuo to give the title compound as a
black solid (508 mg, 60.2%).
[0454] LC-MS (ESI, pos, ion): 282.0 [M+H].sup.+;
[0455] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 8.31 (d,
J=5.6 Hz, 1H), 8.03 (s, 1H), 7.73 (s, 1H), 7.04 (t, J=9.0 Hz, 1H),
6.73 (d, J=5.5 Hz, 1H), 6.54 (dd, J=13.2, 2.4 Hz, 1H), 6.45 (dd,
J=8.7, 2.0 Hz, 1H), 5.55 (s, 2H).
Step 3)
N-(4-((2-carbamoyl-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-
-1-phenyl-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
[0456] To a solution of
2-oxo-1-phenyl-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxylic
acid (476 mg, 1.84 mmol),
4-(4-amino-2-fluorophenoxy)-3-chloropicolinamide (508 mg, 1.8 mmol)
and HOAT (49 mg, 0.36 mmol) in DCM (15 mL) was added EDCI (518 mg,
2.7 mmol). The reaction was allowed to warm up to 50.degree. C. and
stirred overnight. The reaction was cooled down to r.t. and
concentrated in vacuo. The residue was beaten by DCM/EtOAc/H.sub.2O
(20 mL/10 mL/10 mL) for 1 h. The mixture was filtered and the
filter cake was dried in vacuo to afford the title compound as a
yellow solid (318 mg, 33.9%).
[0457] LC-MS (ESI, pos, ion): 521.9 [M+H].sup.+;
[0458] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. (ppm): 10.83 (s,
1H), 8.34 (d, J=5.6 Hz, 1H), 8.06 (s, 1H), 8.02-7.92 (m, 1H), 7.75
(s, 1H), 7.59 (t, J=7.5 Hz, 2H), 7.55-7.44 (m, 3H), 7.43-7.32 (m,
2H), 6.85 (d, J=5.5 Hz, 1H), 3.58 (t, J=5.7 Hz, 2H), 3.20 (t, J=6.2
Hz, 2H), 1.98 (d, J=5.5 Hz, 2H), 1.83 (d, J=5.7 Hz, 2H).
Step 4)
N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo-1-p-
henyl-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
[0459]
N-(4-((2-carbamoyl-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-2-oxo--
1-phenyl-1,2,4,5,6,7-hexahydropyrazolo[1,5-a]pyridine-3-carboxamide
(318 mg, 0.61 mmol) in a mixture of EtOAc (8 mL), MeCN (8 mL) and
H.sub.2O (4 mL) at 0.degree. C. was added PhI(OAc).sub.2 (295 mg,
0.915 mmol). The mixture was stirred at 0.degree. C. for 0.5 h.
Then the reaction was warmed up to r.t. and stirred overnight. The
mixture was filtered. The filtrate was concentrated in vacuo. The
residue was purified by a silica gel column chromatography
(DCM/MeOH (v/v)=50/1) to give the title compound as a white solid
(203.3 mg, 67.6%).
[0460] LC-MS (ESI, pos, ion): 493.9 [M+H].sup.+;
[0461] Q-TOF (ESI, pos, ion): 494.1395 [M+H].sup.+;
[0462] .sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. (ppm): 10.78 (s,
1H), 7.93 (d, J=13.3 Hz, 1H), 7.75 (d, J=5.7 Hz, 1H), 7.59 (t,
J=7.7 Hz, 2H), 7.51 (t, J=7.3 Hz, 1H), 7.46 (d, J=7.8 Hz, 2H), 7.28
(t, J=6.8 Hz, 2H), 6.40 (s, 2H), 5.93 (d, J=5.7 Hz, 1H), 3.57 (t,
J=5.8 Hz, 2H), 3.20 (t, J=6.3 Hz, 2H), 1.99 (dd, J=14.5, 9.1 Hz,
2H), 1.85-1.76 (m, 2H).
Biological Testing
[0463] The LC/MS/MS system used in the analysis consists of an
Agilent 1200 Series vacuum degasser, binary pump, well-plate
autosampler, thermostatted column compartment, the Agilent G6430
Triple Quadrupole Mass Spectrometer with an electrosprayionization
(ESI) source. Quantitative analysis was carried out using MRM mode.
The parameters for MRM transitions are in the Table A.
TABLE-US-00001 TABLE A MRM 490.2.fwdarw.383.1 Fragmentor 230 V CE
55 V Drying Gas Temp 350.degree. C. Nebulize 40 psi Drying Gas Flow
10 L/min
[0464] An Agilent XDB-C18, 2.1.times.30 mm, 3.5 .mu.M column was
used for the analysis. 5 .mu.L of the samples were injected.
Analysis condition: The mobile phase was 0.1% formic acid in water
(A) and 0.1% formic acid in methanol (B). The flow rate was 0.4
mL/min. And the gradient of Mobile phase was in the Table B.
TABLE-US-00002 TABLE B Time Gradient of Mobile Phase B 0.5 min 5%
1.0 min 95% 2.2 min 95% 2.3 min 5% 5.0 min stop
[0465] Alternatively, an Agilent 6330 series LC/MS/MS spectrometer
equipped with G1312A binary pumps, a G1367A autosampler and a
G1314C UV detector were used in the analysis. An ESI source was
used on the LC/MS/MS spectrometer. The analysis was done in
positive ion mode as appropriate and the MRM transition for each
analyte was optimized using standard solution. A Capcell MP-C18
100.times.4.6 mm I.D., 5 .mu.M column (Phenomenex, Torrance,
Calif., USA) was used during the analysis. The mobile phase was 5
mM ammonia acetate, 0.1% MeOH in water (A): 5 mM ammonia acetate,
0.1% MeOH in acetonitrile (B) (70/30, v/v). The flow rate was 0.6
mL/min. Column was maintained at ambient temperature. 20 .mu.L of
the samples were injected.
Example A
Compound Stability in Human and Rat Liver Microsomes
[0466] Human or rat liver microsomes incubations were conducted in
duplicate in polypropylene tubes. The typical incubation mixtures
consisted of human or rat liver microsomes (0.5 mg protein/mL),
compounds of interest (5 .mu.M) and NADPH (1.0 mM) in a total
volume of 200 .mu.L potassium phosphate buffer (PBS, 100 mM, pH
7.4). Compounds were dissolved in DMSO and diluted with PBS such
that the final concentration of DMSO was 0.05%. The enzymatic
reactions were commenced with the addition of protein after a 3-min
preincubation and incubated in a water bath open to the air at
37.degree. C. Reactions were terminated at various time points (0,
5, 10, 15, 30, 60 min) by adding equal volume of ice-cold
acetonitrile. The samples were stored at -80.degree. C. until
LC/MS/MS assays.
[0467] The concentrations of compounds in the incubation mixtures
of human or rat liver microsomes were determined by a LC/MS/MS
method. The ranges of the linearity in the concentration range were
determined for each tested compounds.
[0468] A parallel incubation was performed using denatured
microsomes as the negative control, and reactions were terminated
at various time points (0, 15, 60 min) after incubation at
37.degree. C.
[0469] Dextromethorphan (70 .mu.M) was selected as the positive
control, and reactions were terminated at various time points (0,
5, 10, 15, 30, 60 min) after incubation at 37.degree. C. Both
positive and negative control samples were included in each assay
to ensure the integrity of the microsomal incubation system.
[0470] Alternatively, the stability of some of the compounds
disclosed herein in human (or rat) liver microsomes were also
conducted in the following protocol. The incubations were conducted
in duplicate in polypropylene tubes. The typical incubation
mixtures consisted of liver microsomes (final concentration: 0.5 mg
protein/mL), compounds (final concentration: 1.5 .mu.M) in a total
volume of 30 .mu.L K-buffer (contain 1.0 mM EDTA, 100 mM, pH7.4).
Compounds were dissolved in DMSO and diluted with K-buffer such
that the final concentration of DMSO was 0.2%. The enzymatic
reactions were commenced with the addition of 15 .mu.L of NADPH
(final concentration: 2 mM) after 10 min preincubation and
incubated in a 37.degree. C. incubator. Reactions were terminated
at various time points (0, 15, 30, 60 min) by adding 135 .mu.L
acetonitrile (contain IS). Protein is removed by centrifugation
with 4000 rpm, 10 min. Supernatant was collected for LCMS/MS
analysis
[0471] In the above protocol, ketanserin (1 .mu.M) was selected as
the positive control, and reactions were terminated at various time
points (0, 15, 30, 60 min) after incubation at 37.degree. C. The
positive control sample was included in each assay to ensure the
integrity of the microsomal incubation system.
Data Analysis
[0472] The concentrations of compounds in human or rat liver
microsome incubations were plotted as a percentage of the relevant
zero time point control for each reaction. The in vivo CL.sub.int
were extrapolated (ref: Naritomi, Y.; Terashita, S.; Kimura, S.;
Suzuki, A.; Kagayama, A.; and Sugiyama, Y.; Prediction of human
hepatic clearance from in vivo animal experiments and in vitro
metabolic studies with liver microsomes from animals and humans.
Drug Metab. Dispos., 2001, 29: 1316-1324).
[0473] Table 1 listed the stability of some examples disclosed
herein in human and rat liver microsomes. As showing in Table 1,
the compounds disclosed herein exhibited desirable half-life
(T.sub.1/2) and clearance (CL.sub.int) when the compounds were
incubated in human and rat liver microsomes.
TABLE-US-00003 TABLE 1 Human and rat liver microsomes Stability
Human Rat T.sub.1/2 CL.sub.int T.sub.1/2 CL.sub.int Example (min)
(mL/min/kg) (min) (mL/min/kg) Ex. 1 187.6 9.27 88.86 27.95 Ex. 2
243.9 7.13 191.2 12.99 Ex. 3 1931 0.90 341.9 7.26 Ex. 12 .infin. NA
998.1 2.49 Ex. 14 508.1 3.42 199.2 12.47 Ex. 19 .infin. NA 172.1
14.43
Example B
Evaluation of Pharmacokinetics after Intravenous and Oral
Administration of the Compounds Disclosed Herein in Mice, Rats,
Dogs and Monkeys
[0474] The compounds disclosed herein are assessed in
pharmacokinetic studies in mice, rats, dogs or monkeys. The
compounds are administered as a water solution, 2% HPMC+1%
TWEEN.RTM.80 in water solution, 5% DMSO+5% solutol in saline, 4% MC
suspension or capsule. For the intravenous administration, the
animals are generally given at 1 or 2 mg/kg dose. For the oral
(p.o.) dosing, mice and rats are generally given 5 or 10 mg/kg
dose, and dogs and monkeys are generally given 10 mg/kg dose. The
blood samples (0.3 mL) are drawn at 0.25, 0.5, 1.0, 2.0, 3.0, 4.0,
6.0, 8.0, 12 and 24 h time points or 0.083, 0.25, 0.5, 1.0, 2.0,
4.0, 6.0, 8.0 and 24 h time points and centrifuged at 3,000 or 4000
rpm for 2 to 10 min. The plasma solutions are collected, and stored
at -20.degree. C. or -70.degree. C. until analyzed by LC/MS/MS as
described above.
[0475] Table 2 listed the PK profile of the compounds disclosed
herein in rats. The compounds disclosed herein exhibited optimized
pharmacokinetic properties with desirable clearance (CL), half-life
(T.sub.1/2), exposure (AUC.sub.last) and bioavailability (F).
TABLE-US-00004 TABLE 2 Pharmacokinetic profiles in rats iv dosing
dose T.sub.1/2 AUC.sub.last Cl/F Vss Example mg/kg h ng h/ml L/h/kg
L/kg F % Ex. 1 1 1.51 1190 1.69 1.200 163.3 Ex. 2 1 7.71 27200
0.033 0.313 68.1 Ex. 3 1 3.83 26300 0.039 0.180 94.5 Ex. 12 1 6.68
46700 0.020 0.136 72.1 Ex. 14 1 5.01 12900 0.076 0.359 87.9 Ex. 19
1 7.09 50100 0.018 0.150 66.7
[0476] The efficacy of the compounds disclosed herein as inhibitors
of receptor tyrosine kinases, such as Axl, Mer, c-Met, and/or Ron
related activity and as anti-tumor agents in xenograft animal
models can be evaluated as follows. The assay results can
demonstrate that certain compounds disclosed herein potently
inhibit Axl, Mer, c-Met, and/or Ron phosphorylation, and
demonstrate potent, dose dependent anti-tumor activity in certain
xenograft models.
Example C
Kinase Activity Assay
General Description for Kinase Assays
[0477] Kinase assays can be performed by measurement of
incorporation of .gamma.-.sup.33P ATP into immobilized myelin basic
protein (MBP). High binding white 384 well plates (Greiner) are
coated with MBP (Sigma #M-1891) by incubation of 60 .mu.L/well of
20 .mu.g/mL MBP in Tris-buffered saline (TBS; 50 mM Tris pH 8.0,
138 mM NaCl, 2.7 mM KCl) for 24 h at 4.degree. C. Plates are washed
3.times. with 100 .mu.L TBS. Kinase reactions are carried out in a
total volume of 34 .mu.L in kinase buffer (5 mM Hepes pH 7.6, 15 mM
NaCl, 0.01% bovine gamma globulin (Sigma #I-5506), 10 mM
MgCl.sub.2, 1 mM DTT, 0.02% TritonX-100). Compound dilutions are
performed in DMSO and added to assay wells to a final DMSO
concentration of 1%. Each data point is measured in duplicate, and
at least two duplicate assays are performed for each individual
compound determination. Enzyme is added to final concentrations of
10 nM or 20 nM, for example. A mixture of unlabeled ATP and
.gamma.-.sup.33P ATP is added to start the reaction
(2.times.10.sup.6 cpm of .gamma.-.sup.33P ATP per well (3000
Ci/mmole) and 10 .mu.M unlabeled ATP, typically. The reactions are
carried out for 1 h at room temperature with shaking. Plates are
washed 7.times. with TBS, followed by the addition of 50 .mu.L/well
scintillation fluid (Wallac). Plates are read using a Wallac Trilux
counter. This is only one format of such assays; various other
formats are possible, as known to one skilled in the art.
[0478] The above assay procedure can be used to determine the
IC.sub.50 for inhibition and/or the inhibition constant, K.sub.i.
The IC.sub.50 is defined as the concentration of compound required
to reduce the enzyme activity by 50% under the condition of the
assay. The IC.sub.50 value is estimated by preparing a 10 point
curve using a 1/2 log dilution series (for example, a typical curve
may be prepared using the following compound concentrations: 10
.mu.M, 3 .mu.M, 1 .mu.M, 0.3 .mu.M, 0.1 .mu.M, 0.03 .mu.M, 0.01
.mu.M, 0.003 .mu.M, 0.001 .mu.M and 0 .mu.M).
Axl (h) Assay
[0479] Axl (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 250
.mu.M KKSRGDYMTMQIG, 10 mM MgAcetate and [.gamma.-.sup.33P-ATP]
(specific activity approx. 500 cpm/pmol, concentration as
required). The reaction is initiated by the addition of the MgATP
mix. After incubation for 40 minutes at room temperature, the
reaction is stopped by the addition of 3% phosphoric acid solution.
10 .mu.L of the reaction is then spotted onto a P30 filtermat and
washed three times for 5 minutes in 75 mM phosphoric acid and once
in methanol prior to drying and scintillation counting.
Mer (h) Assay
[0480] Mer (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 30
mM NaCl, 250 .mu.M GGMEDIYFEFMGGKKK, 10 mM MgAcetate and
[.gamma.-.sup.33P-ATP] (specific activity approx. 500 cpm/pmol,
concentration as required). The reaction is initiated by the
addition of the MgATP mix. After incubation for 40 minutes at room
temperature, the reaction is stopped by the addition of 3%
phosphoric acid solution. 10 .mu.L of the reaction is then spotted
onto a P30 filtermat and washed three times for 5 minutes in 75 mM
phosphoric acid and once in methanol prior to drying and
scintillation counting.
c-Met (h) Assay
[0481] Met (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 250
.mu.M KKKSPGEYVNIEFG, 10 mM MgAcetate and [.gamma.-.sup.33P-ATP]
(specific activity approx. 500 cpm/pmol, concentration as
required). The reaction is initiated by the addition of the MgATP
mix. After incubation for 40 minutes at room temperature, the
reaction is stopped by the addition of 3% phosphoric acid solution.
10 .mu.L of the reaction is then spotted onto a P30 filtermat and
washed three times for 5 minutes in 75 mM phosphoric acid and once
in methanol prior to drying and scintillation counting.
[0482] The compound disclosed herein exhibited potent activities in
the Mer (h) and c-Met (h) assays. Table 3 listed the IC.sub.50s of
some example described herein in the Mer (h) and c-Met (h)
assay.
TABLE-US-00005 TABLE 3 IC.sub.50(nM) Example Axl Mer c-Met Ex. 2 /
3 26 Ex. 3 / 35 100 Ex. 5 19 4 15 Ex. 6 36 33 57 Ex. 8 7 3 5 Ex. 9
14 12 27 Ex. 11 5 6 12 Ex. 12 14 74 54 Ex. 14 7 8 21 Ex. 17 5 / 8
Ex. 19 5 5 41 Ex. 20 49 44 / Ex. 21 9 / 14 Ex. 22 16 / 54 Ex. 23 10
14 30
[0483] The kinase assays described herein were performed at
Millipore UK Ltd, Dundee Technology Park, Dundee DD2 1SW, UK.
[0484] Alternatively, the kinase activities of the compounds can be
measured using KINOMEscan.TM., which is based on a competition
binding assay that quantitatively measures the ability of a
compound to compete with an immobilized, active-site directed
ligand. The assay was performed by combining three components:
DNA-tagged kinase; immobilized ligand; and a test compound. The
ability of the test compound to compete with the immobilized ligand
was measured via quantitative PCR of the DNA tag.
[0485] 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.TM. (Pierce), 1% BSA, 0.05% TWEEN.RTM.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.TM., 0.17.times.PBS, 0.05% TWEEN.RTM.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.RTM. 20). The
beads were then re-suspended in elution buffer (1.times.PBS, 0.05%
TWEEN.RTM.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.
[0486] The kinase activity assays described herein were performed
using KINOMEscan.TM. Profiling Service at DiscoveRx Corporation,
42501 Albrae St. Fremont, Calif. 94538, USA.
Example D
Tumor Xenograft Models
[0487] The efficacy of compounds disclosed herein is evaluated in a
standard murine model of tumorigenesis. Human tumor cells (such as
U87MG glioblastoma cells, MKN45 Gastric Adenocarcinoma cells,
MDA-MB-231 breast adenocarcinoma cells, or Caki-1 renal carcinoma
cells, all from ATCC) are expended in culture, harvested, and
injected subcutaneously in the rear flank of 6-7 week old female
athymic nude mice (BALB/cA nu/nu, Shanghai SLAC Laboratory Animal,
Co.) (n=10 for vehicle group, n=8 for each dosing group). When
tumors reach a volume of 100-250 mm.sup.3, animals are randomly
divided into vehicle control (for example, 2% HPMC+1% Tween-80 in
water) and compound groups. Subsequent administration of compound
by oral gavage (for example, 3-50 mpk/dose, dissolved in 2% HPMC+1%
Tween-80 in water) begins anywhere from day 0 to day 15 post tumor
cell challenge and generally continues with once a day for the
duration of the experiment.
Tumor Growth Inhibition (TGI) Analysis
[0488] Progression of tumor growth is assessed by tumor volumes and
recorded as a function of time. The long (L) and short (W) axes of
the subcutaneous tumors are measured with calipers twice weekly,
and the tumor volume (TV) calculated as (L.times.W.sup.2)/2). TGI
is calculated from the difference between the median tumor volumes
of vehicle-treated and drug-treated mice, expressed as a percentage
of the median tumor volume of the vehicle-treated control group, by
the following relation:
% T G I = ( Median Tumor Volume control - Median Tumor Volume drug
- treated Median Tumor Volume control ) .times. 100
##EQU00001##
[0489] Initial statistical analysis is done by repeated measures
analysis of variance (RMANOVA), Followed by Scheffe post hoc
testing for multiple comparisons. Vehicle alone (2% HPMC+1%
Tween-80, or the like) is the negative control.
[0490] Finally, it should be noted that there are alternative ways
of implementing the present invention. Accordingly, the present
embodiments are to be considered as illustrative and not
restrictive and the invention is not be limited to the details
given herein, but may be modified within the scope and equivalents
of the appended claims. All publications and patents cited herein
are incorporated by reference.
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