Patent Application
20060128724
The invention relates generally to novel chemical compounds and methods. More particularly, the invention provides novel pyrazole-substituted aminoheteroaryl compounds, particularly aminopyridines and aminopyrazines, having protein tyrosine kinase activity, and methods of synthesizing and using such compounds. Preferred compounds are c-Met inhibitors useful for the treatment of abnormal cell growth, such as cancers.
The hepatocyte growth factor (HGF) receptor (c-MET or HGFR) receptor tyrosine kinase (RTK) has been shown in many human cancers to be involved in oncogenesis, tumor progression with enhanced cell motility and invasion, as well as metastasis (see, e.g., Ma, P. C., Maulik, G., Christensen, J. & Salgia, R. (2003b). Cancer Metastasis Rev, 22, 309-25; Maulik, G., Shrikhande, A., Kijima, T., Ma, P. C., Morrison, P. T. & Salgia, R. (2002b). Cytokine Growth Factor Rev, 13, 41-59). c-MET (HGFR) can be activated through overexpression or mutations in various human cancers including small cell lung cancer (SCLC) (Ma, P. C., Kijima, T., Maulik, G., Fox, E. A., Sattler, M., Griffin, J. D., Johnson, B. E. & Salgia, R. (2003a). Cancer Res, 63, 6272-6281).
c-MET is a receptor tyrosine kinase that is encoded by the Met proto-oncogene and transduces the biological effects of hepatocyte growth factor (HGF), which is also referred to as scatter factor (SF). Jiang et al., Crit. Rev. Oncol. Hematol. 29: 209-248 (1999). c-MET and HGF are expressed in numerous tissues, although their expression is normally confined predominantly to cells of epithelial and mesenchymal origin, respectively. c-MET and HGF are required for normal mammalian development and have been shown to be important in cell migration, cell proliferation and survival, morphogenic differentiation, and organization of 3-dimensional tubular structures (e.g., renal tubular cells, gland formation, etc.). In addition to its effects on epithelial cells, HGF/SF has been reported to be an angiogenic factor, and c-MET signaling in endothelial cells can induce many of the cellular responses necessary for angiogenesis (proliferation, motility, invasion).
The c-MET receptor has been shown to be expressed in a number of human cancers. c-Met and its ligand, HGF, have also been shown to be co-expressed at elevated levels in a variety of human cancers (particularly sarcomas). However, because the receptor and ligand are usually expressed by different cell types, c-MET signaling is most commonly regulated by tumor-stroma (tumor-host) interactions. Furthermore, c-MET gene amplification, mutation, and rearrangement have been observed in a subset of human cancers. Families with germline mutations that activate c-MET kinase are prone to multiple kidney tumors as well as tumors in other tissues. Numerous studies have correlated the expression of c-MET and/or HGF/SF with the state of disease progression of different types of cancer (including lung, colon, breast, prostate, liver, pancreas, brain, kidney, ovaries, stomach, skin, and bone cancers). Furthermore, the overexpression of c-MET or HGF have been shown to correlate with poor prognosis and disease outcome in a number of major human cancers including lung, liver, gastric, and breast. c-MET has also been directly implicated in cancers without a successful treatment-regimen such as pancreatic cancer, glioma, and hepatocellular carcinoma.
Examples of c-MET (HGFR) inhibitors, their synthesis and use, can be found in U.S. patent application Ser. No. 10/786,610, entitled “Aminoheteroaryl Compounds as Protein Kinase Inhibitors”, filed Feb. 26, 2004, and corresponding international application PCT/US20041005495 of the same title, filed Feb. 26, 2004, the disclosures of which are incorporated herein by reference in their entireties.
It would be desirable to have novel c-MET (HGFR) inhibitors and methods of using such inhibitors for the treatment of abnormal cell growth, such as cancer.
In one embodiment, the invention provides a compound of formula 1
wherein:
In a particular aspect of this embodiments, each R3 is independently halogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 cycloalkyl, C6-12 aryl, 3-12 membered heteroalicyclic, 5-12 membered heteroaryl, —S(O)mR4, —SO2NR4R5, —S(O)2OR4, —NO2, —NR4R5, —(CR6R7)nOR4, —CN, —C(O)R4, —OC(O)R4, —O(CR6R7)nR4, —O(CR6R7)(CR6R7)nNR4R5, —O(CR6R7)(CR6R7)nOR4, —NR4C(O)R5, —(CR6R7)n C(O)OR4, —(CR8R7)nOR4, —(CR6R7)nC(O)NR4R5, —(CR6R7) NCR4R5, —C(═NR6)NR4R5, —NR4C(O)NR5R6, —NR4S(O)pR5 or —C(O)NR4R5, each hydrogen in R3 is optionally substituted by R8, and R3 groups on adjacent atoms may combine to form a C6-12 aryl, 5-12 membered heteroaryl, C3-12 cycloalkyl or 3-12 membered heteroalicyclic group; and
In a particular aspect of this embodiment, A is C6-12 aryl or 5-12 membered heteroaryl, optionally substituted by one or more R3 groups.
In another particular aspect of this embodiment, and in combination with any other particular aspect not inconsistent, A is phenyl substituted by one, two or three R3 groups, preferably one, two or three halogens.
In another particular aspect of this embodiment, and in combination with any other particular aspect not inconsistent, t is 1, R9 is methyl and R10 is hydrogen.
In another particular aspect of this embodiment, and in combination with any other particular aspect not inconsistent, Y is N and R2 is hydrogen.
In another particular aspect of this embodiment, and in combination with any other particular aspect not inconsistent, Y is CR12, R2 is hydrogen, and R12 is hydrogen.
In another particular aspect of this embodiment, and in combination with any other particular aspect not inconsistent, t is 1, R10 is hydrogen, and R9 is combined with a ring atom of A to form a C-12 cycloalkyl ring fused to A.
In another embodiment, the invention provides a compound of formula 2, 3 or 4
wherein:
In a particular aspect of this embodiments, each R3 is independently halogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 cycloalkyl, C6-12 aryl, 3-12 membered heteroalicyclic, 5-12 membered heteroaryl, —S(O)mR4, —SO2NR4R5, —S(O)2OR4, —NO2, —NR4R5, —(CR6R7)nOR4, —CN, —C(O)R4, —OC(O)R4, —O(CR6R7)nR4, —O(CR6R7)(CR6R7)nNR4R5, —O(CR6R7)(CR6R7) OR4, —NR4C(O)R5, —(CR6R7)nC(O)OR4, —(CR6R7) OR4, —(CR6R7)nC(O)NR4R5, —(CR6R7) NCR4R5, —C(═NR6)NR4R5, —NR4C(O)NR5R6, —NR4S(O)pR5 or —C(O)NR4R5, each hydrogen in R3 is optionally substituted by R8, and R3 groups on adjacent atoms may combine to form a C6-12 aryl, 5-12 membered heteroaryl, C3-12 cycloalkyl or 3-12 membered heteroalicyclic group; and
In a particular aspect of this embodiment, A is C6-12 aryl or 5-12 membered heteroaryl, optionally substituted by one or more R3 groups.
In another particular aspect of this embodiment, and in combination with any other particular aspect not inconsistent, A is phenyl substituted by one, two or three R3 groups, preferably one, two or three halogens.
In another particular aspect of this embodiment, and in combination with any other particular aspect not inconsistent, t is 1, R9 is methyl and R10 is hydrogen.
In another particular aspect of this embodiment, and in combination with any other particular aspect not inconsistent, Y is N and R2 is hydrogen.
In another particular aspect of this embodiment, and in combination with any other particular aspect not inconsistent, Y is CR12, R2 is hydrogen, and R12 is hydrogen.
In another particular aspect of this embodiment, and in combination with any other particular aspect not inconsistent, t is 1, R10 is hydrogen, and R9 is combined with a ring atom of A to form a C3-12 cycloalkyl ring fused to A.
In another embodiment, the invention provides a compound of formula 2a
wherein:
In a particular aspect of this embodiments, each R3 is independently halogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 cycloalkyl, C6-12 aryl, 3-12 membered heteroalicyclic, 5-12 membered heteroaryl, —S(O)mR4, —SO2NR4R5, —S(O)2OR4, —NO2, —NR4R5, —(CR6R7)nOR4, —CN, —C(O)R4, —OC(O)R4, —O(CR6R7)nR4, —O(CR6R7)(CR6R7)nNR4R5, —O(CR6R7)(CR6R7)nOR4, —NR4C(O)R5, —(CR6R7)nC(O)OR4, —(CR6R7)nOR4, —(CR6R7)nC(O)NR4R5, —(CR6R7)nNCR4R5, —C(═NR6)NR4R5, —NR4C(O)NR5R6, —NR4S(O)pR5 or —C(O)NR4R5, each hydrogen in R3 is optionally substituted by R8, and R3 groups on adjacent atoms may combine to form a C6-12 aryl, 5-12 membered heteroaryl, C3-12 cycloalkyl or 3-12 membered heteroalicyclic group; and
In a particular aspect of this embodiment, R2 is hydrogen.
In another embodiment, the invention provides a compound of formula 2b
wherein:
In a particular aspect of this embodiments, each R3 is independently halogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 cycloalkyl, C6-2 aryl, 3-12 membered heteroalicyclic, 5-12 membered heteroaryl, —S(O)mR4, —SO2NR4R5, —S(O)2OR4, —NO2, —NR4R5, —(CR6R7)nOR4, —CN, —C(O)R4, —OC(O)R4, —O(CR6R7)R4, —O(CR6R7)(CR6R7)NR4R5, —O(CR6R7)(CR6R7)nOR4, —NR4C(O)R5, —(CR6R7)n C(O)OR4, —(CR6R7)nOR4, —(CR6R7)nC(O)NR4R5, —(CR6R7)nNCR4R5, —C(═NR6)NR4R5, —NR4C(O)NR5R6, —NR4S(O)pR5 or —C(O)NR4R5, each hydrogen in R3 is optionally substituted by R8, and R3 groups on adjacent atoms may combine to form a C6-12 aryl, 5-12 membered heteroaryl, C3-12 cycloalkyl or 3-12 membered heteroalicyclic group; and
In a particular aspect of this embodiment, R2 is hydrogen.
In another particular aspect of this embodiment, R12 is hydrogen.
In another particular aspect of this embodiment, R2 and R12 are hydrogen.
In another embodiment, the invention provides a compound of formula 3a
wherein:
In a particular aspect of this embodiments, each R3 is independently halogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 cycloalkyl, C3-12 aryl, 3-12 membered heteroalicyclic, 5-12 membered heteroaryl, —S(O)mR4, —SO2NR4R5, —S(O)2OR4, —NO2, —NR4R5, —(CR6R7)nOR4, —CN, —C(O)R4, —OC(O)R4, —O(CR6R7)nR4, —O(CR6R7)(CR6R7)nNR4R5, —O(CR6R7)(CR6R7)nOR4, —NR4C(O)R5, —(CR6R7)n C(O)OR4, —(CR6R7)nOR4, —(CR5R7)nC(O)NR4R5, —(CR6R7)nNCR4R5, —C(═NR6)NR4R5, —NR4C(O)NR5R6, —NR4S(O)pR5 or —C(O)NR4R5, each hydrogen in R3 is optionally substituted by R8, and R3 groups on adjacent atoms may combine to form a C6-12 aryl, 5-12 membered heteroaryl, C3-12 cycloalkyl or 3-12 membered heteroalicyclic group; and
In a particular aspect of this embodiment, R2 is hydrogen.
In another embodiment, the invention provides a compound of formula 3b
wherein:
In a particular aspect of this embodiments, each R3 is independently halogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 cycloalkyl, C6-12 aryl, 3-12 membered heteroalicyclic, 5-12 membered heteroaryl, —S(O)mR4, —SO2NR4R5, —S(O)2OR4, —NO2, —NR4R5, —(CR6R7)nOR4, —CN, —C(O)R4, —OC(O)R4, —O(CR6R7)nR4, —O(CR6R7)(CR6R7)nNR4R5, —O(CR6R7)(CR6R7)nOR4, —NR4C(O)R5, —(CR6R7)nC(O)OR4, —(CR6R7)nOR4, —(CR6R7)nC(O)NR4R5, —(CR6R7)nNCR4R5, —C(═NR6)NR4R5, —NR4C(O)NR5R6, —NR4S(O)pR5 or —C(O)NR4R5, each hydrogen in R3 is optionally substituted by R8, and R3 groups on adjacent atoms may combine to form a C6-12 aryl, 5-12 membered heteroaryl, C3-12 cycloalkyl or 3-12 membered heteroalicyclic group; and
In a particular aspect of this embodiment, R2 is hydrogen.
In another particular aspect of this embodiment, R12 is hydrogen.
In another particular aspect of this embodiment, R2 and R12 are hydrogen.
In another embodiment, the invention provides a compound of formula 4a
wherein:
In a particular aspect of this embodiments, each R3 is independently halogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 cycloalkyl, C6-12 aryl, 3-12 membered heteroalicyclic, 5-12 membered heteroaryl, —S(O)mR4, —SO2NR4R5, —S(O)2OR4, —NO2, —NR4R5, —(CR6R7)nOR4, —CN, —C(O)R4, —OC(O)R4, —O(CR6R7)nR4, —O(CR6R7)(CR6R7)nNR4R5, —O(CR6R7)(CR6R7)nOR4, —NR4C(O)R5, —(CR6R7)nC(O)OR4, —(CR6R7)nOR4, —(CR6R7)nC(O)NR4R5, —(CR6R7)nNCR4R5, —C(═NR6)NR4R5, —NR4C(O)NR5R6, —NR4S(O)pR5 or —C(O)NR4R5, each hydrogen in R3 is optionally substituted by R8, and R3 groups on adjacent atoms may combine to form a C6-12 aryl, 5-12 membered heteroaryl, C3-12 cycloalkyl or 3-12 membered heteroalicyclic group; and
In a particular aspect of this embodiment, R2 is hydrogen.
In another embodiment, the invention provides a compound of formula 4b
wherein:
In a particular aspect of this embodiments, each R3 is independently halogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 cycloalkyl, C6-12 aryl, 3-12 membered heteroalicyclic, 5-12 membered heteroaryl, —S(O)mR4, —SO2NR4R5, —S(O)2OR4, —NO2, —NR4R5, —(CR6R7)nOR4, —CN, —C(O)R4, —OC(O)R4, —O(CR6R7)nR4, —O(CR6R7)(CR6R7)nNR4R5, —O(CR6R7)(CR6R7)nOR4, —NR4C(O)R5, —(CR6R7)nC(O)OR4, —(CR6R7)nOR4, —(CR6R7)nC(O)NR4R5, —(CR6R7)nNCR4R5, —C(═NR6)NR4R5, —NR4C(O)NR5R6, —NR4S(O)pR5 or —C(O)NR4R5, each hydrogen in R3 is optionally substituted by R8, and R3 groups on adjacent atoms may combine to form a C6-12 aryl, 5-12 membered heteroaryl, C3-12 cycloalkyl or 3-12 membered heteroalicyclic group; and
In a particular aspect of this embodiment, R2 is hydrogen.
In another particular aspect of this embodiment, R12 is hydrogen.
In another particular aspect of this embodiment, R2 and R12 are hydrogen.
In another embodiment, the invention provides a compound of formula 5a
wherein:
In another embodiment, the invention provides a compound of formula 5b
wherein:
In another embodiment, the invention provides a compound selected from the group consisting of:
or a pharmaceutically acceptable salt, solvate or hydrate thereof.
In another embodiment, the invention provides a compound selected from the group consisting of:
or a pharmaceutically acceptable salt, solvate or hydrate thereof.
In another embodiment, the invention provides a compound selected from the group consisting of:
or a pharmaceutically acceptable salt, solvate or hydrate thereof.
In another embodiment, the invention provides a compound selected from the group consisting of:
or a pharmaceutically acceptable salt, solvate or hydrate thereof.
In another embodiment, the invention provides a compound selected from the group consisting of:
or a pharmaceutically acceptable salt, solvate or hydrate thereof.
In another embodiment, the invention provides a compound selected from the group consisting of:
or a pharmaceutically acceptable salt, hydrate or solvate thereof.
In another embodiment, the invention provides a compound selected from the group consisting of:
or a pharmaceutically acceptable salt, hydrate or solvate thereof.
In another embodiment, the invention provides a compound selected from the group consisting of:
In another embodiment, the invention provides a compound selected from the group consisting of:
or a pharmaceutically acceptable salt, hydrate or solvate thereof.
In another embodiment, the invention provides a compound selected from the group consisting of:
or a pharmaceutically acceptable salt, hydrate or solvate thereof.
In another embodiment, the invention provides a compound selected from the group consisting of:
or a pharmaceutically acceptable salt, hydrate or solvate thereof.
In another embodiment, the invention provides a pharmaceutical composition comprising any of the compounds of the invention and a pharmaceutically acceptable carrier. Examples of such compositions are described below.
Preferred compounds of the invention include those having c-MET inhibitory activity as defined by any one or more of IC50, Ki, or percent inhibition (% I). One skilled in the art can readily determine if a compound has such activity by carrying out the appropriate assay, and descriptions of such assays are shown in the Examples section herein. In one embodiment, particularly preferred compounds have a c-MET Ki of less than 5 μM or less than 2 μM, or less than 1 μM, or less than 500 nM or less than 200 nM or less than 100 nM. In another embodiment, particularly preferred compounds have a c-MET inhibition at 1 μM of at least 10% or at least 20% or at least 30% or at least 40% or at least 50% or at least 60% or at least 70% or at least 80% or at least 90%. Methods for measuring c-MET/HGFR activity are described in the Examples herein.
In another embodiment, the invention provides a method of treating abnormal cell growth in a mammal, including a human, the method comprising administering to the mammal any of the pharmaceutical compositions of the invention.
In a specific embodiment of any of the inventive methods described herein, the abnormal cell growth is cancer, including, but not limited to, lung cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, prostate cancer, chronic or acute leukemia, lymphocytic lymphomas, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of the central nervous system (CNS), primary CNS lymphoma, spinal axis tumors, brain stem glioma, pituitary adenoma, or a combination of one or more of the foregoing cancers. In another embodiment of said method, said abnormal cell growth is a benign proliferative disease, including, but not limited to, psoriasis, benign prostatic hypertrophy or restinosis.
In another embodiment, the invention provides a method of treating an HGFR mediated disorder in a mammal, including a human, the method comprising administering to the mammal any of the pharmaceutical compositions of the invention.
In further specific embodiments of any of the inventive methods described herein, the method further comprises administering to the mammal an amount of one or more substances selected from anti-tumor agents, anti-angiogenesis agents, signal transduction inhibitors, and antiproliferative agents, which amounts are together effective in treating said abnormal cell growth. Such substances include those disclosed in PCT Publication Nos. WO 00/38715, WO 00/38716, WO 00/38717, WO 00/38718, WO 00/38719, WO 00/38730, WO 00/38665, WO 00/37107 and WO 00/38786, the disclosures of which are incorporated herein by reference in their entireties.
Examples of anti-tumor agents include mitotic inhibitors, for example vinca alkaloid derivatives such as vinblastine vinorelbine, vindescine and vincristine; colchines allochochine, halichondrine, N-benzoyltrimethyl-methyl ether colchicinic acid, dolastatin 10, maystansine, rhizoxine, taxanes such as taxol (paclitaxel), docetaxel (Taxotere), 2′-N-[3-(dimethylamino)propyl]glutaramate (taxol derivative), thiocholchicine, trityl cysteine, teniposide, methotrexate, azathioprine, fluorouricil, cytocine arabinoside, 2′2′-difluorodeoxycytidine (gemcitabine), adriamycin and mitamycin. Alkylating agents, for example cis-platin, carboplatin oxiplatin, iproplatin, Ethyl ester of N-acetyl-DL-sarcosyl-L-leucine (Asaley or Asalex), 1,4-cyclohexadiene-1,4-dicarbamic acid, 2,5-bis(1-azirdinyl)-3,6-dioxo-, diethyl ester (diaziquone), 1,4-bis(methanesulfonyloxy)butane (bisulfan or leucosulfan) chlorozotocin, clomesone, cyanomorpholinodoxorubicin, cyclodisone, dianhydroglactitol, fluorodopan, hepsulfam, mitomycin C, hycantheonemitomycin C, mitozolamide, 1-(2-chloroethyl)-4-(3-chloropropyl)-piperazine dihydrochloride, piperazinedione, pipobroman, porfiromycin, spirohydantoin mustard, teroxirone, tetraplatin, thiotepa, triethylenemelamine, uracil nitrogen mustard, bis(3-mesyloxypropyl)amine hydrochloride, mitomycin, nitrosoureas agents such as cyclohexyl-chloroethylnitrosourea, methylcyclohexyl-chloroethylnitrosourea 1-(2-chloroethyl)-3-(2,6-dioxo-3-piperidyl)-1-nitroso-urea, bis(2-chloroethyl)nitrosourea, procarbazine, dacarbazine, nitrogen mustard-related compounds such as mechloroethamine, cyclophosphamide, ifosamide, melphalan, chlorambucil, estramustine sodium phosphate, strptozoin, and temozolamide. DNA anti-metabolites, for example 5-fluorouracil, cytosine arabinoside, hydroxyurea, 2-[(3hydroxy-2-pyrinodinyl)methylene]-hydrazinecarbothioamide, deoxyfluorouridine, 5-hydroxy-2-formylpyridine thiosemicarbazone, alpha-2′-deoxy-6-thioguanosine, aphidicolin glycinate, 5-azadeoxycytidine, beta-thioguanine deoxyriboside, cyclocytidine, guanazole, inosine glycodialdehyde, macbecin II, pyrazolimidazole, cladribine, pentostatin, thioguanine, mercaptopurine, bleomycin, 2-chlorodeoxyadenosine, inhibitors of thymidylate synthase such as raltitrexed and pemetrexed disodium, clofarabine, floxuridine and fludarabine. DNA/RNA antimetabolites, for example, L-alanosine, 5-azacytidine, acivicin, minopterin and derivatives thereof such as N-[2-chloro-5-[[(2,4-diamino-5-methyl-6-quinazolinyl)methyl]amino]benzoyl]-L-aspartic acid, N-[4-[[(2,4-diamino-5-ethyl-6-quinazolinyl)methyl]amino]benzoyl]-L-aspartic acid, N-[2-chloro-4-[[(2,4-diaminopteridinyl)methyl]amino]benzoyl]-L-aspartic acid, soluble Bakers antifol, dichloroallyl lawsone, brequinar, ftoraf, dihydro-5-azacytidine, methotrexate, N-(phosphonoacetyl)-L-aspartic acid tetrasodium salt, pyrazofuran, trimetrexate, plicamycin, actinomycin D, cryptophycin, and analogs such as cryptophycin-52 or, for example, one of the preferred anti-metabolites disclosed in European Patent Application No. 239362 such as N-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L-glutamic acid; growth factor inhibitors; cell cycle inhibitors; intercalating antibiotics, for example adriamycin and bleomycin; proteins, for example interferon; and anti-hormones, for example anti-estrogens such as Nolvadex□ (tamoxifen) or, for example anti-androgens such as Casodex™ (4′-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3′-(trifluoromethyl)propionanilide). Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate dosing of the individual components of the treatment.
Anti-angiogenesis agents include MMP-2 (matrix-metalloprotienase 2) inhibitors, MMP-9 (matrix-metalloprotienase 9) inhibitors, and COX-II (cyclooxygenase II) inhibitors. Examples of useful COX-II inhibitors include CELEBREX™ (alecoxib), valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in WO 96/33172 (published Oct. 24, 1996), WO 96/27583 (published Mar. 7, 1996), European Patent Application No. 97304971.1 (filed Jul. 8, 1997), European Patent Application No. 99308617.2 (filed Oct. 29, 1999), WO 98/07697 (published Feb. 26, 1998), WO 98/03516 (published Jan. 29, 1998), WO 98/34918 (published Aug. 13, 1998), WO 98/34915 (published Aug. 13, 1998), WO 98/33768 (published Aug. 6, 1998), WO 98/30566 (published Jul. 16, 1998), European Patent Publication 606,046 (published Jul. 13, 1994), European Patent Publication 931,788 (published Jul. 28, 1999), WO 90/05719 (published May 331, 1990), WO 99/52910 (published Oct. 21, 1999), WO 99/52889 (published Oct. 21, 1999), WO 99/29667 (published Jun. 17, 1999), PCT International Application No. PCT/IB98/01113 (filed Jul. 21, 1998), European Patent Application No. 99302232.1 (filed Mar. 25, 1999), Great Britain patent application number 9912961.1 (filed Jun. 3, 1999), U.S. Provisional Application No. 60/148,464 (filed Aug. 12, 1999), U.S. Pat. No. 5,863,949 (issued Jan. 26, 1999), U.S. Pat. No. 5,861,510 (issued Jan. 19, 1999), and European Patent Publication 780,386 (published Jun. 25, 1997), all of which are herein incorporated by reference in their entirety. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity inhibiting MMP-1. More preferred, are those that selectively inhibit MMP-2 and/or MMP-9 relative to the other matrix-metalloproteinases (i.e. MMP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).
Examples of MMP inhibitors include AG-3340, RO 32-3555, RS 13-0830, and the following compounds: 3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclopentyl)-amino]-propionic acid; 3-exo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylic acid hydroxyamide; (2R, 3R) 1-[4-(2-chloro-4-fluoro-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylic acid hydroxyamide; 4-[4-(4-fluoro-henoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylic acid hydroxyamide; 3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclobutyl)-amino]-propionic acid; 4-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylic acid hydroxyamide; 3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-3-carboxylic acid hydroxyamide; (2R, 3R) 1-[4-(4-fluoro-2-methyl-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylic acid hydroxyamide; 3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-1-methyl-ethyl)-amino]-propionic acid; 3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(4-hydroxycarbamoyl-tetrahydro-pyran-4-yl)-amino]-propionic acid; 3-exo-3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylic acid hydroxyamide; 3-endo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylic acid hydroxyamide; 3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-furan-3-carboxylic acid hydroxyamide; and pharmaceutically acceptable salts, solvates and hydrates thereof.
Examples of signal transduction inhibitors include agents that can inhibit EGFR (epidermal growth factor receptor) responses, such as EGFR antibodies, EGF antibodies, and molecules that are EGFR inhibitors; VEGF (vascular endothelial growth factor) inhibitors; and erbB2 receptor inhibitors, such as organic molecules or antibodies that bind to the erbB2 receptor, for example, HERCEPTIN™ (Genentech, Inc. of South San Francisco, Calif., USA). EGFR inhibitors are described in, for example in WO 95/19970 (published Jul. 27, 1995), WO 98/14451 (published Apr. 9, 1998), WO 98/02434 (published Jan. 22, 1998), and U.S. Pat. No. 5,747,498 (issued May 5, 1998). EGFR-inhibiting agents include, but are not limited to, the monoclonal antibodies C225 and anti-EGFR 22Mab (ImClone Systems Incorporated of New York, N.Y., USA), the compounds ZD-1839 (AstraZeneca), BIBX-1382 (Boehringer Ingelheim), MDX-447 (Medarex Inc. of Annandale, N.J., USA), and OLX-103 (Merck & Co. of Whitehouse Station, N.J., USA), VRCTC-310 (Ventech Research) and EGF fusion toxin (Seragen Inc. of Hopkinton, Mass.).
VEGF inhibitors, for example SU-5416 and SU-6668 (Sugen Inc. of South San Francisco, Calif., USA), can also be combined or co-administered with the composition. VEGF inhibitors are described in, for example in WO 99/24440 (published May 20, 1999), PCT International Application PCT/IB99/00797 (filed May 3, 1999), in WO 95/21613 (published Aug. 17, 1995), WO 99/61422 (published Dec. 2, 1999), U.S. Pat. No. 5,834,504 (issued Nov. 10, 1998), WO 98/50356 (published Nov. 12, 1998), U.S. Pat. No. 5,883,113 (issued Mar. 16, 1999), U.S. Pat. No. 5,886,020 (issued Mar. 23, 1999), U.S. Pat. No. 5,792,783 (issued Aug. 11, 1998), WO 99/10349 (published Mar. 4, 1999), WO 97/32856 (published Sep. 12, 1997), WO 97/22596 (published Jun. 26, 1997), WO 98/54093 (published Dec. 3, 1998), WO 98/02438 (published Jan. 22, 1998), WO 99/16755 (published Apr. 8, 1999), and WO 98/02437 (published Jan. 22, 1998), all of which are herein incorporated by reference in their entirety. Other examples of some specific VEGF inhibitors are IM862 (Cytran Inc. of Kirkland, Wash., USA); anti-VEGF monoclonal antibody bevacizumab (Genentech, Inc. of South San Francisco, Calif.); and angiozyme, a synthetic ribozyme from Ribozyme (Boulder, Colo.) and Chiron (Emeryville, Calif.).
ErbB2 receptor inhibitors, such as GW-282974 (Glaxo Wellcome plc), and the monoclonal antibodies AR-209 (Aronex Pharmaceuticals Inc. of The Woodlands, Tex., USA) and 2B-1 (Chiron), may be administered in combination with the composition. Such erbB2 inhibitors include those described in WO 98/02434 (published Jan. 22, 1998), WO 99/35146 (published Jul. 15, 1999), WO 99/35132 (published Jul. 15, 1999), WO 98/02437 (published Jan. 22, 1998), WO 97/13760 (published Apr. 17, 1997), WO 95/19970 (published Jul. 27, 1995), U.S. Pat. No. 5,587,458 (issued Dec. 24, 1996), and U.S. Pat. No. 5,877,305 (issued Mar. 2, 1999), each of which is herein incorporated by reference in its entirety. ErbB2 receptor inhibitors useful in the present invention are also described in U.S. Provisional Application No. 60/117,341, filed Jan. 27, 1999, and in U.S. Provisional Application No. 60/117,346, filed Jan. 27, 1999, both of which are herein incorporated by reference in their entirety.
Other antiproliferative agents that may be used include inhibitors of the enzyme farnesyl protein transferase and inhibitors of the receptor tyrosine kinase PDGFr, including the compounds disclosed and claimed in the following United States patent applications: Ser. No. 09/221,946 (filed Dec. 28, 1998); Ser. No. 09/454,058 (filed Dec. 2, 1999); Ser. No. 09/501,163 (filed Feb. 9, 2000); Ser. No. 09/539,930 (filed Mar. 31, 2000); Ser. No. 09/202,796 (filed May 22, 1997); Ser. No. 09/384,339 (filed Aug. 26, 1999); and Ser. No. 09/383,755 (filed Aug. 26, 1999); and the compounds disclosed and claimed in the following United States provisional patent applications: 60/168,207 (filed Nov. 30, 1999); 60/170,119 (filed Dec. 10, 1999); 60/177,718 (filed Jan. 21, 2000); 60/168,217 (filed Nov. 30, 1999), and 60/200,834 (filed May 1, 2000). Each of the foregoing patent applications and provisional patent applications is herein incorporated by reference in their entirety.
Compositions of the invention can also be used with other agents useful in treating abnormal cell growth or cancer, including, but not limited to, agents capable of enhancing antitumor immune responses, such as CTLA4 (cytotoxic lymphocite antigen 4) antibodies, and other agents capable of blocking CTLA4; and anti-proliferative agents such as other farnesyl protein transferase inhibitors. Specific CTLA4 antibodies that can be used in the present invention include those described in U.S. Provisional Application 60/113,647 (filed Dec. 23, 1998) which is herein incorporated by reference in its entirety.
Definitions
Unless otherwise stated, the following terms used in the specification and claims have the meanings discussed below. Variables defined in this section, such as R, X, n and the like, are for reference within this section only, and are not meant to have the save meaning as may be used outside of this definitions section. Further, many of the groups defined herein can be optionally substituted. The listing in this definitions section of typical substituents is exemplary and is not intended to limit the substituents defined elsewhere within this specification and claims.
“Alkyl” refers to a saturated aliphatic hydrocarbon radical including straight chain and branched chain groups of 1 to 20 carbon atoms, preferably 1 to 12 carbon atoms, more preferably 1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 1 to 4 carbon atoms. ‘Lower alkyl’ refers specifically to an alkyl group with 1 to 4 carbon atoms. Examples of alkyl groups include methyl, ethyl, propyl, 2-propyl, n-butyl, iso-butyl, tert-butyl, pentyl, and the like. Alkyl may be substituted or unsubstituted. Typical substituent groups include cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halo, carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy, nitro, silyl, amino and —NRxRy, where Rx and Ry are independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, carbonyl, acetyl, sulfonyl, trifluoromethanesulfonyl and, combined, a five- or six-member heteroalicyclic ring.
“Cycloalkyl” refers to a 3 to 8 member all-carbon monocyclic ring, an all-carbon 5-member/6-member or 6-member/6-member fused bicyclic ring, or a multicyclic fused ring (a “fused” ring system means that each ring in the system shares an adjacent pair of carbon atoms with each other ring in the system) group wherein one or more of the rings may contain one or more double bonds but none of the rings has a completely conjugated pi-electron system. Examples, without limitation, of cycloalkyl groups are cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexadiene, adamantane, cycloheptane, cycloheptatriene, and the like. A cycloalkyl group may be substituted or unsubstituted. Typical substituent groups include alkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halo, carbonyl, thiocarbonyl, C-carboxy, O-carboxy, O-carbamyl, N-carbamyl, C-amido, N-amido, nitro, amino and —NRxRy, with Rx and Ry as defined above. Illustrative examples of cycloalkyl are derived from, but not limited to, the following:
“Alkenyl” refers to an alkyl group, as defined herein, consisting of at least two carbon atoms and at least one carbon-carbon double bond. Representative examples include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl, and the like.
“Alkynyl” refers to an alkyl group, as defined herein, consisting of at least two carbon atoms and at least one carbon-carbon triple bond. Representative examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl, and the like.
“Aryl” refers to an all-carbon monocyclic or fused-ring polycyclic groups of 6 to 12 carbon atoms having a completely conjugated pi-electron system. Examples, without limitation, of aryl groups are phenyl, naphthalenyl and anthracenyl. The aryl group may be substituted or unsubstituted. Typical substituents include halo, trihalomethyl, alkyl, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, nitro, carbonyl, thiocarbonyl, C-carboxy, O-carboxy, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, sulfinyl, sulfonyl, amino and —NRxRy, with Rx and Ry as defined above.
“Heteroaryl” refers to a monocyclic or fused ring group of 5 to 12 ring atoms containing one, two, three or four ring heteroatoms selected from N, O, and S, the remaining ring atoms being C, and, in addition, having a completely conjugated pi-electron system. Examples, without limitation, of unsubstituted heteroaryl groups are pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline, purine, tetrazole, triazine, and carbazole. The heteroaryl group may be substituted or unsubstituted. Typical substituents include alkyl, cycloalkyl, halo, trihalomethyl, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, nitro, carbonyl, thiocarbonyl, sulfonamido, C-carboxy, O-carboxy, sulfinyl, sulfonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, amino and —NRxRy with Rx and Ry as defined above.
A pharmaceutically acceptable heteroaryl is one that is sufficiently stable to be attached to a compound of the invention, formulated into a pharmaceutical composition and subsequently administered to a patient in need thereof.
Examples of typical monocyclic heteroaryl groups include, but are not limited to:
Examples of suitable fused ring heteroaryl groups include, but are not limited to:
“Heteroalicyclic” or “heterocycle” refers to a monocyclic or fused ring group having in the ring(s) of 3 to 12 ring atoms, in which one or two ring atoms are heteroatoms selected from N, O, and S(O)n (where n is 0, 1 or 2), the remaining ring atoms being C. The rings may also have one or more double bonds. However, the rings do not have a completely conjugated pi-electron system. Additionally, one or more of the ring atoms could be substituted by an oxo group. Examples of suitable saturated heteroalicyclic groups include, but are not limited to:
Examples of suitable partially unsaturated heteroalicyclic groups include, but are not limited to:
The heterocycle group is optionally substituted with one or two substituents independently selected from halo, lower alkyl, lower alkyl substituted with carboxy, ester hydroxy, or mono or dialkylamino.
“Hydroxy” refers to an —OH group.
“Alkoxy” refers to both an —O-(alkyl) or an —O-(unsubstituted cycloalkyl) group. Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.
“Haloalkoxy” refers to an —O-(haloalkyl) group. Representative examples include, but are not limited to, trifluoromethoxy, tribromomethoxy, and the like.
“Aryloxy” refers to an —O-aryl or an —O-heteroaryl group, as defined herein. Representative examples include, but are not limited to, phenoxy, pyridinyloxy, furanyloxy, thienyloxy, pyrimidinyloxy, pyrazinyloxy, and the like, and derivatives thereof.
“Mercapto” refers to an —SH group.
“Alkylthio” refers to an —S-(alkyl) or an —S-(unsubstituted cycloalkyl) group. Representative examples include, but are not limited to, methylthio, ethylthio, propylthio, butylthio, cyclopropylthio, cyclobutylthio, cyclopentylthio, cyclohexylthio, and the like.
“Arylthio” refers to an —S-aryl or an —S-heteroaryl group, as defined herein. Representative examples include, but are not limited to, phenylthio, pyridinylthio, furanylthio, thienylthio, pyrimidinylthio, and the like and derivatives thereof.
“Acyl” or “carbonyl” refers to a —C(O)R″ group, where R″ is selected from the group consisting of hydrogen, lower alkyl, trihalomethyl, unsubstituted cycloalkyl, aryl optionally substituted with one or more, preferably one, two, or three substituents selected from the group consisting of lower alkyl, trihalomethyl, lower alkoxy, halo and —NRxRy groups, heteroaryl (bonded through a ring carbon) optionally substituted with one or more, preferably one, two, or three substitutents selected from the group consisting of lower alkyl, trihaloalkyl, lower alkoxy, halo and —NRxRy groups and heteroalicyclic (bonded through a ring carbon) optionally substituted with one or more, preferably one, two, or three substituents selected from the group consisting of lower alkyl, trihaloalkyl, lower alkoxy, halo and —NRxRy groups. Representative acyl groups include, but are not limited to, acetyl, trifluoroacetyl, benzoyl, and the like
“Aldehyde” refers to an acyl group in which R″ is hydrogen.
“Thioacyl” or “thiocarbonyl” refers to a —C(S)R″ group, with R″ as defined above.
A “thiocarbonyl” group refers to a —C(S)R″ group, with R″ as defined above.
A “C-carboxy” group refers to a —C(O)OR″ group, with R″ as defined above.
An “O-carboxy” group refers to a —OC(O)R″ group, with R″ as defined above.
“Ester” refers to a —C(O)OR″ group with R″ as defined herein except that R″ cannot be hydrogen.
“Acetyl” group refers to a —C(O)CH3 group.
“Halo” group refers to fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.
“Trihalomethyl” group refers to a methyl group having three halo substituents, such as a trifluoromethyl group.
“Cyano” refers to a —C═N group.
A “sulfinyl” group refers to a —S(O)R″ group wherein, in addition to being as defined above, R″ may also be a hydroxy group.
A “sulfonyl” group refers to a —S(O)2R″ group wherein, in addition to being as defined above, R″ may also be a hydroxy group.
“S-sulfonamido” refers to a —S(O)2NRxRy group, with Rx and Ry as defined above.
“N-sulfonamido” refers to a —NRxS(O)2Ry group, with Rx and Ry as defined above.
“O-carbamyl” group refers to a —OC(O)NRxRy group with Rx and Ry as defined above.
“N-carbamyl” refers to an RyOC(O)NRx-group, with Rx and Ry as defined above.
“O-thiocarbamyl” refers to a —OC(S)NRxRy group with Rx and Ry as defined above.
“N-thiocarbamyl” refers to a RyOC(S)NRx group, with Ry and Rx as defined above.
“Amino” refers to an —NRxRy group, wherein Rx and Ry are both hydrogen.
“C-amido” refers to a —C(O)NRxRy group with Rx and Ry as defined above.
“N-amido” refers to a RxC(O)NRy group, with Rx and Ry as defined above.
“Nitro” refers to a —NO2 group.
“Haloalkyl” means an alkyl, preferably lower alkyl, that is substituted with one or more same or different halo atoms, e.g., —CH2Cl, —CF3, —CH2CF3, —CH2CCl3, and the like.
“Hydroxyalkyl” means an alkyl, preferably lower alkyl, that is substituted with one, two, or three hydroxy groups; e.g., hydroxymethyl, 1 or 2-hydroxyethyl, 1,2-, 1,3-, or 2,3-dihydroxypropyl, and the like.
“Aralkyl” means alkyl, preferably lower alkyl, that is substituted with an aryl group as defined above; e.g., —CH2phenyl, —(CH2)2phenyl, —(CH2)3phenyl, CH3CH(CH3)CH2phenyl, and the like and derivatives thereof.
“Heteroaralkyl” group means alkyl, preferably lower alkyl, that is substituted with a heteroaryl group; e.g., —CH2pyridinyl, —(CH2)2pyrimidinyl, —(CH2)3imidazolyl, and the like, and derivatives thereof.
“Monoalkylamino” means a radical —NHR where R is an alkyl or unsubstituted cycloalkyl group; e.g., methylamino, (1-methylethyl)amino, cyclohexylamino, and the like.
“Dialkylamino” means a radical —NRR where each R is independently an alkyl or unsubstituted cycloalkyl group; dimethylamino, diethylamino, (1-methylethyl)-ethylamino, cyclohexylmethylamino, cyclopentylmethylamino, and the like.
“Optional” or “optionally” means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not. For example, “heterocycle group optionally substituted with an alkyl group” means that the alkyl may but need not be present, and the description includes situations where the heterocycle group is substituted with an alkyl group and situations where the heterocycle group is not substituted with the alkyl group.
A “pharmaceutical composition” refers to a mixture of one or more of the compounds described herein, or physiologically/pharmaceutically acceptable salts, solvates, hydrates or prodrugs thereof, with other chemical components, such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.
As used herein, a “physiologically/pharmaceutically acceptable carrier” refers to a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.
A “pharmaceutically acceptable excipient” refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a compound. Examples, without limitation, of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.
As used herein, the term “pharmaceutically acceptable salt” refers to those salts which retain the biological effectiveness and properties of the parent compound. Such salts include:
“PK” refers to receptor protein tyrosine kinase (RTKs), non-receptor or “cellular” tyrosine kinase (CTKS) and serine-threonine kinases (STKs).
“Modulation” or “modulating” refers to the alteration of the catalytic activity of RTKs, CTKs and STKs. In particular, modulating refers to the activation of the catalytic activity of RTKs, CTKs and STKs, preferably the activation or inhibition of the catalytic activity of RTKs, CTKs and STKs, depending on the concentration of the compound or salt to which the RTK, CTK or STK is exposed or, more preferably, the inhibition of the catalytic activity of RTKs, CTKs and STKs.
“Catalytic activity” refers to the rate of phosphorylation of tyrosine under the influence, direct or indirect, of RTKs and/or CTKs or the phosphorylation of serine and threonine under the influence, direct or indirect, of STKs.
“Contacting” refers to bringing a compound of this invention and a target PK together in such a manner that the compound can affect the catalytic activity of the PK, either directly, i.e., by interacting with the kinase itself, or indirectly, i.e., by interacting with another molecule on which the catalytic activity of the kinase is dependent. Such “contacting” can be accomplished “in vitro,” i.e., in a test tube, a petri dish or the like. In a test tube, contacting may involve only a compound and a PK of interest or it may involve whole cells. Cells may also be maintained or grown in cell culture dishes and contacted with a compound in that environment. In this context, the ability of a particular compound to affect a PK related disorder, i.e., the IC50 of the compound, defined below, can be determined before use of the compounds in vivo with more complex living organisms is attempted. For cells outside the organism, multiple methods exist, and are well-known to those skilled in the art, to get the PKs in contact with the compounds including, but not limited to, direct cell microinjection and numerous transmembrane carrier techniques.
“In vitro” refers to procedures performed in an artificial environment such as, e.g., without limitation, in a test tube or culture medium.
“In vivo” refers to procedures performed within a living organism such as, without limitation, a mouse, rat or rabbit.
“PK related disorder,” “PK driven disorder,” and “abnormal PK activity” all refer to a condition characterized by inappropriate, i.e., under or, more commonly, over, PK catalytic activity, where the particular PK can be an RTK, a CTK or an STK. Inappropriate catalytic activity can arise as the result of either: (1) PK expression in cells which normally do not express PKs, (2) increased PK expression leading to unwanted cell proliferation, differentiation and/or growth, or, (3) decreased PK expression leading to unwanted reductions in cell proliferation, differentiation and/or growth. Over-activity of a PK refers to either amplification of the gene encoding a particular PK or production of a level of PK activity which can correlate with a cell proliferation, differentiation and/or growth disorder (that is, as the level of the PK increases, the severity of one or more of the symptoms of the cellular disorder increases). Under-activity is, of course, the converse, wherein the severity of one or more symptoms of a cellular disorder increase as the level of the PK activity decreases.
“Treat”, “treating” and “treatment” refer to a method of alleviating or abrogating a PK mediated cellular disorder and/or its attendant symptoms. With regard particularly to cancer, these terms simply mean that the life expectancy of an individual affected with a cancer will be increased or that one or more of the symptoms of the disease will be reduced.
“Organism” refers to any living entity comprised of at least one cell. A living organism can be as simple as, for example, a single eukariotic cell or as complex as a mammal, including a human being.
“Therapeutically effective amount” refers to that amount of the compound being administered which will relieve to some extent one or more of the symptoms of the disorder being treated. In reference to the treatment of cancer, a therapeutically effective amount refers to that amount which has at least one of the following effects:
“Monitoring” means observing or detecting the effect of contacting a compound with a cell expressing a particular PK. The observed or detected effect can be a change in cell phenotype, in the catalytic activity of a PK or a change in the interaction of a PK with a natural binding partner. Techniques for observing or detecting such effects are well-known in the art. The effect is selected from a change or an absence of change in a cell phenotype, a change or absence of change in the catalytic activity of said protein kinase or a change or absence of change in the interaction of said protein kinase with a natural binding partner in a final aspect of this invention.
“Cell phenotype” refers to the outward appearance of a cell or tissue or the biological function of the cell or tissue. Examples, without limitation, of a cell phenotype are cell size, cell growth, cell proliferation, cell differentiation, cell survival, apoptosis, and nutrient uptake and use. Such phenotypic characteristics are measurable by techniques well-known in the art.
“Natural binding partner” refers to a polypeptide that binds to a particular PK in a cell. Natural binding partners can play a role in propagating a signal in a PK-mediated signal transduction process. A change in the interaction of the natural binding partner with the PK can manifest itself as an increased or decreased concentration of the PK/natural binding partner complex and, as a result, in an observable change in the ability of the PK to mediate signal transduction.
As used herein, the terms “optically pure,” “enantiomerically pure,” “pure enantiomer,” and “optically pure enantiomer” mean a composition that comprises one enantiomer of a compound and is substantially free of the opposite enantiomer of the compound. A typical optically pure compound comprises greater than about 80% by weight of one enantiomer of the compound and less than about 20% by weight of the opposite enantiomer of the compound, more preferably greater than about 90% by weight of one enantiomer of the compound and less than about 10% by weight of the opposite enantiomer of the compound, even more preferably greater than about 95% by weight of one enantiomer of the compound and less than about 5% by weight of the opposite enantiomer of the compound, and most preferably greater than about 97% by weight of one enantiomer of the compound and less than about 3% by weight of the opposite enantiomer of the compound.
General schemes for synthesizing the compounds of the invention can be found in the Examples section herein.
Unless indicated otherwise, all references herein to the inventive compounds include references to salts, solvates, hydrates and complexes thereof, and to solvates, hydrates and complexes of salts thereof, including polymorphs, stereoisomers, and isotopically labeled versions thereof.
Pharmaceutically acceptable salts include acid addition and base salts (including disalts). Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include the acetate, aspartate, benzoate, besylate, bicarbonate/carbonate, bisulphate/sulfate, borate, camsylate, citrate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, saccharate, stearate, succinate, tartrate, tosylate and trifluoroacetate salts.
Suitable base salts are formed from bases which form non-toxic salts. Examples include the aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine and zinc salts.
For a review on suitable salts, see “Handbook of Pharmaceutical Salts: Properties, Selection, and Use” by Stahl and Wermuth (Wiley-V C H, Weinheim, Germany, 2002), the disclosure of which is incorporated herein by reference in its entirety.
A pharmaceutically acceptable salt of the inventive compounds can be readily prepared by mixing together solutions of the compound and the desired acid or base, as appropriate. The salt may precipitate from solution and be collected by filtration or may be recovered by evaporation of the solvent. The degree of ionization in the salt may vary from completely ionized to almost non-ionized.
The compounds of the invention may exist in both unsolvated and solvated forms. The term ‘solvate’ is used herein to describe a molecular complex comprising the compound of the invention and one or more pharmaceutically acceptable solvent molecules, for example, ethanol. The term ‘hydrate’ is employed when the solvent is water. Pharmaceutically acceptable solvates in accordance with the invention include hydrates and solvates wherein the solvent of crystallization may be isotopically substituted, e.g. D2O, d6-acetone, d6-DMSO.
Also included within the scope of the invention are complexes such as clathrates, drug-host inclusion complexes wherein, in contrast to the aforementioned solvates, the drug and host are present in stoichiometric or non-stoichiometric amounts. Also included are complexes of the drug containing two or more organic and/or inorganic components which may be in stoichiometric or non-stoichiometrc amounts. The resulting complexes may be ionized, partially ionized, or non-ionized. For a review of such complexes, see J Pharm Sci, 64 (8), 1269-1288 by Haleblian (August 1975), the disclosure of which is incorporated herein by reference in its entirety.
Also within the scope of the invention are polymorphs, prodrugs, and isomers (including optical, geometric and tautomeric isomers) of the inventive compounds
Derivatives of compounds of the invention which may have little or no pharmacological activity themselves but can, when administered to a patient, be converted into the inventive compounds, for example, by hydrolytic cleavage. Such derivatives are referred to as ‘prodrugs’. Further information on the use of prodrugs may be found in ‘Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (T Higuchi and W Stella) and ‘Bioreversible Carriers in Drug Design’, Pergamon Press, 1987 (ed. E B Roche, American Pharmaceutical Association), the disclosures of which are incorporated herein by reference in their entireties.
Prodrugs in accordance with the invention can, for example, be produced by replacing appropriate functionalities present in the inventive compounds with certain moieties known to those skilled in the art as ‘pro-moieties’ as described, for example, in “Design of Prodrugs” by H Bundgaard (Elsevier, 1985), the disclosure of which is incorporated herein by reference in its entirety.
Some examples of prodrugs in accordance with the invention include:
Further examples of replacement groups in accordance with the foregoing examples and examples of other prodrug types may be found in the aforementioned references.
Finally, certain inventive compounds may themselves act as prodrugs of other of the inventive compounds.
Compounds of the invention containing one or more asymmetric carbon atoms can exist as two or more stereoisomers. Where a compound of the invention contains an alkenyl or alkenylene group, geometric cis/trans (or Z/E) isomers are possible. Where the compound contains, for example, a keto or oxime group or an aromatic moiety, tautomeric isomerism (‘tautomerism’) can occur. A single compound may exhibit more than one type of isomerism.
Included within the scope of the invention are all stereoisomers, geometric isomers and tautomeric forms of the inventive compounds, including compounds exhibiting more than one type of isomerism, and mixtures of one or more thereof. Also included are acid addition or base salts wherein the counterion is optically active, for example, D-lactate or L-lysine, or racemic, for example, DL-tartrate or DL-arginine.
Cis/trans isomers may be separated by conventional techniques well known to those skilled in the art, for example, chromatography and fractional crystallization.
Conventional techniques for the preparation/isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC).
Alternatively, the racemate (or a racemic precursor) may be reacted with a suitable optically active compound, for example, an alcohol, or, in the case where the compound contains an acidic or basic moiety, an acid or base such as tartaric acid or 1-phenylethylamine. The resulting diastereomeric mixture may be separated by chromatography and/or fractional crystallization and one or both of the diastereoisomers converted to the corresponding pure enantiomer(s) by means well known to one skilled in the art.
Chiral compounds of the invention (and chiral precursors thereof) may be obtained in enantiomerically-enriched form using chromatography, typically HPLC, on an asymmetric resin with a mobile phase consisting of a hydrocarbon, typically heptane or hexane, containing from 0 to 50% isopropanol, typically from 2 to 20%, and from 0 to 5% of an alkylamine, typically 0.1% diethylamine. Concentration of the eluate affords the enriched mixture.
Stereoisomeric conglomerates may be separated by conventional techniques known to those skilled in the art; see, for example, “Stereochemistry of Organic Compounds” by E L Eliel (Wiley, New York, 1994), the disclosure of which is incorporated herein by reference in its entirety.
The invention also includes isotopically-labeled compounds of the invention, wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds of the invention include isotopes of hydrogen, such as 2H and 3H, carbon, such as 11C, 13C and 14C, chlorine, such as 36Cl, fluorine, such as 18F, iodine, such as 123I and 125I, nitrogen, such as 13N and 15N, oxygen, such as 15O, 17O and 18O, phosphorus, such as 32P, and sulfur, such as 35S. Certain isotopically-labeled compounds of the invention, for example, those incorporating a radioactive isotope, are useful in drug and/or substrate tissue distribution studies. The radioactive isotopes tritium, 3H, and carbon-14, 14C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Substitution with heavier isotopes such as deuterium, 2H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Substitution with positron emitting isotopes, such as 11C, 18F, 15O and 13N, can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy.
Isotopically-labeled compounds of the invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein, using an appropriate isotopically-labeled reagent in place of the non-labeled reagent otherwise employed.
Pharmaceutically acceptable solvates in accordance with the invention include those wherein the solvent of crystallization may be isotopically substituted, e.g. D2O, d6-acetone, d6-DMSO.
Compounds of the invention intended for pharmaceutical use may be administered as crystalline or amorphous products, or mixtures thereof. They may be obtained, for example, as solid plugs, powders, or films by methods such as precipitation, crystallization, freeze drying, spray drying, or evaporative drying. Microwave or radio frequency drying may be used for this purpose.
The compounds can be administered alone or in combination with one or more other compounds of the invention, or in combination with one or more other drugs (or as any combination thereof). Generally, they will be administered as a formulation in association with one or more pharmaceutically acceptable excipients. The term “excipient” is used herein to describe any ingredient other than the compound(s) of the invention. The choice of excipient will to a large extent depend on factors such as the particular mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form.
Pharmaceutical compositions suitable for the delivery of compounds of the invention and methods for their preparation will be readily apparent to those skilled in the art. Such compositions and methods for their preparation can be found, for example, in ‘Remington's Pharmaceutical Sciences’, 19th Edition (Mack Publishing Company, 1995), the disclosure of which is incorporated herein by reference in its entirety.
Oral Administration
The compounds of the invention may be administered orally. Oral administration may involve swallowing, so that the compound enters the gastrointestinal tract, or buccal or sublingual administration may be employed by which the compound enters the blood stream directly from the mouth.
Formulations suitable for oral administration include solid formulations such as tablets, capsules containing particulates, liquids, or powders, lozenges (including liquid-filled), chews, multi- and nano-particulates, gels, solid solution, liposome, films (including muco-adhesive), ovules, sprays and liquid formulations.
Liquid formulations include suspensions, solutions, syrups and elixirs. Such formulations may be used as fillers in soft or hard capsules and typically include a carrier, for example, water, ethanol, polyethylene glycol, propylene glycol, methylcellulose, or a suitable oil, and one or more emulsifying agents and/or suspending agents. Liquid formulations may also be prepared by the reconstitution of a solid, for example, from a sachet.
The compounds of the invention may also be used in fast-dissolving, fast-disintegrating dosage forms such as those described in Expert Opinion in Therapeutic Patents, 11 (6), 981-986 by Liang and Chen (2001), the disclosure of which is incorporated herein by reference in its entirety.
For tablet dosage forms, depending on dose, the drug may make up from 1 wt % to 80 wt % of the dosage form, more typically from 5 wt % to 60 wt % of the dosage form. In addition to the drug, tablets generally contain a disintegrant. Examples of disintegrants include sodium starch glycolate, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, croscarmellose sodium, crospovidone, polyvinylpyrrolidone, methyl cellulose, microcrystalline cellulose, lower alkyl-substituted hydroxypropyl cellulose, starch, pregelatinized starch and sodium alginate. Generally, the disintegrant will comprise from 1 wt % to 25 wt %, preferably from 5 wt % to 20 wt % of the dosage form.
Binders are generally used to impart cohesive qualities to a tablet formulation. Suitable binders include microcrystalline cellulose, gelatin, sugars, polyethylene glycol, natural and synthetic gums, polyvinylpyrrolidone, pregelatinized starch, hydroxypropyl cellulose and hydroxypropyl methylcellulose. Tablets may also contain diluents, such as lactose (monohydrate, spray-dried monohydrate, anhydrous and the like), mannitol, xylitol, dextrose, sucrose, sorbitol, microcrystalline cellulose, starch and dibasic calcium phosphate dihydrate.
Tablets may also optionally include surface active agents, such as sodium lauryl sulfate and polysorbate 80, and glidants such as silicon dioxide and talc. When present, surface active agents are typically in amounts of from 0.2 wt % to 5 wt % of the tablet, and glidants typically from 0.2 wt % to 1 wt % of the tablet.
Tablets also generally contain lubricants such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate, and mixtures of magnesium stearate with sodium lauryl sulphate. Lubricants generally are present in amounts from 0.25 wt % to 10 wt %, preferably from 0.5 wt % to 3 wt % of the tablet.
Other conventional ingredients include anti-oxidants, colorants, flavoring agents, preservatives and taste-masking agents.
Exemplary tablets contain up to about 80 wt % drug, from about 10 wt % to about 90 wt % binder, from about 0 wt % to about 85 wt % diluent, from about 2 wt % to about 10 wt % disintegrant, and from about 0.25 wt % to about 10 wt % lubricant.
Tablet blends may be compressed directly or by roller to form tablets. Tablet blends or portions of blends may alternatively be wet-, dry-, or melt-granulated, melt congealed, or extruded before tableting. The final formulation may include one or more layers and may be coated or uncoated; or encapsulated.
The formulation of tablets is discussed in detail in “Pharmaceutical Dosage Forms: Tablets, Vol. 1”, by H. Lieberman and L. Lachman, Marcel Dekker, N.Y., N.Y., 1980 (ISBN 0-8247-6918-X), the disclosure of which is incorporated herein by reference in its entirety.
Solid formulations for oral administration may be formulated to be immediate and/or modified release. Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.
Suitable modified release formulations are described in U.S. Pat. No. 6,106,864. Details of other suitable release technologies such as high energy dispersions and osmotic and coated particles can be found in Verma et al., Pharmaceutical Technology On-line, 25(2), 1-14 (2001). The use of chewing gum to achieve controlled release is described in WO 00/35298. The disclosures of these references are incorporated herein by reference in their entireties.
Parenteral Administration
The compounds of the invention may also be administered directly into the blood stream, into muscle, or into an internal organ. Suitable means for parenteral administration include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular and subcutaneous. Suitable devices for parenteral administration include needle (including micro needle) injectors, needle-free injectors and infusion techniques.
Parenteral formulations are typically aqueous solutions which may contain excipients such as salts, carbohydrates and buffering agents (preferably to a pH of from 3 to 9), but, for some applications, they may be more suitably formulated as a sterile non-aqueous solution or as a dried form to be used in conjunction with a suitable vehicle such as sterile, pyrogen-free water.
The preparation of parenteral formulations under sterile conditions, for example, by lyophilization, may readily be accomplished using standard pharmaceutical techniques well known to those skilled in the art.
The solubility of compounds of the invention used in the preparation of parenteral solutions may be increased by the use of appropriate formulation techniques, such as the incorporation of solubility-enhancing agents.
Formulations for parenteral administration may be formulated to be immediate and/or modified release. Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted and programmed release. Thus compounds of the invention may be formulated as a solid, semi-solid, or thixotropic liquid for administration as an implanted depot providing modified release of the active compound. Examples of such formulations include drug-coated stents and PGLA microspheres.
Topical Administration
The compounds of the invention may also be administered topically to the skin or mucosa, that is, dermally or transdermally. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, creams, ointments, dusting powders, dressings, foams, films, skin patches, wafers, implants, sponges, fibers, bandages and microemulsions. Liposomes may also be used. Typical carriers include alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethylene glycol and propylene glycol. Penetration enhancers may be incorporated; see, for example, J Pharm Sci, 88 (10), 955-958 by Finnin and Morgan (October 1999). Other means of topical administration include delivery by electroporation, iontophoresis, phonophoresis, sonophoresis and micro needle or needle-free (e.g. Powderject™, Bioject™, etc.) injection. The disclosures of these references are incorporated herein by reference in their entireties.
Formulations for topical administration may be formulated to be immediate and/or modified release. Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.
Inhaled/Intranasal Administration
The compounds of the invention can also be administered intranasally or by inhalation, typically in the form of a dry powder (either alone, as a mixture, for example, in a dry blend with lactose, or as a mixed component particle, for example, mixed with phospholipids, such as phosphatidylcholine) from a dry powder inhaler or as an aerosol spray from a pressurized container, pump, spray, atomizer (preferably an atomizer using electrohydrodynamics to produce a fine mist), or nebulizer, with or without the use of a suitable propellant, such as 1,1,1,2 -tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane. For intranasal use, the powder may include a bioadhesive agent, for example, chitosan or cyclodextrin.
The pressurized container, pump, spray, atomizer, or nebulizer contains a solution or suspension of the compound(s) of the invention comprising, for example, ethanol, aqueous ethanol, or a suitable alternative agent for dispersing, solubilizing, or extending release of the active, a propellant(s) as solvent and an optional surfactant, such as sorbitan trioleate, oleic acid, or an oligolactic acid.
Prior to use in a dry powder or suspension formulation, the drug product is micronized to a size suitable for delivery by inhalation (typically less than 5 microns). This may be achieved by any appropriate comminuting method, such as spiral jet milling, fluid bed jet milling, supercritical fluid processing to form nanoparticles, high pressure homogenization, or spray drying.
Capsules (made, for example, from gelatin or HPMC), blisters and cartridges for use in an inhaler or insufflator may be formulated to contain a powder mix of the compound of the invention, a suitable powder base such as lactose or starch and a performance modifier such as I-leucine, mannitol, or magnesium stearate. The lactose may be anhydrous or in the form of the monohydrate, preferably the latter. Other suitable excipients include dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose and trehalose.
A suitable solution formulation for use in an atomizer using electrohydrodynamics to produce a fine mist may contain from 1 μg to 20 mg of the compound of the invention per actuation and the actuation volume may vary from 1 μL to 100 μL. A typical formulation includes a compound of the invention, propylene glycol, sterile water, ethanol and sodium chloride. Alternative solvents which may be used instead of propylene glycol include glycerol and polyethylene glycol.
Suitable flavors, such as menthol and levomenthol, or sweeteners, such as saccharin or saccharin sodium, may be added to those formulations of the invention intended for inhaled/intranasal administration.
Formulations for inhaled/intranasal administration may be formulated to be immediate and/or modified release using, for example, poly(DL-lactic-coglycolic acid (PGLA). Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.
In the case of dry powder inhalers and aerosols, the dosage unit is determined by means of a valve which delivers a metered amount. Units in accordance with the invention are typically arranged to administer a metered dose or “puff” containing a desired mount of the compound of the invention. The overall daily dose may be administered in a single dose or, more usually, as divided doses throughout the day.
Rectal/Intravaginal Administration
Compounds of the invention may be administered rectally or vaginally, for example, in the form of a suppository, pessary, or enema. Cocoa butter is a traditional suppository base, but various alternatives may be used as appropriate.
Formulations for rectal/vaginal administration may be formulated to be immediate and/or modified release. Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.
Ocular Administration
Compounds of the invention may also be administered directly to the eye or ear, typically in the form of drops of a micronized suspension or solution in isotonic, pH-adjusted, sterile saline. Other formulations suitable for ocular and aural administration include ointments, biodegradable (e.g. absorbable gel sponges, collagen) and non-biodegradable (e.g. silicone) implants, wafers, lenses and particulate or vesicular systems, such as niosomes or liposomes. A polymer such as crossed-linked polyacrylic acid, polyvinylalcohol, hyaluronic acid, a cellulosic polymer, for example, hydroxypropylmethylcellulose, hydroxyethylcellulose, or methyl cellulose, or a heteropolysaccharide polymer, for example, gelan gum, may be incorporated together with a preservative, such as benzalkonium chloride. Such formulations may also be delivered by iontophoresis.
Formulations for ocular/aural administration may be formulated to be immediate and/or modified release. Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted, or programmed release.
Other Technologies
Compounds of the invention may be combined with soluble macromolecular entities, such as cyclodextrin and suitable derivatives thereof or polyethylene glycol-containing polymers, in order to improve their solubility, dissolution rate, taste-masking, bioavailability and/or stability for use in any of the aforementioned modes of administration.
Drug-cyclodextrin complexes, for example, are found to be generally useful for most dosage forms and administration routes. Both inclusion and non-inclusion complexes may be used. As an alternative to direct complexation with the drug, the cyclodextrin may be used as an auxiliary additive, i.e. as a carrier, diluent, or solubilizer. Most commonly used for these purposes are alpha-, beta- and gamma-cyclodextrins, examples of which may be found in PCT Publication Nos. WO 91/11172, WO 94/02518 and WO 98/55148, the disclosures of which are incorporated herein by reference in their entireties.
Dosage
The amount of the active compound administered will be dependent on the subject being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compound and the discretion of the prescribing physician. However, an effective dosage is typically in the range of about 0.001 to about 100 mg per kg body weight per day, preferably about 0.01 to about 35 mg/kg/day, in single or divided doses. For a 70 kg human, this would amount to about 0.07 to about 7000 mg/day, preferably about 0.7 to about 2500 mg/day. In some instances, dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be used without causing any harmful side effect, with such larger doses typically divided into several smaller doses for administration throughout the day.
Kit-of-Parts
Inasmuch as it may desirable to administer a combination of active compounds, for example, for the purpose of treating a particular disease or condition, it is within the scope of the present invention that two or more pharmaceutical compositions, at least one of which contains a compound in accordance with the invention, may conveniently be combined in the form of a kit suitable for coadministration of the compositions. Thus the kit of the invention includes two or more separate pharmaceutical compositions, at least one of which contains a compound of the invention, and means for separately retaining said compositions, such as a container, divided bottle, or divided foil packet. An example of such a kit is the familiar blister pack used for the packaging of tablets, capsules and the like.
The kit of the invention is particularly suitable for administering different dosage forms, for example, oral and parenteral, for administering the separate compositions at different dosage intervals, or for titrating the separate compositions against one another. To assist compliance, the kit typically includes directions for administration and may be provided with a memory aid.
In the following examples, “Et” means ethyl, “Ac” means acetyl, “Me” means methyl, “Ms” means methanesulfonyl (CH3SO2), “iPr” means isopropyl, “HATU” means 2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate, “Ph” means phenyl, “Boc” means tert-butoxycarbonyl, “EtOAc” means ethyl acetate, “HOAc” means acetic acid, “NEt3” or “Et3N” means triethylamine, “THF” means tetrahydrofuran, “DIC” means diisopropylcarbodiimide, “HOBt” means hydroxy benzotriazole, “MeOH” means methanol, “i-PrOAc” means isopropyl acetate, “KOAc” means potassium acetate, “DMSO” means dimethylsulfoxide, “AcCI” means acetyl chloride, “CDCl3” means deuterated chloroform, “MTBE” means methyl t-butyl ether, “DMF” means dimethyl formamide, “Ac2O” means acetic anhydride, “Me3SOI” means trimethylsulfoxonium iodide, “DMAP” means 4-dimethylaminopyridine, “dppf” means diphenylphosphino ferrocene, “DME” means ethylene glycol dimethyl ether (1,2-dimethoxyethane), HOBT means 1-hydroxybenzotriazole, EDC means 1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide.
The following examples are given to illustrate the present invention. It should be understood, however, that the invention is not to be limited to the specific conditions or details described in these examples.
Reagents can be synthesized as shown herein, or are available from commercial sources (e.g., Aldrich, Milwaukee, Wis.; Acros, Morris Plains, N.J.; Biosynth International, Naperville, Ill.; Frontier Scientific, Logan, Utah; TCI America, Portland, Oreg.; Combi-Blocks, San Diego, Calif.; Matrix Scientific, Columbia, S.C.; Acros, Morris Plains, N.J.; Alfa Aesar, Ward Hill, Mass.; Apollo Scientific, UK; etc.) or can be synthesized by procedures known in the art.
The synthesis of several specific reagents is shown in U.S. patent application Ser. No. 10/786,610, entitled “Aminoheteroaryl Compounds as Protein Kinase Inhibitors”, filed Feb. 26, 2004, and corresponding international application PCT/US2004/005495 of the same title, filed Feb. 26, 2004. Other reagents can be synthesized by adapting the procedures therein, and one skilled in the art can readily adapt those procedures to produce the desired compounds. Further, these references contain general procedures and specific examples for the preparation of a large number of heteroarylamino compounds, and one skilled in the art can readily adapt such procedures and examples to the preparation of compounds of the present invention. The disclosures of these references are incorporated herein by reference in their entireties.
When a general or exemplary synthetic procedure is referred to, one skilled in the art can readily determine the appropriate reagents, if not indicated, extrapolating from the general or exemplary procedures. Some of the general procedures are given as examples for preparing specific compounds. One skilled in the art can readily adapt such procedures to the synthesis of other compounds. It should be understood that R groups shown in the general procedures are meant to be generic and non-limiting, and do not correspond to definitions of R groups elsewhere in this document. Each such R group represents one or multiple chemical moieties that can be the same or different from other chemical moieties also represented by the same R symbol. One skilled in the art can readily appreciate the range of R groups suitable in the exemplary syntheses. Moreover, representation of an unsubstituted position in structures shown or referred to in the general procedures is for convenience and does not preclude substitution as described elsewhere herein. For specific groups that can be present, either as R groups in the general procedures or as optional substituents not shown, refer to the descriptions in the remainder of this document, including the claims, summary and detailed description.
Some of the general procedures are shown with reference to synthesis of compounds wherein the 1-(2,6-dichloro-3-fluorophenyl)-ethoxy moiety is the pure (R)-isomer, and some are shown with reference to compounds wherein said moiety is a racemic mixture. It should be understood that the procedures herein can be used to produce racemic compounds or enantiomerically pure (R) isomers by choosing the corresponding racemic or enantiomerically pure starting material.
Select Starting Materials
1. 2,6-Dichloro-3-fluoroacetophenone (15 g, 0.072 mol) was stirred in THF (150 mL, 0.5M) at 0° C. using an ice bath for 10 min. Lithium aluminum hydride (2.75 g, 0.072 mol) was slowly added. The reaction was stirred at ambient temperature for 3 hr. The reaction was cooled in ice bath, and water (3 mL) was added drop wisely followed by adding 15% NaOH (3 mL) slowly. The mixture was stirred at ambient temperature for 30 min. 15% NaOH (9 mL), MgSO4 were added and the mixture filtered to remove solids. The solids were washed with THF (50 mL) and the filtrate was concentrated to give 1-(2,6-dichloro-3-fluoro-phenyl)-ethanol (14.8 gm, 95% yield) as a yellow oil. 1H NMR (400 MHz, DMSO-d6) δ 1.45 (d, 3H), 5.42 (m, 2H), 7.32 (m, 1H), 7.42 (m, 1H).
2. To a stirred solution of triphenyl phosphine (8.2 g, 0.03 mol) and DEAD (13.65 mL of a 40% solution in toluene) in THF (200 mL) at 0° C. was added a solution of 1-(2,6-dichloro-3-fluoro-phenyl)-ethanol (4.55 g, 0.021 mol) and 3-hydroxy-nitropyridine (3.35 g, 0.023 mol) in THF (200 mL). The resulting bright orange solution was stirred under a nitrogen atmosphere at ambient temperature for 4 hours at which point all starting materials had been consumed. The solvent was removed, and the crude material was dry loaded onto silica gel, and eluted with ethyl acetate-hexanes (20:80) to yield 3-(2,6-dichloro-3-fluoro-benzyloxy)-2-nitro-pyridine (6.21 g, 0.021 mol, 98%) as a pink solid. 1H NMR (CDCl3, 300 MHz) 51.8-1.85 (d, 3H), 6.0-6.15 (q, 1H), 7.0-7.1 (t, 1H), 7.2-7.21 (d, 1H), 7.25-7.5 (m, 2H), 8.0-8.05 (d, 1H).
3. To a stirred mixture of AcOH (650 mL) and EtOH (500 mL) was suspended 3-(2,6-dichloro-3-fluoro-benzyloxy)-2-nitro-pyridine (9.43 g, 0.028 mol) and iron chips (15.7 g, 0.28 mol). The reaction was heated slowly to reflux and allowed to stir for 1 hr. The reaction was cooled to room temperature then diethyl ether (500 mL) and water (500 mL) was added. The solution was carefully neutralized by the addition of sodium carbonate. The combined organic extracts were washed with sat'd NaHCO3 (2×100 mL), H2O (2×100 mL) and brine (1×100 mL) then dried (Na2SO4), filtered and concentrated to dryness under vacuum to yield 3-(2,6-dichloro-3-fluoro-benzyloxy)-pyridin-2-ylamine (9.04 g, 0.027 mol, 99%) as a light pink solid. 1H NMR (CDCl3, 300 MHz) δ 1.8-1.85 (d, 3H), 4.9-5.2 (brs, 2H), 6.7-6.84 (q, 1H), 7.0-7.1 (m, 1H), 7.2-7.3 (m, 1H), 7.6-7.7 (m, 1H).
4. A stirring solution of 3-(2,6-dichloro-3-fluoro-benzyloxy)-pyridin-2-ylamine (9.07 g, 0.03 mol) in acetonitrile was cooled to 0° C. using an ice bath. To this solution was added N-bromosuccinimide (NBS) (5.33 g, 0.03 mol) portionwise. The reaction was stirred at 0° C. for 15 min. The reaction was concentrated to dryness under vacuum. The resulting dark oil was dissolved in EtOAc (500 mL), and purified via silica gel chromatography. The solvents were then removed in vacuo to yield 5-bromo-3-(2,6-dichloro-3-fluoro-benzyloxy)-pyridin-2-ylamine (5.8 g, 0.015 mol, 51%) as a white crystalline solid. 1H NMR (CDCl3, 300 MHz) 51.85-1.95 (d, 3H), 4.7-5.0 (brs, 2H), 5.9-6.01 (q, 1H), 6.8-6.95 (d, 1H), 7.01-7.2 (t, 1H), 7.4-7.45 (m, 1H), 7.8-7.85 (d, 1H).
To a solution of 3-[1-(2,6-Dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-2-ylamine (10.0 g, 33.2 mmol) in acetonitrile (600 mL) and acetic acid (120 mL) was added N-iodosuccinimide (11.2 g, 49.8 mmol). The mixture was stirred at room temperature for 4 h and the reaction was quenched with Na2S2O5 solution. After evaporation, the residue was partitioned between ethyl acetate and water. The organic layer was washed with 2N NaOH solution, brine, and dried over Na2SO4. The crude product was purified on a silica gel column to provide 5-iodo-3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-2-ylamine (7.1 g, 50% yield). MS m/z 427 [M+1]. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.74 (d, J=6.57 Hz, 3H) 5.91-5.99 (m, 3H) 6.82 (d, J=1.26 Hz, 1H) 7.46 (t, J=8.72 Hz, 1H) 7.56 (dd, J=8.97, 4.93 Hz, 1H) 7.62 (d, J=1.52 Hz, 1H).
1. 2,6-Dichloro-3-fluoroacetophenone (15 g, 0.072 mol) was stirred in THF (150 mL, 0.5M) at 0° C. using an ice bath for 10 min. Lithium aluminum hydride (from Aldrich, 2.75 g, 0.072 mol) was slowly added. The reaction was stirred at ambient temperature for 3 h. The reaction was cooled in ice bath, and water (3 mL) was added drop wisely followed by adding 15% NaOH (3 mL) slowly. The mixture was stirred at ambient temperature for 30 min. 15% NaOH (9 mL), MgSO4 were added and the mixture filtered to remove solids. The solids were washed with THF (50 mL) and the filtrate was concentrated to give 1-(2,6-dichloro-3-fluoro-phenyl)-ethanol (14.8 gm, 95% yield) as a yellow oil. 1H NMR (400 MHz, DMSO-d6) δ 1.45 (d, 3H), 5.42 (m, 2H), 7.32 (m, 1H), 7.42 (m, 1H).
2. 5-Bromo-3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyrazin-2-ylamine was prepared following procedure 9 below, from 1-(2,6-dichloro-3-fluoro-phenyl)-ethanol and 3,5-dibromo-pyrazin-2-ylamine. 1H NMR (400 MHz, DMSO-d6) δ 1.74 (d, 3H), 6.40 (m, 1H), 6.52 (br s, 2H), 7.30 (m, 1H), 7.48 (m, 1H), 7.56 (s, 1H); MS m/z 382 (M+1).
Enantiomerically Pure Starting Materials
PLE is an enzyme produced by Roche and sold through Biocatalytics Inc. as a crude esterase preparation from pig liver, commonly known as PLE-AS (purchased from Biocatalytics as ICR-123, sold as an ammonium sulfate suspension). The enzyme is classified in the CAS registry as a “carboxylic-ester hydrolase, CAS no. 9016-18-6”. The corresponding enzyme classification number is EC 3.1.1.1. The enzyme is known to have broad substrate specificity towards the hydrolysis of a wide range of esters. The lipase activity is determined using a method based on hydrolysis of ethylbutyrate in a pH titrator. 1 LU (lipase unit) is the amount of enzyme which liberates 1 μmol titratable butyric acid per minute at 22° C., pH 8.2. The preparation reported herein (PLE-AS, as a suspension) is usually shipped as an opaque brown-green liquid with a declared activity of >45 LU/mg (protein content around 40 mg/mL).
(1S)-1-(2,6-dichloro-3-fluorophenyl)ethanol, shown as compound (S-1) in the schemes below, was prepared by a combination of enzymatic hydrolysis of racemic 1-(2,6-dichloro-3-fluorophenyl)ethyl acetate, esterification and chemical hydrolysis with inversion according to Scheme B. Racemic 1-(2,6-dichloro-3-fluorophenyl)ethyl acetate (compound A2) was prepared according to Scheme A.
1-(2,6-dichloro-3-fluorophenyl)ethanol (A1): Sodium borohydride (90 mg, 2.4 mmol) was added to a solution of 2′,6′-dichloro-3′-fluoro-acetophenone (Aldrich, catalog # 52,294-5) (207 mg, 1 mmol) in 2 mL of anhydrous CH3OH. The reaction mixture was stirred at room temperature for 1 h then was evaporated to give a colorless oil residue. The residue was purified by flash chromatography (eluting with 0→10% EtOAc in hexanes) to give compound A1 as a colorless oil (180 mg; 0.88 mmol; 86.5% yield); MS (APCI) (M−H)-208; 1H NMR (400 MHz, chloroform-D) δ ppm 1.64 (d, J=6.82 Hz, 3H) 3.02 (d, J=9.85 Hz, 1H) 6.97-7.07 (m, 1H) 7.19-7.33 (m, 1H).
1-(2,6-dichloro-3-fluorophenyl)ethyl acetate (A2): Acetic anhydride (1.42 mL, 15 mmol) and pyridine (1.7 mL, 21 mmol) were added sequentially to a solution of compound A1 (2.2 g, 10.5 mmol) in 20 mL of CH2Cl2. The reaction mixture was stirred at room temperature for 12 h and then evaporated to give a yellowish oil residue. The residue was purified by flash chromatography (eluting with 7-9% EtOAc in hexanes) to give compound A2 as a colorless oil (2.26 g; 9.0 mmol; 85.6% yield); 1H NMR (400 MHz, chloroform-D) δ ppm 1.88 (d, J=6.82 Hz, 3 H) 2.31 (S, 3H) 6.62 (q, J=6.82 Hz, 1H) 7.25 (t, J=8.46 Hz, 1H) 7.49 (dd, J=8.84, 5.05 Hz, 1H).
To a 50 mL jacketed flask equipped with a pH electrode, an overhead stirrer and a base addition line (1M NaOH), was added 1.2 mL of 100 mM potassium phosphate buffer pH 7.0 and 0.13 mL of PLE AS suspension. Then, compound A2 (0.13 g, 0.5 mmol, 1.00 eq) was added dropwise and the resulting mixture was stirred at room temperature for 20 h, maintaining the pH of the reaction constant at 7.0 using 1 M NaOH. Both the conversion and ee's of the reaction were monitored by RP-HPLC, and stopped after 50% starting material was consumed (approximately 17 hours under these conditions). The mixture was then extracted three times with 10 mL of ethyl acetate to recover both ester and alcohol as a mixture of R-1 and S-2.
Methanesulfonyl chloride (0.06 mL, 0.6 mmol) was added to a solution of a mixture of R-1 and S-2 (0.48 mmol) in 4 mL of pyridine under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 3 h then evaporated to obtain an oil. Water (20 mL) was added to the mixture and then EtOAc (20 mL×2) was added to extract the aqueous solution. The organic layers were combined, dried, filtered, and evaporated to give a mixture of R-3 and S-2. This mixture was used in the next step reaction without further purification. 1H NMR (400 MHz, chloroform-D) δ ppm 1.66 (d, J=7.1 Hz, 3H) 1.84 (d, J=7.1 Hz, 3H) 2.09 (s, 3H) 2.92 (s, 3H) 6.39 (q, J=7.0 Hz, 1H) 6.46 (q, J=6.8 Hz, 1H) 6.98-7.07 (m, 1H) 7.07-7.17 (m, 1H) 7.23-7.30 (m, 1H) 7.34 (dd, J=8.8, 4.80 Hz, 1H).
Potassium acetate (0.027 g, 0.26 mmol) was added to a mixture of R-3 and S-2 (0.48 mmol) in 4 mL of DMF under nitrogen atmosphere. The reaction mixture was heated to 100° C. for 12 h. Water (20 mL) was added to the reaction mixture and EtOAc (20 mL×2) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and evaporated to give an oil of S-2 (72 mg, 61% yield in two steps). Chirality ee: 97.6%. 1H NMR (400 MHz, chloroform-D) δ ppm 1.66 (d, J=7.1 Hz, 3H) 2.09 (s, 3H) 6.39 (q, J=6.8 Hz, 1H) 7.02 (t, J=8.5 Hz, 1H) 7.22-7.30 (m, 1H).
Sodium methoxide (19 mmol; 0.5 M in methanol) was added slowly to compound S-2 (4.64 g, 18.8 mmol) under a nitrogen atmosphere at 0° C. The resulting mixture was stirred at room temperature for 4 hours. The solvent was evaporated and H2O (100 mL) was added. The cooled reaction mixture was neutralized with sodium acetate-acetic acid buffer solution to pH 7. Ethyl acetate (100 mL×2) was added to extract the aqueous solution. The combined organic layers were dried over Na2SO4, filtered, and evaporated to obtain a white solid (4.36 g, 94.9% yield); SFC-MS: 97% ee. 1H NMR (400 MHz, chloroform-D) 3 ppm 1.65 (d, J=6.8 Hz, 3H) 5.58 (q, J=6.9 Hz, 1H) 6.96-7.10 (m, 1H) 7.22-7.36 (m, 1H).
3-Hydroxy-2-nitropyridine (175 mg, 1.21 mmol) and triphenylphosphine (440 mg, 1.65 mmol) were added sequentially to a stirred solution of (1S)-1-(2,6-dichloro-3-fluorophenyl)ethanol (229.8 mg, 1.1 mmol) in THF (10 mL) under a nitrogen atmosphere. The reaction mixture was maintained at room temperature for 1 h and then diisopropyl azo-dicarboxylate (0.34 mL, 1.65 mmol) was added at 0° C. The mixture was stirred for an additional 12 h. The reaction mixture was evaporated under vacuum to give an oil. The residue was purified by flash chromatography (eluting with 20-25% EtOAc in hexanes) to give the title compound as a white solid (321.5 mg; 0.97 mmol; 88.3% yield); MS (APCI) (M+H)+331; SFC-MS: 99.5% ee. 1H NMR (400 MHz, chloroform-D) 8 ppm 1.85 (d, J=6.6 Hz, 3H) 6.10 (q, J=6.6 Hz, 1H) 7.04-7.13 (m, 1H) 7.21 (dd, J=8.5, 1.14 Hz, 1H) 7.30 (dd, J=9.0, 4.9 Hz, 1H) 7.37 (dd, J=8.6, 4.6 Hz, 1H) 8.04 (dd, J=4.6, 1.3 Hz, 1H).
Iron (365 mg) was added to a stirred solution of 3-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]-2-nitropyridine (321 mg, 0.97 mmol) in a mixture of EtOH (2 mL) and 2M HCl (0.2 mL) at 0° C. The resulting solution was heated to 85° C. for 2 h. Celite (0.5 g) was added to the cooled reaction mixture. This mixture was filtered over a bed of celite and evaporated to give the title compound as a dark oil. MS (APCI) (M+H)+ 301.
The enantiomerically pure R isomer was prepared as described above for the racemate, but using the enantiomerically pure starting materials described above. 1H NMR (400 MHz, DMSO-d6) δ 1.74 (d, 3H), 6.40 (m, 1H), 6.52 (br s, 2H), 7.30 (m, 1H), 7.48 (m, 1H), 7.56 (s, 1H); MS m/z 382 (M+1).
Periodic acid (60 mg, 0.24 mmol), iodine (130 mg, 0.5 mmol), and sulfuric acid (0.03 mL) were added sequentially to a stirred solution of 3-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-2-amine (0.97 mmol) in a mixture of acetic acid (3 mL) and H2O (0.5 mL). The resulting solution was heated to 80° C. for 5 h. The cooled reaction mixture was quenched with Na2SO3 (80 mg) and basicified with saturated Na2CO3 (2×100 mL) to pH 7. CH2Cl2 (2×50 mL) was added to extract the aqueous solution. The combined organic layers were dried over Na2SO4 then filtered and concentrated under vacuum. The residue was purified by flash chromatography (eluting with 35→40% EtOAc in hexanes) to give the title compound as a yellow oil (254 mg; 0.6 mmol; 61.6% yield); MS (APCI) (M+H)+426. 1H NMR (400 MHz, chloroform-D) δ ppm 1.81 (d, J=6.8 Hz, 3H) 4.86 (s, 2H) 5.98 (q, J=6.57 Hz, 1H) 6.96 (d, J=1.5 Hz, 1H) 7.08 (dd, J=9.0, 8.0 Hz, 1H) 7.31 (dd, J=8.8, 4.8 Hz, 1H) 7.78 (d, J=1.8 Hz, 1H).
The title compound was prepared according to procedure 2, from (1S)-1-(2,6-dichloro-3-fluorophenyl)ethanol. 1H NMR (400 MHz, DMSO-d6) δ 7.53 (s, 1H), 7.48 (m, 1H), 7.39 (t, 1H), 6.48 (s, 2H), 6.41 (q, 1H), 1.74 (d, 3H); LCMS: 381 [M+1]; c-Met Ki: 0.796 μM.
General Procedure 1
1-(t-butoxycarbonyl)azetidine-3-carboxylic acid (−1)(AXL016917, 1000 mg, 4.97 mmol) was dissolved in MeOH (5 mL)/Toluene (20 mL) and then cooled to 0° C. TMSCHNN (trimethylsilyldiazomethane) (7.45 mmol) was then added drop-wise over 15 minutes with some bubbling observed. The color started clear and slowly turned yellow. The solution was stirred for 10 minutes at 0° C. and then warmed to room temperature over 30 minutes. The solution was then concentrated and pumped on to remove toluene to afford 1.055 g of 1-t-butyl 3-methyl azetidine-1,3-dicarboxylate (−2) that was used directly in the next step without being purified (99% crude yield).
1-tert-butyl 3-methyl azetidine-1,3-dicarboxylate (1055 mg, 4.90 mmol) was dissolved in THF (17 mL) and then cooled to 0° C. MeOH (0.397 mL, 9.80 mmol) and LiBH4 (14.7 mmol) were added sequentially. The reaction was warmed to room temperature over 3 h. Then 10% aqueous potassium sodium tartrate tetrahydrate (Rochelle's Salt) (30 mL) and EtOAc (30 mL) were added and the solution stirred at room temperature over 30 minutes. The organic layer was separated and then dried (Na2SO4) and concentrated to afford 674 mg of t-butyl 3-(hydroxymethyl)azetidine-1-carboxylate (1-3) as a crude product (clear oil). The product was used directly in the next step without purification.
t-butyl 3-(hydroxymethyl)azetidine-1-carboxylate (674 mg, 3.60 mmol) was dissolved in CH2Cl2 (13 mL, 0.25M) and then Et3N (1.0 mL, 7.20 mmol), DMAP (44 mg, 0.360 mmol), and methanesulfonyl chloride (0.31 mL, 3.96 mmol) were added sequentially at 0° C. with the MsCl addition being done slowly. The solution was warmed to rt over 1 h. After 15 h, saturated aqueous NaHCO3 (50 mL) was added and then the product was extracted with CH2Cl2 (2×50 mL) and the combined organic extracts were washed with brine (50 mL), dried (Na2SO4), concentrated, and purified by flash chromatography (Biotage Horizon —10% EtOAc/hexanes—100% EtOAc) to afford 962 mg of (1-4) as an oil (quantitative).
NaH (95%, 96 mg, 3.99 mmol) was combined in DMF (10 mL) under N2 at rt. 4-Bromopyrazole (533 mg, 3.63 mmol) was then added and the mixture stirred at rt. After 30 minutes (1-4) was added and the solution heated to 95° C. After 2 h, saturated aqueous NH4Cl (50 mL) was added and then EtOAc (50 mL). The organic extract was dried (Na2SO4) and concentrated, and then run through a short pad of silica gel with 50% EtOAc/Hexanes to afford 846 mg of crude (1-5) that was used directly in the next step (74% crude yield).
(1-5) (846 mg, 2.68 mmol), (1-6) (815 mg, 3.21 mmol), [1,1′-bis(diphenylphosphino)-ferrocene)dichloropalladium (108 mg, 0.133 mmol), and KOAc (893 mg, 9.10 mmol) were combined in DMSO (10 mL, purged with N2 for 10 minutes) and then the solution was warmed to 80° C. After 16 h, the solution was filtered through Celite and then H2O (50 mL) and EtOAc (50 mL) was added. The organic phase was extracted and dried (Na2SO4), concentrated, and then passed through a silica plug with 50% EtOAc/Hexane. The solvent was concentrated to afford 1.22 g of crude (1-7) used directly in the next step.
The boronic ester (1-7) (4144 mg, 11.4 mmol), (1-8) (2890 mg, 7.60 mmol), dichlorobis(triphenylphosphine)palladium(II) (534 mg, 0.760 mmol), DME (40 mL, De-gassed for 30 minutes with N2), and 1N Na2CO3 (40 mL, De-gassed for 30 minutes with N2) were combined and heated to 80° C. After 16 h, the reaction was cooled to rt and EtOAc (80 mL) was added. The solution was filtered through celite and then water (80 mL) was added. The organic layer was separated, dried (Na2SO4), and concentrated. The product was purified by flash chromatography to afford 1486 mg of (1-9) as a tan solid (36%).
1 gram of DOWEX 50WX2400 ion-exchange resin was prepared by washing it with H2O (500 mL), 1:1H2O/MeOH, MeOH (5×250 mL), CH2Cl2 (500 mL), and hexanes (500 mL). The DOWEX was then dried in a vacuum oven at 40° C. for 1 day. (1-9) was dissolved in MeOH and then DOWEX (588 mg, 1.096 mmol) was added. The solution was stirred at rt for 2 h. The solution was then filtered and the resin was washed with MeOH (3×200 mL) and the wash was discarded. The resin was then washed with 3.5M NH3/MeOH and collected. The solution was then concentrated to afford 374 mg of (1-10) as a gummy solid (78%).
To form compounds of formula (1-11), the following exemplary procedure can be followed. 1 molar equivalent of (1-10) is dissolved in DMF or CH2Cl2 and then base (3 molar equivalents) and/or coupling reagent (1.5 molar equivalents) is added. To the solution is added X—R (1.1 molar equivalent), where X is, for example, Cl, Br, I, OMs, COCl, CO, COOH, ethylene or carbonate and R is a desired group such as those shown in the examples herein or similar groups. The resultant solution is stirred at rt for 4 h. H2O and EtOAc are added and the organic phase extracted, dried (Na2SO4), and concentrated. The crude product can purified by purified by preparative HPLC or other methods well known in the art to afford the product (1-11).
General Procedure 2
3-Azetidinol (2-2): A reaction mixture of N-benzhydrylazetidin-3-ol HCl salt (2.76 g, 10.0 mmol) with palladium hydroxide, 20% Pd (dry base) on C (400 mg) in 50 mL of MeOH was hydrogenated at 55 psi for 48 h. The reaction mixture was filtered through Celite pad and washed well with MeOH. The filtrate was concentrated under vacuum at room temperature water bath. The residue was treated with ether (3×30 ml) and the solvent is decanted. The solid was air dried to give 571 mg of HCl salt product (2-2) as white solid (52% yield). 1H NMR (400 MHz, DMSO-D6) δ ppm 3.33 (s, 1H) 3.63-3.80 (m, 2H) 3.93-4.09 (m, 2H) 4.40-4.58 (m, 1H) 6.18 (d, J=6.32 Hz, 1H).
3-Hydroxy-azetidine-1-carboxlic acid tert-butyl ester (3-3): To a cold (0° C. bath) stirred solution of compound (2-2) (570 mg, 5.20 mmol) in 10 mL of EtOH was added Et3N (1.8 mL, 13.0 mmol) and di-tert-butyldicarbonate (1.702 g, 7.38 mmol). The resulting mixture of clear solution was stirred at room temperature overnight. The reaction mixture was concentrated by vacuum. The residue was portioned between EtOAc (200 mL) and 0.5N citric acid solution (30 mL; brine (30 mL). The organic layer was dried (Na2SO4), then concentrated by vacuum to give 899 mg (2-3) as clear oil (52%). 1H NMR (400 MHz, chloroform-D) δ ppm 1.42 (s, 9H) 3.78 (dd, J=9.47, 4.42 Hz, 2H) 4.13 (dd, J=9.35, 6.57 Hz, 2H) 4.49-4.63 (m, 1H).
3-Methanesulfonyloxy-azetidine-1-carboxylic acid tert-butyl ester (2-4): To a solution of compound (2-3) (466 mg; 2.69 mmol) with Et3N (0.75 mL; 5.38 mmol) and 4-(dimethylamino)-pyridine (33 mg, 0.269 mmol) in 10 mL of CH2Cl2 at 0° C. was added methanesulfonyl chloride (0.25 mL 3.23 mmol). The resulting mixture of brown color solution was stirred at 0° C. to room temperature for overnight. The reaction mixture was quenched with NaHCO3, then partitioned between CH2Cl2 (200 mL) and saturated NaHCO3 solution (50 mL). The organic layer was dried (Na2SO4), then filtered through silica gel pad, eluted with hexane: EtOAc/1:1; the filtrate was concentrated by vacuum to give 614 mg (2-4 as yellow oil (91% yield). 1H NMR (400 MHz, chloroform-D) δ ppm 1.43 (s, 9H) 3.05 (s, 3H) 4.08 (dd, J=10.36, 4.29 Hz, 2H) 4.26 (dd, J=10.36, 6.82 Hz, 2H) 5.11-5.26 (m, 1H).
1-(3-Azetidine-1-carboxylic acid tert-butyl ester)-4-bromoprazole (2-6): A 5 mL microwave tube was charge with compound (2-4) (304 mg, 1.21 mmol); 4-bromopyrazole (2-5, 178 mg, 1.21 mmol) and NaH 60% in mineral oil (73 mg, 1.82 mmol.) with 2 mL of DMF. The resulting mixture was microwaved at 110° C. for 30 minutes. The reaction mixture was partitioned between EtOAc (200 mL) and saturated NaHCO3 solution (2×50 mL); brine (50 mL). The organic layer was dried (Na2SO4), then concentrated by vacuum to afford 360 mg of (2-6) as yellow oil (98%). 1H NMR (400 MHz, DMSO-D6) δ ppm 1.36-1.43 (m, 9H) 4.08 (s, 2H) 4.18-4.31 (m, 2H) 5.12-5.22 (m, 1H) 7.67 (s, 1H) 8.14 (s, 1H).
tert-Butyl 3-[4-(4,4,5,5-tetramethyl-1,3-dioxoborolan-2-yl)-1H-pyrazol-1-yl]azetidine-1-carboxylate (2-8): A reaction mixture of compound (2-6) (225 mg, 0.74 mmol) and bis(pinacolate)diboron (2-7, 227 mg, 0.89 mmol) with KOAc (247 mg, 2.52 mmol) in 3 mL of DMSO was purged with N2 for 15 minutes, then PdCl2(dppf)2CH2Cl2 (30 mg, 2.52 mmol) was added. The resulting mixture was stirred at 80° C. under N2 for overnight. After it cooled down to room temperature, the mixture was filtered through Celite pad and washed well with EtOAc. The filtrate was extracted with H2O (2×50 mL), brine (50 mL). The organic layer was dried (Na2SO4), then concentrated by vacuum. The residue was then filtered through silica gel pad, eluted with hexane:EtOAc/3:2. The filtrate was concentrated by vacuum to give 250 mg of (2-8) as a clear oil (97% yield). 1H NMR (400 MHz, chloroform-D) δ ppm 1.18-1.27 (m, 9H) 1.28-1.34 (m, 6H) 1.41-1.49 (m, 6H) 4.22-4.33 (m, 2H) 4.36 (t, J=8.59 Hz, 2H) 4.98-5.13 (m, 1H) 7.83 (s, 2H).
tert-Butyl 3-(4-{6-amino-5-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-3-yl}-1H-pyrazol-1-yl)azetidine-1-carboxylate (2-10): A reaction mixture of compound (−8) (459 mg; 1.31 mmol) and 3-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]-5-iodopyridin-2-amine (2-9) (374 mg; 0.88 mmol) in 13 mL of ethylene glycol dimethylether, anhydrous (DME) was purged with N2 for 15 minutes, then Pd(II)(PPh3)2Cl2 (46 mg, 0.07 mmol) was added and continued to purge with N2 for another 15 minutes. Another 1.0 N Na2CO3 solution (3.9 mL; 3.9 mmol) was added after purging with N2 for 15 minutes. The resulting mixture was stirred at 85° C. under N2 for overnight. The reaction mixture was filtered through Celite pad and washed well with MeOH. The filtrate was concentrated by vacuum. The residue was partitioned between EtOAc (200 mL) and saturated NaHCO3 solution (2×50 mL); brine (50 mL). The organic layer was dried (Na2SO4), then concentrated by vacuum. The residue was purified by Biotage system (25 M, 100% CH2Cl2; 100% CH2Cl2 to 90% CH2Cl2 with 10% MeOH) to collect the desired fraction to afford 421 mg of (2-10) as a brown color grease (92% yield). 1H NMR (400 MHz, chloroform-D) δ ppm 1.17-1.26 (m, 9 H) 1.80-1.87 (m, 3H) 4.04-4.18 (m, 2H) 4.20-4.33 (m, 2H) 4.34-4.41 (m, 1H) 4.79 (s, 2H) 5.02 (d, J=7.58 Hz, 1H) 7.04 (t, J=8.46 Hz, 1H) 7.33-7.41 (m, 1H) 7.44-7.52 (m, 1H) 7.53-7.58 (m, 1H) 7.59-7.65 (m, 1H) 7.72-7.78 (m, 1H); LCMS calcd for C24H26Cl2FN5O3 (M+H) 523, found 523.
5-(1-Azetidin-3-yl-1H-pyrazol-4-yl)-3-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]yridin-2-amine (2-11): A reaction mixture of compound (2-10) (421 mg; 0.81 mmol) with 4.0 M HCl in dioxane (2.0 mL; 8.1 mmol) in 5 mL of CH2Cl2 was stirred at room temperature for 2.0 hours. The reaction mixture was concentrated by vacuum. The residue was treated with EtOAc. The precipitated solid was filtered off and washed well with EtOAc, hexane, then dried under vacuum to give 275 mg of (2-11) as a sand color solid of HCl salt (81% yield). 1H NMR (400 MHz, DMSO-D6) δ ppm 1.79-1.89 (m, 3H) 3.56 (s, 1H) 4.35 (s, 4H) 5.40 (s, 1H) 6.23 (d, J=6.57 Hz, 2H) 7.09 (s, 1H) 7.40-7.54 (m, 1H) 7.59 (dd, J=8.84, 5.05 Hz, 1H) 7.73-7.83 (m, 1H) 7.86 (s, 1H) 8.12 (s, 1H) 9.20 (s, 1H). LCMS calcd for C19H18Cl2FN5O (M+H) 423, found 423.
Compounds of formula 2-12 can be prepared by the following exemplary procedure: To a reaction mixture of compound (2-11) (1.0 eq.) with Et3N (2.0 eq.) in 2.0 mL of DMF at room temperature is added alkyl bromide (1.1 eq.). The resulting mixture is stirred under N2 at room temperature for overnight. The reaction mixture is partitioned between EtOAc (200 mL) and saturated NaHCO3 solution (2×50 mL); brine (50 mL). The organic layer is dried (Na2SO4), then concentrated by vacuum. The residue is purified by Dionex system (5% to 95% MeCN:H2O w 0.1% HOAc buffer) to collect the desired fraction to afford (2-12).
Alternatively, compounds of formula 2-12 can be prepared by the following exemplary procedure: To a reaction solution of alkyl amine (1.0 eq.) with iPr2EtN (diisopropylethylamine) (3.0 eq.) in 2.0 mL of DMF is added HATU (1.5 eq.). After stirring for 30 minutes, compound (2-11) (1.0 eq.) is added. The resulting mixture is stirred at room temperature for overnight. The reaction mixture is partitioned between EtOAc (200 mL) and saturated NaHCO3 solution (2×50 mL) and brine (50 mL). The organic layer is dried (Na2SO4) and concentrated by vacuum. The residue is purified by Dionex System (5% to 95% McCN:H2O w 0.1% HOAc) to collect the desired product to afford (2-12).
General Procedure 3:
tert-Butyl 1-oxa-6-azaspiro[2.5]octane-6-carboxylate (3-2): A solution of dimethylsulfoxonium methylide was prepared under N2 from NaH 60% dispersion in mineral oil (440 mg; 11.0 mmol) and trimethylsulfoxonium iodide (2.421 g; 11.0 mmol) in 5 ml of anhydrous DMSO. Another solution of 1-Boc-4-oxo-1-piperidincarboxylate (3-1, 1.993 g; 10.0 mmol) in 5 mL of DMSO was added dropwise. The resulting mixture was stirred at 55° C. for 6 hours. The cooled reaction mixture was poured into ice-H2O and extracted with EtOAc (2×200 mL). The combined organic layers were washed with H2O (50 mL); brine (50 mL) and then dried (Na2SO4), then concentrated by vacuum to give 1.4791 g of (3-2) as a yellow oil (69% yield). 1H NMR (400 MHz, chloroform-D) δ ppm 1.37-1.52 (m, 11H) 1.71-1.84 (m, 2H) 2.63-2.72 (m, 2H) 3.35-3.49 (m, 2H) 3.62-3.78 (m, 2H).
tert-Butyl 4-hydroxy-4-{[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]methyl}piperidine-1-carboxylate (3-4): A reaction mixture of compound (3-2) (214 mg; 1.0 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (3-3, 194 mg; 1.0 mmol) with NaH 60% dispersion in mineral oil (60 mg; 1.5 mmol) in 3 mL of DMF was stirred at 90° C. for 3 hours. The reaction mixture was partitioned between EtOAc (200 mL) and saturated NaHCO3 solution (50 mL) and brine (50 mL). The organic layer was dried (Na2SO4) and concentrated by vacuum to give 361 mg of (3-4) as a yellow grease (89% yield). 1H NMR (400 MHz, chloroform-D) δ ppm 1.21-1.34 (m, 12H) 1.39-1.50 (m, 9H) 1.56-1.78 (m, 4H) 3.14 (s, 2H) 3.72-3.91 (m, J=32.34 Hz, 2H) 4.05 (s, 2H) 7.65 (s, 1H) 7.80 (s, 1H) 8.00 (s, 1H). LCMS calcd for C20H34BN3O5 (M+H) 408, found 408. HPLC purity 85%.
tert-Butyl 4-[(4-{6-amino-5-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-3-yl}-1H-pyrazol-1-yl)methyl]-4-hydroxypiperidine-1-carboxylate (3-6): A reaction mixture of compound (3-4) (361 mg; 0.89 mmol) and 3-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]-5-iodopyridin-2-amine (3-5) (378 mg; 0.89 mmol) in 9.0 mL of ethylene glycol dimethylether, anhydrous (DME) was purged with N2 for 15 minutes, then Pd(II)(PPh3)2Cl2 (32 mg, 0.05 mmol) was added and continued to purge with N2 for another 15 minutes. Another 1.0 N Na2CO3 solution (3.9 mL; 3.9 mmol) was added after purging with N2 for 15 minutes. The resulting mixture was stirred at 85° C. under N2 for overnight. The reaction mixture was filtered through Celite pad and washed well with MeOH. The filtrate was concentrated by vacuum. The residue was partitioned between EtOAc (200 mL) and saturated NaHCO3 solution (2×50 mL); brine (50 mL). The organic layer was dried (Na2SO4), then concentrated by vacuum. The residue was purified by Dionex system (25% to 95% MeCN:H2O w 0.1% HOAc buffer) to collect the desired fraction to afford 147 mg of (3-6) as a white solid (28% yield). 1H NMR (400 MHz, DMSO-D6) δ ppm 1.34-1.39 (m, 9H) 1.70-1.77 (m, 2H) 1.79 (d, J=6.57 Hz, 3H) 3.06 (d, J=12.63 Hz, 2H) 3.62 (s, 2H) 4.03 (s, 2H) 4.79 (s, 1H) 5.66 (s, 2H) 6.08 (d, J=6.82 Hz, 1H) 6.86 (d, J=1.52 Hz, 1H) 7.44 (t, J=8.72 Hz, 1H) 7.51-7.58 (m, 2H) 7.58-7.65 (m, 2H) 7.73 (d, J=1.52 Hz, 1H) 7.78 (s, 1H). LCMS calcd for C27H32Cl2FN5O4 (M+H) 581, found 581. HPLC purity 87%.
4-[(4-{6-amino-5-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-3-yl}-1H-pyrazol-1-yl)methyl]piperidin-4-ol (3-7): A reaction mixture of compound (3-6) (145 mg; 0.25 mmol) with 4.0 M HCl in dioxane (2.0 mL; 8.1 mmol) in 5 mL of CH2Cl2 was stirred at room temperature for 2.0 hours. The reaction mixture was concentrated by vacuum. The residue was purified by Dionex system (5% to 95% MeCN:H2O w 0.1% HOAc buffer) to collect the desired fraction to afford 76 mg of (3-7) as a yellow grease (63% yield). 1H NMR (400 MHz, DMSO-D6) δ ppm 1.41-1.55 (m, 2H) 1.59-1.71 (m, 2H) 1.81 (d, J=6.57 Hz, 3H) 2.88-3.00 (m, 2H) 3.02-3.14 (m, 2H) 4.08 (s, 2H) 5.17 (s, 2H) 6.14-6.27 (m, J=6.57 Hz, 1H) 7.05 (s, 1H) 7.40-7.49 (m, J=8.72, 8.72 Hz, 1 H) 7.51-7.60 (m, J=9.09, 4.80 Hz, 1H) 7.63 (s, 1H) 7.76 (s, 1H) 7.91 (s, 1H) 8.51 (s, 1H) 8.81 (s, 1H). LCMS calcd for C22H24Cl2FN5O2 (M+H) 481, found 481. HPLC purity 98%. Anal. (C22H24Cl2FN5O2x2.2HOAcx2.3H2O)C, H, N.
General Procedure 4:
Ethyl 2-[(4-bromo-1H-pyrazol-1-yl)methyl]cyclopropanecarboxylate (4-3): To a reaction solution of ethyl 2-(hydroxymethyl)cyclopropanecarboxylate (4-1) (577 mg; 4.0 mmol) with Et3N (1.1 mL; 8.0 mmol) and DMAP (49 mg; 0.4 mmol) in 12 mL of CH2Cl2 at 0° C. was added methanesulfonyl chloride (0.4 mL; 4.8 mmol). The resulting mixture of brown color suspension was stirred at 0° C. to room temperature under N2 for overnight. The reaction mixture was quenched with NaHCO3, then partitioned between CH2Cl2 (200 mL) and saturated NaHCO3 solution (50 mL); brine (50 mL). The organic layer was dried (Na2SO4), then filtered through silica gel pad, eluted with hexane:EtOAc/1:1. The filtrate was concentrated by vacuum to give 880 mg of ethyl 2-{[(methylsulfonyl)oxy]methyl}cyclopropanecarboxylate as a yellow oil (99% yield). 1H NMR (400 MHz, chloroform-D) δ ppm 0.91-1.02 (m, 1H) 1.26 (q, J=6.99 Hz, 3H) 1.29-1.36 (m, 1H) 1.63-1.74 (m, 1H) 1.79-1.92 (m, 1H) 3.02 (s, 3H) 3.99-4.24 (m, 4H).
A reaction mixture of ethyl 2-{([(methylsulfonyl)oxy]methyl}cyclopropanecarboxylate (880 mg; 4.0 mmol), 4-bromopyrazole (4-2, 588 mg, 4.0 mmol) and NaH 60% in mineral oil (240 mg, 6.0 mmol) with 3.0 mL of DMF was formed. The resulting mixture was stirred at 90° C. under N2 for four hours. The reaction mixture was partitioned between EtOAc (200 mL) and saturated NaHCO3 solution (2×50 mL); brine (50 mL). The organic layer was dried (Na2SO4), then concentrated by vacuum to afford 812 mg of (4-3) as a yellow oil (74%). 1H NMR (400 MHz, chloroform-D) δ ppm 0.85 (dd, J=7.96, 3.16 Hz, 1H) 0.88-0.98 (m, 1H) 1.18-1.29 (m, 3H) 1.56-1.71 (m, 1H) 1.79-1.94 (m, 1H) 3.96-4.08 (m, 2H) 4.07-4.17 (m, 2H) 7.45 (d, J=3.79 Hz, 2H). LCMS calcd for C10H13BrN2O2 (M+H) 274, found 274. HPLC purity 95%.
Ethyl 2-{[4-(4,4,5,5-tetramethyl-1,3-dioxoborolan-2-yl)-1H-pyrazol-1-yl]methyl}cyclopropanecarboxylate (4-4): A reaction mixture of compound (4-3) (812 mg, 2.97 mmol) and bis(pinacolate)diboron (906 mg, 3.57 mmol) with KOAc (991 mg, 10.10 mmol) in 10.0 mL of DMSO was purged with N2 for 15 minutes, then PdCl2(dppf)2CH2Cl2 (122 mg, 0.15 mmol) was added. The resulting mixture was stirred at 80° C. under N2 for overnight. After cooling down to room temperature, the mixture was filtered through Celite pad and washed well with EtOAc. The filtrate was extracted with H2O (2×50 mL), brine (50 mL). The organic layer was dried (Na2SO4), then concentrated by vacuum. The residue was then filtered through silica gel pad, and eluted with hexane:EtOAc/3:1. The filtrate was concentrated by vacuum to give 945 mg of (4-4) as a yellow oil (98% yield). 1H NMR (400 MHz, chloroform-D) δ ppm 0.85 (dd, J=7.83, 3.03 Hz, 1H) 0.90-0.96 (m, 1H) 1.20-1.24 (m, 3H) 1.29-1.34 (m, 12H) 1.62-1.71 (m, 1H) 1.84-1.97 (m, 1H) 3.96-4.07 (m, 1H) 4.06-4.14 (m, 2H) 4.15-4.23 (m, J=14.27, 6.44 Hz, 1H) 7.73 (s, 1H) 7.77 (s, 1H).
Ethyl 2-[(4-{6-amino-5-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-3-yl}-1H-pyrazol-1-yl)methyl]cyclopropanecarboxylate (4-6): A reaction mixture of compound (4-4) (643 mg; 2.01 mmol) and 3-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]-5-iodopyridin-2-amine (4-5) (572 mg; 1.34 mmol) in 20.0 mL of ethylene glycol dimethylether, anhydrous (DME) was purged with N2 for 15 minutes, then Pd(II)(PPh3)2Cl2 (71 mg, 0.1 mmol) was added and continued to purge with N2 for another 15 minutes. Another 1.0 N Na2CO3 solution (6.0 mL; 6.0 mmol) was added after purging with N2 for 15 minutes. The resulting mixture was stirred at 85° C. under N2 for overnight. The reaction mixture was filtered through Celite pad and washed well with MeOH. The filtrate was concentrated by vacuum. The residue was partitioned between EtOAc (200 mL) and saturated NaHCO3 solution (2×50 mL); brine (50 mL). The organic layer was dried (Na2SO4), then concentrated by vacuum. The residue was purified by Biotage system (25 M CH2Cl2 100%; CH2Cl2 100% to 90% CH2Cl2: 10% MeOH) to collect the desired fraction to afford 600 mg of (4-6) as a brown color grease (91% yield). 1H NMR (400 MHz, DMSO-D6) δ ppm 0.96-1.10 (m, 2H) 1.15 (t, J=7.07 Hz, 2H) 1.74 (s, 3H) 1.79 (d, J=6.57 Hz, 3H) 3.95-4.14 (m, 4H) 5.66 (s, 2H) 6.08 (d, J=6.57 Hz, 1H) 6.88 (s, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.49-7.62 (m, 2H) 7.73 (s, 1H) 7.88 (s, 1H). LCMS calcd for C23H23Cl2FN4O3 (M+H) 494, found 494. HPLC purity 95%.
2-[(4-{6-amino-5-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-3-yl}-1H-pyrazol-1-yl)methyl]cyclopropanecarboxylic acid (4-7): To a reaction solution of compound (4-6) (377 mg, 0.76 mmol) in 5.0 mL of MeOH at room temperature under N2 was added another solution of 2.0 N NaOH (2) (1.5 mL, 3.04 mmol). The resulting mixture was stirred at 80° C. for 3 hours. The reaction mixture was concentrated by vacuum to remove most of the MeOH and acidified by 2 M HCl to pH 4.0. The mixture was extracted with CH2Cl2 (2×200 mL); the organic layers were washed with brine (50 mL), and dried (Na2SO4) and concentrated by vacuum to give 324 mg of (4-7) as a yellow solid. (92% yield). 1H NMR (400 MHz, DMSO-D6) δ ppm 0.92-1.04 (m, 2H) 1.57-1.72 (m, 2H) 1.76-1.90 (m, 3H) 3.98-4.18 (m, 2H) 6.46 (s, 2H) 6.89-7.02 (m, 1H) 7.29-7.52 (m, 2H) 7.52-7.63 (m, 2H) 7.73 (d, J=1.52 Hz, 1H) 7.94 (s, 1H) 12.19 (s, 1H). LCMS calcd for C21H19Cl2FN4O3 (M−H) 463, found 463. HPLC purity 87%.
2-[(4-{6-amino-5-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-3-yl}-1H-pyrazol-1-yl)methyl]-N-methylcyclopropanecarboxamide (4-8) (R=Me, R′=H): To a reaction solution of (4-7) (1.0 eq.) with iPr2EtN (2.0 eq.) in 1.0 mL of DMF was added HATU (1.5 eq.). After stirring for 30 minutes, alkylamine (1.1 eq.) was added. The resulting mixture was stirred at room temperature for overnight. The reaction mixture was partitioned between EtOAc (200 mL) and saturated NaHCO3 solution (2×50 mL) and brine (50 mL). The organic layer was dried (Na2SO4) and concentrated by vacuum. The sample was free based by partitioning between EtOAc (200 mL) and saturated NaHCO3 solution (50 mL) and brine (50 mL). The organic layer was dried (Na2SO4) and concentrated by vacuum. The residue was treated with 1.0 mL of H2O and lyophilized to afford (4-8).
General Procedure 5:
To a solution of 4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (5 g, 25.77 mmol) and 2-bromo-2-methyl-propionic acid methyl ester (12.6 g, 27.06 mmol) in DMF (85 mL), was added Cs2CO3 (12.6 g, 38.65 mmol). The reaction mixture was heated to 90° C. in an oil bath overnight. The reaction solution was cooled to room temperature, and partitioned between water and ethyl acetate. The combined ethyl acetate solution was washed with water five times, dried over Na2SO4, and concentrated to give the product 2-methyl-2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyrazol-1-yl]propionic acid methyl ester (4.776 g, 63% yield).
To a solution of 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-iodo-pyridin-2-ylamine (6.363 g, 14.901 mmol) and 2-methyl-2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyrazol-1-yl]propionic acid methyl ester (4.6 g, 15.64 mmol) in DME (27 mL) was added a solution of CsF (6.79 g, 44.7 mmol) in water (9.3 mL). The reaction mixture was degassed 3 times with N2. Pd(dppf)CH2Cl2 was added and the reaction mixture was degassed 3 times with N2. The reaction was heated to 120° C. in the microwave (subsequent Pd was added in intervals of 30 minutes until the reaction was complete). Water was added and the reaction was extracted with EtOAc, dried over Na2SO4, and concentrated to give 2-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-2-methyl-propionic acid methyl ester. The crude product was purified by a silica gel column chromatography with a gradient of 25%-50% EtOAc/hexanes to provide 2-(4-{6-Amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-2-methyl-propionic acid methyl ester (1.46 g, 21% yield) with a Rf 0.11 (50% EtOAc/hexanes).
To a solution of the methyl ester (2.92 g, 6.25 mmol) in MeOH (31 mL) was added a solution of LiOH (450 mg, 18.76 mmol) in water (6.25 mL). The reaction was heated to 60° C. until LCMS showed complete hydrolysis (about 45 minutes). The MeOH was removed in vacuo and MeOH (2.5 mL) and water (1 mL) was added. The pH was adjusted to pH 5 with 1N HCl, in which the product precipitated out. The 2-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-2-methyl-propionic acid product was obtained after filtration (2.825 g, quant.).
To a solution of 2-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-2-methyl-propionic acid (1.00 g, 2.20 mmol) in DMF (5.5 mL) were added HOBT (300 mg, 2.20 mmol), EDC (633 mg, 3.30 mmol), and N,N-dimethyl-propane-1,3-diamine (225 mg, 2.20 mmol). The reaction was stirred overnight at room temperature. The reaction was then purified by reversed phase C-18 prep HPLC eluting with acetonitrile/water with 0.1% acetic acid to afford 2-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-N-(3-dimethylamino-propyl)-isobutyramide (170 mg, 14% yield).
General Procedure 6:
To a solution of 5-bromo-3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-2-ylamine (12.83 g, 33.76 mmol) in anhydrous DMF (100 mL) was added di-tert-butyl dicarbonate (21.25 g, 97.35 mmol) and 4-dimethylaminopyridine (0.793 g, 6.49 mmol). The reaction was stirred at ambient temperature for 18 hours under nitrogen. To the mixture was added saturated NaHCO3 solution (300 mL), and extracted with EtOAc (3×250 mL). The combined extracts were washed with water (5×100 mL), sat. NaHCO3, and brine, then dried over Na2SO4. After filtration, evaporation, and high vacuum drying, di-boc protected 5-bromo-3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-2-ylamine was obtained as an off-white foam solid (19.59 g, 100% yield). 1H NMR (DMSO-d6, 400 MHz) δ 8.18 (d, 1H), 7.83 (d, 1H), 7.59 (dd, 1H), 7.48 (t, 1H), 6.25 (q, 1H), 1.75 (d, 3H), 1.39 (s, 9H), 1.19 (s, 9H).
To a solution of the di-boc protected 5-bromo-3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-2-ylamine (19.58 g, 33.76 mmol) in DMSO (68 mL) was added potassium acetate (11.26 g, 114.78 mmol) and bis(pinacolato)diboron (10.29 g, 40.51 mmol). The mixture was degassed and charged with nitrogen three times, then Pd(dppf)Cl2.CH2Cl2 (1.38 g, 1.69 mmol) was added. The reaction mixture was degassed and charged with nitrogen three times, and then stirred at 80° C. oil bath under nitrogen for 12 hours. The reaction was cooled to ambient temperature, diluted with ethyl acetate (100 mL), and filtered through a celite pad which was washed with ethyl acetate. The combined ethyl acetate solution (700 mL) was washed with water (5×100 mL), brine (100 mL), and dried over Na2SO4. After filtration and concentration, the residue was purified on a silica gel column eluting with EtOAc/Hexane (0%-50%) to provide di-boc protected 3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine as a foam sold (20.59 g, 97% yield). 1H NMR (DMSO-d6, 400 MHz) δ 8.20 (d, 1H), 7.70 (d, 1H), 7.63 (dd, 1H), 7.47 (t, 1H), 6.20 (q, 1H), 1.73 (d, 3H), 1.50-1.13 (m, 30H).
To a solution of di-boc protected 3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine (20.34 g, 32.42 mmol) in CH2Cl2 (80 mL) was added a solution of dry HCl in dioxane (4N, 40.5 mL, 162 mmol). The reaction solution was stirred at 40° C. oil bath under nitrogen for 12 hours. The reaction mixture was cooled to ambient temperature, diluted with EtOAc (400 mL), then washed carefully but quickly with saturated NaHCO3 until the water layer was basic (pH>8). The organic layer was washed with brine, and dried over Na2SO4. After filtration, evaporation, and high vacuum drying, 3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine was obtained as an off-white foam solid (13.48 g, 97% yield). 1H NMR (DMSO-d6, 400 MHz) δ 8.01 (d, 1H), 7.27 (dd, 1H), 7.17 (d, 1H), 7.03 (t, 1H), 6.12 (q, 1H), 5.08 (bs, 2H), 1.81 (d, 3H), 1.30 (s, 6H), 1.28 (s, 6H).
To a stirred solution of 3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine (4.2711 g, 10.0 mmol) and 4-(4-bromo-pyrazol-1-yl)-piperidine-1-carboxylic acid tert-butyl ester (see procedure 11) (3.9628 g, 12.0 mmol) in DME (40 mL) was added a solution of Na2CO3 (3.1787 g, 30.0 mmol) in water (10 mL). The solution was degassed and charged with nitrogen three times. To the solution was added Pd(PPh3)2Cl2 (351 mg, 0.50 mmol). The reaction solution was degassed and charged with nitrogen again three times. The reaction solution was stirred at 87° C. oil bath for about 16 hours (or until consumption of the borane pinacol ester), cooled to ambient temperature and diluted with EtOAc (200 mL). The reaction mixture was filtered through a pad of celite and washed with EtOAc. The EtOAc solution was washed with brine, dried over Na2SO4, and concentrated. The crude product was purified on a silica gel column eluting with EtOAc/hexane system (0% EtOAc to 100% EtOAc) to afford 4-(4-{6-amino-5-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3 -yl}-pyrazol-1-yl)-piperidine-1-carboxylic acid tert-butyl ester (3.4167 g, 65% yield, ˜95% purity) with a Rf of 0.15 (50% EtOAc/Hexanes). MS m/e 550 (M+1)+.
To a solution of 4-(4-{6-amino-5-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-piperidine-1-carboxylic acid tert-butyl ester (566.7 mg, 1.03 mmol) in methanol (5 mL) or dichloromethane (30 mL) was added 4N HCl/dioxane (15 mL). The solution was stirred for about 1 hour or until the de-protection was complete. The solvents were evaporated and the residue was dissolved in methanol and purified on a reversed phase C-18 preparative HPLC eluting with acetonitrile/water with 0.1% acetic acid from 5% to 30% with a linear gradient. After lyophilization, 3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(1-piperidin-4-yl-1H-pyrazol-4-yl)-pyridin-2-ylamine acetate was obtained as a white solid (410 mg, 78% yield, 100% HPLC purity, 96.4% ee). 1H NMR (DMSO-d6, 400 MHz) δ 7.84 (s, 1H), 7.68 (d, 1H), 7.50 (dd, 1H), 7.46 (s, 1H), 7.37 (t, 1H), 6.83 (d, 1H), 6.02 (q, 1H), 5.57 (bs, 2H), 4.09 (m, 1H), 2.98 (m, 2H), 2.53 (m, 2H), 1.88 (m, 2H), 1.82 (s, 3H), 1.73 (d, 3H), 1.70 (m, 2H). MS m/e 450 (M+1)+.
General Procedure 7:
To a suspension of 3-[1-(2,6-Dichloro-3-fluoro-phenyl)-ethoxy]-5-(1-piperidin-4-yl-1H-pyrazol-4-yl)-pyridin-2-ylamine as the HCl salt (procedure 6) (150 mg, 0.288 mmol) in CH2Cl2 (2 mL) was added NEt3 (0.121 mL, 0.863 mmol) and stirred for 30 minutes at room temperature. The reaction was cooled to 0° C. and acetic acid chlorocarbonylmethyl ester was added and stirred for 1 hour at room temperature. The reaction was monitored by LC-MS and after complete conversion to the desired product, water (2 mL) was added. The reaction was extracted with EtOAc (4×10 mL), dried over Na2SO4, and concentrated to give quantitative yield of acetic acid 2-[4-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-piperidin-1-yl]-2-oxo-ethyl ester (164 mg, quant).
To solution of acetic acid 2-[4-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-piperidin-1-yl]-2-oxo-ethyl ester (164 mg, 0.298 mmol) in MeOH (4 mL) was added LiOH (7 mg, 0.298 mmol) dissolved in 1 mL of water. The reaction was stirred for 30 minutes at room temperature in which LC-MS showed complete conversion to the 1-[4-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-piperidin-1-yl]-2-hydroxy-ethanone. The product was purified on a reversed phase C-18 preparative HPLC eluting with acetonitrile/water having 0.1% acetic acid from 10% to 40%.
General Procedure 8:
A 100 mL of flask with a stir bar was dried in an oven and cooled in a dry nitrogen atmosphere. The flask was equipped with a rubber syringe cap. The flask was immersed in an ice-water bath under nitrogen, and 1.6 mL (1.6 mmol) of 1.0 M borane solution in THF was introduced. Then 2-(4-{5-Amino-6-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyrazin-2-yl}-pyrazol-1-yl)-2-methyl-propionic acid (procedure 5) (0.1 g, 0.221 mmol) in anhydrous THF (1.0 mL) was introduced. The resulting mixture was stirred at ambient temperature under nitrogen for 5 hours, and 6 N HCl (1.1 mL) was added slowly, and then H2O (1.1 mL) and MeOH (7.4 mL) were introduced. The reaction mixture was stirred continually overnight. Most of solvents were evaporated in vacuo, and then a 1 N NaOH solution was used to adjust pH to 11. Water was added, and the solution was extracted with EtOAc (3×30 mL) and dried over Na2SO4. After filtration and concentration, the crude product was purified with a reverse phase preparative HPLC eluting with acetonitrile/water containing 0.1% acetic acid from 10% to 60%. After lyophilization of the pure fractions, 2-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-2-methyl-propan-1-ol acetate was obtained as a white solid (21 mg, 22% yield).
General Procedure 9
To an ice cooled solution of substituted benzyl alcohol (1.0 molar equivalent) and anhydrous tetrahydrofuran (0.14 M) is added sodium hydride (1.0 molar equivalent) slowly under nitrogen atmosphere. After stirring for 30 minutes, 3,5-dibromopyrazin-2-ylamine (1.0 molar equivalent) in tetrahydrofuran (0.56 M) is added via an addition funnel at a fast dropwise rate. Once the addition is complete the ice bath is removed and the reaction refluxed under nitrogen and monitored by reversed phase HPLC. After 18 h HPLC typically shows that the majority of the starting 3,5-dibromopyrazin-2-ylamine has been consumed and the reaction is allowed to cool to room temperature. The reaction mixture is concentrated, diluted with ethyl acetate, and washed with brine. The organic layer is dried over anhydrous magnesium sulfate and concentrated in vacuum. The crude product is purified using a silica gel eluting with 1:1 ethyl acetate/dichloromethane to yield the 5-bromo-3-(substituted-benzyloxy)-pyrazin-2-ylamine as a white solid in 60-90% yield.
General Procedure 10
A mixture of 5-bromo-3-(substituted-benzyloxy)-pyridin-2-ylamine or 5-bromo-3-(substituted-benzyloxy)-pyrazin-2-ylamine (1 molar equivalent), substituted 4-pyrazolyl boronic acid or ester (1.2 molar equivalent), bis(triphenylphosphine) palladium II chloride (0.03 molar equivalent) and sodium carbonate (3.0 molar equivalent.) in ethylene glycol dimethyl ether and water (4:1, 0.2 M) is de-gassed and charged with nitrogen three times, and then heated to reflux under nitrogen overnight. The reaction is cooled to ambient temperature and diluted with ethyl acetate. The mixture is washed with water, brine, dried over Na2SO4, and purified on a silica gel column to afford 5-(substituted pyrazol-4-yl)-3-(substituted-benzyloxy)-pyridin-2-ylamine, or 5-(substituted pyrazol-4-yl)-3-(substituted-benzyloxy)-pyrazin-2-ylamine.
General Procedure 11:
To a stirred solution of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (7.94 g, 39.45 mmol) in CH2Cl2 (100 mL), cooled to 0° C., was slowly added NEt3 (5.54 mL, 39.45 mmol) followed by methane sulfonyl chloride (3.06 mL, 39.45 mmol) and DMAP (48 mg, 0.39 mmol). The mixture was stirred at room temperature overnight. To the mixture was added water (30 mL). Extraction with CH2Cl2 (3×30 mL) followed by drying (Na2SO4) and removal of the solvent in vacuo afforded 4-methanesulfonyloxy-piperidine-1-carboxylic acid tert-butyl ester as a white solid (11.00 g, >99% yield). 1H NMR (CDCl3, 400 MHz) δ 4.89 (m, 1H), 3.69 (m, 2H), 3.31 (m, 2H), 3.04 (s, 3H), 1.95 (m, 2H), 1.83 (m, 2H), 1.46 (s, 9H).
To a stirred solution of 4-bromo-pyrazole (10.44 g, 71.03 mmol) in anhydrous DMF (96 mL), cooled to 0° C., was slowly added NaH (60% in mineral oil) (3.13 g, 78.133 mmol). The solution was stirred for 1 hour at 0° C. 4-Methanesulfonyloxy-piperidine-1-carboxylic acid tert-butyl ester (19.82 g, 71.03 mmol) was added slowly and the reaction was heated to 100° C. overnight or until consumption of the pyrazole by NMR. The reaction was cooled to room temperature and water added (20 mL) followed by extraction with EtOAc. The combined extracts were washed with saturated aqueous NaCl (4×20 mL), dried with Na2SO4 and concentrated to afford 4-(4-bromo-pyrazol-1-yl)-piperidine-1-carboxylic acid tert-butyl ester as an orange oil. The oil was purified using silica gel chromatography eluting with 10% EtOAc/hexanes to 25% EtOAc/hexanes to provide 4-(4-bromo-pyrazol-1-yl)-piperidine-1-carboxylic acid tert-butyl ester as a white solid (10.55 g, 45% yield) with a Rf=0.4 (25% EtOAc/hexanes, using iodine as the stain). 1H NMR (CDCl3, 400 MHz) δ 7.46 (s, 1H), 7.43 (s, 1H), 4.23 (m, 3H), 2.88 (m, 2H), 2.10 (m, 2H), 1.88 (m, 2H), 1.47 (s, 9H).
To a solution of 4-(4-bromo-pyrazol-1-yl)-piperidine-1-carboxylic acid tert-butyl ester (500 mg, 1.515 mmol) in CH2Cl2 (3 mL) was added TFA (3 mL). The reaction was stirred at room temperature until LCMS indicated completion of the reaction. The solvents were removed in vacuo, and the residue was dissolved in MeOH (15 mL). The pH of the solution was adjusted to 9 with hydroxide resin to afford 4-(4-bromo-pyrazol-1-yl)-piperidine.
To a solution of 4-(4-bromo-pyrazol-1-yl)-piperidine (375 mg, 1.63 mmol) in DMF (3.26 mL) was added NEt3 (230 μL, 1.63 mmol) and stirred for 5 minutes. Methyliodide (MeI) (1.63 mL, 1M MeI in DMF, freshly made) was added and the reaction was stirred overnight at room temperature. Water was added and the solution was extracted with EtOAc (4×10 mL). The organic solution was washed with brine, dried with Na2SO4, concentrated, and dried in vacuo to afford 4-(4-bromo-pyrazol-1-yl)-1-methyl-piperidine (251 mg, 63% yield).
General Procedure 12:
To a solution of 3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(1H-pyrazol-4-yl)-pyrazin-2-ylamine (295 mg, 0.80 mmol) in anhydrous DMF (4 mL) was added NaH (60% in mineral oil, 30.7 mg, 0.80 mmol). The mixture was stirred at ambient temperature under nitrogen for 0.5 h, and then 4-methanesulfonyloxy-piperidine-1-carboxylic acid tert-butyl ester (223.5 mg, 0.80 mmol) was introduced. The reaction mixture was heated to 90° C. oil bath for 0.5 h under nitrogen, and cooled to ambient temperature. Water was added slowly to the mixture, which was extracted with EtOAc, washed with brine, and dried over Na2SO4. The crude product was purified on a silica gel column to provide 4-(4-{5-amino-6-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyrazin-2-yl}-pyrazol-1-yl)-piperidine-1-carboxylic acid tert-butyl ester as a white solid (265 mg, 59% yield).
To a solution of 4-(4-{5-amino-6-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyrazin-2-yl}pyrazol-1-yl)-piperidine-1-carboxylic acid tert-butyl ester (265 mg, 0.48 mmol) in CH2Cl2 was added 4N HCl/dioxane (4 mL). The mixture was stirred at ambient temperature for one hour. After evaporation, the residue was dissolved in methanol (2.5 mL), and was purified on a reverse phase C-18 reparative HPLC eluting with acetonitrile/water containing 0.1% acetic acid with a linear gradient of 10%-40%. After lyophilization, 3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(1-piperidin-4-yl-1H-pyrazol-4-yl)-pyrazin-2-ylamine acetate was obtained as a white solid (125 mg, 51% yield).
General Procedure 13:
O-(7-azabenzotriazol-1-yl)-N,N, N′,N′-tetramethyluronium phosphorus pentafluoride (HATU) (66 mg, 0.17 mmol) was added to a solution of 2-(4-{6-Amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-propionic acid (69 mg, 0.16 mmol), triethylamine (0.024 mL, 0.17 mmol) and 3-dimethylamino-propylamine (0.022 mL, 0.17 mmol) in 1.6 mL of DMF. After stirring for 3 hours, the reaction was concentrated by rotary evaporation. The residue was purified by silica gel chromatography using gradient elution of dichloromethane, methanol, ammonium hydroxide to afford 2-(4-{6-Amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-N-(3-dimethylamino-propyl)-propionamide. (41 mg, 50%).
General Procedure 14:
Compound 14-1 (1.3 molar equivalent) is added to a solution of aryl halide (0.21 mmol) in 3 mL of DME. The mixture is purged with nitrogen several times and then dichlorobis(triphenylphosphino) palladium (II) (0.05 molar equivalent) is added. Sodium carbonate (3 molar equivalent) in 0.6 mL of H2O is added to the reaction mixture and the resulting solution heated to 85° C. for 12 h. Water is added to the reaction mixture to quench the reaction. EtOAc is then added to extract the aqueous solution. Dry EtOAc layer over Na2SO4. The Na2SO4 is filtered off and the filtrate evaporated to give a dark brown oil residue. The residue is purified by silica gel chromatography (eluting with CH3OH, CH2Cl2, EtOAc, and hexanes) to give desired product, compound 14-2.
General Procedure 15:
This general procedure is illustrated for the specific example of (6-amino-3-aza-bicyclo[3.1.0]hex-3-yl)-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-phenyl)-methanone, but can be adapted to form other compounds by appropriate choice of aryl halide or heteroaryl halide ArX.
To a mixture of [3-(4-iodo-benzoyl)-3-aza-bicyclo[3.1.0]hex-6-yl]-carbamic acid tert-butyl ester (100 mg, 0.234 mmol) and 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine (100 mg, 0.234 mmol) in DME (2 mL) was added Pd(dppf)2Cl2.CH2Cl2 (10 mg, 0.012 mmol) and Cs2CO3 (351 mg, 0.702 mmol). The mixture was bubbled with nitrogen for 10 min then microwaved at 150° C. for 30 min. LCMS checked that the reaction was completed. The crude reaction mixture was diluted with ethyl acetate followed by washings with water and brine. The solution was dried over MgSO4. Purification by prep-HPLC afforded a solid. The solid was stirred with 4 N HCl/dioxane (3 mL) for 3 hr at room temperature. Removal of the volatiles led to a residue that was purified by prep-HPLC to afford (6-amino-3-aza-bicyclo[3.1.0]hex-3-yl)-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-phenyl)-methanone (30 mg, yield 26%).
General Procedure 16:
Diethylazodicarboxylate (0.48 mL, 3.1 mmol) was added to a 0° C. solution of triphenylphosphine (0.80 g, 3.1 mmol) in THF (20 mL). After stirring for 5 minutes, 4-bromo-pyrazole (0.30 mg, 2.0 mmol) was added. After another 5 minutes of stirring, (2-hydroxyethyl)-methyl-carbamic acid tert-butyl ester (0.45 g, 2.6 mmol) was added. The reaction was allowed to warm to room temperature and stir overnight. The reaction was cooled to 0° C. and filtered. The filtrate was concentrated by rotary evaporation. The residue was purified by silica gel chromatography using gradient elution of dichloromethane, ethyl acetate to afford [2-(4-bromo-pyrazol-1-yl)-ethyl]-methyl-carbamic acid tert-butyl ester (541 mg, 87%).
General Procedure 17:
Sodium hydride (0.12 g, 4.9 mmol) was added to a solution of 4-bromo-4H-pyrazole (0.60 g, 4.1 mmol) in DMF (10 mL). After stirring for 10 minutes, a solution of 2-chloro-propionic acid methyl ester in DMF (4 mL) was added. After stirring for 4 hours, the reaction was partitioned between ethyl acetate and water. The phases were separated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were dried over MgSO4 and concentrated by rotary evaporation. The residue was purified by silica gel chromatography using gradient elution of ethyl acetate and hexanes to afford 2-(4-bromo-pyrazol-1-yl)-propionic acid methyl ester (733 mg, 77%).
General Procedure 18:
A solution of LiOH (34 mg, 1.4 mmol) in water (0.4 mL) was added to a solution of 2-(4-{6-Amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-propionic acid methyl ester (70 mg, 0.15 mmol) in a mixture THF (1.5 mL) and MeOH (0.4 mL). After stirring overnight, the reaction was partitioned between dichloromethane and half-saturated brine. A small amount of ethanol was added and the pH was adjusted to 7 with 1 M HCl. The phases were separated and the aqueous phase was extracted with dichloromethane. The combined organic phases were dried over Na2SO4, filtered and concentrated by rotary evaporation to give 2-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-propionic acid (69 mg, 100%).
General Procedure 19:
To a stirred solution of 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-iodo-pyridin-2-ylamine (100 mg, 0.23 mmol) and 3-methyl-1H-pyrazole (59 mg, 0.70 mmol) in DMSO (1 mL was added K3PO4 (101 mg, 0.47 mmol), dodecane (0.015 mL, 0.05 mmol), cyclohexanediamine (0.009 mL, 0.07 mmol) and copper iodide (CuI) (14 mg, 0.07 mmol). The solution was bubbled with nitrogen for 5 minutes, then radiated with microwave at 150° C. for 2 hours, LCMS checked that the reaction was completed, the mixture was purified by prep-HPLC to leave 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(3-methyl-pyrazol-1-yl)-pyridin-2-ylamine (30 mg), yield 34.2%.
General Procedure 20:
To a stirred solution of 4-(3-{6-Amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-pyrrolidine-2-carboxylic acid methyl ester (105 mg, 0.21 mmol) in THF (5 mL) was added 2 M CH3NH2 in THF (1.06 mL, 2.12 mmol), the mixture was stirred and heated at 55° C. for 18 hours, LCMS checked that the reaction was completed, remove THF, the residue was purified by prep-HPLC to leave 4-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-pyrrolidine-2-carboxylic acid methylamide (30 mg), yield 28.6%.
General Procedure 21:
tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-1-carboxylate (21-1): Di-tert-butyl dicarbonate (7.2 molar equivalent), 4-(dimethylamino)pyridine (0.84 molar equivalent) were added to a solution of 4,4,5,5-tetramethyl-2-(1H-pyrazole-4-yl)-1,3,2-dioxaborolane (6 mmol) in 40 mL of DMF. The reaction mixture was stirred at room temperature for 12 h. Water was added to the reaction mixture to quench the reaction. EtOAc was then added to extract the aqueous solution. Dry EtOAc layer over Na2SO4. The Na2SO4 was filtered off and the filtrate was evaporated to give a brown yellow oil residue as compound 21-1 (1.32 g; 4.56 mmol; 76%). 1H NMR (400 MHz, chloroform-D) δ ppm 1.32 (s, 12H) 1.63 (s, 9H) 7.91 (s, 1 H) 8.37 (s, 1H). The residue was used for the next step reaction without further purification.
Compound 21-3, shown with the specific example of 3-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]-5-(1H-pyrazol-4-yl)pyridin-2-amine (21-3a):
Compound 21-1 (1.0 molar equivalent) was added to a solution of compound 21-2a (Compound 21-2, with R substituents to give 2,6-dichloro-3-fluorophenyl) (1.92 mmol) in 20 mL of DME. The mixture was stirred at room temperature under a nitrogen atmosphere for 30 minutes and then dichlorobis(triphenylphosphino) palladium (II) (0.05 molar equivalent) was added. Sodium carbonate (3 molar equivalent) in 4 mL of H2O was added to the reaction mixture and the resulting solution was heated to 85° C. for 12 h. Alternative bases used were CsF and Cs2CO3 in with 1 or 2 equivalents of boronic ester, and at room temperature (CsF) or 80° C. (all). Water was added to the reaction mixture to quench the reaction. EtOAc (150 mL×2) was then added to extract the aqueous solution. Dry EtOAc layer over Na2SO4. The Na2SO4 was filtered off and the filtrated was evaporated to give a dark brown oil residue. The residue was purified by silica gel chromatography (eluting with eluting with 0→10% MeOH in ethyl acetate) to give the desired product, compound 21-3a (2.05 g, 53.6% yield). 1H NMR (400 MHz, chloroform-D) δ ppm 1.60 (s, 1H) 1.84 (d, J=6.57 Hz, 3H) 5.07 (s, 2H) 6.06 (q, J=6.57 Hz, 1H) 6.89 (d, J=1.77 Hz, 1H) 6.96-7.06 (m, 1H) 7.22-7.33 (m, 1H) 7.67 (s, 2H) 7.80 (d, J=1.52 Hz, 1H).
To make compounds of formula 21-4, the following exemplary procedure can be used: sodium hydride (1.2 molar equivalent) is added to a solution of compound 21-3 (0.87 mmol) in 10 mL of DMF. The mixture is stirred at room temperature under a nitrogen atmosphere for 30 min and then compound 21-6 (1 molar equivalent) is added. The resulting solution is heated to 85-90° C. for 12 h. Water (20 mL) is added to the reaction mixture to quench the reaction. EtOAc (50 mL×2) is then added to extract the aqueous solution. Dry EtOAc layer over Na2SO4. The Na2SO4 is filtered off and the filtrate is evaporated. The residue is purified by silica gel chromatography (eluting with EtOAc in hexanes) to give the desired product, compound 21-4 (20-50% yield).
General Procedure 22:
L=Br, Cl, COOH, COCl, OMs, ethylene carbonate, aldehyde
Compounds of formula 22-3 can be prepared by the following exemplary procedure: Compound 22-2 (1.2 molar equivalent) is added to a solution of compound 22-1 (0.24 mmol) and base (3-5 molar equivalent) and/or coupling reagent (1 molar equivalent) in 5 mL of DMF. The mixture is stirred under a nitrogen atmosphere for 12 h. Water (20 mL) is added to the reaction mixture to quench the reaction. EtOAc (50 mL×2) is then added to extract the aqueous solution. Dry EtOAc layer over Na2SO4. The Na2SO4 is filtered off and the filtrate evaporated. The residue is purified by silica gel chromatography (eluting with CH3OH, CH2Cl2, EtOAc, and hexanes) to give the desired product, compound 22-3.
General Procedure 23:
The following procedure can be used to prepare piperidine-pyrazole-2-aminopyridine derivatives.
NaH (1.2 eq., 0.68 mmol) was added portionwise to a stirred solution of 4-iodopyrazole (0.57 mmol) in DMF (2 L) at 4° C. The resulting mixture was stirred for 1 hour at 4° C. and compound 23-4 (1.1 eq., 0.63 mmol) was then added. The resulting mixture was heated to 100° C. for 12 h. The reaction was quenched with H2O and extracted with EtOAc several times. The combined organic layers were dried, filtered, and concentrated to afford an orange oil. The residue was purified by silica gel chromatography (eluting with 5% EtOAc in pentane) to give compound 23-1a as a white solid (140 g, 66%).
Bis(pinacolato)diboron (1.4 eq., 134 g, 0.52 mol) and potassium acetate (4 eq., 145 g, 1.48 mol) were added sequentially to a solution of compound 23-1a (140 g, 0.37 mol) in 1.5 L of DMSO. The mixture was purged with nitrogen several times and dichlorobis(triphenylphosphino) palladium (II) (0.05 eq., 12.9 g, 0.018 mol) was then added. The resulting mixture was heated at 80° C. for 2 h. The reaction mixture was cooled to room temperature and filtered through a bed of celite and washed with EtOAc. The filtrate was washed with saturated NaCl (500 mL×2), dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel chromatography (eluting with 5% EtOAc in hexanes) to give compound 23-1b as a white solid (55 g, 40%).
Compound 23-2 (1.0 molar equivalent) was added to a solution of compound 23-1b (1.3 molar equivalent) in 15 mL of DME. The mixture was purged with nitrogen several times and then dichlorobis(triphenylphosphino) palladium (II) (0.05 molar equivalent) was added. Cesium carbonate (3 molar equivalent) in 4 mL of H2O was added to the reaction mixture and the resulting solution was heated to 85° C. for 12 h. Water (10 mL) was added to the reaction mixture to quench the reaction. EtOAc (150 mL×2) was then added to extract the aqueous solution. Dry EtOAc layer over Na2SO4. The Na2SO4 was filtered off and the filtrated was evaporated to give a dark brown oil residue. The residue was purified by silica gel chromatography (eluting with eluting with 75→100% EtOAc in hexanes) to give compound 23-3a (61% yield).
Hydrochloride (19 eq., 12 mmol) was added to a solution of compound 23-3a (0.63 mmol) in MeOH (4 mL). The mixture was stirred at room temperature for 12 h. The solvent was evaporated and H2O (10 mL) was added. Saturated NaHCO3 (aq) was added to neutralize the solution to pH 7. Ethyl acetate (100 mL×2) was added to extract the aqueous solution. The combined organic layer was dried over Na2SO4, filtered, and evaporated to give compound 23-5a as a solid reside (0.6 mmol, 95% yield).
Compounds of formula 23-7 can be formed according to the following general procedure: Compound 23-8 (1.2 molar equivalent) is added to a solution of compound 23-5a (0.24 mmol) and base (3-5 molar equivalent) and/or coupling reagent (1 molar equivalent) in 5 mL of DMF. The mixture is stirred under a nitrogen atmosphere for 12 h. Water (20 mL) is added to the reaction mixture to quench the reaction. EtOAc (50 mL×2) is then added to extract the aqueous solution. Dry EtOAc layer over Na2SO4. The Na2SO4 is filtered off and the filtrated is evaporated to give an oil residue. The residue is purified by silica gel chromatography (eluting with CH3OH, CH2Cl2, EtOAc, and hexanes) to give the desired product, compound 23-7a.
General Procedure 24:
3-methoxy compounds can be prepared from the corresponding 3-fluoro compounds by the following general procedure. To 4 mL of DMSO is added 0.124 mL ethanol followed by 32 mg NaH. After stirring for 30 minutes 250 mg of 24-1 is added and the reaction heated to 40° C. After three hours the reaction is cooled and poured into water to precipitate. After neutralization to pH 6, the product 24-2 is isolated.
General Procedure 25:
To a stirred solution of 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-[1-(2,2-dimethyl-[1,3]dioxolan-4-ylmethyl)-1H-pyrazol-4-yl]-pyridin-2-ylamine (150 mg, 0.31 mmol) in THF (3 mL) and H2O (2 mL) was added TFA (2 mL) at 0° C., the mixture was stirred and warmed to room temperature, then heated at 50° C. for 5 hours, LCMS checked that the reaction was completed, remove THF, the residue was purified by prep-HPLC to leave 3-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-propane-1,2-diol (102 mg), yield 74.2%.
General Procedure 26:
To a stirred solution of 4-bromo-1H-pyrazole in DMF was added sodium hydride at room temperature. The mixture was stirred for 30 minutes, [1,3]dioxolan-2-one was added, the mixture was stirred and slowly warmed to room temperature. The reaction was monitored by TLC. After the reaction was done, EtOAc was added, washed with saturated NaHCO3, water and brine, dried with Na2SO4, filtered and concentrated. The residue was purified by silica gel, eluants EtOAc and DCM 10%, to give 2-(4-Bromo-pyrazol-1-yl)-ethanol 0.22 g, yield 34%. 1H NMR (400 MHz, chloroform-D) δ ppm 7.49 (s, 1H) 7.46 (s, 1H) 4.18-4.23 (m, 2H) 3.93-3.98 (m, 2H) 3.09 (s, 1 H).
General Procedure 27
In a dry round bottom flask is added the pyrazole (1 eq) and NaH (1 eq) in anhydrous DMF (0.2 M). The desired nucleophile is added (1 eq) and the reaction is heated for 17 hours at 90° C. To the reaction mixture is added water (20 mL) and EtOAc (20 mL). The aqueous layer is extracted with EtOAc (4×20 mL) followed by drying (Na2SO4) and removal of the solvent in vacuo to give the desired crude product. Purify by silica gel column chromatography.
General Procedure 28
To a stirred solution of 2,6-dichloro-3,5-difluoro-phenol (25 g, 125.62 mmol) in CH2Cl2 (250 mL), cooled to 0° C., was slowly added N-methyl morpholine (21 mL, 188.43 mmol) followed by trifluoromethane sulfonic anhydride (32 mL, 188.43 mmol). The mixture was stirred at room temperature overnight. To the mixture was added water (50 mL). Extraction with CH2Cl2 (3×50 mL) followed by drying (Na2SO4) and removal of the solvent in vacuo afforded 28-2 as an oil (42.18 g, >99% yield). 1H NMR (CDCl3, 400 MHz) δ 7.15 (t, 1H)
To a stirred solution of trifluoro-methanesulfonic acid 2,6-dichloro-3,5-difluoro-phenyl ester 28-2 (42.18 g, 127.43 mmol) and 4,4,5,5-tetramethyl-2-vinyl-[1,3,2]dioxaborolane (21.59 g, 140.17 mmol) in DME (200 mL), was added Na2CO3 (40.52 g, 382.29 mmol) dissolved in water (50 mL). The combined solutions were degassed 3× with N2. Palladium dichloro triphenylphosphine (1.78 g, 2.54 mmol) was added and the reaction solution was degassed again 3× with N2. The reaction was stirred for 15 hours at 70° C. The reaction was cooled to room temperature and EtOAc (50 mL) was added. The solution was filtered through a pad of celite and washed with EtOAc and water. The aqueous layer was extracted with EtOAc (3× mL). The combined organics were dried with Na2SO4 and removal of the solvent in vacuo afforded 28-3 as an orange oil. The oil was purified on a pad of silica gel with 5% EtOAc/Hexanes to obtain a clear oil (26.12 g, 99% yield). 1H NMR (CDCl3, 400 MHz) 56.96 (t, 1H), 6.66 (m, 1H), 5.85 (m, 2H).
To a solution of 28-3 (26.63 g, 127.43 mmol), in CH2Cl2 (320 mL) cooled to −78° C. was bubbled in ozone (O3) for 30 minutes until the reaction turned dark blue. The reaction was then purged with N2 for 5 minutes. Dimethyl sulfide (50 mL, 637.15 mmol) was added slowly changing the color of the reaction to yellow. Allow the reaction to warm to room temperature. Wash the organic with water (3×50 mL). The organics were dried with Na2SO4 and removal of the solvent in vacuo afforded 28-4 as an orange oil (17.13 g, 65% yield). 1H NMR (CDCl3, 400 MHz) δ 10.43 (s, 1H), 7.24 (t, 1H).
In a dry round bottom flask purged with N2 was added 28-4 (17.13 g, 81.85 mmol) in THF (109 mL) cooled to 0° C. was added MeMgBr slowly (64.3 mL, 90.04 mmol, 1.4M in THF). The reaction was allowed to stir for 15 hours at room temperature. Saturated aqueous NH4Cl (30 mL) and EtOAc (30 mL) were added and the aqueous layer was then extracted with EtOAc (3×30 mL). The combined organics were dried with Na2SO4 and removal of the solvent in vacuo afforded 28-5 as an orange oil (16.9 g, 92% yield). 1H NMR (CDCl3, 400 MHz) δ 6.98 (t, 1H), 5.62 (m, 1H), 1.65 (d, 3H).
To a stirred solution of 28-5 (16.90 g, 75.169 mmol) in CH2Cl2 (150 mL), cooled to ° C., was slowly added NEt3 (10.7 mL, 75.169 mmol) followed by methane sulfonyl chloride (5.94 mL, 75.169 mmol) and DMAP (92 mg, 0.075 mmol). The mixture was stirred at room temperature overnight. To the mixture was added water (50 mL). Extraction with CH2Cl2 (3×50 mL) followed by drying (Na2SO4) and removal of the solvent in vacuo afforded 28-6 (21.48 g, 95% yield). 1H NMR (CDCl3, 400 MHz) δ 7.07 (t, 1H), 6.48 (m, 1H), 2.96 (s, 3H).
To a stirred solution of 28-6 (1.15 g, 3.78 mmol) and 2-amino-5-bromo-pyridin-3-ol (3.78 g, 3.78 mmol) in DMF (8 mL, 0.5 M) was added Cs2CO3 (1.23 g, 3.78 mmol). The reaction mixture was stirred for 17 hours at 60° C. The reaction was cooled to room temperature and water (20 mL) was added. The solution was extracted with EtOAc (4×20 mL), dried with Na2SO4 and removal of the solvent in vacuo. The crude reaction was purified by silica gel column chromatography to afford a light yellow solid 28-7 (600 mg, 41% yield). 1H NMR (CDCl3, 400 MHz) δ 7.69 (s, 1H), 7.04 (t, 7.04), 6.01 (m, 1H), 4.80 (bs, 2H), 1.82 (d, 3H).
To a stirred solution of 28-7 (395 mg, 1 mmol) and dioxaborolane (565 mg, 1.5 mmol) in DME (200 mL), was added Cs2CO3 (975 mg, 3 mmol) dissolved in water (1 mL). The combined solutions were degassed 3× with N2. Pd(dppf)2CH2Cl2 (41 g, 0.05 mmol) was added and the reaction solution was degassed again 3× with N2. The reaction was stirred for 15 hours at 70° C. The reaction was cooled to room temperature and EtOAc (25 mL) was added. The solution was filtered through a pad of celite and washed with EtOAc and water. The aqueous layer was extracted with EtOAc (3×25 mL). The combined organics were dried with Na2SO4 and removal of the solvent in vacuo afforded 28-8 as brown solid. The crude product was purified by solica gel chromatography (436 mg, 77% yield). The BOC product was dissolved in CH2Cl2 (5 mL) and add 4M HCl/Dioxane (2 mL). The reaction was stirred for 1 hour until LCMS shows complete deprotection. Add water (10 mL) and separate CH2Cl2 layer. Wash organics with water (3×10 mL). Combined aqueous layer was neutralized with Na2CO3 to pH 7 and extracted with CH2Cl2 (4×10 mL), dried with Na2SO4 and remove of the solvent in vacuo (quantitative yield). 1HNMR (400 MHZ, DMSO) δ 7.92 (s, 1H), 7.81 (m, 1H), 7.76 (s, 1H), 7.53 (s, 1H), 6.90 (s, 1H), 6.12 (m, 1H), 5.65 (bs, 2H), 4.12 (m, 1H), 3.00 (m, 2H), 2.66 (m, 2H), 1.90 (m, 2H) 1.80 (d, 3H), 1.70 (m, 2H).
General Procedure 29
In an oven-dried microwave vessel was added a nucleophile (OH, alkoxy, or amine) (0.2 mmol) and base (NaH, 0.2 mmol) in anhydrous DMF. The reaction vessel was capped and heated in the microwave for 30 min at 120° C. After complete displacement, to the reaction mixture was added water (20 mL) and EtOAc (20 mL). The aqueous layer was extracted with EtOAc (4×20 mL). The organics were combined and washed with water (3×20 mL), dried with Na2SO4 and remove of the solvent in vacuo. Purify by HPLC. For, compounds containing a BOC protecting group (0.2 mmol) add CH2Cl2 (5 mL) and add 4M HCl/Dioxane (2 mL). The reaction was stirred for 1 hour until LCMS shows complete deprotection. Add water (10 mL) and separate CH2Cl2 layer. Wash organics with water (3×10 mL). Combined aqueous layer was neutralized with Na2CO3 to pH 7 and extracted with CH2Cl2 (4×10 mL), dried with Na2SO4 and remove of the solvent in vacuo. Purify by HPLC.
The title compound was prepared according to procedure 1 (compound 1-9). 1H NMR (400 MHz, DMSO-D6) δ ppm 1.25-1.33 (m, 9H) 1.69-1.77 (m, 3H) 2.80-2.97 (m, 1H) 3.60 (s, 2H) 3.81 (s, 2H) 4.22 (d, J=7.07 Hz, 2H) 5.59 (s, 2H) 6.01 (q, J=6.32 Hz, 1H) 6.81 (d, J=1.26 Hz, 1H) 7.37 (t, J=8.84 Hz, 1H) 7.43-7.54 (m, 1H) 7.55-7.62 (m, 1H) 7.67 (d, J=1.52 Hz, 1H) 7.84 (s, 1H); LCMS: 537 [M+1]; c-Met Ki: 0.066 μM.
The title compound was prepared according to procedure 1 (compound 1-10). 1H NMR (400 MHz, MeOD) δ ppm 1.78 (d, J=6.57 Hz, 3H) 3.32 (d, J=8.08 Hz, 1H) 3.84-3.95 (m, 2H) 4.00 (t, J=9.73 Hz, 2H) 4.29 (d, J=6.82 Hz, 2H) 6.08 (d, J=6.57 Hz, 1H) 6.83 (s, 1H) 7.14 (t, J=8.59 Hz, 1H) 7.35 (dd, J=8.84, 4.80 Hz, 1H) 7.48 (s, 1H) 7.57 (s, 1H) 7.69 (s, 1H); LCMS: 437 [M+1]; c-Met Ki: 0.044 μM.
The title compound was prepared according to procedure 3 (compound 3-6). 1H NMR (400 MHz, DMSO-D6) δ ppm 1.34-1.39 (m, 9H) 1.70-1.77 (m, 2H) 1.79 (d, J=6.57 Hz, 3H) 3.06 (d, J=12.63 Hz, 2H) 3.62 (s, 2H) 4.03 (s, 2H) 4.79 (s, 1H) 5.66 (s, 2H) 6.08 (d, J=6.82 Hz, 1H) 6.86 (d, J=1.52 Hz, 1H) 7.44 (t, J=8.72 Hz, 1H) 7.51-7.58 (m, 2H) 7.58-7.65 (m, 2H) 7.73 (d, J=1.52 Hz, 1H) 7.78 (s, 1H); LCMS: 581 [M+1]; c-Met Ki: 0.104 μM.
The title compound was prepared according to procedure 3 (compound 37). 1H NMR (400 MHz, DMSO-D6) δ ppm 1.41-1.55 (m, 2H) 1.59-1.71 (m, 2H) 1.81 (d, J=6.57 Hz, 3H) 2.88-3.00 (m, 2H) 3.02-3.14 (m, 2H) 4.08 (s, 2H) 5.17 (s, 2H) 6.14-6.27 (m, J=6.57 Hz, 1H) 7.05 (s, 1 H) 7.40-7.49 (m, J=8.72, 8.72 Hz, 1H) 7.51-7.60 (m, J=9.09, 4.80 Hz, 1H) 7.63 (s, 1H) 7.76 (s, 1H) 7.91 (s, 1H) 8.51 (s, 1H) 8.81 (s, 1H); LCMS: 481 [M+1]; c-Met Ki: 0.062 μM.
The title compound was prepared according to procedure 2 (compound 2-11). 1H NMR (400 MHz, DMSO-D6) δ ppm 1.79-1.89 (m, 3H) 3.56 (s, 1H) 4.35 (s, 4H) 5.40 (s, 1H) 6.23 (d, J=6.57 Hz, 2H) 7.09 (s, 1H) 7.40-7.54 (m, 1H) 7.59 (dd, J=8.84, 5.05 Hz, 1H) 7.73-7.83 (m, 1H) 7.86 (s, 1H) 8.12 (s, 1H) 9.20 (s, 1H); LCMS: 523 [M+1]; c-Met Ki: 0.033 μM.
The title compound was prepared according to procedure 2. 1H NMR (400 MHz, DMSO-D6) δ ppm 0.12 (s, 2H) 0.41 (s, 2H) 0.76 (s, 1H) 1.79 (d, J=6.57 Hz, 3H) 2.27-2.40 (m, J=2.02 Hz, 2 H) 2.44 (d, J=1.77 Hz, 2H) 3.74 (s, 2H) 4.94 (s, 1H) 5.66 (s, 2H) 5.99-6.17 (m, 1H) 6.89 (s, 1 H) 7.43 (t, J=8.72 Hz, 1H) 7.52-7.65 (m, 2H) 7.75 (s, 1H) 8.02 (s, 1H); LCMS: 477 [M+1]; c-Met Ki: 0.038 μM.
The title compound was prepared according to procedure 2. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.78 (d, J=6.82 Hz, 3H) 2.17 (s, 6H) 2.89-2.98 (m, 2H) 4.07-4.16 (m, 1H) 4.30 (t, J=9.09 Hz, 1H) 4.36-4.45 (m, 1H) 4.60 (t, J=8.59 Hz, 1H) 5.15-5.27 (m, 1H) 5.64-5.73 (m, 2H). 6.08 (q, J=6.48 Hz, 1H) 6.90 (s, 1H) 7.38-7.48 (m, J=8.72, 8.72 Hz, 1H) 7.50-7.61 (m, J=8.84, 5.05 Hz, 1H) 7.66 (s, 1H) 7.71-7.79 (m, 1H) 8.05 (s, 1H); LCMS: 508 [M+1]; c-Met Ki: 0.022 μM.
The title compound was prepared according to procedure 2. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.79 (d, J=6.82 Hz, 3H) 3.59 (q, J=6.57 Hz, 2H) 3.68-3.79 (m, 4H) 5.00 (t, J=6.95 Hz, 1 H) 5.72 (s, 2H) 6.03-6.13 (m, J=6.57 Hz, 1H) 6.89 (d, J=1.52 Hz, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.56 (dd, J=9.09, 5.05 Hz, 1H) 7.62 (s, 1H) 7.74 (d, J=1.52 Hz, 1H) 8.01 (s, 1H); LCMS: 462 [M+1]; c-Met Ki: 0.025 μM.
The title compound was prepared according to procedure 4 (compound 4-6). 1H NMR (400 MHz, DMSO-D6) δ ppm 0.96-1.10 (m, 2H) 1.15 (t, J=7.07 Hz, 2H) 1.74 (s, 3H) 1.79 (d, J=6.57 Hz, 3 H) 3.95-4.14 (m, 4H) 5.66 (s, 2H) 6.08 (d, J=6.57 Hz, 1H) 6.88 (s, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.49-7.62 (m, 2H) 7.73 (s, 1H) 7.88 (s, 1H); LCMS: 494 [M+1]; c-Met Ki: 0.042 μM.
The title compound was prepared according to procedure 4 (compound 4-8). 1H NMR (400 MHz, DMSO-D6) δ ppm 0.72-0.82 (m, 1H) 0.86-0.95 (m, 1H) 1.18-1.32 (m, 1H) 1.51-1.66 (m, 2 H) 1.82 (d, J=6.57 Hz, 3H) 2.53-2.58 (m, 3H) 3.98-4.10 (m, 2H) 6.18 (d, J=6.32 Hz, 1H) 7.09 (s, 1H) 7.22 (s, 1H) 7.34 (s, 1H) 7.46 (t, J=8.59 Hz, 1H) 7.54-7.63 (m, 1H) 7.76 (s, 1H) 7.97 (s, 1H) 8.04 (d, J=4.55 Hz, 1H); LCMS: 479 [M+1]; c-Met Ki: 0.071 μM.
The title compound was prepared according to procedure 4, using dimethylamine in the final step. 1H NMR (400 MHz, DMSO-D6) δ ppm 0.78-0.90 (m, 1H) 0.94-1.07 (m, 1H) 1.56 (s, 1H) 1.81-1.90 (m, 3H) 1.97-2.11 (m, 1H) 2.77 (s, 3H) 3.02 (d, J=4.80 Hz, 3H) 4.01-4.08 (m, 1H) 4.08-4.22 (m, 1H) 6.95 (s, 1H) 7.04-7.14 (m, 2H) 7.21 (s, 1H) 7.48 (t, J=8.72 Hz, 1H) 7.57-7.62 (m, J=9.22, 4.93 Hz, 1H) 7.64 (d, J=2.78 Hz, 1H) 7.70 (d, J=1.26 Hz, 1H) 7.98-8.08 (m, 1H); LCMS: 492 [M+1]; c-Met Ki: 0.144 μM.
The title compound was prepared according to procedure 5 (compound 5-2). 1H NMR (400 MHz, DMSO) δ 8.10 (s, 1H), 7.83 (s, 1H), 7.60 (m, 3H), 7.46 (m, 1H), 6.98 (m, 1H), 6.15 (m, 1H), 1.84 (d, 2H), 1.75 (s, 6H); LCMS: 453 [M+1]; c-Met Ki: 0.18 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-bromo-2-methyl-2H-pyrazol-3-ylamine. 1H NMR (400 MHz, CDCl3) δ 7.42 (s, 1H), 7.35 (m, 1H), 7.29 (s, 1H), 6.95 (m, 1H), 6.15 (m, 1H), 3.75 (s, 3H), 1.90 (d, 2H); LCMS: 396 [M+1]; c-Met Ki: 0.13 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-bromo-5-methyl-1H-pyrazole. 1H NMR (400 MHz, CDCl3) δ 7.34 (m, 2H), 7.15 (m, 2H), 6.88 (s, 1H), 6.13 (m, 1H), 2.27 (s, 3H), 1.93 (d, J=8 Hz, 3H); LCMS: 380 [M+1]; c-Met Ki: 0.70 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-bromo-3,5-dimethyl-1H-pyrazole. 1H NMR (400 MHz, CDCl3) δ 7.35 (m, 1H), 7.20 (s, 1H), 7.14 (m, 1H), 6.7 (s, 1H), 6.10 (m, 1H), 2.13 (s, 6H), 1.92 (d, J=8 Hz, 3H); LCMS: 395 [M+1]; c-Met Ki: 0.15 μM.
The title compound was prepared according to procedure 5 (compound 5-3). 1H NMR (400 MHz, CDCl3) δ 7.71 (s, 1H), 7.63 (s, 1H), 7.46 (s, 1H), 7.35 (m, 1H), 7.13 (m, 2H), 7.00 (m, 1H), 6.18 (m, 1H), 3.32 (m, 2H), 3.01 (m, 2H), 2.78 (s, 6H), 1.96 (m, 2H), 1.94 (m, 2H), 1.84 (s, 6H); LCMS: 537 [M+1]; c-Met Ki: 0.06 μM.
The title compound was prepared according to procedure 5 using piperazine-1-carboxylic acid tert-butyl ester as the amine in the final step followed with conventional method to remove the Boc-protecting group using 20% TFA in dichloromethane. 1H NMR (400 MHz, CDCl3) δ 7.63 (s, 1H), 7.58 (s, 1H), 7.51 (m, 1H), 7.35 (m, 1H), 7.13 (m, 1H), 7.02 (s, 1H), 6.17 (m, 1H), 2.90 (m, 4H), 1.93 (d, J=8 Hz, 2H), 1.80 (s, 6H); LCMS: 521 [M+1]; c-Met Ki: 0.44 μM.
The title compound was prepared according to procedure 5 using (R)-pyrrolidin-3-ol as the amine in the final step. 1H NMR (400 MHz, CDCl3) δ 7.60 (m 2H), 7.50 (s, 1H), 7.31 (m, 1H), 7.14 (m, 1H), 7.04 (s, 1H), 6.17 (m, 1H), 4.37 (m, 1H), 3.70 (m, 2H), 2.65 (m, 2H0, 1.94 (d, J=8 Hz, 3H), 1.81 (m, 8H); LCMS: 522 [M+1]; c-Met Ki: 0.27 μM.
The title compound was prepared according to procedure 5 using (2-morpholin-4-ylethyl)amine as the amine in the final step. 1H NMR (400 MHz, CDCl3) δ 7.69 (m, 2H), 7.58 (s, 1H), 7.47 (s, 1H), 7.34 (m, 1H), 7.13 (m, 1H), 7.08 (s, 1H), 6.18 (m, 1H), 3.95 (m, 4H), 3.65 (m, 2H), 3.50 (m, 2H), 3.19 (m, 2H), 2.90 (m, 2H), 1.94 (d, J=8 Hz, 3H), 1.83 (s, 6H); LCMS: 565 [M+1]; c-Met Ki: 0.14 μM.
The title compound was prepared according to procedure 5 The title compound was prepared according to procedure 5 using 2-iodo-propane as alkylation reagent. 1H NMR (400 MHz, CDCl3) δ 7.53 (s, 1H), 7.48 (s, 1H), 7.43 (s, 1H), 7.35 (m, 1H), 7.13 (m, 1H), 7.02 (s, 1H), 6.15 (m, 1H), 4.50 (m, 1H), 1.93 (d, J=8 Hz, 3H), 1.52 (d, J=4 Hz, 6H); LCMS: 409 [M+1]; c-Met Ki: 0.04 μM.
The title compound was prepared according to procedure 5 using (2-pyrrolidin-1-ylethyl)amine as the amine in the final step. 1H NMR (400 MHz, CDCl3) δ 7.70 (s, 1H), 7.61 (s, 1H), 7.46 (m, 1H), 7.35 (m, 1H), 7.13 (m, 1H), 7.09 (m, 1H), 6.18 (m, 1H), 3.79 (m, 2H), 3.62 (m, 1H), 3.24 (m, 1H), 2.81 (m, 1H), 2.10 (m, 4H), 1.93 (d, J=8 Hz, 3H), 1.84 (s, 6H); LCMS: 549 [M+1]; c-Met Ki: 0.06 μM.
The title compound was prepared according to procedure 5 using 1-Iodo-2-methyl-propane as alkylation reagent. 1H NMR (400 MHz, CDCl3) δ 7.52 (m, 1H), 7.48 (m, 1H), 7.40 (m, 1H), 7.35 (m, 1H), 7.11 (m, 1H), 7.00 (m, 1H), 6.15 (m, 1H), 3.91 (d, J=8 Hz, 2H), 1.92 (d, J=8 Hz, 3H), 0.92 (d, J=8 Hz, 6H); LCMS: 423 [M+1]; c-Met Ki: 0.10 μM.
The title compound was prepared according to procedure 5 using 1-iodo-2,2-dimethyl-propane as alkylation reagent. 1H NMR (400 MHz, CDCl3) δ 7.51 (s, 1H), 7.54 (s, 1H), 7.39 (s, 1H), 7.35 (m, 1H), 7.14 (m, 1H), 7.01 (m, 1H), 6.16 (m, 1H), 3.91 (s, 2H), 1.93 (d, J=8 Hz, 3H), 0.97 (s, 9H); LCMS: 437 [M+1]; c-Met Ki: 0.12 μM.
The title compound was prepared according to procedure 5 using N,N-diethylethane-1,2-diamine as the amine in the final step. 1H NMR (400 MHz, CDCl3) δ 7.69 (s, 1H), 7.65 (m, 1H), 7.52 (s, 1H), 7.40 (m, 1H), 7.31 (s, 1H), 7.21 (m, 1H), 7.15 (m, 1H), 7.09 (m, 1H), 7.00 (m, 1H), 6.19 (m, 1H), 3.69 (m, 2H), 3.40 (m, 2H), 3.21 (m, 4H), 2.81 (m, 6H), 1.93 (d, 2H), 1.80 (s, 6H); LCMS: 551 [M+1]; c-Met Ki: 0.22 μM.
The title compound was prepared according to procedure 5 using N,N-dimethylethane-1,2-diamine as the amine in the final step. 1H NMR (400 MHz, CDCl3) δ 7.68 (m, 2H), 7.63 (s, 1H), 7.30 (m, 1H), 7.07 (m, 1H), 6.95 (m, 1H), 6.86 (m, 1H), 6.07 (m, 1H), 3.45 (m, 2H), 2.79 (m, 2H), 2.50 (s, 6H), 2.05 (s, 3H), 1.85 (m, 9H); LCMS: 523 [M+1]; c-Met Ki: 0.06 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-(4-bromo-pyrazol-1-ylmethyl)-tetrahydro-pyran-4-ol (prepared according to general procedure 11 starting from 1,6-dioxa-spiro[2.5]octane). 1H NMR (400 MHz, CDCl3) δ 7.67 (s, 1H), 7.59 (s, 1H), 7.45 (s, 1H), 7.31 (m, 1H), 7.08 (m, 1H), 6.86 (s, 1H), 6.07 (m, 1H), 5.43 (m, 2H), 4.08 (s, 2H), 3.77 (m, 5H), 2.09 s, 3H), 1.86 (d, 2H), 1.62 (m, 3H), 1.37 (m, 2H); LCMS: 481 [M+1]; c-Met Ki: 0.04 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-Bromo-1-(tetrahydro-furan-3-yl)-1H-pyrazole (prepared according to general procedure 11 starting from tetrahydro-furan-3-ol). 1H NMR (400 MHz, CDCl3) δ 7.75 (s, 1H), 7.55 (m, 2H), 7.30 (m, 1H), 7.05 (m, 1H), 6.86 (m, 1H), 6.09 (m, 1H), 5.00 (m, 1H), 7.84 (m, 1H), 4.10 (m, 2H), 4.06 (m, 1H), 2.50 (m, 1H), 2.30 (m, 1H), 1.86 (d, J=8 Hz, 3H); LCMS: 437 [M+1]; c-Met Ki: 0.04 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-Bromo-1-(7-oxa-bicyclo[2.2.1]hept-2-ylmethyl)-1H-pyrazole (prepared according to general procedure 11 starting from (7-oxa-bicyclo[2.2.1]hept-2-yl)-methanol). 1H NMR (400 MHz, CDCl3) δ 7.73 (s, 1H), 7.65 (s, 1H), 7.55 (s, 1H), 7.30 (m, 1H), 7.05 (m, 1H), 6.85 (m, 1H), 6.10 (m, 1H), 4.91 (m, 2H), 4.60 (m, 1H), 4.40 (m, 1H), 4.10 (m, 2H), 2.65 (m, 1H), 2.10 (m, 1H), 2.00 (m, 4H), 1.86 (d, 2H), 1.56 (m, 1H), 1.10 (m, 1H); LCMS: 477 [M+1]; c-Met Ki: 0.06 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-bromo-1-(tetrahydro-pyran-4-yl)-1H-pyrazole (prepared according to general procedure 11 starting from tetrahydro-pyran-4-ol). 1H NMR (400 MHz, CDCl3) δ 7.55 (m, 1H), 7.50 (m, 1H), 7.47 (m, 1H), 7.40 (m, 1H), 7.14 (m, 1H), 7.03 (m, 1H), 6.17 (m, 1H), 4.35 (m, 1H), 4.12 (m, 2H), 3.57 (m, 2H), 2.10 (m, 4H), 1.94 (d, 3H); LCMS: 451 [M+1]; c-Met Ki: 0.04 μM.
The title compound was prepared according to procedure 6. 1H NMR (400 MHz, CDCl3) δ 7.65 (s, 1H), 7.54 (s, 1H), 7.50 (s, 1H), 7.40 (m, 1H), 7.15 (m, 1H), 7.02 (s, 1H), 6.15 (m, 1H), 4.45 (m, 1H), 3.56 (m, 2H), 3.15 (m, 2H), 2.35 (m, 4H), 1.93 (d, J=8 Hz, 3H); LCMS: 450 [M+1]; c-Met Ki: 0.02 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 3-((R)-4-bromo-pyrazol-1-yl)-pyrrolidine-1-carboxylic acid tert-butyl ester (prepared according to general procedure 11 starting from (R)-3-Hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester). 1H NMR (400 MHz, CDCl3) δ 7.64 (s, 1H), 7.55 (s, 1H), 7.45 (s, 1H), 7.31 (m, 1H), 7.14 (m, 1H), 7.00 (s, 1H), 6.16 (m, 1H), 5.09 (m, 1H), 3.78 (m, 2H), 3.65 (m, 2H), 2.60 (m, 1H), 2.41 (m, 1H), 1.94 (d, J=8 Hz, 3H); LCMS: 436 [M+1]; c-Met Ki: 0.03 μM.
The title compound was prepared according to procedure 5 using 3-aminopropan-1-ol as the amine in the final step. 1H NMR (400 MHz, CDCl3) δ 7.72 (s, 1H), 7.69 (s, 1H), 7.67 (s, 1H), 7.33 (m, 1H), 7.08 (m, 1H), 6.90 (s, 1H), 6.49 (m, 1H), 6.09 (m, 1H), 3.54 (m, 2H), 3.34 (m, 2H), 2.10 (s, 3H), 1.87 (m, 9H), 1.62 (m, 2H); LCMS: 510 [M+1]; c-Met Ki: 0.07 μM.
The title compound was prepared according to procedure 5 (compound 5-2). 1H NMR (400 MHz, CDCl3) δ 7.54 (s, 1H), 7.51 (s, 1H), 7.50 (m, 1H), 7.49 (m, 2H), 6.01 (m, 1H), 5.03 (bs, 2H), 1.95 (s, 3H), 1.75 (d, 3H), 1.63 (s, 6H); LCMS: 467 [M+1]; c-Met Ki: 0.13 μM.
The title compound was prepared according to procedure 5 using (2-methoxyethyl)amine as the amine in the final step. 1H NMR (400 MHz, CDCl3) δ 7.73 (s, 1H), 7.69 (s, 1H), 7.66 (s, 1H), 7.31 (m, 1H), 7.08 (m, 1H), 7.05 (s, 1H), 6.45 (m, 1H), 6.10 (m, 1H), 5.10 (m, 2H), 3.36 (s, 3H), 3.25 (m, 4H), 1.87 (m, 9H); LCMS: 510 [M+1]; c-Met Ki: 0.09 μM.
The title compound was prepared according to procedure 5 using 2-aminoethanol as the amine in the final step. 1H NMR (400 MHz, CDCl3) δ 7.75 (s, 1H), 7.70 (s, 1H), 7.67 (s, 1H), 7.31 (m, 1H), 7.09 (m, 1H), 6.89 (s, 1H), 6.56 (m, 1H), 6.08 (m, 1H), 5.0 (m, 2H), 3.65 (m, 2H), 3.36 (m, 2H), 1.87 (m, 9H); LCMS: 495 [M+1]; c-Met Ki: 0.06 μM.
The title compound was prepared according to procedure 5 using N,N-dimethylpropane-1,3-diamine as the amine in the final step. 1H NMR (400 MHz, CDCl3) δ 7.60 (s, 1H), 7.48 (s, 1H), 7.30 (m, 1H), 7.08 (m, 1H), 6.73 (s, 1H), 6.04 (m, 1H), 6.15 (m, 2H), 3.28 (m, 2H), 2.59 (m, 4H), 2.38 (s, 6H), 2.20 (s, 3H), 2.05 (s, 3H), 1.86 (d, J=8 Hz, 3H), 1.81 (s, 6H); LCMS: 551 [M+1]; c-Met Ki: 0.09 μM.
The title compound was prepared according to procedure 5 using (2-pyrrolidin-1-ylethyl)amine as the amine in the final step. 1H NMR (400 MHz, CDCl3) δ 7.59 (s, 1H), 7.48 (s, 1H), 7.30 (m, 1H), 7.07 (m, 2H), 6.04 (m, 1H), 5.10 (m, 2H), 3.48 (m, 2H), 3.00 (m, 6H), 2.18 (s, 3H), 2.06 (s, 3H), 1.93 (m, 4H), 1.84 (d, J=8 Hz, 3H), 1.82 (s, 6H); LCMS: 563 [M+1]; c-Met Ki: 0.07 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-(4-Bromo-pyrazol-1-yl)-1-isopropyl-piperidine (prepared according to general procedure 11 using 2-iodo-propane as alkylation reagent). 1H NMR (400 MHz, CDCl3) δ 7.70 (s, 1H), 7.55 (s, 1H), 7.50 (s, 1H), 7.31 (m, 1H), 7.06 (m, 1H), 6.87 (s, 1H), 5.13 (bs, 2H), 4.16 (m, 1H), 3.13 (m, 2H), 2.94 (m, 1H), 2.30 (m, 9H), 2.07 (s, 3H), 1.86 (d, J=8 Hz, 3H), 1.13 (d, J=8 Hz, 6H); LCMS: 492 [M+1]; c-Met Ki: 0.01 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-(4-bromo-pyrazol-1-yl)-1-methy-piperidine (prepared according to general procedure 11). 1H NMR (400 MHz, CDCl3) δ 7.70 (s, 1H), 7.56 (s, 1H), 7.49 (s, 1H), 7.29 (m, 1H), 7.06 (m, 1H), 6.87 (s, 1H), 6.08 (m, 1H), 5.17 (bs, 2H), 4.15 (m, 1H), 3.06 (m, 2H), 2.08 (s, 3H), 2.38 (s, 3H), 2.24 (m, 6H), 1.86 (d, J=8 Hz, 3H); LCMS: 464 [M+1]; c-Met Ki: 0.01 μM.
The title compound was prepared according to procedure 7. 1H NMR (400 MHz, CDCl3) δ 7.74 (s, 1H), 7.57 (s, 1H), 7.47 (s, 1H), 7.30 (m, 1H), 7.06 (m, 1H), 6.86 (s, 1H), 6.06 (m, 1H), 4.60 (m, 1H), 4.32 (m, 1H), 4.21 (s, 2H), 3.65 (m, 1H), 3.17 (m, 1H), 2.97 (m, 1H), 2.23 (m, 2H), 2.01 (m, 2H), 1.86 (d, J=8 Hz, 3H); LCMS: 508 [M+1]; c-Met Ki: 0.01 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 1-[4-(4-bromo-pyrazol-1-yl)-piperidin-1-yl]-ethanone (prepared from 4-(4-bromo-pyrazol-1-yl)-piperidine and acetyl chloride in the presence of triethylamine in dichloromethane solvent). 1H NMR (400 MHz, CDCl3) δ 7.71 (s, 1H), 7.57 (s, 1H), 7.47 (s, 1H), 7.30 (m, 1H), 7.07 (m, 1H), 6.87 (s, 1H), 6.07 (m, 1H), 5.08 (bs, 2H), 4.76 (m, 1H), 4.33 (m, 1H), 3.95 (m, 1H), 3.25 (m, 1H), 2.78 (m, 1H), 2.25 (m, 2H), 2.15 (s, 3H), 1.95 (m, 2H), 1.86 (d, J=8 Hz, 3H); LCMS: 492 [M+1]; c-Met Ki: 0.01 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and [4-(4-Bromo-pyrazol-1-yl)-piperidin-1-yl]-acetonitrile (prepared according to general procedure 11 using 2-bromo-acetonitile as alkylation reagent). 1H NMR (400 MHz, CDCl3) δ 7.75 (s, 1H), 7.57 (s, 1H), 7.49 (s, 1H), 7.30 (m, 1H), 7.06 (m, 1H), 6.87 (s, 1H), 6.08 (m, 1H), 4.85 (bs, 2H), 4.13 (m, 1H), 3.60 (s, 2H), 2.94 (m, 2H), 2.56 (m, 2H), 2.20 (m, 2H), 2.13 (m, 2H), 1.86 (d, J=8 Hz, 3H); LCMS: 489 [M+1]; c-Met Ki: 0.02 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-(4-Bromo-pyrazol-1-yl)-1-ethyl-piperidine (prepared according to general procedure 11 using 1-iodoethane as alkylation reagent). 1H NMR (400 MHz, CDCl3) δ 7.71 (s, 1H), 7.55 (s, 1H), 7.50 (s, 1H), 7.31 (m, 1H), 7.05 (m, 1H), 6.87 (s, 1H), 6.07 (m, 1H), 5.08 (bs, 2H), 4.15 (m, 1H), 3.14 (m, 2H), 2.51 (m, 2H), 2.19 (m, 6H), 2.08 (s, 3H), 1.86 (d, J=8 Hz, 3H), 1.14 (m, 3H); LCMS: 478 [M+1]; c-Met Ki: 0.01 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 1-[4-(4-Bromo-pyrazol-1-yl)-piperidin-1-yl]-2-dimethylamino-ethanone (prepared from 4-(4-bromo-pyrazol-1-yl)-piperidine and dimethylamino-acetic acid using HOBt/EDC/Triethylamine in DMF solvent). 1H NMR (400 MHz, CDCl3) δ 7.67 (s, 1H), 7.56 (s, 1H), 7.47 (s, 1H), 7.30 (m, 1H), 7.07 (m, 1H), 6.67 (s, 1H), 6.07 (m, 1H), 5.31 (bs, 2H), 4.70 (m, 1H), 4.33 (m, 2H), 3.20 (m, 4H), 2.79 (m, 1H), 2.33 (s, 6H), 2.20 (m, 2H), 2.08 (s, 3H), 1.92 (m, 1H), 1.86 (d, J=8 Hz, 3H); LCMS: 535 [M+1]; c-Met Ki: 0.01 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-(4-bromo-pyrazol-1-yl)-1-cyclopropylmethyl-piperidine (prepared according to general procedure 11 using iodomethyl-cyclopropane as alkylation reagent). 1H NMR (400 MHz, CDCl3) δ 7.62 (s, 1H), 7.54 (m, 2H), 7.37 (m, 1H), 7.16 (m, 1H), 7.06 (s, 1H), 7.01 (s, 1H), 6.18 (m, 1H), 4.50 (m, 1H), 3.98 (m, 1H), 3.69 (m, 1H), 3.50 (m, 1H), 3.02 (m, 2H), 2.95 (m, 1H), 2.65 (m, 2H), 2.44 (m, 1H), 2.31 (m, 1H), 1.94 (m, 3H), 1.14 (m, 1H), 0.80 (m, 2H), 0.42 (m, 2H); LCMS: 504 [M+1]; c-Met Ki: 0.01 μM.
The title compound was prepared according to procedure 6 using 3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-(4-bromo-pyrazol-1-yl)-1-methy-piperidine (prepared according to general procedure 11. 1H NMR (400 MHz, CDCl3) δ 7.65 (s, 1H), 7.55 (s, 1H), 7.50 (s, 1H), 7.31 (m, 1H), 7.06 (m, 1H), 6.87 (s, 1H), 6.08 (m, 1H), 5.50 (bs, 2H), 4.18 (m, 1H), 3.11 (m, 2H), 2.40 (s, 3H), 2.30 (m, 2H), 2.20 (m, 4H), 2.07 (s, 3H), 1.86 (d, J=8 Hz, 3H); LCMS: 464 [M+1]; c-Met Ki: 0.01 μM.
The title compound was prepared according to procedure 7. 1H NMR (400 MHz, CDCl3) δ 7.72 (s, 1H), 7.57 (s, 1H), 7.47 (s, 1H), 7.31 (m, 1H), 7.06 (m, 1H), 6.86 (s, 1H), 6.08 (m, 1H), 5.00 (bs, 2H), 4.70 (m, 1H), 4.36 (m, 1H), 4.21 (s, 1H), 3.70 (m, 1H), 3.18 (m, 1H), 3.00 (m, 1H), 2.223 (m, 2H), 2.01 (m, 2H), 1.86 (d, J=8 Hz, 3H); LCMS: 508 [M+1]; c-Met Ki: 0.004 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-(4-bromo-pyrazol-1-yl)-1-cyclopentyl-piperidine (prepared according to general procedure 11 using bromocyclopentane as alkylation reagent). 1H NMR (400 MHz, CDCl3) δ 7.75 (s, 1H), 7.55 (s, 1H), 7.48 (s, 1H), 7.28 (m, 1H), 7.05 (m, 1H), 6.87 (s, 1H), 6.07 (m, 1H), 4.81 (bs, 2H), 4.64 (m, 1H), 2.18 (m, 2H), 2.05 (m, 2H), 1.86 (m, 3H), 1.71 (m, 4H); LCMS: 435 [M+1]; c-Met Ki: 0.04 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-(4-bromo-pyrazol-1-yl)-1-cyclobutyl-piperidine (prepared according to general procedure 11 using bromocyclopentane as alkylation reagent). 1H NMR (400 MHz, CDCl3) δ 7.74 (s, 1H), 7.57 (s, 1H), 7.50 (s, 1H), 7.28 (m, 1H), 7.05 (m, 1H), 6.87 (s, 1H), 6.08 (m, 1H), 4.90 (bs, 2H), 4.75 (m, 1H), 2.51 (m, 4H), 1.88 (m, 5H); LCMS: 421 [M+1]; c-Met Ki: 0.02 μM.
The title compound was prepared according to procedure 7 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(1-piperidin-4-yl-1H-pyrazol-4-yl)-pyridin-2-ylamine and methylsulfonyl chloride. 1H NMR (400 MHz, CDCl3) δ 7.75 (s, 1H), 7.57 (s, 1H), 7.49 (s, 1H), 7.31 (m, 1H), 7.06 (m, 1H), 6.08 (m, 1H), 4.82 (bs, 2H), 4.26 (m, 1H), 3.94 (m, 2H), 2.98 (m, 2H), 2.85 (s, 3H), 2.26 (m, 2H), 2.18 (m, 2H), 1.86 (d, J=8 Hz, 3H); LCMS: 528 [M+1]; c-Met Ki: 0.01 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-(4-bromo-pyrazol-1-yl)-[1,3′]bipiperidinyl-1′-carboxylic acid tert-butyl ester (prepared according to general procedure 11 using 3-hydroxy-piperidine-1-carboxylic acid tert-butyl ester). 1H NMR (400 MHz, CDCl3) δ 7.69 (s, 1H), 7.55 (s, 1H), 7.52 (s, 1H), 7.31 (m, 1H), 7.07 (m, 1H), 6.86 (s, 1H), 6.07 (m, 1H), 5.21 (bs, 2H), 4.26 (m, 1H), 3.40 (m, 1H), 3.07 (m, 2H), 2.77 (m, 1H), 2.11 (m, 4H), 2.08 (s, 3H), 1.86 (d, J=8 Hz, 3H); LCMS: 450 [M+1]; c-Met Ki: 0.01 μM.
The title compound was prepared according to procedure 6 using 3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-(4-bromo-pyrazol-1-yl)-1-cyclopentyl-piperidine (prepared according to general procedure 11 using bromocyclopentane as alkylation reagent). 1H NMR (400 MHz, CDCl3) δ 7.73 (s, 1H), 7.55 (s, 1H), 7.48 (s, 1H), 7.31 (m, 1H), 7.07 (m, 1H), 6.88 (s, 1H), 6.08 (m, 1H), 4.64 (m, 1H), 2.04 (m, 2H), 1.98 (m, 2H), 1.86 (d, J=8 Hz, 3H), 1.73 (m, 2H); LCMS: 435 [M+1]; c-Met Ki: 0.02 μM.
The title compound was prepared according to procedure 6. 1H NMR (400 MHz, CDCl3) δ 7.69 (s, 1H), 7.56 (s, 1H), 7.50 (s, 1H), 7.32 (m, 1H), 7.07 (m, 1H), 6.87 (m, 1H), 6.07 (m, 1H), 5.25 (bs, 2H), 4.30 (m, 1H), 3.41 (m, 2H), 2.96 (m, 2H), 2.26 (m, 2H), 2.12 (m, 2H), 1.86 (d, J=8 Hz, 3H); LCMS: 450 [M+1]; c-Met Ki: 0.003 μM.
The title compound was prepared according to procedure 6 using 5-bromo-3-(3-fluoro-6,7,8,9-tetrahydro-5H-benzocyclohepten-5-yloxy)-pyridin-2-ylamine and 4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole. 1H NMR (400 MHz, DMSO-d6) δ 7.09 (s, 2H), 6.86 (s, 1H), 6.66 (s, 1H), 6.38 (t, 1H), 6.25 (dd, 1H), 6.08 (dt, 1H), 5.03 (d, 1H), 2.16 (t, 2H), 1.42 (m, 1H), 1.33 (br, 1H), 1.19 (br, 2H), 1.07 (br, 1H), 0.66 (br, 1H); LCMS: 339 [M+1]; c-Met % Inhibition: 21% μM.
The title compound was prepared according to procedure 5 using diethylamine as the amine in the final step. 1H NMR (400 MHz, DMSO-d6) δ 7.96 (s, 1H), 7.78 (d, 1H), 7.61 (s, 1H), 7.45 (q, 1H), 7.42 (t, 1H), 6.90 (d, 1H), 6.09 (q, 1H), 5.68 (s, 2H), 4.09 (q, 1H), 3.20 (m, 1H), 3.16 (d, 2H), 2.81 (m, 1H), 1.80 (d, 3H), 1.66 (s, 6H), 1.18 (m, 1H), 1.10 (t, 1H), 1.01 (br, 2H), 0.60 (br, 2H); LCMS: 508 [M+1]; c-Met Ki: 0.3964 μM.
The title compound was prepared according to procedure 5 using (3S)-pyrrolidin-3-ol as the amine in the final step. 1H NMR (400 MHz, DMSO-d6) δ 8.32 (d, 1H), 7.97 (br, 1H), 7.81 (s, 1H), 7.72 (s, 1H), 7.59 (q, 1H), 7.47 (t, 1H), 7.18 (t, 1H), 6.29 (d, 1H), 4.10 (br, 1H), 4.00 (br, 1H), 3.59 (m, 2H), 3.42 (m, 2H), 2.31 (m, 1H), 1.85 (d, 3H), 1.75 (m, 3H), 1.70 (d, 5H), 1.63 (m, 2H); LCMS: 522 [M+1]; c-Met Ki: 0.47 μM.
The title compound was prepared according to procedure 5 using pyrrolidine as the amine in the final step. 1H NMR (400 MHz, DMSO-d6) δ 8.04 (s, 1H), 7.80 (s, 1H), 7.61 (s, 1H), 7.55 (q, 1H), 7.43 (t, 1H), 6.92 (s, 1H), 6.10 (q, 1H), 5.67 (s, 2H), 2.37 (m, 1H), 1.79 (d, 3H), 1.66 (s, 6H), 1.59 (m, 4H); LCMS: 506 [M+1]; c-Met Ki: 0.425 μM.
The title compound was prepared according to procedure 5 using dimethylamine as the amine in the final step. 1H NMR (400 MHz, DMSO-d6) δ 8.16 (d, 1H), 7.79 (d, 1H), 7.70 (d, 1H), 7.45 (q, 1H), 7.11 (s, 1H), 6.25 (t, 1H), 1.80 (d, 3H), 1.66 (s, 6H); LCMS: 480 [M+1]; c-Met Ki: 0.125 μM.
The title compound was prepared according to procedure 8. 1H NMR (400 MHz, DMSO-d6) δ 7.90 (s, 1H), 7.75 (s, 1H), 7.52 (m, 1H), 7.51 (s, 1H), 7.43 (t, 1H), 6.87 (s, 1H), 6.06 (q, 1H), 5.61 (s, 2H), 4.93 (br, 1H), 3.54 (s, 2H), 1.80 (d, 3H), 1.44 (s, 6H); LCMS: 439 [M+1]; c-Met Ki: 0.014 μM.
The title compound was prepared according to procedure 8 using 2-(4-{6-Amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-N,N-dimethyl-isobutyramide for the reduction. 1H NMR (400 MHz, DMSO-d6) δ 7.88 (s, 1H), 7.76 (s, 1H), 7.53 (m, 2H), 7.43 (t, 1H), 6.87 (s, 1H), 6.06 (q, 1H), 5.62 (s, 2H), 1.80 (s, 6H), 1.78 (d, 3H), 1.48 (s, 6H); LCMS: 466 [M+1]; c-Met Ki: 0.084 μM.
The title compound was prepared according to procedure 8 using 2-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-1-((S)-3-hydroxy-pyrrolidin-1-yl)-2-methyl-propan-1-one for the reduction. 1H NMR (400 MHz, DMSO-d6) δ 7.88 (s, 1H), 7.76 (s, 1H), 7.54 (m, 2H), 7.43 (t, 1H), 6.87 (s, 1H), 6.08 (q, 1H), 5.62 (s, 2H), 4.52 (br, 1H), 4.00 (br, 1H), 2.75 (br, 2H), 2.34 (br, 1H), 2.20 (br, 1H), 2.06 (br, 1H), 1.90 (d, 2H), 1.80 (d, 3H), 1.49 (s, 6H), 1.39 (br, 1H); LCMS: 508 [M+1]; c-Met Ki: 0.07 μM.
The title compound was prepared according to procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 4-bromo-1-[1,1-dimethyl-2-(2-pyrrolidin-1-yl-ethoxy)-ethyl]-1H-pyrazole, which were prepared as follows:
To a solution of 2-(4-bromo-pyrazol-1-yl)-2-methyl-propionic acid methyl ester (5.00 g, 20.2 mmol) in anhydrous THF (67 mL) was added LiAlH4 (1.0M in THF, 22.3 mL) drop by drop under nitrogen. The reaction mixture was stirred under nitrogen at ambient temperature for overnight, and quenched with HCl solution (2N) in ice bath to pH 5. The product was extracted with ethyl acetate. The combined extracts were washed with water, brine, and dried over Na2SO4 to provide 2-(4-bromo-pyrazol-1-yl)-2-methyl-propan-1-ol (3.022 g, 68% yield).
To a solution of 2-(4-bromo-pyrazol-1-yl)-2-methyl-propan-1-ol (0.50 g, 2.28 mmol) and 1-(2-chloro-ethyl)-pyrrolidine (0.3072 g, 2.28 mmol) was added NaH (60% in mineral oil, 27.4 mmol). The reaction was stirred under nitrogen at ambient temperature for 0.5 h, and then at 70° C. for over-night. The reaction was quenched after cooling down to ambient temperature with the addition of water. The product was extracted with ethyl acetate, and the extracts were washed with water, brine, and dried over Na2SO4. The crude product was purified on a reversed phase column to provide 4-bromo-1-[1,1-dimethyl-2-(2-pyrrolidin-1-yl-ethoxy)-ethyl]-1H-pyrazole (97.8 mg, 18% yield).
3-[1-(2,6-Dichloro-3-fluoro-phenyl)-ethoxy]-5-{1-[1,1-dimethyl-2-(2-pyrrolidin-1-yl-ethoxy)-ethyl]-1H-pyrazol-4-yl}-pyridin-2-ylamine: 1H NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.56 (dd, 2H), 7.41 (m, 1H), 7.22 (t, 1H), 7.08 (s, 1H), 6.26 (q, 1H), 3.68 (s, 2H), 3.56 (t, 2H), 3.35 (m, 2H), 2.87 (m, 2H), 2.57 (d, 2H), 1.85 (m, 6H), 1.53 (s, 6H), 1.20 (d, 1H); LCMS: 536 [M+1]; c-Met Ki: 0.229 μM.
The title compound was prepared according to procedure 11 followed by procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 2-[(R)4-(4-bromo-pyrazol-1-yl)-piperidin-1-ylmethyl]-pyrrolidine-1-carboxylic acid tert-butyl ester (prepared according to general procedure 11 using (R)-2-hydroxymethyl-pyrrolidine-1-carboxylic acid tert-butyl ester). 1H NMR (400 MHz, DMSO-d6) δ 9.95 (br, 2H), 8.68 (d, 1H), 8.50 (br, 1H), 8.34 (s, 1H), 8.27 (d, 1H), 8.12 (m, 1H), 7.99 (t, 1H), 7.65 (s, 1H), 6.76 (q, 1H), 6.26 (s, 2H), 5.05 (br, 2H), 4.40 (br, 1H), 3.74 (m, 2H), 2.56 (m, 1H), 2.36 (d, 3H), 2.16 (m, 1H); LCMS: 450 [M+1]; c-Met Ki: 0.072 μM.
The title compound was prepared according to procedure 11 followed by procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 2-[(S)4-(4-bromo-pyrazol-1-yl)-piperidin-1-ylmethyl]-pyrrolidine-1-carboxylic acid tert-butyl ester (prepared according to general procedure 11 using (S)-2-hydroxymethyl-pyrrolidine-1-carboxylic acid tert-butyl ester). 1H NMR (400 MHz, DMSO-d6) δ 7.86 (s, 1H), 7.72 (s, 1H), 7.56 (m, 1H), 7.51 (d, 1H), 6.86 (s, 1H), 6.06 (q, 1H), 5.64 (s, 2H), 3.96 (d, 2H), 2.76 (m, 2H), 1.79 (d, 6H), 1.60 (m, 3H), 1.33 (m, 1H); LCMS: 450 [M+1]; c-Met Ki: 0.0415 μM.
The title compound was prepared according to procedure 11 followed by procedure 6 using 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine and 3-((S)-4-bromo-pyrazol-1-yl)-pyrrolidine-1-carboxylic acid tert-butyl ester (prepared according to general procedure 11 starting from (S)-3-Hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester). 1H NMR (400 MHz, DMSO-d6) δ 7.93 (s, 1H), 7.74 (d, 1H), 7.55 (m, 1H), 7.51 (s, 1H), 7.43 (t, 1H), 6.87 (d, 1H), 6.06 (q, 1H), 5.64 (s, 2H), 4.75 (m, 1H), 3.11 (m, 1H), 3.01 (m, 1H), 2.94 (m, 1H), 2.85 (m, 1H), 2.15 (m, 1H), 1.99 (m, 1H), 1.87 (s, 3H), 1.78 (d, 3H); LCMS: 436 [M+1]; c-Met Ki: 0.298 μM.
The title compound was prepared according to procedure 12. 1H NMR (400 MHz, DMSO-d6) δ 7.86 (s, 1H), 7.76 (s, 1H), 7.63 (m, 2H), 7.54 (m, 1H), 7.37 (t, 1H), 6.46 (q, 1H), 6.15 (s, 1H), 4.10 (m, 1H), 3.01 (m, 2H), 1.95 (m, 2H), 1.85 (s, 2H), 1.75 (d, 3H), 1.67 (dd, 1H); LCMS: 451 [M+1]; c-Met Ki: 0.010 μM.
The title compound was prepared according to procedure 10 using 5-bromo-3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyrazin-2-ylamine and 4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyrazole-1-carboxylic acid tert-butyl ester. 1H NMR (400 MHz, DMSO-d6) δ 12.81 (s, 1H), 7.79 (s, 1H), 7.48 (m, 1H), 7.36 (t, 1H), 6.48 (q, 1H), 6.12 (s, 2H), 1.75 (d, 3H); LCMS: 368 [M+1]; c-Met Ki: 0.065 μM.
The title compound was prepared according to procedures 6 and 7, using 5-bromo-3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyrazin-2-ylamine as the starting material. 1H NMR (400 MHz, DMSO-d6) δ 7.91 (s, 1H), 7.76 (s, 1H), 7.64 (s, 1H), 7.49 (m, 1H), 7.36 (t, 1H), 6.46 (q, 1H), 6.15 (s, 2H), 4.57 (br, 1H), 4.40 (m, 2H), 4.12 (br, 2H), 3.77 (m, 1H), 3.35 (m, 2H), 3.43 (m, 1H), 3.16 (m, 2H), 1.75 (d, 3H); LCMS: 509 [M+1]; c-Met Ki: 0.015 μM.
The title compound was prepared according to procedure 6 using 5-bromo-3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyrazin-2-ylamine and 4-(4-bromo-pyrazol-1-yl)-1-methy-piperidine (prepared according to general procedure 11). 1H NMR (400 MHz, DMSO-d6) δ 7.88 (s, 1H), 7.76 (s, 1H), 7.64 (s, 1H), 7.49 (m, 1H), 7.36 (t, 1H), 6.46 (q, 1H), 6.15 (s, 2H), 4.02 (m, 1H), 2.84 (m, 2H), 2.19 (s, 3H), 2.00 (m, 4H), 1.85 (m, 3H), 1.75 (d, 3H); LCMS: 465 [M+1]; c-Met Ki: 0.03 μM.
The title compound was prepared according to procedure 7 using 3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(1-piperidin-4-yl-1H-pyrazol-4-yl)-pyrazin-2-ylamine coupled with dimethylamino-acetic acid in the presence of HOBt/EDC/triethylamine in DMF as described in procedure 5 using 5-bromo-3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyrazin-2-ylamine as the starting material. 1H NMR (400 MHz, DMSO-d6) δ 7.90 (s, 1H), 7.76 (s, 1H), 7.65 (s, 1H), 7.49 (m, 1H), 7.36 (t, 1H), 6.47 (q, 1H), 6.15 (s, 2H), 4.39 (m, 1H), 4.16 (m, 1H), 3.16 (m, 2H), 3.02 (m, 1H), 2.75 (m, 1H), 2.19 (s, 6H), 2.01 (m, 2H), 1.88 (s, 1H), 1.75 (d, 3H); LCMS: 536 [M+1]; c-Met Ki: 0.015 μM.
The title compound was prepared according to procedure 5 using 5-bromo-3-[(R)-1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-2-ylamine and 1-methanesulfonyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (prepared from 4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole reacted with methylsulfonyl chloride in the presence of triethylamine in dichloromethane solvent. 1H NMR (400 MHz, CDCl3) δ 8.41 (s, 1H), 8.13 (s, 1H), 7.89 (d, 1H), 7.51 (dd, 1H), 7.38 (t, 1H), 7.07 (d, 1H), 6.09 (q, 1H), 5.79 (bs, 2H), 3.49 (s, 3H), 1.74 (d, 3H); LCMS: 445 [M+1]; c-Met Ki: 0.17 μM.
The title compound was prepared according to procedure 6 using 5-bromo-3-[(R)-1-(2-chloro-3,6-difluoro-phenyl)-ethoxy]-pyridin-2-ylamine as starting material (according to the methods for the synthesis of 5-bromo-3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-2-ylamine from (S)-1-(2-chloro-3,6-difluoro-phenyl)-ethanol, obtained from SynChem, Inc.). 1H NMR (400 MHz, DMSO-d6) δ 7.88 (s, 1H), 7.70 (s, 1H), 7.50 (s, 1H), 7.38 (m, 1H), 7.25 (m, 1H), 6.99 (s, 1H), 5.88 (m, 1H), 5.48 (bs, 2H), 4.08 (m, 1H), 2.96 (m, 2H), 2.53 (m, 1H), 2.45 (m, 1H), 1.89 (m, 1H), 1.80 (m, 4H), 1.67 (m, 4H); LCMS: 434 [M+1]; c-Met Ki: 0.09 μM.
The title compound was prepared according to procedure 16, followed by 14, using [2-(4-Bromo-pyrazol-1-yl)-ethyl]-methyl-carbamic acid tert-butyl ester as aryl halide, and 13. 1H NMR (400 MHz, chloroform-D) δ ppm 7.75 (d, J=1.77 Hz, 1H) 7.58 (s, 1H) 7.50 (s, 1H) 7.30 (dt, J=9.16, 4.64 Hz, 1H) 7.05 (dd, J=8.84, 7.83 Hz, 1H) 6.86 (d, J=1.77 Hz, 1H) 6.07 (q, J=6.74 Hz, 1H) 4.83 (s, 2H) 4.22-4.30 (m, 2H) 3.08-3.13 (m, 2H) 2.49 (s, 3H) 1.86 (d, J=6.82 Hz, 3H); LCMS: 424 [M+1]; c-Met Ki: 0.055 μM.
The title compound was prepared according to procedure 17, followed by 14 using 2-(4-Bromo-pyrazol-1-yl)-N-(3-dimethylamino-propyl)-propionamide as aryl halide, then 18, then 13. 1H NMR (400 MHz, chloroform-D) δ ppm 7.78 (s, 1H) 7.66 (s, 1H) 7.55 (s, 1H) 7.46-7.53 (m, 1H) 7.31 (dd, J=8.72, 4.93 Hz, 1H) 7.06 (t, J=8.46 Hz, 1H) 6.87 (s, 1H) 6.07 (q, J=6.48 Hz, 1H) 4.90 (q, J=7.41 Hz, 1H) 4.80 (s, 2H) 3.31 (q, J=5.64 Hz, 2H) 2.27 (t, J=5.81 Hz, 2H) 2.02 (s, 6H) 1.86 (d, J=6.57 Hz, 3H) 1.80 (d, J=7.33 Hz, 3H) 1.57 (dt, J=12.06, 5.97 Hz, 2H); LCMS: 523 [M+1]; c-Met Ki: 0.04 μM.
The title compound was prepared according to procedure 14 using 4-(3-iodo-pyrazole-1-yl)-piperdine-1-carboxylic acid tert-butyl ester followed by removal of the protecting group. To 70 mg 4-(3-{6-Amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-piperidine-1-carboxylic acid tert-butyl was added 3 ml of dichloromethane followed by 1 mL trifluoroacetic acid at room temperature. After stirring for 2 hours, the reaction was done. Purification by reverse phase HPLC gave 80 mg white hygroscopic solid. Conversion to the free base was accomplished by dissolving in dichloromethane and washing with sodium bicarbonate which led to 17 mg of a white solid, 3-[1-(2,6-Dichloro-3-fluoro-phenyl)-ethoxy]-5-(1-piperidin-4-yl-1H-pyrazol-3-yl)-pyridin-2-ylamine. 1H NMR (400 MHz, DMSO-D6) δ ppm 7.87 (d, J=1.77 Hz, 1H) 7.68 (d, J=2.27 Hz, 1 H) 7.53 (m, 1H) 7.42 (m, 1H) 7.09 (d, J=1.77 Hz, 1H) 6.34 (d, J=2.27 Hz, 1H) 6.04 (q, J=6.82 Hz, 1H) 5.77 (s, 2H) 4.10 (m, 1H) 3.03 (m, 2H) 2.58 (m, 2H) 1.91 (m, 2H) 1.79 (d, J=6.82 Hz, 3H) 1.75 (m, 2H); LCMS: 450 [M+1]; c-Met Ki: 0.038 μM.
The starting material, 4-(3-iodo-pyrazole-1-yl)-piperdine-1-carboxylic acid tert-butyl ester was prepared as follows:
1-(2-Trimethylsilanyl-ethoxymethyl)-1H-Pyrazole: To 4.9 g 95% NaH in mineral oil was added 200 mL THF. Pyrazole (12 g) was added in 50 mL THF dropwise and stirred at rt for 1 hour. After cooling to 0° C., a solution of 34.32 mL SEMCI in 50 mL THF was added dropwise. The cooling bath was removed, and the reaction was stirred at room temperature for overnight. The reaction was quenched with water, concentrated, diluted with diethylether and washed with ammonium chloride, brine, and dried over sodium sulfate. Chromatography with 20-30% EtOAc/hexanes led to 12 g of product. 1H NMR (400 MHz, chloroform-d) δ ppm 7.55 (dd, J=5.81, 1.77 Hz, 2H) 6.33 (t, J=2.02 Hz, 1H) 5.44 (s, 2H) 3.50-3.58 (m, 2H) 0.85-0.95 (m, 2H) 0.04 (s, 9H).
5-iodo-1-(2-trimethylsilanyl-ethoxymethyl)-1H-pyrazole: To 2.0 g 1-(2-trimethylsilanyl-ethoxymethyl)-1H-pyrazole in 40 mL THF at −78° C. was added 4.43 mL 2.5 N nBuLi dropwise. The solution was stirred for an additional 45 min. Then iodine (7.67 g) in 20 mL THF was added dropwise. After all the iodine was added the cooling bath was removed, and the mixture was allowed to warm to room temperature over 2 hours. The reaction was quenched by addition of a few mLs of aqueous NH4Cl (saturated). The solution was concentrated, diluted with diethyl ether, washed with aqueous sodium sulfite, water, brine, and dried over sodium sulfate. Chromatography with 5-10% EtOAc/hexanes gave 1.66 g of product as an oil. 1H NMR (400 MHz, chloroform-d) δ ppm 7.53 (d, J=1.77 Hz, 1H) 6.49 (d, J=1.77 Hz, 1H) 5.50 (s, 2H) 3.53-3.61 (m, 2H) 0.86-0.95 (m, 2H) 0.04 (m, 9H).
5-iodo-1H-pyrazole: To 1.0 g 5-iodo-1-(2-trimethylsilanyl-ethoxymethyl)-1H-pyrazole was added a mixture of 1.72 mL triethylsilane and 4 mL TFA at 0° C. After addition the cooling bath was removed. At 1.5 hours the solvents were removed leaving 570 mg of a white solid. 1H NMR (400 MHz, CHLOROFORM-D) δ ppm 7.53 (m, 1H) 6.51 (m, 1H).
4-(3-Iodo-pyrazol-1-yl)-piperidine-1-carboxylic acid tert-butyl ester: To 500 mg 5-iodo-1H-pyrazole in 3 mL DMF was added 54 mg 95% NaH. This was stirred at room temperature for 5 minutes followed by heating for 30 minutes at 80° C. The solution was cooled to room temperature and 417 mg BOC piperidine mesylate was added. The reaction was heated to 90° C. overnight. Still some iodo-pyrazole by TLC so added 100 mg more mesylate and heated another 18 hours. The reaction was cooled and some of the DMF was removed in vacuo. The solution was diluted with ethyl acetate and washed with sodium bicarbonate, brine, and dried over sodium sulfate. Removal of solvent led to a clear oil Chromatography with 5, 10, 20% EtOAc/CH2Cl2 led to 180 mg of product as a clear oil. 1H NMR (400 MHz, CHLOROFORM-D) δ ppm 7.23 (d, J=2.27 Hz, 1H) 6.40 (d, J=2.27 Hz, 1H) 4.24 (m, 3H) 2.84 (s, 2H) 2.02-2.12 (m, 2H) 1.87 (m, 2H) 1.40-1.47 (m, 9H).
The title compound was prepared according to procedure 10 using 3-iodo-1-(2-trimethylsilanyl-ethoxymethyl)-1H-pyrazole followed by removal of the 2-trimethylsilanyl-ethoxymethyl. To 60 mg 3-[1-(2,6-Dichloro-3-fluoro-phenyl)-ethoxy]-5-[1-(2-trimethylsilanyl-ethoxymethyl)-1H-pyrazol-3-yl]-pyridin-2-ylamine in 1 mL dichloromethane was added 60 μL triethylsilane and 0.5 mL trifluoroacetic acid. After stirring for 4 hours at room temperature 3 mL of toluene was added, and the solvent was removed in vacuo. The residue was dissolved in EtOAc and washed with sodium bicarbonate. Chromatography with 10% MeOH/40% EtOAc/50% CH2Cl2 gave 20 mg of a white solid. 1H NMR (400 MHz, DMSO-D6) δ ppm 7.92 (d, J=1.52 Hz, 1H) 7.68 (s, 1H) 7.49-7.59 (m, 1H) 7.43 (t, J=8.72 Hz, 2H) 7.12 (s, 1H) 6.41 (s, 1H) 6.06 (s, 1H) 5.90 (s, 2H) 3.32 (s, 4H) 1.78 (d, J=6.57 Hz, 3H); LCMS: 367 [M+1]; c-Met Ki: 0.035 μM.
The title compound was prepared according to procedure 19. 1H NMR (400 MHz, DMSO-D6) δ ppm 8.07 (d, J=2.27 Hz, 1H) 7.90 (d, J=2.02 Hz, 1H) 7.57 (dd, J=9.09, 5.05 Hz, 1H) 7.46 (t, J=8.72 Hz, 1H) 7.33 (d, J=1.77 Hz, 1H) 6.27 (d, J=2.27 Hz, 1H) 6.17 (q, J=6.48 Hz, 1H) 2.20 (s, 3H) 1.82 (d, J=6.57 Hz, 3H); LCMS: 381 [M+1]; c-Met Ki: 1.92 μM.
The title compound was prepared according to procedure 19 using 1H-indazole in place of 3-methyl-1H-pyrazole. 1H NMR (400 MHz, DMSO-D6) δ ppm 8.29 (s, 1H) 7.83-7.89 (m, 2H) 7.58-7.63 (m, 1H) 7.52 (t, J=8.72 Hz, 1H) 7.39 (t, J=7.71 Hz, 1H) 7.17-7.26 (m, 2H) 7.00 (d, J=2.02 Hz, 1H) 6.14 (q, J=6.40 Hz, 1H) 1.82 (d, J=6.57 Hz, 3H); LCMS: 417 [M+1]; c-Met Ki: 1.56 μM.
The title compound was prepared according to procedure 19 using pyrazole in place of 3-methyl-1H-pyrazole. 1H NMR (400 MHz, DMSO-D6) δ ppm 8.21 (d, J=2.53 Hz, 1H) 7.95 (d, J=2.02 Hz, 1H) 7.67 (d, J=1.77 Hz, 1H) 7.57 (dd, J=8.97, 4.93 Hz, 1H) 7.45 (t, J=8.72 Hz, 1H) 7.29 (d, J=1.77 Hz, 1H) 6.46-6.52 (m, 1H) 6.16 (t, J=6.57 Hz, 1H) 1.81 (d, J=6.57 Hz, 3H); LCMS: 367 [M+1]; c-Met Ki: 0.87 μM.
The title compound was prepared according to procedure 19 using ethyl 1H-pyrazole-4-carboxylate in place of 3-methyl-1H-pyrazole. 1H NMR (400 MHz, DMSO-D6) δ ppm 8.81 (s, 1H) 8.02-8.08 (m, 2H) 7.56 (dd, J=9.09, 5.05 Hz, 1H) 7.45 (t, J=8.84 Hz, 1H) 7.29 (d, J=2.02 Hz, 1 H) 6.14 (q, J=6.57 Hz, 1H) 4.24 (q, J=7.07 Hz, 3H) 1.80 (d, J=6.57 Hz, 3H) 1.26 (q, J=7.58 Hz, 4H); LCMS: 439 [M+1]; c-Met Ki: 2.05 μM.
The title compound was prepared according to procedure 15 using 4-(4-bromo-pyrazol-1-ylmethyl)-piperidine-1-carboxylic acid tert-butyl ester (prepared by using General Procedure 11) and 3-[1-(2,6-Dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine, followed by de-protection (General Procedure 3). 1H NMR (400 MHz, DMSO-D6) δ ppm 8.51 (s, 1H) 8.20 (s, 1H) 7.99 (s, 1H) 7.72 (d, J=1.52 Hz, 1H) 7.65 (s, 1H) 7.59 (dd, J=8.97, 4.93 Hz, 1H) 7.47 (t, J=8.72 Hz, 1H) 7.06 (s, 1H) 6.23 (q, J=6.48 Hz, 1H) 4.04 (d, J=6.82 Hz, 3H) 3.68-3.78 (m, 1H) 3.44-3.54 (m, 2H) 3.19-3.29 (m, 3H) 2.78-2.88 (m, 2H) 2.06 (s, 1H) 1.84 (d, J=6.57 Hz, 3H) 1.63 (d, J=14.40 Hz, 2H) 1.26-1.37 (m, 2H); LCMS: 464 [M+1]; c-Met Ki: 0.056 μM.
The title compound was prepared according to procedure 15 using 4-[2-(4-Bromo-pyrazol-1-yl)-ethyl]-piperazine-1-carboxylic acid tert-butyl ester (prepared by using General Procedure 11) and 3-[1-(2,6-Dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine, followed by de-protection (General Procedure 3). 1H NMR (400 MHz, DMSO-D6) δ ppm 9.39 (s, 1H) 8.09 (s, 1H) 7.79 (s, 1H) 7.70 (s, 1H) 7.61 (dd, J=8.97, 4.93 Hz, 1H) 7.49 (t, J=8.59 Hz, 1H) 7.13 (s, 1H) 6.27 (q, J=6.48 Hz, 1H) 4.47 (s, 2H) 3.27 (s, 6H) 3.11 (s, 3 H) 1.85 (d, J=6.82 Hz, 3H); LCMS: 479 [M+1]; c-Met Ki: 0.047 μM.
The title compound was prepared according to procedure 15 using 2-(4-bromo-pyrazol-1-yl)-ethanol (prepared using Procedure 26) and 3-[1-(2,6-Dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine. 1H NMR (400 MHz, DMSO-D6) δ ppm 7.96 (s, 1H) 7.72 (s, 1H) 7.56-7.63 (m, 2H) 7.43-7.52 (m, 2H) 7.07 (d, J=1.52 Hz, 1H) 6.24 (q, J=6.57 Hz, 1H) 4.08-4.17 (m, 2H) 3.70 (dt, J=8.78, 5.46 Hz, 3H) 1.84 (d, J=6.57 Hz, 3H); LCMS: 411 [M+1]; c-Met Ki: 0.046 μM.
The title compound was prepared according to procedure 15 using 4-bromo-1-[1,3]dioxolan-4-ylmethyl-1H-pyrazole (prepared by using General Procedure 11) and 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine. 1H NMR (400 MHz, DMSO-D6) δ ppm 7.99 (s, 1H) 7.71-7.79 (m, 2H) 7.67 (d, J=3.28 Hz, 1H) 7.55-7.64 (m, 2H) 7.49 (td, J=8.65, 2.65 Hz, 2H) 7.08 (s, 1H) 6.25 (dd, J=6.95, 2.15 Hz, 1H) 4.91 (d, J=4.04 Hz, 1H) 4.80 (d, J=2.53 Hz, 1H) 4.33 (d, J=16.67 Hz, 1H) 4.20-4.29 (m, 2H) 4.06-4.15 (m, 1 H) 3.94 (dd, J=8.34, 6.82 Hz, 2H) 3.65 (ddd, J=8.21, 6.19, 6.06 Hz, 4H) 1.85 (d, J=6.57 Hz, 3H) 1.80 (d, J=6.57 Hz, 1H); LCMS: 453 [M+1]; c-Met Ki: 0.061 μM.
The title compound was prepared according to procedure 15 using 1-[4-(4-bromo-pyrazol-1-ylmethyl)-piperidin-1-yl]-2-hydroxy-ethanone (prepared by using General Procedure 11) and 3-[1-(2,6-ichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine. 1H NMR (400 MHz, DMSO-D6) δ ppm 7.94-8.00 (m, 1H) 7.78 (d, J=7.58 Hz, 1H) 7.72 (d, J=1.26 Hz, 1H) 7.64 (s, 1H) 7.60 (dd, J=8.97, 4.93 Hz, 1H) 7.45-7.54 (m, 1H) 7.09 (s, 1H) 6.26 (q, J=6.48 Hz, 1H) 4.31 (d, J=12.38 Hz, 1H) 3.97-4.09 (m, 4H) 3.63 (s, 1H) 2.89 (t, J=12.13 Hz, 1H) 2.56 (t, J=12.13 Hz, 1H) 2.03 (ddd, J=11.05, 7.39, 3.79 Hz, 1H) 1.85 (d, J=6.57 Hz, 3H) 1.48 (d, J=13.14 Hz, 2H) 1.10-1.18 (m, 1H) 0.98-1.10 (m, 1H); LCMS: 522 [M+1]; c-Met Ki: 0.03 μM.
The title compound was prepared according to procedure 15 using 4-bromo-1-(2,2-dimethyl-[1,3]dioxolan-4-ylmethyl)-1H-pyrazole (prepared by using General Procedure 11) and 3-[1-(2,6-Dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine. 1H NMR (400 MHz, DMSO-D6) δ ppm 7.82 (s, 1H) 7.73 (d, J=1.77 Hz, 1H) 7.56-7.59 (m, 1H) 7.52-7.56 (m, 1H) 7.40-7.47 (m, 1H) 6.85 (d, J=1.77 Hz, 1H) 6.02-6.11 (m, 1H) 5.69 (s, 2H) 4.32-4.39 (m, 1H) 4.13-4.25 (m, 2H) 4.00 (ddd, J=8.34, 6.57, 1.26 Hz, 1H) 3.72 (ddd, J=8.65, 5.49, 3.54 Hz, 1H) 1.76-1.84 (m, 4H) 1.28 (d, J=16.67 Hz, 3H) 1.25 (s, 3H); LCMS: 481 [M+1]; c-Met Ki: 0.062 μM.
The title compound was prepared according to procedure 21. 3-[1-(2,6-Dichloro-3-fluoro-phenyl)-ethoxy]-5-[1-(2,2-dimethyl-[1,3]dioxolan-4-ylmethyl)-1H-pyrazol-4-yl]-pyridin-2-ylamine was de-protected with TFA and water. 1H NMR (400 MHz, DMSO-D6) δ ppm 7.96 (d, J=3.54 Hz, 2H) 7.76 (s, 1H) 7.64 (d, J=2.78 Hz, 1H) 7.60 (dd, J=8.97, 4.93 Hz, 1H) 7.47 (t, J=8.72 Hz, 1H) 7.11 (s, 1H) 6.23-6.31 (m, 1H) 4.22 (d, J=3.79 Hz, 1H) 4.19 (d, J=3.54 Hz, 1H) 3.96 (dd, J=13.64, 7.83 Hz, 2H) 3.72-3.82 (m, 2H) 3.26-3.37 (m, 2H) 1.85 (d, J=6.57 Hz, 3H); LCMS: 441 [M+1]; c-Met Ki: 0.028 μM.
The title compound was prepared according to procedure 15 using 4-(4-bromo-pyrazol-1-yl)-pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (prepared by using General Procedure 11) and 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine, followed by de-protection (General Procedure 3). 1H NMR (400 MHz, DMSO-D6) δ ppm 8.09-8.15 (m, 2H) 7.74-7.79 (m, 3H) 7.69 (s, 1H) 7.56-7.62 (m, 2H) 7.47 (t, J=8.72 Hz, 2H) 7.06 (s, 2H) 6.18-6.26 (m, 2H) 5.25 (s, 1H) 5.20 (s, 1H) 4.67-4.79 (m, 2H) 3.77-3.84 (m, 5H) 3.74 (d, J=2.78 Hz, 3H) 3.70 (s, 1H) 3.58 (s, 3H) 2.83 (s, 1H) 2.53-2.62 (m, 3H) 2.44 (s, 1H) 1.84 (d, J=6.57 Hz, 6H); LCMS: 494 [M+1]; c-Met Ki: 0.034 μM.
The title compound was prepared according to procedure 15 using 4-(4-bromo-pyrazol-1-ylmethyl)-1-methyl-piperidine (prepared by using General Procedure 11) and 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-2-ylamine. 1H NMR (400 MHz, DMSO-D6) δ ppm 8.58 (s, 1H) 8.25 (s, 1H) 8.00 (s, 2H) 7.75 (s, 2H) 7.66 (s, 2H) 7.59 (dd, J=9.09, 5.05 Hz, 2H) 7.48 (t, J=8.72 Hz, 2H) 7.09 (s, 2H) 6.20-6.31 (m, 2H) 4.05 (d, J=6.82 Hz, 3H) 3.40 (d, J=12.88 Hz, 1H) 3.19-3.29 (m, 3H) 2.78-2.90 (m, 3H) 2.72 (d, J=4.55 Hz, 1H) 2.07 (s, 1H) 1.85 (d, J=6.57 Hz, 4H) 1.59-1.71 (m, 3H) 1.29-1.40 (m, 2H); LCMS: 478 [M+1]; c-Met Ki: 0.024 μM.
The title compound was prepared according to procedure 20 using 4-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-pyrrolidine-2-carboxylic acid methyl ester and methylamine. 1H NMR (400 MHz, DMSO-D6) δ ppm 8.96 (s, 1H) 8.45 (d, J=4.55 Hz, 1 H) 8.11 (s, 2H) 7.72-7.78 (m, 3H) 7.70 (s, 1H) 7.58 (dd, J=8.97, 4.93 Hz, 2H) 7.47 (td, J=8.65, 1.39 Hz, 2H) 7.05 (d, J=6.06 Hz, 2H) 6.16-6.24 (m, 2H) 5.14-5.22 (m, J=7.58, 7.26, 7.11, 7.11 Hz, 1H) 4.42-4.50 (m, 1H) 4.31 (t, J=8.59 Hz, 1H) 3.76-3.83 (m, 1H) 3.68-3.75 (m, 2 H) 3.51-3.63 (m, 3H) 2.86 (dt, J=13.39, 7.83 Hz, 1H) 2.67 (t, J=4.42 Hz, 5H) 2.28-2.39 (m, 2 H) 1.83 (d, J=6.57 Hz, 5H); LCMS: 493 [M+1]; c-Met Ki: 0.034 μM.
The title compound was prepared according to procedure 20 using 4-(4-{6-amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-pyrazol-1-yl)-pyrrolidine-2-carboxylic acid methyl ester and amine (2 M ammonia) in methanol. 1H NMR (400 MHz, DMSO-D6) δ ppm 9.90 (s, 1H) 8.85 (s, 1H) 8.15 (s, 2H) 7.99 (s, 2H) 7.71-7.81 (m, 6H) 7.59 (dd, J=8.97, 4.93 Hz, 2H) 7.48 (t, J=8.72 Hz, 2H) 7.26 (s, 1H) 7.13 (s, 1H) 7.09 (d, J=2.27 Hz, 2H) 6.24 (q, J=6.48 Hz, 2H) 5.25 (s, 1H) 4.42-4.51 (m, 1H) 4.32 (t, J=8.46 Hz, 1H) 3.78 (s, 1H) 3.57 (s, 2H) 2.53-2.63 (m, 1H) 2.32-2.41 (m, 1H) 1.84 (d, J=6.57 Hz, 5H); LCMS: 479 [M+1]; c-Met Ki: 0.044 μM.
The title compound was prepared according to procedure 24, from 3-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-5-(1-methyl-1H-pyrazol-4-yl)-pyridin-2-ylamine. The starting material can be obtained according to Example I-615 of U.S. patent application Ser. No. 10/786,610, entitled “Aminoheteroaryl Compounds as Protein Kinase Inhibitors”, filed Feb. 26, 2004, and corresponding international application PCT/US2004/005495 of the same title, filed Feb. 26, 2004, the disclosures of which are incorporated herein by reference in their entireties. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.77 (d, J=6.57 Hz, 3H) 3.80 (s, 3H) 3.81-3.85 (m, 3H) 5.60 (s, 2 H) 6.08 (q, J=6.57 Hz, 1H) 6.86 (d, J=1.52 Hz, 1H) 7.10 (d, J=9.09 Hz, 1H) 7.43 (d, J=8.84 Hz, 1H) 7.47 (s, 1H) 7.69 (d, J=1.77 Hz, 1H) 7.79 (s, 1H); LCMS: 394 [M+1]; c-Met Ki: 0.012 μM.
The title compound was prepared according to procedure 21 using 4-(chloromethyl)-3,5-dimethylisoxazole as the alkylating agent. 1H NMR (400 MHz, chloroform-D) δ ppm 1.85 (d, J=6.57 Hz, 3H) 2.20 (s, 3H) 2.43 (s, 3H) 5.03-5.05 (m, 2H) 6.04 (q, J=6.57 Hz, 1H) 6.79 (d, J=1.77 Hz, 1H) 6.97-7.13 (m, 1H) 7.29 (q, J=2.86 Hz, 2H) 7.57 (s, 1H) 7.73 (d, J=1.77 Hz, 1 H); LCMS: 477 [M+1]; c-Met Ki: 0.069 μM.
The title compound was prepared according to procedure 21 using tert-butyl-4-methanesulfonate-1-piperidine carboxylate (procedure 11) as the alkylating agent. 1H NMR (400 MHz, chloroform-D) δ ppm 1.47 (s, 9H) 1.85 (d, J=6.82 Hz, 3H) 1.87-1.98 (m, 2H) 2.05-2.20 (m, 2H) 2.82-2.92 (m, 2H) 4.14-4.34 (m, 3H) 4.75 (s, 2H) 6.06 (q, J=6.82 Hz, 1H) 6.86 (d, J=1.52 Hz, 1H) 7.00-7.08 (m, 1H) 7.29 (dd, J=8.84, 4.80 Hz, 1H) 7.47 (s, 1H) 7.56 (s, 1H) 7.75 (d, J=1.77 Hz, 1H); LCMS: 551 [M+1]; c-Met Ki: 0.035 μM.
The title compound was prepared according to procedures 21 and 22 using ethylene carbonate as the acylating agent. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.79 (d, J=6.57 Hz, 3H) 1.82 (d, J=4.04 Hz, 2H) 1.92-2.08 (m, 2H) 2.96 (s, 1H) 3.97-4.04 (m, 2H) 4.09 (d, J=13.39 Hz, 2H) 4.27-4.40 (m, 1H) 4.78 (t, J=5.56 Hz, 1H) 5.64 (s, 2H) 6.07 (q, J=6.74 Hz, 1H) 6.89 (d, J=1.77 Hz, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.52 (s, 1H) 7.56 (dd, J=8.97, 4.93 Hz, 1H) 7.74 (d, J=1.77 Hz, 1H) 7.97 (s, 1H); LCMS: 539 [M+1]; c-Met Ki: 0.01 μM.
The title compound was prepared according to procedure 21. 1H NMR (400 MHz, chloroform-D) δ ppm 1.60 (s, 1H) 1.84 (d, J=6.57 Hz, 3H) 5.07 (s, 2H) 6.06 (q, J=6.57 Hz, 1H) 6.89 (d, J=1.77 Hz, 1H) 6.96-7.06 (m, 1H) 7.22-7.33 (m, 1H) 7.67 (s, 2H) 7.80 (d, J=1.52 Hz, 1H); LCMS: 368 [M+1]; c-Met Ki: 0.035 μM.
The title compound was prepared according to procedure 2 using 2-iodopropane as the alkylating agent. 1H NMR (400 MHz, DMSO-D6) δ ppm 0.88 (d, J=6.06 Hz, 6H) 1.79 (d, J=6.82 Hz, 3H) 2.35-2.42 (m, 1H) 3.28 (t, 2H) 3.62 (t, J=7.58 Hz, 2H) 4.78-4.91 (m, 1H) 5.67 (s, 2H) 6.08 (q, J=6.57 Hz, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.55 (dd, J=8.97, 4.93 Hz, 1H) 7.58 (s, 1H) 7.75 (d, J=1.77 Hz, 1H) 8.03 (s, 1H); LCMS: 465 [M+1]; c-Met Ki: 0.03 μM.
The title compound was prepared according to procedures 21 and 22 using 2-bromoacetamide as the alkylating agent. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.79 (d, J=6.57 Hz, 3H) 1.92-2.08 (m, 4H) 2.16-2.31 (m, 2H) 2.83-2.88 (m, 2H) 2.89 (s, 2H) 4.04-4.17 (m, 1H) 5.64 (s, 2H) 6.07 (q, J=6.74 Hz, 1H) 6.88 (d, J=1.52 Hz, 1H) 7.08-7.16 (m, 1H) 7.20 (s, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.52 (s, 1H) 7.57 (dd, J=8.97, 4.93 Hz, 1H) 7.74 (d, J=1.77 Hz, 1H) 7.93 (s, 1H); LCMS: 508 [M+1]; c-Met Ki: 0.007 μM.
The title compound was prepared according to procedure 21 using 3-methanesulfonyloxy-8-aza-bicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester as the alkylating agent. The starting material was obtained by reducing tert-butyl 3-oxo-8-azabicyclo[3.2.1]octane-8-carboxylate (commercially available from Fluka) with NaBH4/ethanol to obtain tert-butyl 3-hydroxy-8-azabicyclo[3.2.1]octane-8-carboxylate, from which the corresponding methanesulfonyloxy compound was prepared. 1H NMR (400 MHz, MeOD) δ ppm 1.85 (d, J=6.57 Hz, 3H) 1.87-2.11 (m, 7H) 3.69-3.78 (m, 2H) 4.08 (d, J=7.07 Hz, 1H) 4.51-4.67 (m, 1H) 6.14 (q, J=6.57 Hz, 1 H) 6.90 (s, 1H) 7.20 (t, J=8.59 Hz, 1H) 7.43 (dd, J=8.97, 4.93 Hz, 1H) 7.51 (s, 1H) 7.65 (s, 1H) 7.79 (s, 1H); LCMS: 477 [M+1]; c-Met Ki: 0.005 μM.
The title compound was prepared according to procedure 21 using 5-chloromethyl-1(2)-tetrazole as the alkylating agent. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.84 (d, J=6.57 Hz, 3H) 5.74 (s, 2 H) 6.25 (q, J=6.48 Hz, 1H) 7.10 (d, J=1.01 Hz, 1H) 7.47 (t, J=8.72 Hz, 1H) 7.53-7.63 (m, 1H) 7.71 (s, 1H) 7.76 (d, J=1.01 Hz, 1H) 8.14 (s, 1H); LCMS: 450 [M+1]; c-Met Ki: 0.04 μM.
The title compound was prepared according to procedures 21 and 22 using ethyl chloroformate as the acylating agent. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.19 (t, J=7.07 Hz, 3H) 1.71-1.79 (m, 2H) 1.82 (d, J=6.57 Hz, 3H) 1.97-2.06 (m, 2H) 2.93-3.04 (m, 2H) 4.05 (q, J=7.07 Hz, 4 H) 4.29-4.47 (m, 1H) 6.17 (q, J=6.32 Hz, 1H) 6.50-6.88 (m, 2H) 7.00 (s, 1H) 7.46 (t, J=8.72 Hz, 1H) 7.58 (q, J=4.55 Hz, 1H) 7.58 (s, 1H) 7.72 (d, J=1.52 Hz, 1H) 8.04 (s, 1H); LCMS: 523 [M+1]; c-Met Ki: 0.019 μM.
The title compound was prepared according to procedure 2 using methyl bromoacetate as the alkylating agent then using LiBH4 as the reducing agent to reduce the methyl ester to alcohol. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.83 (d, J=6.57 Hz, 3H) 3.02 (t, J=5.81 Hz, 2H) 3.75 (q, J=5.73 Hz, 2H) 3.88-3.99 (m, 2H) 4.18-4.28 (m, 2H) 4.78 (t, J=5.18 Hz, 1H) 5.31-5.42 (m, J=7.96, 7.96 Hz, 1H) 6.19 (q, J=6.57 Hz, 1H) 7.01 (d, J=1.26 Hz, 1H) 7.43-7.51 (m, 1H) 7.59 (dd, J=8.97, 4.93 Hz, 1H) 7.76 (d, J=1.26 Hz, 1H) 7.78 (s, 1H) 8.15 (s, 1H); LCMS: 467 [M+1]; c-Met Ki: 0.05 μM.
The title compound was prepared according to procedure 2 using 2-bromoethyl methyl ether as the alkylating agent. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.79 (d, J=6.57 Hz, 3H) 2.60-2.67 (m, 2H) 3.10-3.18 (m, 2H) 3.22 (s, 3H) 3.35-3.42 (m, 2H) 3.68 (t, J=7.45 Hz, 2H) 4.85-4.97 (m, 1H) 5.67 (s, 2H) 6.08 (q, J=6.40 Hz, 1H) 6.89 (d, J=1.52 Hz, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.56 (dd, J=8.97, 4.93 Hz, 1H) 7.58 (s, 1H) 7.75 (d, J=1.52 Hz, 1H) 8.03 (s, 1H); LCMS: 481 [M+1]; c-Met Ki: 0.07 μM.
compound was prepared according to procedure 2. The last alkylation procedure was performed by dissolving 2-11 (1 molar equivalent) in DMF (3 mL). Sulfonyl chloride (1 molar equivalent) and triethyl amine (3 molar equivalents) were added and the reaction stirred at rt for 16 h. The general work-up conditions were then followed (procedure 2) to afford title compound in 30% yield. 1H NMR (400 MHz, chloroform-D) δ ppm 1.85 (d, J=6.82 Hz, 3H) 3.03 (s, 3H) 4.35-4.45 (m, 4H) 4.83 (s, 2H) 5.06 (ddd, J=14.02, 7.83, 6.44 Hz, 1H) 6.06 (q, J=6.82 Hz, 1H) 6.84 (d, J=1.77 Hz, 1H) 7.02-7.08 (m, 1H) 7.31 (dd, J=8.84, 4.80 Hz, 1H) 7.53 (s, 1H) 7.64 (s, 1H) 7.74 (d, J=1.77 Hz, 1H); LCMS: 499.85 [M+1]; c-Met Ki: 0.036 μM.
The title compound was prepared according to procedure 2. The last alkylation procedure was performed by dissolving 2-11 (1 molar equivalent) in DMF (3 mL). 2-chloro-2-oxoethyl acetate (1 molar equivalent) and triethyl amine (5 molar equivalents) were added and the reaction stirred at rt for 16 h. The general work-up conditions were then followed (procedure 2) to afford 2-[3-(4-{6-amino-5-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-3-yl}-1H-pyrazol-1-yl)azetidin-1-yl]-2-oxoethyl acetate in 27% yield. The compound was dissolved in MeOH/H2O (4:1) and then potassium hydroxide (1 molar equivalent) was added. After stirring at rt for 3 h, H2O and EtOAc were added and the organic phase extracted, dried (Na2SO4), concentrated, and then purified by preparative HPLC to afford the title compound in 68% yield. 1H NMR (400 MHz, DMSO-D6) 5 ppm 1.84 (d, J=6.57 Hz, 3H) 3.95 (s, 2H) 4.08-4.18 (m, 1H) 4.32-4.44 (m, 2H) 4.64 (t, J=8.72 Hz, 1H) 5.24-5.32 (m, 1H) 6.23 (q, J=6.57 Hz, 1H) 7.11 (s, 1H) 7.47 (t, J=8.72 Hz, 1H) 7.59 (dd, J=9.09, 5.05 Hz, 1H) 7.74-7.77 (m, 2H) 8.17 (s, 1H); LCMS: 479.90 [M+1]; c-Met Ki: 0.022 μM.
The title compound was prepared according to procedure 1. The last alkylation procedure was performed by dissolving 1-10 (1 molar equivalent) in dichloromethane (3 mL). Acetic anhydride (1.1 molar equivalent) and triethyl amine (3 molar equivalents) were added and the reaction stirred at rt for 16 h. The reaction was then concentrated and purified by flash chromatography (gradient 50% EtOAc/Hexanes—100% EtOAc) to afford the title compound in 89% yield. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.71 (d, J=2.78 Hz, 3H) 1.79 (d, J=6.57 Hz, 3H) 2.92-3.03 (m, 1H) 3.62 (dd, J=9.60, 5.56 Hz, 1H) 3.82-3.93 (m, 2H) 4.13 (t, J=8.46 Hz, 1H) 4.30 (d, J=7.33 Hz, 2H) 5.67 (s, 2H) 6.07 (q, J=6.57 Hz, 1H) 6.86 (s, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.53-7.58 (m, 2H) 7.73 (d, J=1.77 Hz, 1H) 7.90 (d, J=1.77 Hz, 1H); LCMS: 477.90 [M+1]; c-Met Ki: 0.029 μM.
The title compound was prepared according to procedure 1. The last alkylation procedure was performed by dissolving 1-10 (1 molar equivalent) in DMF (3 mL). Sulfonyl chloride (1 molar equivalent) and triethyl amine (3 molar equivalents) were added and the reaction stirred at rt for 16 h. The general work-up conditions were then followed (procedure 1) to afford title compound in 54% yield. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.85 (d, J=6.57 Hz, 3H) 2.99 (s, 3H) 3.01-3.12 (m, 1H) 3.77 (dd, J=8.08, 6.06 Hz, 2H) 3.97 (t, J=8.34 Hz, 2H) 4.37 (d, J=7.07 Hz, 2H) 5.74 (s, 2H) 6.14 (q, J=6.40 Hz, 1H) 6.94 (d, J=1.52 Hz, 1H) 7.50 (t, J=8.72 Hz, 1H) 7.58-7.65 (m, 2H) 7.79 (d, J=1.77 Hz, 1H) 8.01 (s, 1H); LCMS: 514.10 [M+1]; c-Met Ki: 0.044 μM.
The title compound was prepared according to procedure 1. The last alkylation procedure was performed by dissolving 1-10 (1 molar equivalent) in DMF (3 mL). 2-iodopropane (1 molar equivalent) and triethyl amine (3 molar equivalents) were added and the reaction stirred at 50° C. for 16 h. The general work-up conditions were then followed (procedure 1) to afford title compound in 15% yield. 1H NMR (400 MHz, DMSO-D6) δ ppm 0.80 (d, J=6.06 Hz, 6H) 1.79 (d, J=6.32 Hz, 3H) 2.18 (ddd, J=12.25, 6.32, 6.19 Hz, 1H) 2.65 (td, J=13.45, 6.19 Hz, 1H) 2.83 (t, J=5.31 Hz, 2H) 3.13 (t, J=6.69 Hz, 2H) 4.22 (d, J=7.33 Hz, 2H) 5.65 (s, 2H) 6.07 (q, J=6.65 Hz, 1H) 6.86 (d, J=1.52 Hz, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.51 (s, 1H) 7.55 (dd, J=8.97, 4.93 Hz, 1H) 7.72 (d, J=1.52 Hz, 1H) 7.85 (s, 1H); LCMS: 478.20 [M+1]; c-Met Ki: 0.057 μM.
The title compound was prepared according to procedure 2. The last alkylation procedure was performed by dissolving 2-11 (1 molar equivalent) in DMF (3 mL). 2-iodopropane (1 molar equivalent) and triethyl amine (5 molar equivalents) were added and the reaction stirred at rt for 16 h. The general work-up conditions were then followed (procedure 2) to afford title compound in 23% yield. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.79 (d, J=6.82 Hz, 3H) 3.09 (s, 2H) 3.42-3.51 (m, 2H) 3.78 (t, J=7.58 Hz, 2H) 4.95 (qd, J=6.86, 6.69 Hz, 1H) 5.67 (s, 2H) 6.08 (q, J=6.65 Hz, 1H) 6.90 (d, J=1.52 Hz, 1H) 7.08 (s, 1H) 7.19 (s, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.54-7.59 (m, 2H) 7.76 (d, J=1.77 Hz, 1H) 8.11 (s, 1H); LCMS: 478.90 [M+1]; c-Met Ki: 0.021 μM.
The title compound was prepared according to procedure 2. The last alkylation procedure was performed by dissolving 2-11 (1 molar equivalent) in DMF (3 mL). Acetic anhydride (1 molar equivalent) and triethyl amine (3 molar equivalents) were added and the reaction stirred at rt for 16 h. The general work-up conditions were then followed (procedure 2) to afford title compound in 9% yield. 1H NMR (400 MHz, DMSO-D6) 5 ppm 1.79 (d, J=6.57 Hz, 3H) 1.80 (s, 3H) 4.08 (dt, J=9.35, 4.67 Hz, 1H) 4.27 (t, J=9.09 Hz, 1H) 4.36 (ddd, J=8.97, 4.42, 4.29 Hz, 1H) 4.54 (t, J=8.46 Hz, 1H) 5.20 (ddd, J=13.26, 7.96, 5.31 Hz, 1H) 5.69 (s, 2H) 6.08 (q, J=6.57 Hz, 1H) 6.91 (d, J=1.77 Hz, 1H) 7.44 (t, J=8.72 Hz, 1H) 7.56 (dd, J=9.09, 5.05 Hz, 1H) 7.66 (s, 1H) 7.76 (d, J=1.77 Hz, 1H) 8.07 (s, 1H); LCMS: 464.10 [M+1]; c-Met Ki: 0.032 μM.
The title compound was prepared according to procedure 1. The last alkylation procedure was performed by dissolving 1-10 (1 molar equivalent) in dichloromethane (3 mL). Bromoacetonitrile (1 molar equivalent) and triethyl amine (3 molar equivalents) were added and the reaction stirred at rt for 16 h. The general work-up conditions were then followed (procedure 1) to afford title compound in 51% yield. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.79 (d, J=6.57 Hz, 3H) 2.80 (ddd, J=13.14, 7.33, 5.81 Hz, 1H) 3.08 (t, J=6.44 Hz, 2H) 3.32 (t, J=7.20 Hz, 2H) 3.59 (s, 2H) 4.25 (d, J=7.33 Hz, 2H) 5.66 (s, 2H) 6.07 (q, J=6.57 Hz, 1H) 6.86 (d, J=1.77 Hz, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.52 (s, 1H) 7.56 (dd, J=8.97, 4.93 Hz, 1H) 7.72 (d, J=1.52 Hz, 1H) 7.88 (s, 1 H); LCMS: 475.10 [M+1]; c-Met Ki: 0.037 μM.
The title compound was prepared according to procedure 1 The last alkylation procedure was performed by dissolving 1-10 (1 molar equivalent) in DMF (3 mL). Bromocyclopropane (1 molar equivalent) and triethyl amine (3 molar equivalents) were added and the reaction stirred at rt for 16 h. The general work-up conditions were then followed (procedure 1) to afford title compound in 36% yield. 1H NMR (400 MHz, DMSO-D6) δ ppm 0.03 (q, J=4.80 Hz, 2H) 0.34 (ddd, J=8.08, 5.68, 4.17 Hz, 2H) 0.62-0.72 (m, 1H) 1.79 (d, J=6.82 Hz, 3H) 2.18 (d, J=6.57 Hz, 2H) 2.69-2.78 (m, 1H) 2.90 (t, J=5.81 Hz, 2H) 3.19 (t, J=7.20 Hz, 2H) 4.23 (d, J=7.33 Hz, 2H) 5.65 (s, 2 H) 6.07 (q, J=6.57 Hz, 1H) 6.86 (d, J=1.52 Hz, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.51 (s, 1H) 7.55 (dd, J=8.97, 4.93 Hz, 1H) 7.72 (d, J=1.77 Hz, 1H) 7.86 (s, 1H); LCMS: 490.10 [M+1]; c-Met Ki: 0.047 μM.
The title compound was prepared according to procedure 1. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.79 (d, J=6.82 Hz, 3H) 2.69-2.80 (m, 1H) 2.91 (s, 2H) 3.02 (t, J=5.68 Hz, 2H) 3.29 (td, J=7.07, 2.53 Hz, 2H) 4.28 (d, J=7.33 Hz, 2H) 5.66 (s, 2H) 6.07 (q, J=6.74 Hz, 1H) 6.85 (d, J=1.77 Hz, 1H) 7.06 (d, J=19.45 Hz, 2H) 7.43 (t, J=8.72 Hz, 1H) 7.52 (s, 1H) 7.56 (dd, J=8.97, 4.93 Hz, 1H) 7.72 (d, J=1.77 Hz, 1H) 7.85 (s, 1H); LCMS: 493 [M+1]; c-Met Ki: 0.035 μM.
The title compound was prepared according to procedure 1. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.79 (d, J=6.57 Hz, 3H) 2.13 (d, J=2.02 Hz, 6H) 2.85 (d, J=2.78 Hz, 2H) 2.95-3.05 (m, 1 H) 3.67 (dd, J=9.60, 5.31 Hz, 1H) 3.89 (t, J=8.97 Hz, 1H) 3.96 (dd, J=9.09, 5.31 Hz, 1H) 4.19 (t, J=8.59 Hz, 1H) 4.29 (d, J=7.07 Hz, 2H) 5.67 (s, 2H) 6.07 (q, J=6.57 Hz, 1H) 6.86 (s, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.49-7.59 (m, 2H) 7.73 (d, J=1.52 Hz, 1H) 7.90 (d, J=2.02 Hz, 1H); LCMS: 521.10[M+1].
The title compound was prepared according to procedures 21 and 22. 1H NMR (400 MHz, MeOD) δ ppm 1.72-1.84 (m, 2H) 1.86 (d, J=6.82 Hz, 3H) 1.90-2.02 (m, 2H) 2.08-2.25 (m, 4 H) 2.38 (s, 3H) 3.32-3.40 (m, 2H) 4.43-4.60 (m, 1H) 6.17 (q, J=6.57 Hz, 1H) 6.92 (d, J=1.77 Hz, 1H) 7.11-7.29 (m, 1H) 7.45 (dd, J=8.84, 4.80 Hz, 1H) 7.50 (s, 1H) 7.78 (s, 1H); LCMS: 491 [M+1]; c-Met Ki: 0.009 μM.
The title compound was prepared according to procedure 21. 1H NMR (400 MHz, MeOD) δ ppm 1.94 (d, J=6.57 Hz, 3H) 2.44 (d, J=13.90 Hz, 2H) 2.53-2.70 (m, 2H) 4.53-4.66 (m, 1H) 4.89-4.95 (m, 2H) 6.35 (q, J=6.15 Hz, 1H) 7.15 (s, 1H) 7.28 (t, J=8.59 Hz, 1H) 7.50 (dd, J=8.59, 4.55 Hz, 1H) 7.60 (s, 1H) 7.65 (s, 1H) 7.95 (s, 1H); LCMS: 500 [M+1]; c-Met Ki: 0.017 μM.
The title compound was prepared according to procedure 21. 1H NMR (400 MHz, MeOD) δ ppm 1.94 (d, J=6.57 Hz, 3H) 2.03-2.13 (m, 2H) 2.18-2.32 (m, 2H) 3.07-3.21 (m, 2H) 3.42-3.50 (m, 1H) 3.61 (t, J=12.76 Hz, 1H) 4.73-4.82 (m, 1H) 6.35 (q, J=6.57 Hz, 1H) 7.15 (s, 1H) 7.28 (t, J=8.59 Hz, 1H) 7.51 (dd, J=8.97, 4.93 Hz, 1H) 7.59 (s, 2H) 7.66 (s, 2H) 7.96 (s, 2H); LCMS: 500 [M+1]; c-Met Ki: 0.014 μM.
The title compound was prepared according to procedure 23. 1H NMR (400 MHz, DMSO-D6) δ ppm 1.65-1.76 (m, 2H) 1.79 (d, J=6.57 Hz, 3H) 1.85-1.95 (m, 2H) 2.51-2.64 (m, 2H) 3.01 (d, J=12.38 Hz, 2H) 4.03-4.23 (m, 1H) 5.63 (s, 2H) 6.07 (q, J=6.57 Hz, 1H) 6.88 (d, J=1.52 Hz, 1H) 7.43 (t, J=8.72 Hz, 1H) 7.51 (s, 1H) 7.56 (dd, J=8.97, 4.93 Hz, 1H) 7.74 (d, J=1.77 Hz, 1H) 7.90 (s, 1H); LCMS: 451 [M+1].
The title compound was prepared according to procedure 23. 1H NMR (400 MHz, chloroform-D) δ ppm 1.83 (d, J=6.57 Hz, 3H) 2.01-2.21 (m, 4H) 2.32-2.49 (m, 2H) 3.05 (d, J=11.87 Hz, 2H) 3.28 (s, 2H) 3.72 (s, 3H) 3.98-4.23 (m, 1H) 4.76 (s, 2H) 6.05 (q, J=6.82 Hz, 1H) 6.84 (d, J=1.77 Hz, 1H) 6.96-7.09 (m, 1H) 7.29 (dd, J=8.84, 4.80 Hz, 1H) 7.47 (s, 1H) 7.54 (s, 1H) 7.74 (d, J=1.77 Hz, 3H); LCMS: 523 [M+1].
It will be appreciated that, in any given series of compounds, a range of biological activities will be observed. In its presently preferred aspects, this invention relates to novel compounds capable of modulating, regulating and/or inhibiting protein kinase activity. The following assays may be employed to select those compounds demonstrating the optimal degree of the desired activity.
Assay Procedures
The following in vitro assay may be used to determine the level of activity and effect of the different compounds of the present invention on one or more of the PKs. Similar assays can be designed along the same lines for any PK using techniques well known in the art. A literature reference is provided (Technikova-Dobrova Z, Sardanelli A M, Papa S FEBS Lett. 1991 Nov. 4; 292: 69-72).
The general procedure is as follows: compounds and kinase assay reagents are introduced into test wells. The assay is initiated by addition of the kinase enzyme. Enzyme inhibitors reduce the measured activity of the enzyme.
In the continuous-coupled spectrophotometric assay the time-dependent production of ADP by the kinase is determined by analysis of the rate of consumption of NADH by measurement of the decrease in absorbance at 340 nm. As the PK produces ADP it is re-converted to ATP by reaction with phosphoenol pyruvate and pyruvate kinase. Pyruvate is also produced in this reaction. Pyruvate is subsequently converted to lactate by reaction with lactate dehydrogenase, which simultaneously converts NADH to NAD. NADH has a measurable absorbance at 340 nm whereas NAD does not.
The presently preferred protocol for conducting the continuous-coupled spectrophotometric experiments for specific PKs is provided below. However, adaptation of this protocol for determining the activity of compounds against other RTKs, as well as for CTKs and STKs, is well within the scope of knowledge of those skilled in the art.
HGFR Continuous-Coupled Spectrophotometric Assay
This assay analyzes the tyrosine kinase activity of HGFR on the Met-2 substrate peptide, a peptide derived from the activation loop of the HGFR.
Materials and Reagents:
Prep Dilution Buffer (DB) for Enzyme (For 30 mL prep)
1. Final concentration in assay:
1. Spectrophotometer settings:
The following in vitro assays may be used to determine the level of activity and effect of the different compounds of the present invention on one or more of the PKs. Similar assays can be designed along the same lines for any PK using techniques well known in the art.
Several of the assays described herein are performed in an ELISA (Enzyme-Linked Immunosorbent Sandwich Assay) format (Voller, et al., 1980, “Enzyme-Linked Immunosorbent Assay,” Manual of Clinical Immunology, 2d ed., Rose and Friedman, Am. Soc. Of Microbiology, Washington, D.C., pp. 359-371). General procedure is as follows: a compound is introduced to cells expressing the test kinase, either naturally or recombinantly, for a selected period of time after which, if the test kinase is a receptor, a ligand known to activate the receptor is added. The cells are lysed and the lysate is transferred to the wells of an ELISA plate previously coated with a specific antibody recognizing the substrate of the enzymatic phosphorylation reaction. Non-substrate components of the cell lysate are washed away and the amount of phosphorylation on the substrate is detected with an antibody specifically recognizing phosphotyrosine compared with control cells that were not contacted with a test compound.
The presently preferred protocols for conducting the ELISA experiments for specific PKs is provided below. However, adaptation of these protocols for determining the activity of compounds against other RTKs, as well as for CTKs and STKs, is well within the scope of knowledge of those skilled in the art.
Other assays described herein measure the amount of DNA made in response to activation of a test kinase, which is a general measure of a proliferative response. General procedure for this assay is as follows: a compound is introduced to cells expressing the test kinase, either naturally or recombinantly, for a selected period of time after which, if the test kinase is a receptor, a ligand known to activate the receptor is added. After incubation at least overnight, a DNA labeling reagent such as 5-bromodeoxyuridine (BrdU) or H3-thymidine is added. The amount of labeled DNA is detected with either an anti-BrdU antibody or by measuring radioactivity and is compared to control cells not contacted with a test compound.
MET Transphosphorylation Assay
This assay is used to measure phosphotyrosine levels on a poly(glutamic acid: tyrosine, 4:1) substrate as a means for identifying agonists/antagonists of met transphosphorylation of the substrate.
Materials and Reagents:
The following assays use cells engineered to express a selected receptor and then evaluate the effect of a compound of interest on the activity of ligand-induced DNA synthesis by determining BrdU incorporation into the DNA. The following materials, reagents and procedure are general to each of the following BrdU incorporation assays. Variances in specific assays are noted.
General Materials and Reagents:
A549 cells (ATCC) were used in this assay. Cells were seeded in the growth media (RPMI+10% FBS) into 96 well plates and cultured overnight at 37° C. for attachment. Cells were exposed to the starvation media (RPMI+0.05% BSA). Dilutions of the inhibitors were added to the plates and incubated at 37° C. for 1 hour. Cells were then stimulated by adding 40 ng/mL HGF for 15 minutes. Cells were washed once with 1 mM Na3VO4 in HBSS and then lysed. The lysates were diluted with 1 mM Na3VO4 in HBSS and transferred to a 96 well goat ant-rabbit coated plate (Pierce) which was pre-coated with anti-HGFR antibody (Zymed Laboratories). The plates were incubated overnight at 4° C. and washed with 1% Tween 20 in PBS for seven times. HRP-PY20 (Santa Cruz) was diluted and added to the plates for 30 minutes incubation. Plates were then washed again and TMB peroxidase substrate (Kirkegaard & Perry) was added and incubated for 10 minutes. The reaction was then stopped by adding 0.09N H2SO4. Plates were measured at OD-450 nm using a spectrophotometer. IC50 values were calculated by curve fitting using a four-parameter analysis.
Compounds of the invention were measured for HGFR inhibition activity; the data are shown in each Example. Ki data were obtained using the HGFR Continuous-Coupled Spectrophotometric Assay, and IC50 data were obtained using the Cellular HGFR Autophosphorylation Assay, both of which are described above.
While the invention has been illustrated by reference to specific and preferred embodiments, those skilled in the art will recognize that variations and modifications may be made through routine experimentation and practice of the invention. Thus, the invention is intended not to be limited by the foregoing description, but to be defined by the appended claims and their equivalents.
All references cited herein, including any priority documents, are hereby incorporated by reference in their entireties.
This application claims the benefit of U.S. Provisional Application No. 60/605,244 filed Aug. 28, 2004, and U.S. Provisional Application No. 60/690,803 filed Jun. 14, 2065, the contents of which are hereby incorporated by reference in their entireties.
| Number | Date | Country | |
|---|---|---|---|
| 60605244 | Aug 2004 | US | |
| 60690803 | Jun 2005 | US |
