TRK INHIBITORS USEFUL AS ANTICANCER DRUGS

Abstract

A group of tropomyosin receptor kinase inhibitors including an amido-phenoxy-indazole structure fragment. These compounds show potent inhibitory effects on various oncogenic kinases, especially TRK, and thus can be used to treat tumors or cancers.

Claims

1. A compound of formula I: ##STR00175## wherein: R.sup.1 is C.sub.1-6 alkyl or C.sub.3-6 cycloalkyl, optionally substituted by one or more halogens, —OR.sup.a, C.sub.1-6 alkyl or amino; R.sup.5 is H, halogen, —OR.sup.a or optionally substituted C.sub.1-6 alkyl, and the substituent is selected from one or more halogen, —OR.sup.a, C.sub.1-6 alkyl or amino; or R.sup.1 and R.sup.5 together with the atoms to which they are attached form a cyclic structure of the following formula: ##STR00176## where X is —CR.sup.7R.sup.8—, ═CR.sup.7—, —CR.sup.7R.sup.8—CR.sup.7R.sup.8— or —CR.sup.7═CR.sup.8—; R.sup.6, R.sup.7, R.sup.8 and R.sup.9 are independently selected from H, halogen, —OR.sup.a, optionally substituted C.sub.1-6 alkyl or C.sub.3-6 cycloalkyl, wherein the substituent is selected from one or more halogens, —OR.sup.a, C.sub.1-6 alkyl or amino; or R.sup.6 and R.sup.9 or R.sup.7 and R.sup.8 attached to the same carbon atom can form ═O or ═S together, or form a C.sub.3-6 cycloalkyl group together with the carbon atom to which they are attached; R.sup.2 is C.sub.1-6 alkyl optionally substituted by one or more halogens, C.sub.1-6 alkyl, C.sub.1-6 alkoxy or amino; C.sub.3-6 cycloalkyl or C.sub.3-6 heterocyclic group, optionally substituted by one or more halogens, —OR.sup.a, C.sub.1-6 alkyl or amino; nitro, cyano, acyl, halogen, mercapto, C.sub.1-6 alkylthio, C.sub.1-6 alkylsulfonyl, C.sub.1-6 alkyl sulfinyl or carboxy; R.sup.3 and R.sup.4 are each independently selected from H, halogen, nitro, cyano, acyl or carboxy; R.sup.a is selected from H or C.sub.1-6 alkyl; provided that R.sup.1 and R.sup.2 are not methyl at the same time, or isomers thereof, or their pharmaceutically acceptable salts or solvates.

2. The compound of claim 1, isomers thereof or their pharmaceutically acceptable salts or solvates, wherein R.sup.1 is C.sub.1-6 alkyl, preferably methyl, ethyl or isopropyl, or R.sup.1 is C.sub.3-6 cycloalkyl, preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

3. The compound of claim 1, isomers thereof, or their pharmaceutically acceptable salts or solvates, wherein R.sup.5 is H.

4. The compound of claim 1, isomers thereof or their pharmaceutically acceptable salts or solvates, wherein R.sup.1 and R.sup.5 together with the atoms to which they are attached form a cyclic structure of formula (a), wherein —X— is —CR.sup.7R.sup.8— or —CR.sup.7R.sup.8—CR.sup.7R.sup.8—, and R.sup.6, R.sup.7, R.sup.8 and R.sup.9 are each independently H, halogen, hydroxy, C.sub.1-6 alkyl or C.sub.3-6 cycloalkyl.

5. The compound of claim 1, isomers thereof or their pharmaceutically acceptable salts or solvates, wherein R.sup.1 and R.sup.5 together with the atoms to which they are attached form a cyclic structure of formula (a), wherein —X— is ═CR.sup.7— or —CR.sup.7═CR.sup.8—, and R.sup.6, R.sup.7, R.sup.8 and R.sup.9 are each independently H, halogen, C.sub.1-6 alkyl or C.sub.3-6 cycloalkyl.

6. The compound of claim 1, isomers thereof or their pharmaceutically acceptable salts or solvates, wherein R.sup.1 and R.sup.5 together with the atoms to which they are attached form a cyclic structure of formula (a), wherein R.sup.6 and R.sup.9 or R.sup.7 and R.sup.8 attached at the same carbon atom together form ═O, ═S or C.sub.3-6 cycloalkyl.

7. The compound of claim 1, isomers thereof or their pharmaceutically acceptable salts or solvates, wherein R.sup.2 is C.sub.1-6 alkyl, preferably methyl, ethyl, isopropyl or trifluoromethyl.

8. The compound of claim 1, isomers thereof or their pharmaceutically acceptable salts or solvates, wherein R.sup.2 is C.sub.3-6 cycloalkyl or C.sub.3-6 heterocyclyl, preferably cyclopropyl.

9. The compound of claim 1, isomers thereof or their pharmaceutically acceptable salts or solvates, wherein R.sup.3 and R.sup.4 are each independently selected from H or halogen, preferably F.

10. A compound of formula III, isomers thereof or their pharmaceutically acceptable salts or solvates, ##STR00177## wherein R.sup.1 is C.sub.1-6 alkyl or C.sub.3-6 cycloalkyl, optionally substituted by one or more halogen, —OR.sup.a, —SR.sup.a, C.sub.1-6 alkyl or amino; R.sup.5 is H, halogen, —OR.sup.a, —SR.sup.a, or optionally substituted C.sub.1-6 alkyl, and the substituent is selected from one or more halogen, —OR.sup.a, —SR.sup.a, C.sub.1-6 alkyl or amino; or R.sup.1 and R.sup.5 together with the atoms to which they are attached form a cyclic structure of the following formula: ##STR00178## where X is —CR.sup.7R.sup.8—, ═CR.sup.7—, —CR.sup.7R.sup.8—CR.sup.7R.sup.8— or —CR.sup.7═CR.sup.8—; R.sup.6, R.sup.7, R.sup.8 and R.sup.9 are each independently H, halogen, —OR.sup.a, —SR.sup.a, optionally substituted C.sub.1-6 alkyl or optionally substituted C.sub.3-6 cycloalkyl, wherein the substituent is selected from one or more halogens, —OR.sup.a, —SR.sup.a, C.sub.1-6 alkyl or amino; or R.sup.6 and R.sup.9 or R.sup.7 and R.sup.8 attached to the same carbon atom can form ═O or ═S together, or form a C.sub.3-6 cycloalkyl group together with the carbon atom to which they are attached; R.sup.2 is —NR.sup.aR.sup.b or —OR.sup.b; R.sup.3 and R.sup.4 are each independently selected from H, halogen, nitro, cyano, acyl or carboxy; R.sup.10 is hydrogen, halogen, cyano, nitro, —NR.sup.aR.sup.b, —OR.sup.b, —SR.sup.b, C(O)R.sup.b, CO.sub.2R.sup.b; C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 Cycloalkyl, C.sub.3-6 heterocycloalkyl, 6-10 membered aryl, 5-10 membered heteroaryl; C.sub.1-6 alkyl or C.sub.2-6 alkenyl or C.sub.2-6 alkynyl, optionally substituted by one or more halogens, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylthio or amino; C.sub.3-6 cycloalkyl or C.sub.3-6 heterocycloalkyl, optionally substituted by one or more hydroxy, halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylthio or amino; 6-10 membered aryl or 5-10 membered heteroaryl, optionally substituted by one or more hydroxy, halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylthio or amino; R.sup.a and R.sup.b are each independently H, C.sub.1-6 alkyl optionally substituted by one or more halogens, CN or nitro, or C.sub.3-6 cycloalkyl optionally substituted by one or more halogens, CN or nitro, or R.sup.a and R.sup.b together with the N to which they are attached form a 3-6 membered ring optionally substituted by one or more halogens, CN or nitro; provided that R.sup.1 and R.sup.2 are not methyl at the same time.

11. The compound of formula III of claim 10, isomers thereof or their pharmaceutically acceptable salts or solvates, which is of formula III-a, ##STR00179## wherein R.sup.1 is C.sub.1-6 alkyl; R.sup.2 is —NR.sup.aR.sup.b or —OR.sup.b; R.sup.3 and R.sup.4 are each independently selected from H or halogen; R.sup.a and R.sup.b are each independently selected from H, C.sub.1-6 alkyl optionally substituted by one or more halogens, or C.sub.3-6 cycloalkyl optionally substituted by one or more halogens, or R.sup.a and R.sup.b together with the N to which they are attached form a 3-6 membered ring optionally substituted by one or more halogens; and provided that R.sup.1 and R.sup.2 are not methyl at the same time.

12. The compound of claim 11, isomers or their pharmaceutically acceptable salts or solvates, wherein R.sup.1 is C.sub.1-6 alkyl, preferably methyl; R.sup.2 is —OR.sup.b, wherein R.sup.b is selected from C.sub.1-6 alkyl or C.sub.3-6 cycloalkyl, optionally substituted by one or more halogens, preferably F, R.sup.2 is preferably methoxy, ethoxy, propoxy, cyclopropoxy, isopropoxy, trifluoroethoxy, trifluoromethoxy, difluoromethoxy; R.sup.3 and R.sup.4 are each independently selected from halogen, preferably F.

13. The compound of formula III of claim 10, isomers thereof or their pharmaceutically acceptable salts or solvates, which is of formula III-b, ##STR00180## wherein: R.sup.2 is —NR.sup.aR.sup.b or —OR.sup.b; X is —CR.sup.7R.sup.8—, ═CR.sup.7—, —CR.sup.7R.sup.8—CR.sup.7R.sup.8— or —CR.sup.7═CR.sup.8—; R.sup.6 and R.sup.9 are each independently H, halogen, —OH, C.sub.1-6 alkyl or C.sub.3-6 cycloalkyl; R.sup.7 and R.sup.8 are each independently H, halogen, —OH, C.sub.1-6 alkyl optionally substituted with one or more halogens or C.sub.3-6 cycloalkyl optionally substituted with one or more halogens, or R.sup.7 and R.sup.8 attached to the same carbon atom together form a C.sub.3-6 cycloalkyl; R.sup.3 and R.sup.4 are each independently selected from H or halogen; R.sup.a and R.sup.b are each independently selected from H, C.sub.1-6 alkyl optionally substituted by one or more halogens, or C.sub.3-6 cycloalkyl optionally substituted by one or more halogens, or R.sup.a and R.sup.b together with N to which they are attached form a 3-6 membered ring optionally substituted with one or more halogens; provided that R.sup.1 and R.sup.2 are not methyl at the same time.

14. The compound of claim 13, its isomers or their pharmaceutically acceptable salts or solvates, wherein X is —CR.sup.7R.sup.8— or —CR.sup.7R.sup.8—CR.sup.7R.sup.8—; R.sup.2 is —OR.sup.b, wherein R.sup.b is selected from C.sub.1-6 alkyl optionally substituted by one or more halogens, preferably F, or C.sub.3-6 cycloalkyl optionally substituted by one or more halogens, preferably F, R.sup.2 is preferably methoxy, ethoxy, propoxy, cyclopropoxy, isopropoxy, trifluoroethoxy, trifluoromethoxy, difluoromethoxy; R.sup.3 and R.sup.4 are each independently selected from halogen, preferably F.

15. The compound of claim 13, its isomers or their pharmaceutically acceptable salts or solvates, wherein X is ═CR.sup.7— or —CR.sup.7═CR.sup.8—; R.sup.2 is —OR.sup.b, wherein R.sup.b is selected from C.sub.1-6 alkyl optionally substituted by one or more halogens, preferably F, or C.sub.3-6 cycloalkyl optionally substituted with one or more halogens, preferably F, R.sup.2 is preferably methoxy, ethoxy, propoxy, cyclopropoxy, isopropoxy, trifluoroethoxy, trifluoromethoxy, difluoromethoxy; R.sup.3 and R.sup.4 are each independently selected from halogen, preferably F.

16. The compound, isomers thereof or their pharmaceutically acceptable salts or solvates, selected from: ##STR00181## ##STR00182## ##STR00183## ##STR00184## ##STR00185## ##STR00186## ##STR00187## ##STR00188## ##STR00189## ##STR00190## ##STR00191## ##STR00192## ##STR00193## ##STR00194## ##STR00195## ##STR00196## ##STR00197##

17. The compound of claim 1, isomers thereof or their pharmaceutically acceptable salts or solvates, wherein the pharmaceutically acceptable salt is p-toluenesulfonate, benzenesulfonate, or methanesulfonate.

18. A pharmaceutical composition comprising the compound of claim 1, isomers thereof or their pharmaceutically acceptable salts or solvates, and a pharmaceutically acceptable carrier, diluent or excipient, and optionally other therapeutically active agents selected from: other Trk inhibitors, kinase inhibitors, antibody drugs, immunotherapeutics, anticancer agents, anti-inflammatory drugs, analgesics, cardiovascular drugs, lipid-lowering drugs, antibacterial agents, antiviral agents, antidiabetic agents, antiproliferative agents, antiangiogenic agents or antiallergic agents.

19. A method comprising use of the compound of claim 1, isomers thereof or their pharmaceutically acceptable salts or solvates, or a pharmaceutical composition comprising the compound, in the manufacture of medicaments for the treatment or prevention of diseases or disorders wherein the activity of Trk plays a role or is involved or diseases or disorders wherein the kinases shown in Table 10 of the specification play a role or are involved.

20. The method of claim 19, wherein the diseases or disorders are treated or prevented by inhibiting Trk or the kinases shown in Table 10 of the specification, and said diseases or disorders are tumors or cancers selected from the group consisting of sarcoma (especially infantile fibrosarcoma carcinoma), skin cancer, neuroblastoma, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, salivary gland cancer, multiple myeloma, astrocytoma and medulloblastoma, neuroglioma, melanoma, thyroid cancer, lung adenocarcinoma, large cell neuroendocrine tumors, head and neck cancer and colorectal cancer, cholangiocarcinoma, glioblastoma, glioma, secretory breast cancer, mammary secretory carcinoma, acute myeloid leukemia, congenital mesoderm nephroma, congenital fibrosarcoma, acute lymphoblastic leukemia, colon adenocarcinoma, gastrointestinal stromal tumor; preferably lung adenocarcinoma, intrahepatic cholangiocarcinoma, colorectal cancer, papillary thyroid carcinoma, spitzoid neoplasm, glioblastoma, sarcoma, astrocytoma, head and neck cancer, low-grade glioma, secretory breast cancer, acute myeloid leukemia, congenital mesodermal nephroma, congenital fibrosarcoma, acute lymphoblastic leukemia, colorectal adenocarcinoma, thyroid carcinoma, cutaneous melanoma and pediatric glioma sarcoma, skin cancer, neuroblastoma, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, multiple myeloma, astrocytoma, and medulloblastoma, neuroglioma, melanoma, thyroid cancer, lung adenocarcinoma, large cell neuroendocrine tumor, head and neck cancer, and colorectal cancer.

21. The method for preparing the compound of claim 1, isomers thereof or their pharmaceutically acceptable salts or solvates thereof, which comprises: (a) reacting a compound of formula A ##STR00198## with a compound of formula B in the presence of a condensing reagent to form an amide, ##STR00199## to obtain a compound of formula C ##STR00200## or (a′) reacting a compound of formula A ##STR00201## with a compound of formula D in the presence of a condensing reagent to form an amide, ##STR00202## to obtain a compound of formula E, ##STR00203## the compound of formula E is subjected to a Suzuki coupling reaction to obtain the compound of formula C; and (b) deprotecting the compound of formula C to obtain the compound of formula I ##STR00204## wherein R.sup.1 to R.sup.5 are as defined in claim 1.

Description

DESCRIPTION OF THE FIGURES

[0241] FIG. 1: Anti-tumor activity of the compounds of the present disclosure in a KM12 xenograft mouse model. FIG. 1-A shows the effect of the compounds of the present disclosure on tumor volume; FIG. 1-B shows the concentrations of the compounds of the present disclosure in plasma and tumor tissue 12 days after administration; FIG. 1-C shows the concentrations of the compounds of the disclosure in plasma and tumor tissue.

[0242] FIG. 2: Comparison of the activity profile of the compound of Example 12 of the present disclosure and the prior art Merestinib compound.

EXAMPLES

[0243] The present disclosure will be further illustrated in conjunction with the examples as follows. It should be noted that the following examples cannot be recognized as a limitation to the protection scope of the present disclosure.

[0244] The experimental methods that do not indicate specific conditions in the following examples usually follow the conventional conditions for this type of reaction or the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are percentages by weight and parts by weight. Unless otherwise specified, ratios of the liquid are by volume.

[0245] The experimental materials and reagents used in the following examples can be obtained from commercial channels, prepared according to the methods of prior art, or prepared according to a method similar to that disclosed in the present application, unless otherwise specified.

[0246] In the following examples, .sup.1H-NMR spectra are recorded with Bruker 400 MHz NMR and Agilent 500 MHz NMR nuclear magnetic resonance instruments, and chemical shifts are expressed in δ (ppm); mass spectroscopy is recorded using Agilent 1290 liquid chromatography+6120B mass spectrometry LCMS instrument.

Abbreviations

[0247] ACN(MeCN) acetonitrile [0248] Ar argon [0249] BBr.sub.3 boron tribromide [0250] tBuONO tert-butyl nitrite [0251] CDCl.sub.3 deuterated chloroform [0252] CS.sub.2CO.sub.3 cesium carbonate [0253] CuCl cuprous chloride [0254] DCM dichloromethane [0255] DIBAL-H diisobutyl aluminum hydride [0256] DIPEA N,N-diisopropylethylainine [0257] DMF dimethylfonnamide [0258] DMF-DMA N,N-dimethylformamide dimethyl acetal [0259] DMSO dimethyl sulfoxide [0260] DMSO-d.sup.6 deuterated dimethyl sulfoxide [0261] ESI electrospray ionization [0262] EA(EtOAc) ethyl acetate [0263] EDCI 1-ethyl-(3-dimethylaminopropyl)carbodiimide [0264] EtOH ethanol [0265] FaSSGF fasted state simulated gastric fluids [0266] FaSSIF fasted state simulated intestinal fluid [0267] FeSSIF fed state simulated intestinal fluid [0268] h hour [0269] HATU O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate [0270] HCl hydrochloric acid or hydrogen chloride [0271] HOBt 1-hydroxybenzotriazole [0272] HPLC high performance liquid chromatography [0273] H.sub.2O water [0274] K.sub.3PO.sub.4 potassium phosphate [0275] LCMS liquid chromatography-mass spectrometry [0276] LiOH lithium hydroxide [0277] MeOH methanol [0278] MHz megahertz [0279] MS mass spectrometry [0280] MsCl methanesulfonyl chloride [0281] NaH sodium hydride [0282] NaHCO.sub.3 sodium bicarbonate [0283] NaOH sodium hydroxide [0284] NaOMe sodium methoxide [0285] Na.sub.2SO.sub.4 sodium sulfate [0286] NMR nuclear magnetic resonance [0287] PE petroleum ether [0288] Pd/C palladimn/carbon [0289] PdCl.sub.2(dbpf) [1,1′-bis(tert-butylphosphino)ferrocene]palladium dichloride [0290] PdCl.sub.2(dppf) [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride [0291] o/n overnight [0292] rt room temperature [0293] SOCl.sub.2 thionyl chloride [0294] SiO.sub.2 silica, silica gel [0295] TBAF tetra-n-butylammonium fluoride [0296] TEA triethylamine [0297] TEMPO tetramethylpiperidine nitrogen oxide [0298] TFA trifluoroacetic acid [0299] TMS-CN trimethylsilyl cyanide [0300] TFAA trifluoroacetic anhydride [0301] THF tetrahydrofuran [0302] TLC thin layer chromatography [0303] TMSCHN.sub.2 trimethylsilyldiazomethane

SYNTHESIS EXAMPLES

[0304] Each abbreviation in the following synthesis examples has the meaning commonly understood by one skilled in the art. Unless otherwise stated, all temperatures are given in degrees Celsius; all reagents are commercially available and used without further purification.

Preparation of Intermediate 1: 2,4-dibromo-5-hydroxybenzaldehyde

[0305] Bromine (6.72 g, 84 mmol) was slowly added dropwise to a solution of 3-hydroxybenzaldehyde (4.48 g, 40 mmol) in dichloromethane (80 mL) at 20° C. The resulting reaction mixture was stirred at room temperature for 24 hours, and then 15% sodium thiosulfate aqueous solution (40 mL) was added at 40° C. to quench the reaction. The precipitated product was washed with water and dried to obtain 4.6 g of solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.17 (s, 1H), 10.16-10.06 (m, 1H), 8.01 (d, J=2.2 Hz, 1H), 7.41 (d, J=2.2 Hz, 1H), MS [M+H].sup.+: 279.0.

Preparation of Intermediate 2: 2,4-dibromo-5-(2-fluoro-4-nitrophenoxy) benzaldehyde

[0306] To a solution of 2,4-dibromo-5-hydroxybenzaldehyde (Intermediate 1, 4.5 g, 16 mmol) in N,N-dimethylformamide (50 mL) was added potassium carbonate powder (3.26 g, 24 mmol) and 1,2-difluoro-4-nitrobenzene (2.68 g, 16.9 mmol), the resulting reaction mixture was stirred at 50° C. for 6 hours, TLC showed that the reaction was complete. The reaction mixture was extracted three times with ethyl acetate (30 mL each time), washed with water five times, dried, and evaporated under reduced pressure to remove the solvent to obtain 5.6 g of a yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 10.18 (s, 1H), 8.08 (dd, J=10.1, 2.5 Hz, 1H), 7.99 (d, J=9.2 Hz, 1H), 7.97 (s, 1H), 7.44 (s, 1H), 6.95 (t, J=8.4 Hz, 1H).

Preparation of Intermediate 3: (E)-1-(2,4-dibromo-5-(2-fluoro-4-nitrophenoxy) benzylidene)-2-isopropylhydrazine

[0307] To a solution of 2,4-dibromo-5-(2-fluoro-4-nitrophenoxy)benzaldehyde (Intermediate 2, 2.2 g, 5.3 mmol) in N,N-dimethylformamide (50 mL) was added isopropylhydrazine (0.641 g, 5.8 mmol), and the resulting reaction mixture was stirred at 20° C. for 0.5 hours. TLC showed that the reaction was complete. The reaction mixture was extracted three times with ethyl acetate (30 mL each time), washed with water 5 times, dried and evaporated under reduced pressure to remove the solvent to obtain 2.4 g of a yellow solid, which was directly used in the next reaction.

Preparation of Intermediate 4: 6-bromo-5-(2-fluoro-4-nitrophenoxy)-1-isopropyl-1H-indazole

[0308] To a N.sub.2 protected solution of (E)-1-(2,4-dibromo-5-(2-fluoro-4-nitrophenoxy)benzylidene)-2-isopropylhydrazine (Intermediate 3, 2.4 g, 5 mmol) in N,N-dimethylformamide was added potassium carbonate powder (1.1 g, 8 mmol) and cuprous chloride (0.08 g, 0.8 mmol), the resulting reaction mixture was stirred at 100° C. for 3 hours, TLC showed that the reaction was complete. The reaction mixture was cooled to room temperature and extracted three times with ethyl acetate (30 mL each time), washed with water five times, dried, and evaporated under reduced pressure to remove the solvent. The erode product was purified by a silica gel column chromatography, eluted with ethyl acetate/petroleum ether (1:20 to 1:10) to obtain 0.7 g of a yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.13 (d, J=10.3 Hz, 1H), 8.01 (s, 1H), 7.83 (s, 1H), 7.45 (s, 1H), 6.74 (t, J=8.5 Hz, 1H), 4.83 (dt, J=13.0, 6.5 Hz, 1H), 1.64 (d, J=6.6 Hz, 6H). MS [M+H].sup.+: 394.0.

Preparation of Intermediate 5: 6-bromo-5-(2 fluoro 4 nitrophenoxy)-1-methyl-1H-indazole

[0309] The intermediate was prepared according to the synthetic route of Intermediate 4, using Intermediate 2 and methylhydrazine. MS [M+H].sup.+: 366.0.

Preparation of Intermediate 6: 6 bromo-5-(2-fluoro-4-nitrophenoxy)-1-ethyl-1H-indazole

[0310] The intermediate was prepared according to the synthetic route of Intermediate 4, using Intermediate 2 and ethylhydrazine. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.98 (dd, J=10.3, 2.5 Hz, 1H), 7.84 (s, 1H), 7.79 (d, J=9.1 Hz, 1H), 7.66 (s, 1H), 7.39 (s, 1H), 6.58 (t, J=8.5 Hz, 1H), 4.30 (q, J=7.2 Hz, 2H), 1.42 (t, J=7.2 Hz, 3H).

Preparation of Intermediate 7: N-(4-fluorophenyl)-2-cyanoacetamide

[0311] To a solution of 4-fluoroaniline (6.6 g, 56 mmol) in dichloromethane (200 mL) was added HATU (25 g, 75 mmol), N,N-diisopropylethylamine (21.6 g, 166 mmol) and 2-cyanoacetic acid (5.6 g, 75 mmol), the resulting reaction mixture was stirred at room temperature for 2 hours, then washed with saturated aqueous ammonium chloride solution, and dried. The erode product obtained by concentration was purified with a silica gel column and eluted with ethyl acetate-petroleum ether (1:5 to 2:1) to obtain 5 g of a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.38 (s, 1H), 7.56 (dd, J=8.8 Hz, 4.9 Hz, 2H), 7.18 (t, J=8.8 Hz, 2H), 3.90 (s, 2H). MS [M+H].sup.+: 179.1.

Preparation of Intermediate 8: 1-trimethylsilyl-pent-1-yn-3-one

[0312] To a solution of bistrimethylsilylacetylene (5.6 g, 35 mmol) in dichloromethane at 0° C. was added propionyl chloride (2.79 g, 30 mmol). The resulting reaction mixture was stirred for 10 minutes and then added with aluminum trichloride (4.8 g, 36 mmol), further stirred at 0° C. for 2 hours, then naturally warmed to room temperature, and stirred for another 2 hours. The reaction mixture was poured into a 10% hydrochloric acid (70 mL) solution in ice-water and stirred for 10 minutes. After the two phases were separated, the aqueous phase was extracted three times with dichloromethane (25 mL each time), dried and evaporated under reduced pressure to remove the solvent to obtain a crude brown oil product, which was directly used in the next reaction.

Preparation of Intermediate 9: (E)-1-methoxy-pent-1-en-3-one

[0313] To a solution of 1-trimethylsilyl-pent-1-yn-3-one (Intermediate 8, all erode products) in methanol (20 mL) was slowly added 1,4-diazabicyclo[2,2,2]Octane (6.72 g, 60 mmol), the resulting reaction mixture was stirred at room temperature for 20 minutes and then concentrated, then diluted with ethyl acetate (20 mL), washed twice with saturated brine (10 mL each time), dried, and evaporated under reduced pressure to remove solvent. The crude product was purified by a silica gel column and eluted with ethyl acetate-petroleum ether (1:10) to obtain 2.1 g of the product. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.62 (d, J=12.7 Hz, 1H), 5.60 (d, J=12.7 Hz, 2.4 Hz, 1H), 3.71 (q, J=7.8 Hz, 3H), 1.11 (t, J=7.8 Hz, 3H).

Preparation of Intermediate 10: (E)-1-methoxy-4-methyl-pent-1-en-3-one

[0314] The intermediate was prepared from isobutyryl chloride according to the synthetic route of Intermediate 9. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.61 (d, J=12.5, 1H), 5.62 (d, J=12.5, 1H), 2.72-2.56 (m, 1H), 1.12-1.09 (d, 6H).

Preparation of Intermediate 11: (E)-1-cyclopropyl-3-methoxy-prop-2-en-1-one

[0315] The intermediate was prepared from cyclopropylformyl chloride according to the synthetic route of Intermediate 9. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.64 (d, J=12.6 Hz, 1H), 5.72 (d, J=12.6 Hz, 1H), 3.72 (d, J=4.7 Hz, 3H), 1.93 (d, J=4.5 Hz, 1H), 1.10-1.02 (m, 2H), 0.91-0.81 (m, 2H).

Preparation of Intermediate 12: 1-(4-fluorophenyl)-6-methyl-3-cyano-2-oxo-1,2-dihydropyridine

[0316] To a solution of N-(4-fluorophenyl)-2-cyanoacetamide (Intermediate 7, 1 g, 5.6 mmol) in diethylene glycol monoethyl ether (20 mL) was added 1,4-diazabicyclo[2,2,2]octane (0.62 g, 5.6 mmol) and (E)-1-methoxy-but-1-en-3-one (0.84 g, 8.4 mmol), the resulting reaction mixture was stirred at 120° C. for 14 hours. After cooling to room temperature, it was washed with hydrochloric acid and saturated sodium chloride aqueous solution. The crude product was purified with a silica gel column and eluted with ethyl acetate-petroleum ether (1:1) to obtain 0.7 g of yellow solid product. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.81 (d, J=7.4 Hz, 1H), 7.26 (t, J=7.7 Hz, 2H), 7.22-7.16 (m, 2H), 6.27 (d, J=7.4 Hz, 1H), 2.09 (s, 3H). MS [M+H].sup.+: 229.1.

Preparation of Intermediate 13: 1-(4-fluorophenyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0317] To 1-(4-fluorophenyl)-6-methyl-3-cyano-2-oxo-1,2-dihydropyridine (Intermediate 12, 0.7 g) was added concentrated sulfuric acid/water (1 mL/1 mL), the resulting reaction mixture was stirred at 100° C. for 15 hours, cooled to room temperature, and extracted with ethyl acetate three times (20 mL each time), and then washed with a saturated aqueous sodium chloride solution. After the solvent was evaporated under reduced pressure, the crude product was dissolved in ethanol (2 mL), added with 2N sodium hydroxide solution, and washed twice with ethyl acetate (10 mL each time). The aqueous solution was acidified with IN hydrochloric acid, precipitated, filtered, and dried to obtain 0.5 g of the product. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.43 (d, J=7.5 Hz, 1H), 7.54 (dd, J=8.8 Hz, 5.1 Hz, 2H), 7.46 (t, J=8.7 Hz, 2H), 6.82 (d, J=7.6 Hz, 1H), 2.11 (s, 3H). MS [M+H].sup.+: 248.1.

Preparation of Intermediate 14: 1-(4-fluorophenyl)-6-ethyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0318] The intermediate was prepared according to the synthetic route of Intermediate 13 using Intermediate 7 and Intermediate 9. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 14.23 (s, 1H), 8.45 (dd. J=7.7, 2.6 Hz, 1H), 7.58-7.49 (m, 2H), 7.45 (t, J=10.0 Hz, 2H), 6.76 (dd, J=7.8, 2.5 Hz, 1H), 2.31 (q, J=7.4 Hz, 2H), 1.05 (t, J=7.4 Hz, 3H), MS [M+H].sup.+: 262.3.

Preparation of Intermediate 15: 1-(4-fluorophenyl)-6-isopropyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0319] The intermediate was prepared according to the synthetic route of Intermediate 13 using Intermediate 7 and Intermediate 10. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.47 (d, J=7.8 Hz, 1H), 7.58 (dd, J=8.8, 4.9 Hz, 2H), 7.46 (t, J=8.7 Hz, 2H), 6.86 (d, J=7.8 Hz, 1H), 2.46 (m, 1H), 1.13 (t, J=7.2 Hz, 6H). MS [M+H].sup.+: 276.3.

Preparation of Intermediate 16: 1-(4-fluorophenyl)-6-cyclopropyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0320] The intermediate is prepared according to the synthetic route of Intermediate 13 using Intermediate 7 and Intermediate 11. .sup.1H NMR (400 MHz. DMSO-d.sub.6) δ 8.37 (d, J=7.8 Hz, 1H), 7.56 (d, J=4.3 Hz, 2H), 7.45 (t, J=8.6 Hz, 2H), 6.54 (d, J=8.0 Hz, 1H), 1.35 (m, 1H), 0.93 (m, 2H), 0.85 (m, 2H). MS [M+H].sup.+: 274.3.

Preparation of Intermediate 17: 1-(4-Fluorophenyl)-6-trifluoromethyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0321] This intermediate is prepared according to the synthetic route of Intermediate 13, using Intermediate 7 and (E)-1-methoxy-4,4,4-trifluoro-1-buten-3-one (obtained by a method similar to the synthesis of Intermediate 9). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.66 (s, 1H), 8.42 (d, J=7.4 Hz, 1H), 7.60-7.51 (m, 2H), 7.42 (t. J=8.7 Hz, 2H), 7.25 (d, J=7.5 Hz, 1H). MS [M+H].sup.+: 302.1.

Preparation of Intermediate 18: 6-bromo-5-(2-fluoro-4-aminophenoxy)-1-isopropyl-1H-indazole

[0322] In a 100 mL round bottom flask was added 6-bromo-5-(2-fluoro-4-nitrophenoxy)-1-isopropyl-1H-indazole (Intermediate 4, 0.2 g, 0.5 mmol), 10% palladium on carbon (0.1 g) and ethyl acetate. The gas in the flask was replaced with hydrogen 5 times, and then stirred overnight at room temperature. TLC and LCMS showed that the reaction was complete to obtain single product. The palladium/carbon catalyst was removed by filtration, the filter cake was rinsed with ethyl acetate (25 mL), and the product obtained was directly used in the next reaction. MS [M+H].sup.+: 364.0.

Preparation of Intermediate 19: N-(3-fluoro-4-(1-isopropyl-6-bromo-1H-indazol-5-yloxy)phenyl)-6-methyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0323] In a 25 mL round bottom flask was added 6-bromo-5-(2-fluoro-4-aminophenoxy)-1-isopropyl-1H-indazole (Intermediate 18, 0.2 g, 0.5 mmol), 1-(4-fluorophenyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid (Intermediate 13, 0.15 g, 0.6 mmol), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (0.215 g, 1.12 mmol), 1-hydroxybenzotriazole (0.105 g, 0.75 mmol), N,N-diisopropylethyl amine (0.325 g, 2.5 mmol) and N,N-dimethylformamide (4.5 mL), the mixture was stirred overnight at room temperature, then distributed in ethyl acetate (25 mL) and saturated aqueous sodium chloride solution (25 mL). The organic phase was separated and washed four times with saturated sodium chloride aqueous solution (25 mL each time), the aqueous phase was extracted three times with ethyl acetate (25 mL each time), the combined organic phase was dried, concentrated, and the crude product was purified with a silica gel column, eluted with ethyl acetate-petroleum ether (1:5 to 1:3) to obtain a yellow solid product. MS [M+H].sup.+: 593.1.

Preparation of Intermediate 20: N-(3-fluoro-4-(1-isopropyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0324] In a 25 mL round bottom flask was added N-(3-fluoro-4-(1-isopropyl-6-bromo-1H-indazol-5-yloxy)phenyl)-6-methyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide (Intermediate 19, 0.5 mmol), tripotassium phosphate (0.223 g, 1.05 mmol), 1-Boc-pyrazole-4-boronic acid pinacol ester (0.182 g, 0.62 mmol), di-tert-butyl dicarbonate (0.022 g, 0.1 mmol), [1,1-bis(diphenylphosphino)ferrocene]palladium dichloride (0.005 g, 0.0075 mmol), water (1 mL) and tetrahydrofuran (15 mL), the mixture was replaced with nitrogen 8 times, and then stirred at 45° C. overnight. The solvent was evaporated under reduced pressure, and the crude product was purified with a silica gel column, eluted with ethyl acetate-petroleum ether (1:3 to 1:2) to obtain 0.263 g of a yellow solid product. MS [M+H].sup.+: 680.2.

Preparation of Intermediate 21: N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0325] Intermediate 21 was prepared according to the synthetic route of Intermediate 20, using Intermediate 6 (6-bromo-5-(2-fluoro-4-nitrophenoxy)-1-ethyl-1H-indazole) and Intermediate 13 (1-(4-fluorophenyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS [M+H].sup.+: 667.2.

Preparation of Intermediate 22: N-(3-Fluoro-4-(1-isopropyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-isopropyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0326] Intermediate 22 was prepared according the synthetic route of Intermediate 20, using Intermediate 4 (6-bromo-5-(2-fluoro-4-nitrophenoxy)-1-isopropyl-1H-indazole) and Intermediate 15 (1-(4-fluorophenyl)-6-isopropyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid).

Preparation of Intermediate 23: N-(3-fluoro-4-(1-isopropyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-trifluoromethyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0327] Intermediate 23 was prepared according the synthetic route of Intermediate 20, using Intermediate 4 (6-bromo-5-(2-fluoro-4-nitrophenoxy)-1-isopropyl-1H-indazole) and Intermediate 17 (1-(4-fluorophenyl)-6-trifluoromethyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid).

Preparation of Intermediate 24: 6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-nitrophenoxy)-1-methyl-1H-indazole

[0328] In a 100 mL round bottom flask was added 6-bromo-5-(2-fluoro-4-nitrophenoxy)-1-methyl-1H-indazole (Intermediate 5, 2.334 g, 6.37 mmol), tripotassium phosphate (2.84 g, 13.4 mmol), 1-Boc-pyrazole-4-boronic acid pinacol ester (2.81 g, 9.6 mmol), di-tert-butyl dicarbonate (0.278 g, 1.27 mmol), [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (0.25 g, 0.383 mmol), water (10 mL) and tetrahydrofuran (150 mL), the mixture was replaced with nitrogen 8 times, and then stirred at 45° C. overnight. The solvent was evaporated under reduced pressure, and the crude product was purified with a silica gel column, eluted with ethyl acetate-petroleum ether (1:3 to 1:2) to obtain 3.78 g of a yellow solid product. MS [M+H].sup.+: 454.2.

Preparation of Intermediate 25: 6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole

[0329] In a 250 mL round bottom flask was added 6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-nitrophenoxy)-1-methyl-1H-indazole (Inter-mediate 24, 1.89 g, 4.17 mmol), 10% palladium/carbon (1 g) and ethyl acetate (75 mL), replaced the gas in the flask with hydrogen 5 times, and then stirred at 40° C. for 1 hour. TLC and LCMS showed that the reaction was complete with a single product. The palladium/carbon was removed by filtration, and the solvent was evaporated under reduced pressure to obtain 1.2 g of the product which was directly used in the next reaction. MS [M+H].sup.+: 424.2.

Preparation of Intermediate 26: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-ethyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0330] In a 25 mL round bottom flask was added 6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole (Intermediate 25, 0.212 g, 0.5 mmol), 1-(4-fluorophenyl)-6-ethyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid (Intermediate 14, 0.131 g, 0.5 mmol), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (0.215 g, 1.12 mmol), 1-hydroxybenzotriazole (0.105 g, 0.75 mmol), N,N-diisopropylethylamine (0.325 g, 2.5 mmol) and N,N-dimethylformamide (4.5 mL), the mixture was stirred at room temperature overnight, and then distributed in ethyl acetate (25 mL) and saturated sodium chloride aqueous solution (25 mL). The organic phase was separated and washed four times with saturated sodium chloride aqueous solution (25 mL each time), the aqueous phase was extracted three times with ethyl acetate (25 mL each time), the combined organic phase was dried and concentrated, and the crude product was purified with silica gel column, eluted with ethyl acetate-petroleum ether (1:1 to 2:1) to obtain a yellow solid product. MS [M+H].sup.+: 667.2.

Preparation of Intermediate 27: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-isopropyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0331] Intermediate 27 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 15 (1-(4-fluorophenyl)-6-isopropyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS [M+H].sup.+: 681.2.

Preparation of Intermediate 28: N-(3 Fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-cyclopropyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0332] Intermediate 28 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 16 (1-(4-fluorophenyl)-6-cyclopropyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS [M+H].sup.+: 679.2.

Preparation of Intermediate 29: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-trifluoromethyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0333] Intermediate 29 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 17 (1-(4-fluorophenyl)-6-trifluoromethyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS [M+H].sup.+: 707.2.

Preparation of intermediate 30: 3-((4-fluorophenyl)amino) 3 oxo-propionic acid ethyl ester

[0334] The mixture of monoethyl malonate (10 g, 8.93 mL, 75.69 mmol), 4-fluoroaniline (7.17 mL, 75.69 mmol), EDCI HCl (14.51 g, 75.69 mmol), HOBt (10.23 g, 75.69 mmol), TEA (21.04 mL, 151.38 mmol) and DCM (120 mL) was stirred at room temperature for 16 hours. The reaction was quenched with water (200 mL), acidified to pH=7 with concentrated hydrochloric acid, and extracted with ethyl acetate (3×200 mL). The organic phase was washed with saturated brine (200 mL) and dried over anhydrous Na.sub.2SO.sub.4. After filtration and concentration, a white solid (17.2 g, 75.7% yield) was obtained. The crude product was used directly in the next reaction without purification. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.26 (s, 1H), 7.54-7.50 (m, 2H), 7.03 (dd, J=11.9, 5.4 Hz, 2H), 4.26 (q, J=7.1 Hz, 2H), 3.47 (s, 2H), 1.33 (t, J=7.2 Hz, 3H). MS-ESI [M+H].sup.+: 226.2.

Preparation of Intermediate 31: ethyl 3-dimethylamino-2-((4-fluorophenyl)carboxamido) acrylate

[0335] To a solution of 3-((4-fluorophenyl)amino)-3-oxo-propionic acid ethyl ester (3 g, 13.32 mmol) in toluene (50 mL) was added DMF-DMA (2.65 mL, 19.98 mmol). The mixture was stirred at 90° C. for 2 hours. After the reaction mixture was concentrated, the crude product was recrystallized with ethyl acetate/petroleum ether (1:10, 22 mL) to obtain a white solid (1.71 g, 45.8% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.03 (s, 1H), 7.65-7.62 (m, 2H), 7.50-7.43 (m, 1H), 7.11 (t, J=8.8 Hz, 2H), 4.07 (q, J=6.9 Hz, 2H), 2.99 (s, 6H), 1.17 (t, J=7.0 Hz, 3H). MS-ESI [M+H].sup.+: 281.2.

Preparation of Intermediate 32: 6-amino-1-(4-fluorophenyl)-5-(methylthio)-2-oxo-1,2-dihydropyridine-3-carboxylic add ethyl ester

[0336] To a solution of cyanomethyl dimethylsulfonium bromide (16.29 g, 89.48 mmol) in DMSO (250 mL) was added Cs.sub.2CO.sub.3 (72.88 g, 223.69 mmol), and stirred at room temperature for 0.5 hour, and then was added ethyl 3-dimethylamino-2-((4-fluorophenyl)carboxamido)acrylate. The mixture was heated to 100° C. and stirred for 16 hours. The reaction was quenched by adding water (300 mL), filtered, washed with water (2*100 mL), and the crude product was recrystallized with ethyl acetate/petroleum ether (1:2.90 mL) to obtain a white solid (18 g, 74.9% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.15 (s, 1H), 7.39-7.31 (m, 2H), 7.30-7.26 (m, 2H), 7.00 (br, 2H), 4.09 (q, J=7.1 Hz, 2H), 2.20 (s, 3H), 1.19 (t, J=7.2 Hz, 3H). MS-ESI [M+H].sup.+: 323.0.

Preparation of Intermediate 33: ethyl 6-amino-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylate

[0337] To a solution of 6-amino-1-(4-fluorophenyl)-5-(methylthio)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester (1 g, 3.1 mmol) in THF (30 mL) was added PdCl.sub.2 (55 mg, 0.31 mmol) and triethylsilane (0.99 mL, 6.20 mmol), and stirred at room temperature for 16 horns. After filtration and concentration, the reaction was washed with ethyl acetate (2×10 mL) to obtain a white solid 33 (0.5 g, 58.3% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.92 (d, J=8.8 Hz, 1H), 7.40-7.35 (m, 2H), 7.28-7.25 (m, 2H), 6.92 (br, 2H), 5.61 (d, J=8.8 Hz, 1H), 4.07 (q, J=7.2 Hz, 2H), 1.18 (t, J=7.2 Hz, 3H). MS-ESI [M+H].sup.+: 277.2.

Preparation of Intermediate 34: ethyl 6-chloro-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylate

[0338] To a mixture of 6-amino-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester (6 g, 21.7 mmol) and CuCl (2.6 g, 26.1 mmol) in ACN (50 mL) was added t-BuONO (4.8 mg, 35.8 mmol) dropwise, and stirred overnight at room temperature. After the reaction solution was concentrated, the crude product was purified with a silica gel column, eluted with a gradient of 0-20% ethyl acetate/dichloromethane to obtain 34 as a yellow solid (1.6 g, 25% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.122 (d, J=8.0 Hz, 1H), 7.46-7.39 (m, 4H), 6.70 (d, J=8.0 Hz, 1H), 4.22 (q, J=7.2 Hz, 2H), 1.25 (t, J=7.2 Hz, 3H). MS-ESI [M+H].sup.+: 295.9.

Preparation of Intermediate 35: 1-(4-fluorophenyl)-6-methoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0339] To a mixture of 6-chloro-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester (0.5 g, 1.69 mmol) in THE (20 mL) was added NaOMe (915 mg, 16.9 mmol), and stirred at room temperature for 2 horns. The reaction solution was quenched with 1N HCl (100 mL) and extracted with ethyl acetate (2×100 mL). The organic phase was washed with saturated brine (100 mL) and dried over anhydrous sodium sulfate. After filtration and concentration, the crude product was prepared by HPLC. A white solid (0.178 g, 40% yield) was obtained. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.77 (s, 1H), 8.48 (d, J=8.8 Hz, 1H), 7.47-7.36 (m, 4H), 6.43 (d, J=8.8 Hz, 1H), 3.93 (s, 3H). MS-ESI [M+H].sup.+: 264.1.

Preparation of Intermediate 36: 1-(4-fluorophenyl)-6-ethoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0340] Intermediate 36 was prepared according to the synthetic route of Intermediate 35, from Intermediate 34 (6-chloro-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester) and sodium ethoxide. .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ 13.80 (s, 1H), 8.46 (d, J=8.4 Hz, 1H), 7.48-7.39 (m, 4H), 6.43 (d, J=8.8 Hz, 1H), 4.29 (q, 7=6.8 Hz, 2H), 1.16 (t, J=7.0 Hz, 3H). MS-ESI [M+H].sup.+: 278.1.

Preparation of Intermediate 37: 1-(4-fluorophenyl)-6-cyclopropoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0341] Intermediate 37 was prepared according to the synthetic route of Intermediate 35, from Intermediate 34 (6-chloro-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester) and sodium hydride and cyclopropanol. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.78 (s, 1H), 8.52 (d, J=8.8 Hz, 1H), 7.44-7.37 (m, 4H), 6.65 (d, J=8.4 Hz, 1H), 4.23-4.18 (m, 1H), 0.831 (t, J=6.4 Hz, 2H), 0.371 (t, J=6.4 Hz, 7.2 Hz, 2H). MS-ESI [M+H].sup.+: 290.0.

Preparation of Intermediate 38: 1-(4-fluorophenyl)-6-isopropoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0342] Intermediate 38 was prepared according to the synthetic route of Intermediate 35, from Intermediate 34 (6-chloro-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester) and sodium hydride and isopropanol. .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ 13.801 (s, 1H), 8.436 (d, J=8.8 Hz, 1H), 7.442-7.354 (m, 4H), 6.468 (d, J=8.8 Hz, 1H), 4.967-4.876 (m, 1H), 1.180 (d, J=8.8 Hz, 6H). MS-ESI [M+H].sup.+: 292.0.

Preparation of Intermediate 39: 1-(4-fluorophenyl)-6-(2,2,2-trifluoroethoxy)-2-oxo-1,2 dihydropyridine-3-carboxylic acid

[0343] Intermediate 39 was prepared according to the synthetic route of Intermediate 35, from Intermediate 34 (6-chloro-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester) and sodium hydride and 2,2,2-trifluoroethanol. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.68 (s, 1H), 8.52 (d, J=8.5 Hz, 1H), 7.54-7.32 (m, 4H), 6.51 (d, J=8.6 Hz, 1H), 5.05 (q, J=8.4 Hz, 2H). MS-ESI [M+H].sup.+: 332.0.

Preparation of Intermediate 40: 1-(4-fluorophenyl)-6-methylthio-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0344] Intermediate 40 was prepared according to the synthetic route of Intermediate 35, from Intermediate 34 (6-chloro-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester) and sodium thiomethoxide. The crude product was directly used in the next reaction. MS-ESI [M+H].sup.+: 280.0.

Preparation of Intermediate 41: 6-amino-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0345] To 6-amino-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester (1.0 g, 3.62 mmol) and ethanol (15 mL) was added NaOH (2.90 g, 72.39 mmol) and water (8 mL), and stirred at 60° C. for 24 hours. The reaction solution was concentrated and dissolved in water (30 mL), and extracted with DCM (3*50 mL). The aqueous solution was neutralized to pH 6.0 with concentrated HCl, and extracted with ethyl acetate (3×50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow solid (0.450 g, 50% yield). The crude product was directly used in the next reaction. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.75 (s, 1H), 8.00 (d, J=8.8 Hz, 1H), 7.45 (s, 2H), 7.43 (d, J=3.0 Hz, 2H), 7.22 (dd, J=50.1, 31.4 Hz, 2H), 5.95 (d, J=8.8 Hz, 1H). MS-ESI [M+H].sup.+: 249.0.

Preparation of Intermediate 42: 1-(4-fluorophenyl)-6-chloro-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0346] To a mixture of 6-amino-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid (0.45 g, 1.81 mmol) and CuCl (0.179 g, 1.81 mmol) in MeCN (10 mL) was added t-BuONO (0.62 ml, 2.99 mmol) dropwise, and stirred at room temperature for 3 days. The reaction was quenched with aqueous HCl (1N, 20 mL) and extracted with ethyl acetate (5×30 mL). After the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, the crude product was subjected to HPLC to obtain a yellow solid (0.080 g, 16.5% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 13.31 (s, 1H), 8.55 (d, J=7.9 Hz, 1H), 7.32-7.27 (m, 4H), 6.80 (d, J=7.9 Hz, 1H). MS-ESI [M+H].sup.+: 267.9.

Preparation of Intermediate 43: 6 (Boc-amino)-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester

[0347] To a mixture of ethyl 6-amino-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylate (0.32 g, 1.16 mmol) in DCM (10 mL) was added TEA (0.32 mL, 2.32 mmol) and Boc.sub.2O (0.53 mL, 2.32 mmol) dropwise, and stirred at room temperature for 16 hours. The reaction was quenched with water (10 mL) and extracted with ethyl acetate (3×20 mL). After the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, the crude product was purified with a silica gel column, eluted with a gradient of 1/10 to ⅕ to ½ ethyl acetate/dichloromethane to obtain a yellow solid (1.6 g, 25% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.32 (d, J=8.6 Hz, 1H), 7.34-7.26 (m, 4H), 7.12 (d, J=8.4 Hz, 1H), 6.17 (s, 1H), 4.33 (q, J=7.2 Hz, 2H), 1.44 (s, 9H), 1.35 (t, J=7.1 Hz, 3H). MS-ESI [M+H].sup.+: 377.1.

Preparation of Intermediate 44: 6-(Boc-amino)-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0348] To a mixture of 6-(Boc-amino)-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester (0.18 g, 0.48 mmol) in EtOH (2 mL) was added NaOH (0.382 ml, 9.56 mmol) and water (2 mL) dropwise and stirred at room temperature for 64 hours. The reaction solution was diluted with water (10 mL) and extracted with DCM (2×10 mL). The aqueous phase was acidified with concentrated HCl to pH 6, and then extracted with ethyl acetate (4×10 mL). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow solid (0.12 g, 72% yield). The crude product was used directly in the next reaction without purification. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 13.36 (br s, 1H), 8.56 (d, J=8.8 Hz, 1H), 7.44-7.39 (m, 3H), 7.35-7.28 (m, 2H), 6.29 (s, 1H), 1.46 (s, 9H). MS-ESI [M+H].sup.+: 349.1.

Preparation of Intermediate 45: 6-(methylsulfoxide)-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0349] To a mixture of 6-(methylthio)-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester (0.15 g, 0.54 mmol) in MeCN (3 mL) was added 30% H.sub.2O.sub.2 (0.12 mL, 1.08 mmol) and concentrated HCl (0.06 mL) dropwise, and stirred at room temperature for 16 hours. After the reaction solution was concentrated, it was subjected to HPLC to obtain a white solid (0.08 g, 50.4% yield). MS-ESI [M+H].sup.+: 296.0.

Preparation of Intermediate 46: (E)-N-(5-bromo-4-methoxy-2-methylphenyl)-2-oxime acetamide

[0350] A mixture of trichloroacetaldehyde hydrate (11.0 g, 66.6 mmol), sodium sulfate (16.0 g, 113 mmol), 5-bromo-4-methoxy-2-methylaniline (13 g, 51.8 mmol), concentrated hydrochloric acid (8 mL), hydroxylamine hydrochloride (13.4 g, 195 mmol) and water (500 mL) was stirred at 100° C. for 6 hours, then cooled to 0° C. The resulting precipitate was collected by filtration, washed with water, and dried to obtain a brown solid (9.0 g, 61% yield). MS-ESI [M+H].sup.+: 286.9.

Preparation of Intermediate 47: 4 bromo-5-methoxy-7-methylindoline 2,3 dione

[0351] A mixture of (E)-N-(5-bromo-4-methoxy-2-methylphenyl)-2-oximeacetamide (9.0 g, 31 mmol) and concentrated sulfuric acid (26 mL) was heated to 80° C. and stirred for 2 hours, then cooled to room temperature, slowly poured on ice, and stirred for 1 hour. The resulting precipitate was collected by filtration, dispersed in ethanol (100 mL) at 80° C. and stirred for 1 hour. After cooling to room temperature, a solid precipitated, filtered, and dried to obtain a red solid (6.3 g, 75% yield). MS-ESI [M+H].sup.+: 269.9.

Preparation of Intermediate 48: 2-amino-6˜bromo-5-methoxy-3-methylbenzoic acid

[0352] At 80° C., to the solution of 4-bromo-5-methoxy-7-methylindoline-2,3-dione (6.0 g, 22 mmol) in 3N NaOH (0.7 mL) was slowly add 30% of aqueous H.sub.2O.sub.2 (6 mL) dropwise and kept stirring for 2 hours. Then the reaction was cooled to room temperature, adjusted the solution to pH 3-5 with concentrated hydrochloric acid, and stirred for another 1 hour at room temperature. The resulting precipitate was collected by filtration and dried to obtain a light brown solid (4.5 g, 78% yield). MS-ESI [M+H].sup.+: 259.9.

Preparation of Intermediate 49: methyl 2-amino-6-bromo-5-methoxy-3-methylbenzoate

[0353] To a solution of 2-amino-6-bromo-5-methoxy-3-methylbenzoic acid (4.5 g, 17 mmol) in THF/MeOH (4:1.50 mL) was added dropwise trimethyl silicon diazomethane solution (2M in hexane, 34 mL) at room temperature, and kept stirring for 2 hours. Then the reaction was treated with 3 mL of glacial acetic acid, the solvent was evaporated, and the residue was purified by silica gel column chromatography, eluted with 3:1 petroleum ether/ethyl acetate, to obtain a light yellow solid (3.0 g, 63% yield). MS-ESI [M+H].sup.+: 273.9.

Preparation of Intermediate 50: methyl 6-bromo-5-methoxy-1H-indazole-7-carboxylate

[0354] To a mixture of methyl 2-amino-6-bromo-5-methoxy-3-methylbenzoate (3.0 g, 10.98 mmol) and glacial acetic acid (50 mL) was added a solution of NaNO.sub.2 (0.91 g, 13 mmol) in water (10 mL), heated to 50° C. and kept stirring for 16 hours. The solvent was evaporated, and the residue was washed with water and dried in vacuo to obtain a pale yellow solid (2.84 g, 91% yield). MS-ESI [M+H].sup.+: 285.0.

Preparation of Intermediate 51: methyl 6 bromo-5-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-7-carboxylate

[0355] Methyl 6-bromo-5-methoxy-1H-indazole-7-carboxylate (10 g, 35.2 mmol) and DHP (20 mL, 70.4 mmol) were dissolved in THF/CHCl.sub.3 (1:2, 240 mL), and TsOH (1.0 g, 5.26 mmol) was added. The mixture was stirred at 50° C. overnight. After cooling to room temperature, it was concentrated, and the residue was purified by silica gel column chromatography, eluted with a gradient of 0-20% ethyl acetate/petroleum ether, to obtain a yellow solid (10 g, 77% yield). MS-ESI [M+H].sup.+: 369.1.

Preparation of Intermediate 52: 6 bromo-5-methoxy-1-(tetrahydro-2H-pyran-2-y 1)-1H-indazole-7-methanol

[0356] At −20° C., to a toluene solution of methyl 6-bromo-5-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-7-carboxylate (10 g, 27.1 mmol) was added DIBAL-H (70 mL, 108.4 mmol, 1.5M in toluene). The mixture was slowly warmed to room temperature and kept stirring for 1 hour. The reaction was quenched with water and extracted with ethyl acetate. The organic phase was washed with water, saturated brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by silica gel column chromatography, eluted with a gradient of 0-20% ethyl acetate/petroleum ether, to obtain a white solid (9 g, 97% yield). MS-ESI [M+H].sup.+: 341.0.

Preparation of Intermediate 53: (6-bromo-5-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-7-methyl methanesulfonate

[0357] At 0° C., to a solution of 6-bromo-5-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-7-methanol (10.0 g, 29.4 mmol) in DCM (200 mL) were added TEA (16 mL, 117.6 mmol) and MsCl (0.45 mL, 5.88 mmol). The mixture was slowly warned to room temperature and stirred for 1 hour. The reaction was quenched with water and extracted with DCM. The organic phase was washed with water, saturated brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was used directly in the next reaction. MS-ESI [M+H].sup.+: 356.9.

Preparation of Intermediate 54: 2-(6-bromo-5-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-7-yl)acetonitrile

[0358] To a solution of (6-bromo-5-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-7-yl)methyl methanesulfonate (10.0 g, 29.4 mmol) in ACN (20 mL) were added TMS-CN (5.9 mL, 44.1 mmol) and TBAF (1M, 44 mL, 117.6 mmol). After the mixture was stirred at room temperature for 3 hours, the reaction was diluted with water and extracted with ethyl acetate. The organic phase was washed with water, saturated brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by silica gel column chromatography, eluted with a gradient of 0-20% ethyl acetate/petroleum ether, to obtain a white solid (3.5 g, 35% yield). MS-ESI [M+H].sup.+: 350.1.

Preparation of Intermediate 55: 2-(6-bromo-5-methoxy-1-(tetrahydro-1H-pyran-2-yl)-1H-indazol-7-yl)acetaldehyde

[0359] At 0° C., to a solution of 2-(6-bromo-5-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-7-yl)acetonitrile (2.50 g, 10 mmol) in toluene (35 mL) was added DIBAL-H (7.16 mL, 15 mmol, 1.5M in toluene). The mixture was slowly warmed to room temperature and stirred for 2 hours. The reaction was quenched with water, extracted with ethyl acetate, the organic phase was washed with water, saturated brine, dried over anhydrous Na.sub.2SO.sub.4, and concentrated. The crude product was purified by silica gel column chromatography, eluted with a gradient of 0-20% ethyl acetate/petroleum ether, to obtain a white solid (1.5 g, 59% yield). MS-ESI [M+H].sup.+: 269.0.

Preparation of Intermediate 56: 2 (6 bromo-5-methoxy-1-(tetrahydro-2H-pyran-2 yl)-1H-indazol-7-yl)ethanol

[0360] To a solution of 2-(6-bromo-5-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-7-yl)acetaldehyde (1.5 g, 2.6 mmol) in methanol (30 mL) was added NaBH.sub.4 (0.66 g, 5.2 mmol). The mixture was stirred at room temperature for 2 hours. The reaction was quenched with water, extracted with DCM, the organic phase was washed with water, saturated brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by silica gel column chromatography, eluted with a gradient of 0-20% ethyl acetate/petroleum ether, to obtain a white solid (1.3 g, 86% yield). MS-ESI [M+H].sup.+: 255.0.

Preparation of Intermediate 57: 6 bromo 7 (2 chloroethyl)-5-methoxy-1H-indazole

[0361] At 0° C., to a solution of 2-(6-bromo-5-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-7-yl)ethanol (0.35 g, 0.99 mmol) in ACN (35 mL) were added DMF (2 drops) and SOCl.sub.2 (3.5 mL). The mixture was warmed to room temperature and stirred for 48 hours. The reaction solution was concentrated to obtain the crude product as a brown solid, which was used directly in the next reaction. MS-ESI [M+H].sup.+: 291.0.

Preparation of Intermediate 58: 5-bromo-4-methoxy-6,7-dihydropyrrolo[3,2,1-hi]-indazole

[0362] At room temperature, to a solution of 6-bromo-7-(2-chloroethyl)-5-methoxy-1H-indazole (0.6 g, 2.07 mmol) in DMF (60 mL) was added Cs.sub.2CO.sub.3 (3.38 g, 10.36 mmol). The mixture was heated to 100° C. and stirred for 4 hours. The reaction mixture was filtered and concentrated, and the crude product was purified by silica gel column chromatography, eluted with a gradient of 0-40% ethyl acetate/petroleum ether to obtain a yellow solid 58 (0.11 g, 44% yield in two steps). MS-ESI [M+H].sup.+: 255.0.

Preparation of Intermediate 59: 5-bromo-6,7-dihydropyrrolo[3,2,1-hi]-indazol-4-ol

[0363] At 0° C., to a solution of 5-bromo-4-methoxy-6,7-dihydropyrrolo[3,2,1-hi]-indazole (0.15 g, 0.592 mmol) in DCM (5 mL) was added BBr.sub.3 Solution (17% wt in DCM, 5.93 mL, 10 mmol). The mixture was warmed to room temperature and stirred for 12 hours. The reaction was quenched with ice water and extracted with DCM. The organic phase was concentrated to obtain crude product as a brown solid. MS-ESI [M+H].sup.+: 241.1.

Preparation of Intermediate 60: 5-bromo-4-(2-fluoro-4-nltrophenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazole

[0364] To a solution of 5-bromo-6,7-dihydropyrrole[3,2,1-hi]-indazol-4-ol (0.25 g) in DMF (10 ml) were added NaHCO.sub.3 (0.263 g, 3.14 mmol) and 1,2-difluoro-4-nitrobenzene (0.166 g, 1.05 mmol). The mixture was heated to 60° C. and stirred for 12 hours. The reaction mixture was filtered and concentrated, and the crude product was purified by silica gel column chromatography, eluted with a gradient of 0-30% ethyl acetate/petroleum ether to obtain a solid (0.08 g, 36% yield in two steps). MS-ESI [M+H].sup.+: 380.0.

Preparation of Intermediate 61: tert-butyl 4-(4-(2-fluoro-4-nitrophenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-5-yl)-1H-pyrazole-1-carboxylate

[0365] To a solution of 5-bromo-4-(2-fluoro-4-nitrophenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazole (60 mg, 0.158 mmol) in THF (6 mL) and water (0.6 mL) were added K.sub.3PO.sub.4 (67 mg, 0.316 mmol) and Pd(dbpf)Cl.sub.2 (4.9 mg, 0.08 mmol). The mixture was heated to 60° C. and stirred for 12 hours. The reaction solution was washed with water, extracted with ethyl acetate, the organic phase was concentrated, and the crude product was purified by silica gel column chromatography, eluted with a gradient of 0-50% ethyl acetate/petroleum ether, to obtain a solid (40 mg, 54% yield). MS-ESI [M+H].sup.+: 466.2.

Preparation of Intermediate 62: tert-butyl 4-(4-(4-amino-2-fluorophenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-5-yl)-1H-pyrazole-1-carboxylate

[0366] To a solution of 4-(4-(2-fluoro-4-nitrophenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-5-yl)-1H-pyrazole-1-carboxylic acid tert-butyl ester (50 mg, 0.1 mmol) in THF (5 ml) was added Pd/C (10%, 30 mg). The mixture was stirred at room temperature for 12 hours under hydrogen atmosphere. The hydroxylamine intermediate was detected by LCMS, the reaction mixture was filtered, added additional Pd/C (10%, 30 mg), and stirred for another 8 horns at room temperature under hydrogen atmosphere, filtered and concentrated to give the crude product as a solid (68 mg). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 13.31 (s, 1H), 8.55 (d, J=7.9 Hz, 1H), 7.32-7.27 (m, 4H), 6.80 (d, J=7.9 Hz, 1H). MS-ESI [M+H].sup.+: 267.9.

Preparation of Intermediate 63: 6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-ethyl-1H-indazole

[0367] Intermediate 63 was prepared according to the synthetic route of Intermediate 25 from Intermediate 14 6-bromo-5-(2-fluoro-4-nitrophenoxy)-1-ethyl-1H-indazole. MS-ESI[M+H].sup.+: 438.3.

Preparation of Intermediate 64: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-Chloro-2-oxy-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0368] Intermediate 64 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 42 (1-(4-fluorophenyl)-6-chloro-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 673.3.

Preparation of Intermediate 65: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-(Boc amino)-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0369] Intermediate 65 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 44 (1-(4-fluorophenyl)-6-(Boc amino)-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 754.3,

Preparation of Intermediate 66: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methylthio-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0370] Intermediate 66 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 40 (1-(4-fluorophenyl)-6-methylthio-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 685.2.

Preparation of Intermediate 67: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methylsulfoxide-2-oxo-1-(4-fluorophenyl)-1,2-dihydro pyridine-3-carboxamide

[0371] Intermediate 67 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 45 (1-(4-fluorophenyl)-6-methylsulfoxide-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 701.2.

Preparation of Intermediate 68: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0372] Intermediate 68 was prepared according to the synthetic route of Intermediate 26, starting from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 35 (1-(4-fluorophenyl)-6-methoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 669.1.

Preparation of Intermediate 69: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-ethoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0373] Intermediate 69 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 36 (1-(4-fluorophenyl)-6-ethoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 683.2.

Preparation of Intermediate 70: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-cyclopropoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0374] Intermediate 70 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 37 (1-(4-fluorophenyl)-6-cyclopropoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 695.2.

Preparation of Intermediate 71: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-isopropoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0375] Intermediate 71 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 38 (1-(4-fluorophenyl)-6-isopropoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 697.2.

Preparation of Intermediate 72: N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-(2,2,2-trifluoroethoxy)-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0376] Intermediate 72 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-methyl-1H-indazole) and Intermediate 39 (1-(4-fluorophenyl)-6-(2,2,2-trifluoroethoxy)-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 737.1.

Preparation of Intermediate 73: N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-ethyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0377] Intermediate 73 was prepared according to the synthetic route of Intermediate 26, from Intermediate 63 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-ethyl-1H-indazole) and Intermediate 14 (1-(4-fluorophenyl)-6-ethyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 681.0.

Preparation of Intermediate 74: N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-cyclopropyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0378] Intermediate 74 was prepared according to the synthetic route of Intermediate 26, from Intermediate 63 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-ethyl-1H-indazole) and Intermediate 16 (1-(4-fluorophenyl)-6-cyclopropyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 693.0.

Preparation of Intermediate 75: N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-chloro-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0379] Intermediate 75 was prepared according to the synthetic route of Intermediate 26, from Intermediate 63 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-ethyl-1H-indazole) and Intermediate 42 (1-(4-fluorophenyl)-6-chloro-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 687.1.

Preparation of Intermediate 76: N-(3-fluoro-4 (1-ethyl 6 (1-Boc-pyrazol-4 yl)-1H-indazol-5-yloxy)phenyl)-6-Boc amino-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0380] Intermediate 76 was prepared according to the synthetic route of Intermediate 26, from Intermediate 63 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-ethyl-1H-indazole) and Intermediate 44 (1-(4-fluorophenyl)-6-Bocamino-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 766.4.

Preparation of Intermediate 77: N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methylthio-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0381] Intermediate 75 was prepared according to the synthetic route of Intermediate 26, from Intermediate 63 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-ethyl-1H-indazole) and Intermediate 40 (1-(4-fluorophenyl)-6-methylthio-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 699.1.

Preparation of Intermediate 78: N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0382] Intermediate 78 was prepared according to the synthetic route of Intermediate 26, starting from Intermediate 63 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-ethyl-1H-indazole) and Intermediate 35 (1-(4-fluorophenyl)-6-methoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 683.3.

Preparation of Intermediate 79: N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-ethoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0383] Intermediate 79 was prepared according to the synthetic route of Intermediate 26, from Intermediate 63 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-ethyl-1H-indazole) and Intermediate 36 (1-(4-fluorophenyl)-6-ethoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 697.3.

Preparation of Intermediate 80: N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-cyclopropoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0384] Intermediate 80 was prepared according to the synthetic route of Intermediate 26, from Intermediate 63 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-ethyl-1H-indazole) and Intermediate 37 (1-(4-fluorophenyl)-6-cyclopropoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 709.1.

Preparation of Intermediate 81: N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-isopropoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0385] Intermediate 81 was prepared according to the synthetic route of Intermediate 26, from Intermediate 63 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-ethyl-1H-indazole) and Intermediate 38 (1-(4-fluorophenyl)-6-isopropoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 711.3.

Preparation of Intermediate 82: N-(3 fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-(2,2,2-trifluoroethoxy)-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide

[0386] Intermediate 82 was prepared according to the synthetic route of Intermediate 26, from Intermediate 63 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1-ethyl-1H-indazole) and Intermediate 39 (1-(4-fluorophenyl)-6-(2,2,2-trifluoroethoxy)-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 751.2.

Preparation of Intermediate 83: tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-3-yl)-1H-pyrazole-1-carboxylate

[0387] Intermediate 83 was prepared according to the synthetic route of Intermediate 26, from Intermediate 62 (tert-butyl 4-(4-(4-amino-2-fluorophenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-5-yl)-1H-pyrazole-1-carboxylate) and Intermediate 13 (1-(4-fluorophenyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 665.5.

Preparation of Intermediate 84: tert butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-methoxy-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1-Ar]-indazol-3-yl)-1H-pyrazole-1-carboxylate

[0388] Intermediate 84 was prepared according to the synthetic route of Intermediate 26, from Intermediate 62 (tert-butyl 4-(4-(4-amino-2-fluorophenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-5-yl)-1H-pyrazole-1-carboxylate) and Intermediate 35 (1-(4-fluorophenyl)-6-methoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 681.2.

Preparation of Intermediate 85: tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-ethoxy-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1-hi]indazol-3-yl)-1H-pyrazole-1-carboxylate

[0389] Intermediate 85 was prepared according to the synthetic route of Intermediate 26, from Intermediate 62 (tert-butyl 4-(4-(4-amino-2-fluorophenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-5-yl)-1H-pyrazole-1-carboxylate) and Intermediate 36 (1-(4-fluorophenyl)-6-ethoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 695.2.

Preparation of Intermediate 86: tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-cyclopropoxy-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1-hi]indazol-3-yl)-1H-pyrazol-1-carboxylate

[0390] Intermediate 86 was prepared according to the synthetic route of Intermediate 26, from Intermediate 62 (tert-butyl 4-(4-(4-amino-2-fluorophenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-5-yl)-1H-pyrazole-1-carboxylate) and Intermediate 37 (1-(4-fluorophenyl)-6-cyclopropoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 707.2.

Preparation of Intermediate 87: tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-isopropoxy-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-3-yl)-1H-pyrazole-1-carboxylate

[0391] Intermediate 87 was prepared according to the synthetic route of Intermediate 26, from Intermediate 62 (tert-butyl 4-(4-(4-amino-2-fluorophenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-5-yl)-1H-pyrazole-1-carboxylate) and Intermediate 38 (1-(4-fluorophenyl)-6-isopropoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 709.3.

Preparation of Intermediate 88: methyl 6-difluoromethoxynicotinate

[0392] To a solution of methyl 6-hydroxynicotinate (1.5 g, 9.8 mmol) in ACN (50 ml) was added NaH (0.979 g, 24.49 mmol). After stirring at room temperature for 0.5 hours, 2,2-difluoro-2-fluorosulfonylacetic acid (2.62 g, 14.69 mmol) was added. The reaction mixture was stirred at 25° C. for 2 hours, quenched with water (50 mL), and extracted with ethyl acetate (5*50 mL). The organic phase was washed with saturated brine (50 mL) and dried over anhydrous Na.sub.2SO.sub.4. After Alteration and concentration, the crude product was purified by silica gel column, eluted with a gradient of 0-10% ethyl acetate/petroleum ether, to obtain a white solid (1.70 g, 85.4% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.84 (d, J=2.3 Hz, 1H), 8.32 (dd, J=8.6, 2.3 Hz, 1H), 7.53 (t, J=72.3 Hz, 1H), 6.95 (d, J=8.6 Hz, 1H), 3.94 (s, 3H). MS (m/z): 204.0[M+H].

Preparation of Intermediate 89:6-difluoromethoxy-5-methoxycarbonylpyridine-1-N-oxide

[0393] To a solution of methyl 6-difluoromethoxynicotinate (0.8 g, 3.94 mmol) in DCM (40 mL) was added urea-hydrogen peroxide (3.70 g, 39.38 mmol) and TFAA (5.5 mL, 39.38 mmol). The reaction mixture was stirred at 25° C. for 16 hours, quenched with water (50 mL), and extracted with ethyl acetate (5×50 mL). The organic phase was washed sequentially with saturated sodium thiosulfate (50 mL), saturated NaHCO.sub.3 (3*50 mL) and saturated brine (50 mL), and dried over anhydrous Na.sub.2SO.sub.4. After Alteration and concentration, the crude product was obtained as a white solid (0.735 g, 85% yield), which was used directly in the next reaction. MS(m/z): 220.0[M+H].

Preparation of Intermediate 90: 6-difluoromethoxy-2-oxo-1,2-dihydropyridine-3-carboxylic add methyl ester

[0394] To a solution of 6-difluoromethoxy-5-methoxycarbonylpyridine-1-N-oxide (0.5 g, 2.28 mmol) in THF (20 mL) were added TFAA (1.29 mL, 9.13 mmol) and TEA (1.27 mL, 9.13 mmol). The reaction mixture was stirred at 25° C. for 2 hours. After concentration, the crude product was purified by a silica gel column, eluted with a gradient of 0-5% ethyl acetate/petroleum ether, to obtain a white solid (0.360 g, 72% yield). MS(m/z): 220.0 [M+H].

Preparation of Intermediate 91: 6-difluoromethoxy-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic add methyl ester

[0395] Under the protection of Ar, to a solution of methyl 6-difluoromethoxy-2-oxo-1,2-dihydropyridine-3-carboxylate (150 mg, 0.684 mmol) in 1,4-dioxane (3 mL) was added 4-fluorophenylboronic acid (95.8 mg, 0.684 mmol), copper acetate (248.7 mg, 1.36 mmol), TEMPO (128.4 mg, 0.821 mmol) and pyridine (541.7 mg, 6.84 mmol). The reaction mixture was stirred at 80° C. for 0.5 hours, LCMS showed 10% product, 4-fluorophenylboronic acid (0.5 equivalent) was added into the reaction mixture every half hour, totally 8 times, and the LCMS yield reached to 69%. After filtration and concentration, the crude product was purified by a silica gel column, eluted with THF/petroleum ether (1:10), to obtain 1.0 g of a yellow solid, which was further purified by reverse phase preparative HPLC to obtain a white solid (50 mg, 23% yield). MS (m/z): 314.0 [M+H].

Preparation of Intermediate 92: 6 difluoromethoxy-1-(4-fluorophenyl)-2-oxo-1,2 dihydropyridine-3-carboxylic acid

[0396] To a solution of 6-difluoromethoxy-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid methyl ester (150 mg, 0.477 mmol) in THF/water (4 mL/0.8 mL) was added LiOH H.sub.2O (22 mg, 0.525 mmol) at 0° C., the reaction mixture was stirred overnight at room temperature under the protection of Ar. Then the reaction was adjusted to pH 3-4 with 1lNHCl, extracted with ethyl acetate (3×20 mL). The organic phase was washed with saturated brine (3×50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was rectystallized from petroleum ether to obtain a white solid (100 mg. 70% yield). 1H NMR (400 MHz, DMSO-d6) δ13.16 (s, 1H), 8.37 (d, J=8.0 Hz, 1H), 7.35 (s, 0.28 H), 7.26-7.22 (m, 4H), 7.17 (s, 0.49 H), 6.99 (s, 0.23H), 6.85 (d, J=8.4 Hz, 1H). MS-ESI [M+H].sup.+: 300.0.

Preparation of Intermediate 93: tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-(2,2,2-trifluoroethoxy)-2-oxo-1,2-dihydropropine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1hi]-indazol-3-yl)-1H-pyrazole-1carboxylate

[0397] Intermediate 93 was prepared according to the synthetic route of Intermediate 26, from Intermediate 62 (tert-butyl 4-(4-(4-amino-2-fluorophenoxy)-6,7-dihydropyrrolo[3,2,1-hi]- indazol-5-yl)-1H-pyrazole-1-carboxylate) and Intermediate 39 (1-(4-fluorophenyl)-6-(2.2,2-trifluoroethoxy)-2-oxo-1,2-dihydropyridine-3-carboxylic acid), MS-ESI [M+H].sup.+:749.2.

Preparation of Intermediate 94:N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazole-4-yl)-1H-indazol-5-yloxy)phenyl)-6-difluoromethoxy-2-oxo-1-(4-flurophenyl)-1,2-dihydro pyridine-3-carboxamide

[0398] Intermediate 94 was prepared according to the synthetic route of Intermediate 26, from Intermediate 25 (6-(1-Boc-pyrazol-4-yl)-5-(2-fluoro-4-aminophenoxy)-1methyl-1H indazole) and Intermediate 92 (1-(4-fluorophenyl)-6-difluoromethoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid). MS-ESI [M+H].sup.+: 705.2.

Preparation of Intermediate 95: tert-butyl 4-(4-(2- fluoro-4- (1-(4-fluorophenyl)-6-difluoromethoxy-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)-6.7-dihydropyrrolo[3,2,1-hi]-indazole-3-yl)-1H-pyrazole-1-carboxylate

[0399] Intermediate 95 was prepared according to the synthetic route of Intermediate 26, from Intermediate 62 (tert-butyl 4-(4-(4-amino-2-fluorophenoxy)-6,7- dihydropyrrolo[3,2,1hi]-indazol-5yl)-1H-pyrazole-1carboxylate) and Intermediate 92 (1-(4-fluorophenyl)-6-difluoromethoxy-2-oxo-1,2-dihydropyridine-3-carboxylic acid) MS-ES [M+H].sup.+:717.2

Example 1: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazole-4-yl)-1H-indazole-5-yloxy)-1-(4-fluorophenyl)-6-trifluoromethyl-2-oxo-1,2-dihydropyridine-3-carboximide monomethanesulfonate

[0400] ##STR00122##

[0401] To a solution of 7V-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-trifluoromethyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide (Intermediate 29, 255 mg, 0.33 mmol) in dichloromethane (15 mL) was added trifluoroacetic acid (3 mL), the resulting reaction mixture was stirred at room temperature for 1 hour, and the solvent was evaporated under reduced pressure. The crude product was dissolved in ethyl acetate (50 mL), neutralized by adding saturated sodium bicarbonate aqueous solution (25 mL), then washed twice with saturated sodium chloride aqueous solution (25 mL each time), dried and concentrated to obtain free base solid 200 mg as shown by the chemical structure above.

[0402] The above free base was dissolved in anhydrous methanol (5 mL) and dichloromethane (5 mL), and a methanol (5 mL) solution of methanesulfonic acid (32 mg, 0.33 mmol) was added. After mixing thoroughly, the solvent was evaporated under reduced pressure and dried to obtain about 230 mg of a yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.56 (s, 1H), 8.55 (d, J=7.6 Hz, 1H), 8.04 (s, 2H), 7.94 (s, 1H), 7.90 (dd, J=13.2, 2.5 Hz, 1H), 7.85 (s, 1H), 7.51 (dd, J=8.6, 4.9 Hz, 2H), 7.35 (t, J=8.6 Hz, 2H), 7.30-7.20 (m, 3H), 6.79 (t, J=9.1 Hz, 1H), 4.73 (s, b, 2H), 4.01 (s, 3H), 2.29 (s, 3H). MS [M+H].sup.+: 607.1.

Example 2: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-cyclopropyl-2-oxo-1,2-dihydropyridine-3-carboxamide

[0403] ##STR00123##

[0404] This compound was prepared according to the synthetic route of Example 1, from Intermediate 28 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazole)-5-yloxy)phenyl)-6-cyclopropyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). MS [M+H].sup.+: 579.2 .sup.1H NMR (500 MHz, chloroform-d) δ 11.82 (s, 1H), 8.62 (d, J=7.8 Hz, 1H), 8.23 (s, 2H), 7.89 (dd, J=12.7, 2.5 Hz, 1H), 7.84 (s, 1H), 7.57 (s, 1H), 7.34-7.28 (m, 4H), 7.20 (d, J=9.1 Hz, 1H), 7.17 (s, 1H), 6.88 (t, J=8.8 Hz, 1H), 6.22 (d, J=7.8 Hz, 1H), 4.11 (s, 3H), 3.54 (s, b, 3H), 2.95 (s, 3H), 1.41-1.13 (m, 2H), 1.00-0.64 (m, 2H).

Example 3: Preparation of N-(3-fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carboxamide

[0405] ##STR00124##

[0406] This compound was prepared according to the synthetic route of Example 1, from Intermediate 21 (N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazole-5-yloxy)phenyl)-6-methyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.87 (s, 1H), 8.40 (d, J=7.5 Hz, 1H), 8.06 (s, 2H), 7.97 (s, 1H), 7.90 (dd, J=13.4, 2.4 Hz, 1H), 7.86 (s, 1H), 7.52-7.27 (m, 4H), 7.25-7.10 (m, 2H), 6.80 (t, J=9.1 Hz, 1H), 6.63 (d, J=7.6 Hz, 1H), 5.29 (s, b, 2H), 4.40 (q, J=7.2 Hz, 2H), 2.30 (s, 4H), 2.00 (s, 3H), 1.35 (t, J=7.2 Hz, 3H). MS [M+H].sup.+: 567.2.

Example 4: Preparation of N-(3-fluoro-4-(1-isopropyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carboxamide

[0407] ##STR00125##

[0408] This compound was prepared according to the synthetic route of Example 1, from Intermediate 20 (N-(3-fluoro-4-(1-isopropyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.87 (s, 1H), 8.40 (d, J=7.6 Hz, 1H), 8.06 (s, 2H), 7.98 (s, 1H), 7.90 (dd, J=13.3, 2.5 Hz, 1H), 7.87 (s, 1H), 7.47-7.29 (m, 4H), 7.23-7.10 (m, 2H), 6.79 (t, J=9.1 Hz, 1H), 6.63 (d, J=7.6 Hz, 1H), 5.07 (s, b, 2H), 5.01 (h, J=6.7 Hz, 1H), 2.30 (s, 3H), 2.00 (s, 3H), 1.43 (d, J=6.5 Hz, 6H). MS [M+H].sup.+: 581.2.

Example 5: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-ethyl-2-oxo-1,2-dihydropyridine-3-carboxamide

[0409] ##STR00126##

[0410] This compound was prepared according to the synthetic route of Example 1, from Intermediate 26 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-ethyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.94 (s, 1H), 8.52 (d, J=7.7 Hz, 1H), 8.12 (s, 2H), 8.01 (s, 1H), 7.97 (dd, J=13.2, 2.4 Hz, 1H), 7.92 (s, 1H), 7.58-7.34 (m, 4H), 7.30-7.16 (m, 2H), 6.86 (t, J=9.1 Hz, 1H), 6.67 (d, J=7.7 Hz, 1H), 5.27 (s, b, 2H), 4.08 (s, 3H), 2.37 (s, 3H), 2.30 (q, J=7.4 Hz, 2H), 1.05 (t, J=7.4 Hz, 3H). MS [M+H].sup.+: 567.2.

Example 6: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-isopropyl-2-oxo-1,2-dihydropyridine-3-carboxamide

[0411] ##STR00127##

[0412] This compound was prepared according to the synthetic route of Example 1, from Intermediate 27 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-isopropyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.86 (s, 1H), 8.46 (d, J=7.8 Hz, 1H), 8.05 (s, 2H), 7.94 (s, 1H), 7.90 (dd, J=13.5, 2.4 Hz, 1H), 7.84 (s, 1H), 7.46 (dd, J=8.7, 4.9 Hz, 2H), 7.36 (t, J=8.6 Hz, 2H), 7.19 (d, J=7.7 Hz, 2H), 6.79 (t, J=9.1 Hz, 1H), 6.69 (d, J=7.9 Hz, 1H), 5.05 (s, b, 2H), 4.01 (s, 3H), 2.39 (h, 1H), 2.29 (s, 3H), 1.05 (d, J=6.8 Hz, 6H). MS [M+H].sup.+: 581.2.

Example 7: Preparation of N-(3-fluoro-4-(1-isopropyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-isopropyl-2-oxo-1,2-dihydropyridine-3-carboxamide

[0413] ##STR00128##

[0414] This compound was prepared according to the synthetic route of Example 1, from Intermediate 22 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-isopropyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (500 MHz, chloroform-d) δ 11.85 (s, 1H), 8.69 (d, J=7.8 Hz, 1H), 8.49 (s, 2H), 7.93-7.85 (m, 2H), 7.63 (s, 1H), 7.31 (t, J=8.4 Hz, 2H), 7.30-7.21 (m, 3H), 7.12 (s, 1H), 6.97 (t, J=8.7 Hz, 1H), 6.56 (d, J=7.8 Hz, 1H), 6.12 (s, b, 2H), 4.88 (s, 1H), 2.96 (s, 3H), 2.60 (hept, J=6.8 Hz, 1H), 1.61 (d, J=4.7 Hz, 6H), 1.18 (d, J=6.8 Hz, 6H).

Example 8: Preparation of N-(3-fluoro-4-(1-isopropyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-trifluoromethyl-2-oxo-1,2-dihydropyridine-3-carboxamide

[0415] ##STR00129##

[0416] This compound was prepared according to the synthetic route of Example 1, from Intermediate 23 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-isopropyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (500 MHz, chloroform-d) δ 11.62 (s, 1H), 8.78 (d, J=7.6 Hz, 1H), 8.53 (s, 2H), 7.89 (s, 1H), 7.88 (dd, J=10.1 Hz, 2.5 Hz, 1H), 7.64 (s, 1H), 7.33-7.22 (m, 5H), 7.14 (s, 1H), 7.09 (d, J=7.6 Hz, 1H), 6.98 (t, J=8.8 Hz, 1H), 5.60 (s, b, 2H), 4.88 (hept, J=6.7 Hz, 1H), 2.95 (s, 3H), 1.61 (d, J=6.6 Hz, 6H).

Example 9: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-chloro-2-oxo-1,2-dihydropyridine-3-carboxamide

[0417] ##STR00130##

[0418] This compound was prepared according to the synthetic route of Example 1, from Intermediate 64 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-chloro-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.97 (s, 1H), 11.57 (s, 1H), 8.51 (d, J=7.9 Hz, 1H), 8.15 (s, 1H), 8.02 (s, 1H), 8.00 (s, 1H), 7.95 (d, J=14.5 Hz, 1H), 7.91 (s, 1H), 7.56 (s, 2H), 7.44 (t, J=8.5 Hz, 2H), 7.28 (s, 2H), 7.01 (d, J=8.0 Hz, 1H), 6.85 (t, J=9.1 Hz, 1H), 4.07 (s, 3H). MS-ESI [M+H].sup.+: 573.1.

Example 10: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-amino-2-oxo-1,2-dihydropyridine-3-carboxamide

[0419] ##STR00131##

[0420] This compound was prepared according to the synthetic route of Example 1, from Intermediate 65 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-(Bocamino)-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.67 (s, 1H), 11.73 (s, 1H), 8.15 (d, J=8.8 Hz, 1H), 8.10 (s, 2H), 7.98 (s, 1H), 7.94 (dd, J=13.6, 2.4 Hz, 1H), 7.89 (d, J=0.6 Hz, 1H), 7.41 (d, J=1.8 Hz, 2H), 7.40 (s, 2H), 7.21 (s, 1H), 7.14 (d, J=10.0 Hz, 1H), 6.84 (t, J=9.2 Hz, 1H), 5.88 (d, J=8.8 Hz, 1H), 4.07 (s, 3H). MS-ESI [M+H].sup.+: 554.2.

Example 11: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-methylthio-2-oxo-1,2-dihydropyridine-3-carboxamide

[0421] ##STR00132##

[0422] This compound was prepared according to the synthetic route of Example 1, from Intermediate 66 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methylthio-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 11.66 (s, 1H), 8.62 (d, J=8.4 Hz, 1H), 8.32 (s, 2H), 7.90 (dd, J=12.4, 2.4 Hz, 1H), 7.84 (s, 1H), 7.54 (s, 1H), 7.30-7.27 (m, 4H), 7.25-7.20 (m, 1H), 7.14 (s, 1H), 6.94 (t, J=8.8 Hz, 1H), 6.33 (d, J=8.4 Hz, 1H), 4.11 (s, 3H), 2.47 (s, 3H). MS-ESI [M+H].sup.+: 585.1.

Example 12: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-methoxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0423] ##STR00133##

[0424] This compound was prepared according to the synthetic route of Example 1, from Intermediate 68 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.95 (br s, 1H), 11.69 (s, 1H), 8.56 (d, 8.6 Hz, 1H), 8.09 (br s, 2H), 7.99 (s, 1H), 7.95 (dd, J=13.4, 2.4 Hz, 1H), 7.91 (s, 1H), 7.46-7.33 (m, 4H), 7.25 (s, 1H), 7.23 (d, J=9.0 Hz, 1H), 6.86 (t, J=9.1 Hz, 1H), 6.34 (d, J=8.7 Hz, 1H), 4.07 (s, 3H), 3.90 (s, 3H). MS-ESI [M+H].sup.+: 569.1.

Example 13: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-methylsulfoxide-2-oxo-1,2-dihydropyridine-3-carboxamide

[0425] ##STR00134##

[0426] This compound was prepared according to the synthetic route of Example 1, from Intermediate 67(N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methylsulfoxide-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.96 (br, 1H), 11.71 (s, 1H), 8.75 (d, J=7.6 Hz, 1H), 8.06 (br, 2H), 8.00 (s, 1H), 7.99-7.94 (m, 1H), 7.92 (s, 1H), 7.77-7.71 (m, 1H), 7.67 (d, J=5.0 Hz, 1H), 7.46 (t. J=8.8 Hz, 2H), 7.30 (d, J=9.8 Hz, 2H), 7.22 (d, J=7.7 Hz, 1H), 6.86 (t, J=9.1 Hz, 1H), 4.07 (s, 3H), 2.60 (s, 3H). MS-ESI [M+H].sup.+: 601.0.

Example 14: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-ethoxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0427] ##STR00135##

[0428] This compound was prepared according to the synthetic route of Example 1, starting from Intermediate 69 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-ethoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.94 (br s, 1H), 11.70 (s, 1H), 8.53 (d, J=8.6 Hz, 1H), 8.09 (br s, 2H), 7.99 (s, 1H), 7.95 (d, J=13.5 Hz, 1H), 7.90 (s, 1H), 7.46-7.33 (m, 4H), 7.27-7.17 (m, 2H), 6.85 (t, J=9.2 Hz, 1H), 6.33 (d, J=8.6 Hz, 1H), 4.30-4.20 (q, 6.4 Hz, 2H), 4.07 (s, 3H), 1.14 (t, J=6.3 Hz, 3H). MS-ESI [M+H].sup.+: 583.1.

Example 15: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-cyclopropoxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0429] ##STR00136##

[0430] This compound was prepared according to the synthetic route of Example 1, from Intermediate 70 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-cyclopropoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 11.62 (s, 1H), 8.72 (d, J=8.4 Hz, 1H), 8.27 (br s, 2H), 7.89 (dd, J=12.8, 2.0 Hz, 1H), 7.84 (s, 1H), 7.55 (s, 1H), 7.24-7.09 (m, 6H), 6.91 (t, J=8.8 Hz, 1H), 6.35 (d, J=8.8 Hz, 1H), 4.11 (s, 3H), 3.93-3.90 (m, 1H), 0.90-0.85 (m, 2H), 0.76-0.72 (m, 2H). MS-ESI [M+H].sup.+: 595.2.

Example 16: Preparation of N-(3-fluoro 4 (1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol 5-yloxy)phenyl)-1-(4-fluorophenyl)-6-isopropoxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0431] ##STR00137##

[0432] This compound was prepared according to the synthetic route of Example 1, from Intermediate 71 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-isopropoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 11.60 (s, 1H), 8.68 (d, J=8.8 Hz, 1H), 8.15 (br s, 2H), 7.88 (dd, J=12.8, 2.2 Hz, 1H), 7.83 (s, 1H), 7.56 (s, 1H), 7.22 (d, J=8.2 Hz, 2H), 7.20-7.12 (m, 4H), 6.84 (t, J=8.9 Hz, 1H), 5.96 (d, J=8.8 Hz, 1H), 4.73-4.67 (m, 1H), 4.10 (s, 3H), 1.27 (d, J=6.0 Hz, 6H). MS-ESI [M+H].sup.+: 597.2.

Example 17: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4 yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-(2,2,2-trifluoroethoxy)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0433] ##STR00138##

[0434] This compound was prepared according to the synthetic route of Example 1, from Intermediate 72(N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-(2,2,2-trifluoroethoxy)-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.63 (s, 1H), 8.59 (d, J=8.8 Hz, 1H), 8.09 (s, 2H), 7.99 (s, 1H), 7.95 (dd, J=13.2, 2.4 Hz, 1H), 7.91 (s, 1H), 7.46-7.35 (m, 4H), 7.28-7.21 (m, 2H), 6.86 (t, J=9.1 Hz, 1H), 6.45 (d, J=8.8 Hz, 1H), 5.02 (q, J=8.4 Hz, 2H), 4.07 (s, 3H). MS-ESI [M+H].sup.+: 637.0.

Example 18: Preparation of N-(3-fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-ethyl-2-oxo-1,2-dihydropyridine-3-carboxamide

[0435] ##STR00139##

[0436] This compound was prepared according to the synthetic route of Example 1, from Intermediate 73 (N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-ethyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, Methanol-d.sub.4) δ 8.58 (d, J=7.6 Hz, 1H), 8.10 (s, 2H), 7.93-7.81 (m, 3H), 7.41-7.28 (m, 4H), 7.23 (s, 1H), 7.17-7.11 (m, 1H), 6.86 (t, J=9.0 Hz, 1H), 6.67 (d, J=7.8 Hz, 1H), 4.49 (q, J=7.2 Hz, 2H), 2.39 (q, J=7.4 Hz, 2H), 1.49 (t, J=7.2 Hz, 3H), 1.14 (t, J=1.4 Hz, 3H). MS-ESI [M+H].sup.+: 581.3.

Example 19: Preparation of N-(3-fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-cyclopropyl-2-oxo-1,2-dihydropyridine-3-carboxamide

[0437] ##STR00140##

[0438] This compound was prepared according to the synthetic route of Example 1, from Intermediate 74 (N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-cyclopropyl-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, Methanol-d.sub.4) δ 8.54 (d, J=7.6 Hz, 1H), 8.20 (s, 2H), 7.89 (s, 2H), 7.86 (dd, J=12.8, 2.4 Hz, 1H), 7.43-7.30 (m, 4H), 7.24 (s, 1H), 7.18-7.14 (m, 1H), 6.88 (d, J=9.2 Hz, 1H), 6.43 (d, J=8.0 Hz, 1H), 4.50 (q, J=7.2 Hz, 2H), 1.47 (t, J=7.2 Hz, 3H), 1.45-1.41 (m, 1H), 0.92-0.89 (m, 4H). MS-ESI [M+H].sup.+: 593.2.

Example 20: Preparation of N-(3-fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-chloro-2-oxo-1,2-dihydropyridine-3-carboxamide

[0439] ##STR00141##

[0440] This compound was prepared according to the synthetic route of Example 1, from Intermediate 75 (N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-chloro-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.96 (br s, 1H), 11.57 (s, 1H), 8.51 (d, J=8.0 Hz, 1H), 8.02 (br s, 3H), 7.96 (dd, J=13.2, 2.5 Hz, 1H), 7.93 (d, J=0.8 Hz, 1H), 7.61-7.52 (m, 2H), 7.49-7.40 (m, 2H), 7.31-7.24 (m, 2H), 7.01 (d, J=8.0 Hz, 1H), 7.01-6.86 (m, 1H), 4.47 (q, J=7.2 Hz, 2H), 1.42 (t, J=7.2 Hz, 3H). MS-ESI [M+H].sup.+: 587.2.

Example 21: Preparation of N-(3-Fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-amino-2-oxo-1,2-dihydropyridine-3-carboxamide

[0441] ##STR00142##

[0442] This compound was prepared according to the synthetic route of Example 1, from Intermediate 76 (N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-Boc-amino-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.89 (brs, 1H), 11.72 (s, 1H), 8.16 (d, J=8.8 Hz, 1H), 8.10 (brs, 2H), 8.01 (s, 1H), 7.94 (dd, J=13.6, 2.4 Hz, 1H), 7.91 (s, 1H), 7.42-7.40 (m, 4H), 7.21 (s, 1H), 7.13 (d, J=10.0 Hz, 1H), 6.85 (t, J=9.2 Hz, 1H), 5.88 (d, J=8.8 Hz, 1H), 4.46 (q, J=7.1 Hz, 2H), 1.42 (t, J=7.2 Hz, 3H). MS-ESI [M+H].sup.+: 568.2.

Example 22: Preparation of N-(3-fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-methylthio-2-oxo-1,2-dihydropyridine-3-carboxamide

[0443] ##STR00143##

[0444] This compound was prepared according to the synthetic route of Example 1, from Intermediate 77 (N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methylthio-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.95 (br s, 1H), 11.70 (s, 1H), 8.43 (d, J=8.0 Hz, 1H), 8.09 (hr, 2H), 8.02 (s, 1H)), 7.95 (d, J=15.2 Hz, 1H), 7.92 (s, 1H), 7.55-7.46 (m, 2H), 7.42 (t, J=8.8 Hz, 2H), 7.24 (d, J=12.8 Hz, 2H), 6.85 (t, J=9.2 Hz, 1H), 6.61 (d, J=8.4 Hz, 1H), 4.46 (q, J=7.2 Hz, 2H), 2.50 (s, 3H), 1.42 (t, J=7.2 Hz, 3H). MS-ESI [M+H].sup.+: 599.1.

Example 23: Preparation of N-(3-fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-methoxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0445] ##STR00144##

[0446] This compound was prepared according to the synthetic route of Example 1, from Intermediate 78 (N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-methoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.94 (br s, 1H), 11.69 (s, 1H), 8.56 (d, J=8.6 Hz, 1H), 8.09 (br s, 2H), 8.02 (s, 1H), 7.96 (dd, J=13.4, 2.4 Hz, 1H), 7.92 (s, 1H), 7.47-7.33 (m, 4H), 7.25 (s, 1H), 7.22 (d, J=10.1 Hz, 1H), 6.86 (t, J=9.2 Hz, 1H), 6.34 (d, J=8.7 Hz, 1H), 4.47 (q, J=7.1 Hz, 2H), 3.90 (s, 3H), 1.42 (t, J=7.2 Hz, 3H). MS-ESI [M+H].sup.+: 583.1.

Example 24: Preparation of N-(3-fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-ethoxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0447] ##STR00145##

[0448] This compound was prepared according to the synthetic route of Example 1, from Intermediate 79 (N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-ethoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.97 (br s, 1H), 11.70 (s, 1H), 8.53 (d, J=8.4 Hz, 1H), 8.09-8.04 (m, 2H), 8.01 (s, 1H), 7.95 (dd, J=13.4, 2.3 Hz, 1H), 7.92 (s, 1H), 7.48-7.32 (m, 4H), 7.25 (s, 1H), 7.21 (d, J=9.2 Hz, 1H), 6.86 (t, J=9.2 Hz, 1H), 6.32 (d, J=8.8 Hz, 1H), 4.46 (q, J=7.2 Hz, 2H), 4.25 (q, J=7.2 Hz, 2H), 1.42 (t, J=7.0 Hz, 3H), 1.14 (t, J=7.0 Hz, 3H). MS-ESI [M+H].sup.+: 597.1.

Example 25: Preparation of N-(3-fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-cyclopropoxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0449] ##STR00146##

[0450] This compound was prepared according to the synthetic route of Example 1, from Intermediate 80 (N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-cyclopropoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.95 (br s, 1H), 11.69 (s, 1H), 8.59 (d, J=8.4 Hz, 1H), 8.09 (br s, 2H), 8.02 (s, 1H), 7.96 (dd, J=13.3, 2.4 Hz, 1H), 7.92 (s, 1H), 7.43-7.31 (m, 4H), 7.27-7.18 (m, 2H), 6.86 (t, J=9.1 Hz, 1H), 6.56 (d, J=8.4 Hz, 1H), 4.47 (q, J=7.2 Hz, 2H), 4.17-4.13 (m, 1H), 1.42 (t, J=7.2 Hz, 3H), 0.84-0.76 (m, 2H), 0.69-0.60 (m, 2H). MS-ESI [M+H].sup.+: 609.1.

Example 26: Preparation of N-(3-fluoro-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-isopropoxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0451] ##STR00147##

[0452] This compound was prepared according to the synthetic route of Example 1, from Intermediate 81 (N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-isopropoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.96 (br, 1H), 11.71 (s, 1H), 8.52 (d, J=8.8 Hz, 1H), 8.09 (br s, 2H), 8.02 (s, 1H), 7.96 (dd, J=13.4, 2.4 Hz, 1H), 7.92 (s, 1H), 7.43-7.30 (m, 4H), 7.25 (s, 1H), 7.21 (d, 7=8.9 Hz, 1H), 6.86 (t, J=9.1 Hz, 1H), 6.37 (d, J=8.8 Hz, 1H), 4.89-4.84 (m, 1H), 4.47 (q, J=7.2 Hz, 2H), 1.42 (t, J=7.2 Hz, 3H), 1.17 (d, J=6.0 Hz, 6H). MS-ESI [M+H].sup.+: 611.2.

Example 27: Preparation of N-(3-fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-(2,2,2-trifluoroethoxy)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0453] ##STR00148##

[0454] This compound was prepared according to the synthetic route of Example 1, from Intermediate 82(N-(3-fluoro-4-(1-ethyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-(2,2,2-trifluoroethoxy)-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.63 (s, 1H), 8.59 (d, J=8.4 Hz, 1H), 8.09 (s, 2H), 8.02 (s, 1H), 7.96 (dd, J=13.3, 2.4 Hz, 1H), 7.92 (s, 1H), 7.48-7.35 (m, 4H), 7.24 (d, J=11.9 Hz, 2H), 6.86 (t, 7=9.1 Hz, 1H), 6.45 (d, J=8.4 Hz, 1H), 5.02 (q, J=8.4 Hz, 2H), 4.47 (q, J=7.2 Hz, 2H), 1.42 (t, J=7.2 Hz, 3H). MS-ESI [M+H].sup.+: 651.1.

Example 28: Preparation of N-(4-((3-(1H-pyrazol-4-yl)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-4-yl)oxy)-3-fluorophenyl)-1-(4-fluorophenyl)-6-methoxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0455] ##STR00149##

[0456] This compound was prepared according to the synthetic route of Example 1, from Intermediate 84 (tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-methoxy-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-3-yl)-1H-pyrazole-1-carboxylate). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.65 (d, J=8.8 Hz, 1H), 8.07 (s, 2H), 7.86-7.69 (m, 2H), 7.19 (m, 4H), 7.06 (d, J=8.8 Hz, 1H), 6.96 (s, 1H), 6.76 (t, J=8.8 Hz, 1H), 5.99 (d, J=8.4 Hz, 1H), 4.71 (t, J=6.4 Hz, 2H), 4.04 (t, J=6.4 Hz, 2H), 3.88 (s, 3H). MS-ESI [M+H].sup.+: 581.1.

Example 29: Preparation of N-(3-fluoro-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-hydroxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0457] ##STR00150##

[0458] To a solution of Example 9 (N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-chloro-2-oxo-1,2-dihydropyridine-3-carboxamide) (0.130 g, 0.19 mmol) in THF (2 mL) were added NaOH (0.076 g, 1.90 mmol) and water (0.8 mL). The mixture was stirred at room temperature for 16 hours, then diluted with water (5 mL), neutralized to pH 7 with concentrated HCl, and extracted with ethyl acetate (5×10 mL). The organic phase was dried and concentrated and purified by HPLC to obtain a yellow solid (3 mg, 2.8% yield). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.95 (br s, 1H), 12.19 (s, 1H), 8.14-8.06 (m, 2H), 8.00-7.92 (m, 2H), 7.91-7.87 (m, 2H), 7.27-7.16 (m, 3H), 7.15-7.03 (m, 3H), 6.95 (s, 1H), 6.85 (t, J=9.2 Hz, 1H), 5.34 (d, J=9.2 Hz, 1H), 4.06 (s, 3H). MS-ESI [M+H].sup.+: 553.1.

Example 30: Preparation of N-(3-fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-hydroxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0459] ##STR00151##

[0460] This compound was prepared according to the synthetic route of Example 29, from Example 20 (N-(3-fluoro-4-(1-ethyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-chloro-2-oxo-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.94 (br s, 1H), 12.19 (s, 1H), 8.11 (br s, 2H), 7.99-7.92 (m, 2H), 7.90 (s, 1H), 7.83 (d, J=8.1 Hz, 1H), 7.26-7.08 (m, 6H), 6.95 (s, 1H), 6.85 (t, J=9.2 Hz, 1H), 5.49 (d, J=9.1 Hz, 1H), 4.46 (q, J=7.2 Hz, 2H), 1.41 (t, J=7.2 Hz, 3H). MS-ESI [M+H].sup.+: 567.1.

Example 31: Preparation of N-(4-((3-(1H-pyrazol-4-yl)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-4-yl)oxy)-3-fluorophenyl)-1-(4-fluorophenyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carboxamide

[0461] ##STR00152##

[0462] This compound was prepared according to the synthetic route of Example 1, from Intermediate 83 (tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-3-yl)-1H-pyrazole-1-carboxylate). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 11.63 (s, 1H), 8.62-8.46 (m, 1H), 8.05-7.28 (m, 6H), 7.21 (s, 2H), 7.04-6.73 (m, 2H), 6.45 (d, J=4.3 Hz, 2H), 4.71 (s, 2H), 4.06 (s, 2H), 2.08 (s, 3H). MS-ESI [M+H].sup.+: 565.3.

Example 32: Preparation of N-(4-((3-(1H-pyrazol-4-yl)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-4-yl)oxy)-3-fluorophenyl)-6-ethoxy-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0463] ##STR00153##

[0464] This compound was prepared according to the synthetic route of Example 1, from Intermediate 85 (tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-ethoxy-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1-/?/]-indazol-3-yl)-1H-pyrazole-1-carboxylate). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.66 (s, 1H), 8.52 (d, 8.7 Hz, 1H), 8.02-7.90 (m, 3H), 7.82 (s, 1H), 7.45-7.32 (m, 4H), 7.14 (d, J=9.3 Hz, 1H), 7.04 (s, 1H), 6.73 (t, 9.2 Hz, 1H), 6.32 (d, J=8.8 Hz, 1H), 4.71 (t, J=6.5 Hz, 2H), 4.24 (q, J=7.0 Hz, 2H), 4.07 (t, J=6.6 Hz, 2H), 1.13 (t, J=7.0 Hz, 3H). MS-ESI [M+H].sup.+: 595.2.

Example 33: Preparation of N-(4-((3-(1H-pyrazol-4-yl)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-4-yl)oxy)-3-fluorophenyl)-6-cyclopropoxy-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0465] ##STR00154##

[0466] This compound was prepared according to the synthetic route of Example 1, from Intermediate 86 (tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-cyclopropoxy-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-3-yl)-1H-pyrazole-1-carboxylate). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.65 (s, 1H), 8.57 (d, J=8.6 Hz, 1H), 8.00-7.89 (m, 3H), 7.82 (s, 1H), 7.44-7.28 (m, 4H), 7.21-7.12 (m, 1H), 7.05 (s, 1H), 6.73 (t, J=9.2 Hz, 1H), 6.55 (d, J=8.6 Hz, 1H), 4.71 (t, J=6.6 Hz, 2H), 4.14 (dd, J=6.0, 3.2 Hz, 1H), 4.07 (t, J=6.5 Hz, 2H), 0.88-0.81 (m, 2H), 0.64 (s, 2H). MS-ESI [M+H].sup.+: 607.2.

Example 34: Preparation of N-(4-((3-(1H-pyrazol-4-yl)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-4-yl)oxy)-3-fluorophenyl)-1-(4-fluorophenyl)-6-isopropoxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0467] ##STR00155##

[0468] This compound was prepared according to the synthetic route of Example 1, from Intermediate 87 (tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-isopropoxy-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-3-yl)-1H-pyrazole-1-carboxylate). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.67 (s, 1H), 8.50 (d, J=8.6 Hz, 1H), 8.03-7.88 (m, 3H), 7.82 (s, 1H), 7.45-7.30 (m, 4H), 7.20-7.10 (m, 1H), 7.04 (s, 1H), 6.73 (t, J=9.2 Hz, 1H), 6.36 (d, J=8.9 Hz, 1H), 4.93-4.79 (m, 1H), 4.71 (t, J=6.5 Hz, 2H), 4.07 (t, J=6.6 Hz, 2H), 1.16 (d, J=6.1 Hz, 6H). MS-ESI [M+H].sup.+: 609.2.

Example 35: Preparation of N-(4-((3-(1H-pyrazol-4-yl)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-4-yl)oxy)-3-fluorophenyl)-1-(4-fluorophenyl)-2-oxo-6-(2,2,2-trifluoroethoxy)-1,2-dihydropyridine-3-carboxamide

[0469] ##STR00156##

[0470] This compound was prepared according to the synthetic route of Example 1, from Intermediate 93 (tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-2-oxo-6-(2,2,2-trifluoroethoxy)-1,2-dihydropyridine-3-carboxamido)phenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-3-yl)-1H-pyrazole-1-carboxylate). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.03 (s, 1H), 11.60 (s, 1H), 8.57 (d, J=8.6 Hz, 1H), 7.94 (dd, 0.7=13.4, 2.6 Hz, 3H), 7.83 (s, 1H), 7.50-7.32 (m, 4H), 7.21-7.13 (m, 1H), 7.05 (s, 1H), 6.74 (t, J=9.2 Hz, 1H), 6.44 (d, J=8.7 Hz, 1H), 5.00 (t, J=8.5 Hz, 2H), 4.71 (t, J=6.6 Hz, 2H), 4.07 (t, J=6.6 Hz, 2H). MS-ESI [M+H].sup.+: 649.1.

Example 36: Preparation of N-(3-fluoro-4-(1-methyl-6-(1H-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-1-(4-fluorophenyl)-6-difluoromethoxy-2-oxo-1,2-dihydropyridine-3-carboxamide

[0471] ##STR00157##

[0472] This compound was prepared according to the synthetic route of Example 1, from Intermediate 94 (N-(3-fluoro-4-(1-methyl-6-(1-Boc-pyrazol-4-yl)-1H-indazol-5-yloxy)phenyl)-6-difluoromethoxy-2-oxo-1-(4-fluorophenyl)-1,2-dihydropyridine-3-carboxamide). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ10.53 (s, 1H), 8.24 (d, J=8.1 Hz, 1H), 8.09 (s, 2H), 7.99 (s, 1H), 7.92-7.84 (m, 2H), 7.44-7.18 (m, 7H), 7.07-6.88 (m, 2H), 4.07 (s, 3H). MS-ESI [M+H].sup.+: 605.1.

Example 37: Preparation of N-(4-((3-(1H-pyrazol-4-yl)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-4-yl)oxy)-3-fluorophenyl)-6-(difluoromethoxy)-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0473] ##STR00158##

[0474] This compound was prepared according to the synthetic route of Example 1, from Intermediate 95 (tert-butyl 4-(4-(2-fluoro-4-(1-(4-fluorophenyl)-6-(difluoromethoxy)-2-oxo-1,2-dihydropyridine-3-carboxamido) phenoxy)-6,7-dihydropyrrolo[3,2,1-hi]-indazol-3-yl)-1H-pyrazole-1-carboxylate), 1H NMR (400 MHz, DMSO-d.sub.6) δ 13.03 (s, 1H), 10.47 (s, 1H), 8.23 (d, J=8.1 Hz, 1H), 7.84 (d, J=12.2 Hz, 4H), 7.44-7.16 (m, 6H), 7.04 (d, J=8.2 Hz, 1H), 6.91 (d, J=8.1 Hz, 1H), 6.84 (t, J=9.2 Hz, 1H), 4.72 (t, J=6.6 Hz, 2H), 4.07 (t, J=6.5 Hz, 2H). MS-ESI [M+H].sup.+: 617.1.

[0475] The following compounds were essentially made by a similar method of Example 37 above.

TABLE-US-00001 Example number Structure MS data Example 38 [00159] 623.2 Example 39 [00160] 635.2 Example 40 [00161] 564.2 Example 41 [00162] 576.2 Example 42 [00163] 563.2 Example 43 [00164] 579.2 Example 44 [00165] 593.2 Example 45 [00166] 605.2 Example 46 [00167] 607.2 Example 47 [00168] 615.2 Example 48 [00169] 633.2 Example 49 [00170] 647.2 Example 50 [00171] 574.2

ACTIVITY EXAMPLES

Example 1: Inhibitory Activity of the Compounds of the Present Disclosure on the Proliferation of KM12 Cells

[0476] KM12 is a human colon cancer cell line in which NTRK1 is highly expressed, and gene fusion occurs with TPM3 to form TPM3-NTRK1.

[Test method]: Promega's CellTiter-Glo® Luminescent Cell Viability Assay kit was used to evaluate the inhibitory activity of the compounds on the proliferation of KM12 cells.
[Instrument]: Spectramax M3 multifunctional microplate reader from Molecular Devices.
[Test materials]: KM12 cell line (ATCC, item number CRL-12496), 96-well transparent flat-bottomed black-walled cell culture plate (Corning, item number #3603), RPMI-1640 media (GE, cat #SH30027.01), Fetal Bovine Serum FBS (Thermo Fisher, cat #10099-141), CellTiter-Glo® Luminescent Cell Viability Assay Kit (Promega, cat #G7572), PBS (Solarbio, cat #P1020), Trypsin (Thermo Fisher, cat #25200072), DMSO (Sigma, cat #D2650).
[Experimental procedure]: When resuscitating KM12 cells, the freezing tube was quickly shaken in a 37° C. water bath to melt within 1 minute. The thawed cell suspension was mixed uniformly with RPMI1640 medium containing 10% FBS, centrifuged at 1000 rpm for 5 minutes, and the supernatant discarded, 5 mL of complete medium (RPMI1640 medium with 10% FBS) was taken to resuspend the cell pellets, charged into a cell culture flask with a bottom area of 25 cm.sup.2, and incubated in a cell incubator at 37° C., 95% humidity and 5% CO.sub.2. Cells were allowed to passage when the cell confluence rate reached about 80%. For cell passage, the old medium was discarded, and the cells were washed twice with PBS, and 0.5 mL trypsin was added to digest the cells. Upon dispersing into single cells and exfoliating from the bottom of the cell culture flask, 4.5 mL of new complete medium was added to terminate the digestion. After pipetting the cell suspension evenly, 1/5 of the cell suspension was kept and added with 4 mL of new complete medium, and after pipetting evenly, the cell flask was placed in the cell incubator for further culturing. Cell plating was performed when the ceil confluence rate reached about 80% again. For cell plating, ⅕ of the cell suspension was kept for further culturing as in the cell passage method, and the remaining ⅘ of the cell suspension was placed in a 15 mL centrifuge tube, detected for cell viability by Trypan blue exclusion method to ensure that cell viability was above 90%. A cell suspension with a density of 3.33×10.sup.4 viable cells/mL was prepared with complete medium, and 90 μL of such cell suspension was added to a 96-well cell culture plate to obtain a cell density of 3000 viable cells/mL in the cell culture plate. A control group containing no cells, no compound and only complete medium (i.e. culture medium control) and a control group containing no compound but containing cells (i.e. cell control) were set. The cell plate was placed in a cell incubator overnight.

[0477] The 10 mM compound stock solution in DMSO was first serially diluted with DMSO at a dilution factor of 3.16 times, to obtain 9 concentrations, and the 10th concentration was set as DMSO control without compound. Then, PBS was used to dilute the DMSO solutions of compounds of different concentrations, at a dilution factor of 100 times, so that the DMSO concentration in the compound solution of each concentration was 1%. Finally, 10 μL of each of the above solutions was added to the corresponding cell culture plate, so that the initial compound concentration was 10 μM, with the remaining concentrations being successively diluted at a dilution factor of 3.16 times, and the DMSO content in the cell culture plate was 0.1%.

[0478] The cell plate was placed in a cell incubator and cultured for 72 hours. For endpoint detection, the CellTiter-Glo reagent was melted, and the cell plate was moved to room temperature to equilibrate for 30 minutes, added with 100 μL of CellTiter-Glo for each well, and shaken on an orbital shaker for 5 minutes to frilly lyse the cells. The plate was kept at room temperature for 20 minutes to stabilize the luminescence signal, and the luminescence value of each well was scanned with a multi-functional microplate reader at full wavelength.

[Test samples] The compounds of Example 1-6 and Cisplatin (positive control compound)

LOXO-101:

[0479] ##STR00172## [0480] (Doebele, R. C. et al, Cancer Discov. 2015, 5, 1049-1057)

RXDX-101:

[0481] ##STR00173## [0482] (C. Rolfo et al., Expert Opin. Investig. Drugs 2015, 24 (11), 1493-1500)

Merestinib:

[0483] ##STR00174##

[Data analysis] The following formula was used to calculate the cell survival rate under the action of each concentration of the compounds:

Cell survival rate(%)=(Lum.sub.drug to be tested−Lum.sub.medium control)/(Lum.sub.cell control−Lum.sub.medium control)×100%

[0484] GraphPad Prism 5.0 software was used to analyze the data, and nonlinear S-curve regression was used to fit the data to obtain a dose-response curve, and the IC50 values were calculated therefrom.

TABLE-US-00002 TABLE 1 The inhibitory activity of the compounds of Examples 1-6 on the proliferation of KM12 cells (IC50, μM) Compound KM-12, IC50 (μM) Example 1 0.0156 Example 2 0.0091 Example 3 0.0167 Example 4 0.0310 Example 5 0.0051 Example 6 0.0063 Merestinib 0.0087 LOXO-101 0.0083 RXDX-101 0.0051 Cisplatin 27.3059

[0485] The inhibitory activity of the compounds of the following examples on the proliferation of KM12 cells was investigated in separate tests as compared with merestinib using experimental methods, conditions, and procedures basically analogous to the above. The results are shown in the following tables.

TABLE-US-00003 TABLE 1-1 Compound KM-12, IC50 (μM) Merestinib 0.033 Example 18 0.032 Example 19 0.032 Example 12 0.016 Example 23 0.032

TABLE-US-00004 TABLE 1-2 Compound KM-12, IC50 (μM) Merestinib 0.059 Example 14 0.024 Example 15 0.017 Example 16 0.007 Example 17 0.019 Example 11 0.035 Example 25 0.029

TABLE-US-00005 TABLE 1-3 Compound KM-12, IC50 (μM) Merestinib 0.027 Example 24 0.023 Example 26 0.018

Example 2: Inhibitory Activity of the Compounds of the Present Disclosure on the Proliferation of Ba/F3 ETV6-NTRK2 Cells

[0486] Ba/F3 ETV6-NTRK2 cell is a stable transgenic cell line constructed by KYinno Biotechnology Co., Ltd. on mouse primary B cell Ba/F3 wherein NTRK2 is highly expressed and fused with ETV6 to form ETV6-NTRK2.

[Test method]: Promega's CellTiter-Glo® Luminescent Cell Viability Assay kit was used to evaluate the inhibitory activity of the compounds on the proliferation of Ba/F3 ETV6-NTRK2C cells.
[Instrument]: Spectramax M3 multifunctional microplate reader from Molecular Devices.
[Test materials]: Ba/F3 ETV6-NTRK2 cell line (constructed by KYinno Biotechnology Co., Ltd. (Beijing)), 96-well transparent flat-bottomed black-walled cell culture plate (Corning, cat #3603), RPMI-1640 media (GE, cat #SH30027.01), fetal bovine serum FBS (Thermo Fisher, cat #10099-141), CellTiter-Glo® Luminescent Cell Viability Assay kit (Promega, cat #G7572), PBS (Solarbio, cat #P1020), DMSO (Sigma, cat #D2650).
[Experimental procedure]: When resuscitating Ba/F3 ETV6-NTRK2 cells, the freezing tube was quickly shaken in a 37° C. water bath to melt within 1 minute. The thawed cell suspension was mixed uniformly with RPMI1640 medium containing 10% FBS, centrifuged at 1000 rpm for 5 minutes, and the supernatant discarded, 5 mL of complete medium (RPMI1640 medium with 10% FBS) was taken to resuspend the cell pellets, charged into a cell culture flask with a bottom area of 25 cm.sup.2, and incubated in a cell incubator at 37° C., 95% humidity and 5% CO.sub.2. Cells were allowed to passage when the number of cells reached about 10.sup.6 cells/mL. For cell passage, the old cell suspension was directly pipetted evenly, ⅕ of the cell suspension was kept, 4 mL of new complete medium was added, and after pipetting evenly, the cell flask was placed in the cell incubator for further culturing. Cell plating was performed when the number of cells reached about 10.sup.6 cells/mL again. For cell plating, ⅕ of the cell suspension was kept for further culturing as in the cell passage method, and the remaining ⅘ of the cell suspension was placed in a 15 mL centrifuge tube, detected for cell viability by Trypan blue exclusion method to ensure that cell viability was above 90%. A cell suspension with a density of 5.56×10.sup.4 viable cells/mL was prepared with complete medium, and 90 μL of such cell suspension was added to a 96-well cell culture plate to obtain a cell density of 5000 viable cells/mL in the cell culture plate. A control group containing no cells, no compound and only complete medium (i.e, culture medium control) and a control group containing no compound but containing cells (i.e, cell control) were set. The cell plate was placed in a cell incubator overnight. The 10 mM compound stock solution in DMSO was first serially diluted with DMSO at a dilution factor of 3.16 times, to obtain 9 concentrations, and the 10th concentration was set as DMSO control without compound. Then, PBS was used to dilute the DMSO solutions of compounds of different concentrations, at a dilution factor of 100 times, so that the DMSO concentration in the compound solution of each concentration was 1%. Finally, 10 μL of each of the above solutions was added to the corresponding cell culture plate, so that the initial compound concentration was 10 μM, with the remaining concentrations being successively diluted at a dilution factor of 3.16 times, and the DMSO content in the cell culture plate was 0.1%. The cell plate was placed in a cell incubator and cultured for 72 hours. For endpoint detection, the CellTiter-Glo reagent was melted, and the cell plate was moved to room temperature to equilibrate for 30 minutes, added with 100 μL of CellTiter-Glo for each well, and shaken on an orbital shaker for 5 minutes to fully lyse the cells. The plate was kept at room temperature for 20 minutes to stabilize the luminescence signal, and the luminescence value of each well was scanned with a multi-functional microplate reader at full wavelength.
[Test sample] Compounds of Example 1-6 and LOXO-101 (positive control compound), Merestinib (comparative compound)
[Data analysis] The following formula was used to calculate the cell survival rate under the action of each concentration of the compounds:

Cell survival rate(%)=(Lum.sub.drug to be tested−Lum.sub.medium control)/(Lum.sub.cell control−Lum.sub.medium control)×100%

[0487] GraphPad Prism 5.0 software was used to analyze the data, and nonlinear S-curve regression was used to fit the data to obtain a dose-response curve, and the IC50 values were calculated therefrom.

TABLE-US-00006 TABLE 2 The inhibitory activity of the example compounds on the proliferation of Ba/F3 ETV6-NTRK2 cells (IC50, μM) Compound Ba/F3 ETV6-NTRK2, IC50 (μM) Example 1 0.428 Example 2 0.269 Example 3 0.341 Example 4 0.876 Example 5 0.128 Example 6 0.209 Merestinib 0.211 LOXO-101 0.069

Example 3: Inhibitory Activity of the Compounds of Present Disclosure on the Proliferation of Ba/F3 ETV6-NTRK3 Cells

[0488] Ba/F3 ETV6-NTRK3 is a stable transgenic cell line constructed by KYinno Biotechnology Co., Ltd. on mouse primary B cell Ba/F3 wherein NTRK3 is highly expressed and fused with ETV6 to form ETV6-NTRK3.

[Test method]: Promega's CellTiter-Glo® Luminescent Cell Viability Assay kit was used to evaluate the inhibitory activity of the compounds on the proliferation of Ba/F3 ETV6-NTRK3 cells.
[Instrument]: Spectramax M3 multifunctional microplate reader from Molecular Devices.
[Test materials]: Ba/F3 ETV6-NTRK3 cell line (constructed by KYinno Biotechnology Co., Ltd. (Beijing)), 96-well transparent flat-bottomed black-walled cell culture plate (Corning, cat #3603), RPMI-1640 media (GE, cat #SH30027.01), fetal bovine serum FBS (Thermo Fisher, cat #10099-141), CellTiter-Glo® Luminescent Cell Viability Assay kit (Promega, cat #G7572), PBS (Solarbio, cat #P1020), DMSO (Sigma, cat #D2650).
[Experimental procedure]: When resuscitating Ba/F3 ETV6-NTRK3 cells, the freezing tube was quickly shaken in a 37° C. water bath to melt within 1 minute. The thawed cell suspension was mixed uniformly with RPMI1640 medium containing 10% FBS, centrifuged at 1000 rpm for 5 minutes, and the supernatant discarded, 5 mL of complete medium (RPMI1640 medium with 10% FBS) was taken to resuspend the cell pellets, charged into a cell culture flask with a bottom area of 25 cm.sup.2, and incubated in a cell incubator at 37° C., 95% humidity and 5% CO.sub.2. Cells were allowed to passage when the number of cells reached about 10.sup.6 cells/mL. For cell passage, the old cell suspension was directly pipetted evenly, ⅕ of the cell suspension was kept, 4 mL of new complete medium was added, after pipetting evenly, the cell flask was placed in the cell incubator for further culturing. Cell plating was performed when the number of cells reached about 10.sup.6 cells/mL again. For cell plating, ⅕ of the cell suspension was kept for further culturing as in the cell passage method, and the remaining ⅘ of the cell suspension was placed in a 15 mL centrifuge tube, detected for cell viability by Trypan blue exclusion method to ensure that cell viability was above 90%. A cell suspension with a density of 5.56×10.sup.4 viable cells/mL was prepared with complete medium, and 90 μL of such cell suspension was added to a 96-well cell culture plate to obtain the cell density of 5000 viable cells/mL in the cell culture plate. A control group containing no cells, no compound and only complete medium (i.e. culture medium control) and a control group containing no compound but containing cells (i.e. cell control) were set. The cell plate was placed in a cell incubator overnight. The 10 mM compound stock solution in DMSO was first serially diluted with DMSO at a dilution factor of 3.16 times, to obtain 9 concentrations, and the 10th concentration was set as DMSO control without compound. Then, PBS was used to dilute the DMSO solutions of compounds of different concentrations, at a dilution factor of 100 times, so that the DMSO concentration in the compound solution of each concentration was 1%. Finally, 10 μL of each of the above solutions was added to the corresponding cell culture plate, so that the initial compound concentration was 10 μM, with the remaining concentrations being successively diluted at a dilution factor of 3.16 times, and the DMSO content in the cell culture plate was 0.1%. The cell plate was placed in a cell incubator and cultured for 72 hours. For endpoint detection, the CellTiter-Glo reagent was melted, and the cell plate was moved to room temperature to equilibrate for 30 minutes, added with 100 μL of CellTiter-Glo for each well, and shaken on an orbital shaker for 5 minutes to fully lyse the cells. The plate was kept at room temperature for 20 minutes to stabilize the luminescence signal, and the luminescence value of each well was scanned with a multi-functional microplate reader at full wavelength.
[Test samples] Example compounds, LOXO-101 (positive control compound), Merestinib (comparative compound)
[Data analysis] The following formula was used to calculate the cell survival rate under the action of each concentration of the compounds:

Cell survival rate(%)=(Lum.sub.drug to be tested−Lum.sub.medium control)/(Lum.sub.cell control−Lum.sub.medium control)×100%

[0489] GraphPad Prism 5.0 software was used to analyze the data, and nonlinear S-curve regression was used to fit the data to obtain a dose-response curve, and the IC50 values were calculated therefrom.

TABLE-US-00007 TABLE 3 The inhibitory activity (IC50, μM) of the examples on the proliferation of Ba/F3 ETV6-NTRK3 cells Compound Ba/F3 ETV6-NTRK3, IC50 (μM) Example 1 0.435 Example 2 0.094 Example 3 0.663 Example 4 0.762 Example 5 0.060 Example 6 0.284 Merestinib 0.684 LOXO-101 0.005

Example 4: Inhibitory Activity of the Compounds of the Present Disclosure on the Proliferation of Ba/F3 LMNA-NTRK1-G667C Cells

[0490] Ba/F3 LMNA-NTRK1-G667C is a stable transgenic cell line constructed by KYinno Biotechnology Co., Ltd, on mouse primary B cell Ba/F3 wherein NTRK1 has a mutation of G to Cat position 667 and is highly expressed and fused with LMNA to form LMNA-NTRK1-G667C.

[Test method]: Promega's CellTiter-Glo® Luminescent Cell Viability Assay kit was used to evaluate the inhibitory activity of the compounds on the proliferation of Ba/F3 LMNA-NTRK1-G667C cells.
[Instrument]: Spectramax M3 multifunctional microplate reader from Molecular Devices.
[Test materials]: Ba/F3 LMNA-NTRK1-G667C cell line (constructed by KYinno Co., Ltd. (Beijing)), 96-well transparent flat-bottomed black-walled cell culture plate (Corning, cat #3603), RPMI-1640 media (GE, cat #SH30027.01), fetal bovine serum FBS (Thermo Fisher, cat #10099-141), CellTiter-Glo® Luminescent Cell Viability Assay kit (Promega, cat #G7572), PBS (Solarbio, cat #P1020), DMSO (Sigma, cat #D2650).
[Experimental procedure]: When resuscitating Ba/F3 LMNA-NTRK1-G667C cells, the freezing tube was quickly shaken in a 37° C. water bath to melt within 1 minute. The thawed cell suspension was mixed uniformly with RPMI1640 medium containing 10% FBS, centrifuged at 1000 rpm for 5 minutes, and the supernatant discarded. 5 mL of complete medium (RPMI1640 medium with 10% FBS) was taken to resuspend the cell pellets, charged into a cell culture flask with a bottom area of 25 cm.sup.2, and incubated in a cell incubator at 37° C., 95% humidity and 5% CO.sub.2. Cells were allowed to passage when the number of cells reached about 10.sup.6 cells/mL. For cell passage, the old cell suspension was directly pipetted evenly, ⅕ of the cell suspension was kept, 4 mL of new complete medium was added, after pipetting evenly, the cell flask was placed in the cell incubator for further culturing. Cell plating was performed when the number of cells reached about 10.sup.6 cells/mL again. For cell plating, ⅕ of the cell suspension was kept for Anther culturing as in the cell passage method, and the remaining ⅘ of the cell suspension was placed in a 15 mL centrifuge tube, detected for cell viability by Trypan blue exclusion method to ensure that cell viability was above 90%. A cell suspension with a density of 5.56×10.sup.4 viable cells/mL was prepared with complete medium, and 90 μL of such cell suspension was added to a 96-well cell culture plate to obtain the cell density of 5000 viable cells/mL in the cell culture plate. A control group containing no cells, no compound and only complete medium (i.e. culture medium control) and a control group containing no compound but containing cells (i.e. cell control) were set. The cell plate was placed in a cell incubator overnight. The 10 mM compound stock solution in DMSO was first serially diluted with DMSO at a dilution factor of 3.16 times, to obtain 9 concentrations, and the 10th concentration was set as DMSO control without compound. Then, PBS was used to dilute the DMSO solutions of compounds of different concentrations, at a dilution factor of 100 times, so that the DMSO concentration in the compound solution of each concentration was 1%. Finally, 10 μL of each of the above solutions was added to the corresponding cell culture plate, so that the initial compound concentration was 10 μM, with the remaining concentrations being successively diluted at a dilution factor of 3.16 times, and the DMSO content in the cell culture plate was 0.1%. The cell plate was placed in a cell incubator and cultured for 72 hours. For endpoint detection, the CellTiter-Glo reagent was melted, and the cell plate was moved to room temperature to equilibrate for 30 minutes, added with 100 μL of CellTiter-Glo for each well, and shaken on an orbital shaker for 5 minutes to fully lyse the cells. The plate was kept at room temperature for 20 minutes to stabilize the luminescence signal, and the luminescence value of each well was scanned with a multi-functional microplate reader at full wavelength.
[Test samples] Example compounds and LOXO-101 (positive control compound)
[Data analysis] The following formula was used to calculate the cell survival rate under the action of each concentration of the compounds:

Cell survival rate(%)=(Lum.sub.drug to be tested−Lum.sub.medium control)/(Lum.sub.cell control−Lum.sub.medium control)×100%

[0491] GraphPad Prism 5.0 software was used to analyze the data, and nonlinear S-curve regression was used to fit the data to obtain a dose-response curve, and the IC50 values were calculated therefrom.

TABLE-US-00008 TABLE 4 The inhibitory activity (IC50, μM) of the example compounds on Ba/F3 LMNA-NTRK1-G667C cell proliferation Compound Ba/F3 LMNA-NTRK1-G667C, IC50 (μM) Example 1 0.0020 Example 2 0.0011 Example 3 0.0016 Example 4 0.0028 Example 5 <0.001 Example 6 0.00066 Merestinib 0.0011 LOXO-101 2.409

[0492] The inhibitory activity of some compounds of the present disclosure on the proliferation of Ba/F3 LMNA-NTRK1-G667C cells was investigated in separate tests as compared with merestinib using experimental methods, conditions, and procedures basically analogous to the above. The results are shown in the following tables.

TABLE-US-00009 TABLE 4-1 Compound Ba/F3 LMNA-NTRK1-G667C, IC50 (μM) Merestinib 0.001 Example 18 0.001 Example 19 0.002 Example 12 <0.001 Example 23 0.001

TABLE-US-00010 TABLE 4-2 Compound Ba/F3 LMNA-NTRK1-G667C, IC50 (μM) Merestinib 0.0141 Example 14 0.0086 Example 15 0.0108 Example 16 0.0107 Example 11 0.0105 Example 24 0.0173 Example 26 0.0175

Example 5: Inhibitory Activity of the Compounds of the Present Disclosure on the Proliferation of Ba/F3 LMNA NTRK1 G595R Cells

[0493] Ba/F3 LMNA-NTRK1-G595R is a stable transgenic cell line constructed by KYinno Biotechnology Co., Ltd. on mouse primary B cell Ba/F3 wherein NTRK1 is highly expressed with a mutation of G at position 595 to R, and fused with LMNA to form LMNA-NTRK1.

[Test method]: Promega's CellTiter-Glo® Luminescent Cell Viability Assay kit was used to evaluate the inhibitory activity of the compounds on the proliferation of Ba/F3 LMNA-NTRK1-G595R cells.
[Instrument]: Spectramax M3 multifunctional microplate reader from Molecular Devices.
[Test materials]: Ba/F3 LMNA-NTRK1-G595R cell line (constructed by KYinno Biotechnology Co., Ltd. (Beijing)), 96-well transparent flat-bottomed black-walled cell culture plate (Corning, cat #3603), RPMI-1640 media (GE, cat #SH30027.01), fetal bovine serum FBS (Thermo Fisher, cat #10099-141), CellTiter-Glo® Luminescent Cell Viability Assay kit (Promega, cat #G7572), PBS (Solarbio, cat #P1020), DMSO (Sigma, cat #D2650).
[Experimental procedure]: When resuscitating Ba/F3 LMNA-NTRK1-G595R cells, the freezing tube was quickly shaken in a 37° C. water bath to melt within 1 minute. The thawed cell suspension was mixed uniformly with RPMI1640 medium containing 10% FBS, centrifuged at 1000 rpm for 5 minutes, and the supernatant discarded. 5 mL of complete medium (RPMI1640 medium with 10% FBS) was taken to resuspend the cell pellets, charged into a cell culture flask with a bottom area of 25 cm.sup.2, and incubated in a cell incubator at 37° C., 95% humidity and 5% CO.sub.2. Cells were allowed to passage when the number of cells reached about 10.sup.6 cells/mL. For cell passage, the old cell suspension was directly pipetted evenly, ⅕ of the cell suspension was kept, 4 mL of new complete medium was added, after pipetting evenly, the cell flask was placed in the cell incubator for further culturing. Cell plating was performed when the number of cells reached about 10.sup.6 cells/mL again. For cell plating, ⅕ of the cell suspension was kept for further culturing as in the cell passage method, and the remaining ⅘ of the cell suspension was placed in a 15 mL centrifuge tube, detected for cell viability by Trypan blue exclusion method to ensure that cell viability was above 90%. A cell suspension with a density of 5.56×10.sup.4 viable cells/mL was prepared with complete medium, and 90 μL of such cell suspension was added to a 96-well cell culture plate to obtain the cell density of 5000 viable cells/mL in the cell culture plate. A control group containing no cells, no compound and only complete medium (i.e. culture medium control) and a control group containing no compound but containing cells (i.e. cell control) were set. The cell plate was placed in a cell incubator overnight. The 10 mM compound stock solution in DMSO was first serially diluted with DMSO at a dilution factor of 3.16 times, to obtain 9 concentrations, and the 10th concentration was set as DMSO control without compound. Then, PBS was used to dilute the DMSO solutions of compounds of different concentrations, at a dilution factor of 100 times, so that the DMSO concentration in the compound solution of each concentration was 1%. Finally, 10 μL of each of the above solutions was added to the corresponding cell culture plate, so that the initial confound concentration was 10 μM, with the remaining concentrations being successively diluted at a dilution factor of 3.16 times, and the DMSO content in the cell culture plate was 0.1%. The cell plate was placed in a cell incubator and cultured for 72 hours. For endpoint detection, the CellTiter-Glo reagent was melted, and the cell plate was moved to room temperature to equilibrate for 30 minutes, added with 100 μL of CellTiter-Glo for each well, and shaken on an orbital shaker for 5 minutes to fully lyse the cells. The plate was kept at room temperature for 20 minutes to stabilize the luminescence signal, and the luminescence value of each well was scanned with a multi-functional microplate reader at full wavelength.
[Test samples] Example compounds and LOXO-101 (positive control compound)
[Data analysis] The following formula was used to calculate the cell survival rate under the action of each concentration of the compounds:

Cell survival rate(%)=(Lum.sub.drug to be tested−Lum.sub.medium control)/(Lum.sub.cell control−Lum.sub.medium control)×100%

[0494] GraphPad Prism 5.0 software was used to analyze the data, and nonlinear S-curve regression was used to fit the data to obtain a dose-response curve, and the IC50 values were calculated therefrom.

TABLE-US-00011 TABLE 5 The inhibitory activity of the example compounds on the Ba/F3 LMNA-NTRK1-G595R cell proliferation (IC50, μM) Compound Ba/F3 LMNA-NTRK1-G595R, IC50 (μM) Example 1 1.435 Example 2 0.752 Example 3 3.152 Example 4 4.654 Example 5 0.799 Example 6 0.747 Merestinib 1.015 LOXO-101 3.657

Example 6: Inhibitory Activity of the Compounds of the Present Disclosure on the Proliferation of Ba/F3 ETV6 M RK2 G639R Cells

[0495] Ba/F3 ETV6-NTRK2-G639R is a stable transgenic cell line constructed by KYinno Biotechnology Co., Ltd. on mouse primary B cell Ba/F3 wherein NTRK2 is highly expressed with a mutation of G at position 639 to R., and fused with ETV6 to form ETV6-NTRK2.

[Test method]: Promega's CellTiter-Glo® Luminescent Cell Viability Assay kit was used to evaluate the inhibitory activity of the compounds on the proliferation of Ba/F3 ETV6-NTRK2-G639R cells.
[Instrument]: Spectramax M3 multifunctional microplate reader from Molecular Devices.
[Test materials]: Ba/F3 ETV6-NTRK2-G639R cell line (constructed by KYinno Biotechnology Co., Ltd. (Beijing)), 96-well transparent flat-bottomed black-walled cell culture plate (Corning, catalog #3603), RPMI-1640 media (GE, cat #SH30027.01), fetal bovine serum FBS (Thermo Fisher, cat #10099-141), CellTiter-Glo® Luminescent Cell Viability Assay kit (Promega, cat #G7572), PBS (Solarbio, cat #P1020), DMSO (Sigma, cat #D2650).
[Experimental procedure]: When resuscitating Ba/F3 ETV6-NTRK2-G639R cells, the freezing tube was quickly shaken in a 37° C. water bath to melt within 1 minute. The thawed cell suspension was mixed uniformly with RPMI1640 medium containing 10% FBS, centrifuged at 1000 rpm for 5 minutes, and the supernatant discarded. 5 mL of complete medium (RPMI1640 medium with 10% FBS) was taken to resuspend the cell pellets, charged into a cell culture flask with a bottom area of 25 cm.sup.2, and incubated in a cell incubator at 37° C., 95% humidity and 5% CO.sub.2. Cells were allowed to passage when the number of cells reached about 10.sup.6 cells/mL. For cell passage, the old cell suspension was directly pipetted evenly, ⅕ of the cell suspension was kept, 4 mL of new complete medium was added, after pipetting evenly, the cell flask was placed in the cell incubator for further culturing. Cell plating was performed when the number of cells reached about 10.sup.6 cells/mL again. For cell plating, ⅕ of the cell suspension was kept for further culturing as in the cell passage method, and the remaining ⅘ of the cell suspension was placed in a 15 mL centrifuge tube, detected for cell viability by Trypan blue exclusion method to ensure that cell viability was above 90%. A cell suspension with a density of 5.56×10.sup.4 viable cells/mL was prepared with complete medium, and 90 μL of such cell suspension was added to a 96-well cell culture plate to obtain the cell density of 5000 viable cells/mL in the cell culture plate. A control group containing no cells, no compound and only complete medium (i.e. culture medium control) and a control group containing no compound but containing cells (i.e. cell control) were set. The cell plate was placed in a cell incubator overnight. The 10 mM compound stock solution in DMSO was first serially diluted with DMSO at a dilution factor of 3.16 times, to obtain 9 concentrations, and the 10th concentration was set as DMSO control without compound. Then, PBS was used to dilute the DMSO solutions of compounds of different concentrations, at a dilution factor of 100 times, so that the DMSO concentration in the compound solution of each concentration was 1%. Finally, 10 μL of each of the above solutions was added to the corresponding cell culture plate, so that the initial compound concentration was 10 μM, with the remaining concentrations being successively diluted at a dilution factor of 3.16 times, and the DMSO content in the cell culture plate was 0.1%. The cell plate was placed in a cell incubator and cultured for 72 hours. For endpoint detection, the CellTiter-Glo reagent was melted, and the cell plate was moved to room temperature to equilibrate for 30 minutes, added with 100 μL of CellTiter-Glo for each well, and shaken on an orbital shaker for 5 minutes to fully lyse the cells. The plate was kept at room temperature for 20 minutes to stabilize the luminescence signal, and the luminescence value of each well was scanned with a multi-functional microplate reader at frill wavelength.
[Test samples] Example compounds and LOXO-101 (positive control compound)
[Data analysis] The following formula was used to calculate the cell survival rate under the action of each concentration of the compounds:

Cell survival rate(%)=(Lum.sub.drug to be tested−Lum.sub.medium control)/(Lum.sub.cell control−Lum.sub.medium control)×100%

[0496] GraphPad Prism 5.0 software was used to analyze the data, and nonlinear S-curve regression was used to fit the data to obtain a dose-response curve, and the IC50 values were calculated therefrom.

TABLE-US-00012 TABLE 6 The inhibitory activity of example compounds on Ba/F3 ETV6-NTRK2-G639R cell proliferation (IC50, μM) Compound Ba/F3 ETV6-NTRK2-G639R, IC50 (μM) Example 1 3.116 Example 2 1.878 Example 3 9.866 Example 4 >10 Example 5 2.691 Example 6 2.792 Merestinib 3.038 LOXO-101 >10

Example 7: Inhibitory Activity of the Compounds of the Present Disclosure on the Proliferation of Ba/F3 ETV6 NTRK3-G696C Cells

[0497] Ba/F3 ETV6-NTRK3-G696C is a stable transgenic cell line constructed by KYinno Biotechnology Co., Ltd. on mouse primary B cell Ba/F3 wherein NTRK3 is highly expressed with a mutation of G at position 639 to C and fused with ETV6 to form ETV6-NTRK3.

[Test method]: Promega's CellTiter-Glo® Luminescent Cell Viability Assay kit was used to evaluate the inhibitory activity of the compounds on the proliferation of Ba/F3 ETV6-NTRK3-G696C cells.
[Instrument]: Spectramax M3 multifunctional microplate reader from Molecular Devices.
[Test materials]: Ba/F3 ETV6-NTRK3-G696C cell line (constructed by KYinno Biotechnology Co., Ltd. (Beijing)), 96-well transparent flat-bottomed black-walled cell culture plate (Corning, cat #3603), RPMI-1640 media (GE, product number #SH30027.01), fetal bovine serum FBS (Thermo Fisher, cat #10099-141), CellTiter-Glo® Luminescent Cell Viability Assay kit (Promega, cat #G7572), PBS (Solarbio, cat #P1020), DMSO (Sigma, cat #D2650).
[Experimental procedure]: When resuscitating Ba/F3 ETV6-NTRK3-G696C cells,
the freezing tube was quickly shaken in a 37° C. water bath to melt within 1 minute. The thawed cell suspension was mixed uniformly with RPMI1640 medium containing 10% FBS, centrifuged at 1000 rpm for 5 minutes, and the supernatant discarded. 5 mL of complete medium (RPMI1640 medium with 10% FBS) was taken to resuspend the cell pellets, charged into a cell culture flask with a bottom area of 25 cm.sup.2, and incubated in a cell incubator at 37° C., 95% humidity and 5% CO.sub.2. Cells were allowed to passage when the number of cells reached about 10.sup.6 cells/mL. For cell passage, the old cell suspension was directly pipetted evenly, ⅕ of the cell suspension was kept, 4 mL of new complete medium was added, after pipetting evenly, the cell flask was placed in the cell incubator for further culturing. Cell plating was performed when the number of cells reached about 10.sup.6 cells/mL again. For cell plating, ⅕ of the cell suspension was kept for further culturing as in the cell passage method, and the remaining ⅘ of the cell suspension was placed in a 15 mL centrifuge tube, detected for cell viability by Trypan blue exclusion method to ensure that cell viability was above 90%. A cell suspension with a density of 5.56×10.sup.4 viable cells/mL was prepared with complete medium, and 90 μL of such cell suspension was added to a 96-well cell culture plate to obtain the cell density of 5000 viable cells/mL in the cell culture plate. A control group containing no cells, no compound and only complete medium (i.e. culture medium control) and a control group containing no compound but containing cells (i.e. cell control) were set. The cell plate was placed in a cell incubator overnight. The 10 mM compound stock solution in DMSO was first serially diluted with DMSO at a dilution factor of 3.16 times, to obtain 9 concentrations, and the 10th concentration was set as DMSO control without compound. Then, PBS was used to dilute the DMSO solutions of compounds of different concentrations, at a dilution factor of 100 times, so that the DMSO concentration in the compound solution of each concentration was 1%. Finally, 10 μL of each of the above solutions was added to the corresponding cell culture plate, so that the initial compound concentration was 10 μM, with the remaining concentrations being successively diluted at a dilution factor of 3.16 times, and the DMSO content in the cell culture plate was 0.1%. The cell plate was placed in a cell incubator and cultured for 72 hours. For endpoint detection, the CellTiter-Glo reagent was melted, and the cell plate was moved to room temperature to equilibrate for 30 minutes, added with 100 μL of CellTiter-Glo for each well, and shaken on an orbital shaker for 5 minutes to fully lyse the cells. The plate was kept at room temperature for 20 minutes to stabilize the luminescence signal, and the luminescence value of each well was scanned with a multi-functional microplate reader at full wavelength.
[Test samples] Example compounds and LOXO-101 (positive control compound)
[Data analysts] The following formula was used to calculate the cell survival rate under the action of each concentration of the compounds:

Cell survival rate(%)=(Lum.sub.drug to be tested−Lum.sub.medium control)/(Lum.sub.cell control−Lum.sub.medium control)×100%

[0498] GraphPad Prism 5.0 software was used to analyze the data, and nonlinear S-curve regression was used to fit the data to obtain a dose-response curve, and the IC50 values were calculated therefrom.

TABLE-US-00013 TABLE 7 The inhibitory activity of example compounds on Ba/F3 ETV6-NTRK3-G696C cell proliferation (IC50, μM) Compound Ba/F3 ETV6-NTRK3-G696C, IC50 (μM) Example 1 0.033 Example 2 0.020 Example 3 0.018 Example 4 0.034 Example 5 0.0068 Example 6 0.016 Merestinib 0.010 LOXO-101 1.891

[0499] The inhibitory activity of some compounds of the present disclosure on the proliferation of Ba/F3 ETV6-NTRK3-G696C cells was investigated in separate tests as compared with merestinib using experimental methods, conditions, and procedures basically analogous to the above. The results are shown in the following tables.

TABLE-US-00014 TABLE 7-1 Compound Ba/F3 ETV6-NTRK3-G696C, IC50 (μM) Merestinib 0.013 Example 18 0.012 Example 19 0.014 Example 12 0.003 Example 23 0.011

TABLE-US-00015 TABLE 7-2 Compound Ba/F3 ETV6-NTRK3-G696C, IC50 (μM) Merestinib 0.0017 Example 14 0.0016 Example 15 0.0019 Example 16 0.0012 Example 17 0.0037 Example 11 0.0018 Example 25 0.0063 Example 24 0.0035 Example 27 0.0030 Example 26 0.0031

Example 8: Inhibitory Activity of the Compounds of the Present Disclosure on the Proliferation of MKN45 Cells

[0500] MKN45 is a human poorly differentiated gastric cancer cell line with high MET expression. Cell proliferation experiments were performed according to literature methods (Zhang, D. et al. Bioorg. Med. Chem. 2013, 21, 6804; Riss, T. L.; Moravec, R. A.; Niles, A. L.; et al. Cell Viability Assays 2013 May 1 [Updated 2016 Jul. 1], In; Sittampalam, G. S.; Grossman, A.; Brimacombe, K.; et al., editors. Assay Guidance Manual [Internet]. Bethesda (Md.): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004-. Available from; https://www.ncbi.nlm.nih.gov/books/NBK144065/).

TABLE-US-00016 TABLE 8 Inhibitory activity of some example compounds on the proliferation of MKN45 cells (IC50, μM) Compound MKN45, IC50 (μM) Example 12 0.004 Example 14 0.011 Example 16 0.009 Example 17 0.015 Example 11 0.012 Merestinib 0.019 LOXO-101 >10

[0501] The experimental results show that the compounds of the present disclosure have a more effective inhibitory effect on MKN45 cells which highly express the main target of Merestinib, namely Met.

Example 9: Inhibitory Activity of Selected Compounds of the Present Disclosure on the Proliferation of KM12, Ba/F3 LMNA NTRK1G667C, Ba/F3 ETV6 NTRK3 G696C Cells

[0502] Using experimental methods and conditions analogous to the above activity examples 1, 4 and 7, the inhibitory activity of the following selected compounds of the disclosure on each of the cell lines was investigated, and the results are shown in the following table:

TABLE-US-00017 TABLE 9 Cell proliferation inhibitory activity (IC50, μM) of some example compounds Ba/F3 LMNA- Ba/F3 ETV6- Compound KM12 NTRK1-G667C NTRK3-G696C Example 28 0.056 0.0198 0.0036 Example 31 0.071 0.0330 0.0053 Example 32 0.067 0.0190 0.0047 Example 34 0.037 0.0237 0.0048

Example 10: Inhibitory Activity of the Compounds of the Present Disclosure on Other Oncogenic Kinases

[0503] The experiment was performed according to the standard kinase activity determination protocol by Eurofins DiscoverX Corporation (11180 Roselle St. Suite D, San Diego, Calif., USA, www.discoverX.com). Specifically, the experiment was conducted using the KINOMEscan™ screening platform, which adopts an active site-directed competition binding assay to quantitatively measure the interaction between test compounds and more than hundreds of human kinases and disease-related mutants. The working principle of the screening platform and kinase assay methods are well known in the art, for example, see www.discoverX.com

[0504] The compound Example 12 of the present disclosure showed >90% inhibitory activity on the kinases listed in Table 10 at a concentration of 2 μM.

TABLE-US-00018 TABLE 10 Kinases % inhibition CDK11 100 CDKL2 100 DDR2 100 FLT3(N841I) 100 KIT(L576P) 100 MYO3B 100 AXL 100 DDR1 100 KIT(V559D) 100 TRKA 100 KIT 100 LOK 100 CSF1R 100 MKNK2 100 MET(M1250T) 99 FLT3 99 MET(Y1235D) 99 TIE1 99 TIE2 99 KIT(V559D, T670I) 99 MST1R 98 PDGFRB 98 FLT3(ITD) 98 MUSK 98 IKK-alpha 97 TRKC 97 IKK-beta 97 HIPK4 97 LCK 97 MERTK 97 ROS1 97 RET 97 EPHA8 96 CDK7 96 RET(M918T) 95 MKNK1 95 EPHB6 95 NEK9 95 BLK 95 TRKB 94 FLT3(K663Q) 93 MET 93 FLT3(D835V) 93 RAF1 92 FLT4 92 PLK4 90

[0505] Experimental results show that, in addition to inhibiting the activities of TRKA, TRKB and TRKC kinases, the compounds of the present disclosure also inhibit other confirmed, under-research, and emerging anticancer drug kinase targets (ROS1, RAF1, PDGFRB, CSF1R, LCK, IKKa, IKKb, PLK, AXL, TIE, LOK, TIE1, DDR), as well as those anticancer drug targets in applications and their drug-resistant mutant targets (FLT3, FLT3 (N841I), FLT3 (K663Q), FLT3 (D835V), FLT3 (ITD), KIT, KIT (L576P), KIT (V559D), KIT (V559D, T670I), MET, MET (Y1235D), MET (Y1250T), RET, RET (M918T)), etc., therefore are believed to show a broad-spectrum anti-cancer effect in clinical practice.

Example 11: Anti-Tumor Activity of the Compounds of the Present Disclosure in a KM12 Xenograft Mouse Model

[Instrument]: API-400-Qtrap Shimadzu 20 AC

[0506] [Test materials and conditions]: KM12 cell line was provided by Via Biotechnology, BALB/c nude mice, female, purchased from Shanghai Lingchang Biotechnology Co., Ltd.
[Experimental procedure]: 7-week-old athymic female nude mice were injected subcutaneously with 5×10.sup.6 KM12 cells on the right side of the ribs. The tumors were measured every 3 days at 3 dimensions to calculate the volume V=D×d×d/2, and the mouse body weights were measured at the same time. When the average tumor size reached 180 mm.sup.3 about 6 days after tumor inoculation, the mice were dosed for 12 days (or 18 days) prior sacrifice. For pharmacokinetic and pharmacodynamic analysis, plasma samples and tumor samples were taken from animals at 4 hours and 8 horns after the last dose.

[0507] The tumor samples from the animals treated as above were weighed, 9 volumes of ice water were added, and the resulting tumor tissue was homogenized by a tissue homogenizer. 50 μL of plasma or tumor homogenate sample was transferred to a 96-well plate, and added with 250 μL of ACN (containing 260 ng/ml dexamethasone as an internal standard) to precipitate protein. The plate was centrifuged at 4000 rpm at 4° C. for 20 minutes, 150 μL of supernatant was transferred to another new 96-well plate, mixed with 150 μL of 0.1% FA water, and 10 μL was injected into LC-MS/MS to collect data.

[Experiment Results] The data on the 12th day after administration show that, the tested example compounds of the present disclosure significantly inhibited the growth of tumors, for example, some of the compounds showed superior tumor growth inhibition activity than Merestinib (FIG. 1-A). The drug concentrations in plasma and tumor tested on the 12th and 18th days after administration show that, the tested compounds of the present disclosure can be enriched in plasma and tumor tissue at a higher concentration than Merestinib (FIG. 1-B, FIG. 1-C), which is expected to exert stronger anti-tumor activity.

Example 12: Comparison of Kinase Profile of the Compounds of the Present Disclosure and Merestinib

[0508] The inhibitory activity of Example 12 of the present disclosure on various kinases at a concentration of 2 μM was compared with that of Merestinib. The experiment was performed according to the standard kinase activity determination protocol by Eurofins DiscoverX Corporation (11180 Roselle St. Suite D, San Diego, Calif. USA, www.discoverX.com). At a concentration of 2 μM, Example 12 of the present disclosure shows a significantly different trend in the kinase activity profile, as compared with Merestinib, as shown in FIG. 2 (kinases with a difference of inhibition rate >20%).

[0509] The physicochemical properties of a compound determine its druggability. The property parameters that affect the druggability of a compound and the established experimental methods can refer to Kerns, Edward H. and Di Li (2008). Drug-like Properties: Concepts, Structure Design and Methods: from ADME to Toxicity Optimization. San Diego: Academic Press. The experimental results of the present disclosure show that the compound of the present disclosure is significantly superior to the prior art inhibitors such as Merestinib in many properties related to druggability. The following examples illustrate the significant advantages of the overall druggability of the compound of the present disclosure over the prior art by the metabolic stability in liver microsomes, P450 enzyme inhibition, and solubility.

Example 13: Metabolic Stability Test in Liver Microsomes of the Compound of the Present Disclosure

[0510] According to standard methods conventional in the art for in vitro metabolic stability studies, for example those protocols described in Kerns, Edward H. and Di Li (2008). Drug-like Properties: Concepts, Structure Design and Methods: from ADME to Toxicity Optimization. San Diego: Academic Press; Di, Li et al., Optimization of a Higher Throughput Microsomal Stability Screening Assay for Profiling Drug Discovery Candidates, J. Biomol. Screen. 2003, 5(4), 453., the metabolic stability test in liver microsomes of the present compound was performed analogously as follows.

[0511] The in vitro metabolic stability of the compound in 5 species (mouse, rat, dog, monkey, and human) was evaluated by liver microsomal stability test. 0.1M potassium phosphate buffer solution (pH 7.4, containing 1.0 mM EDTA) was prepared. The test compound and the control compound were prepared into a 10 mM DMSO solution, and then 10 μL of the 10 mM DMSO compound solution was added to 190 μL of acetonitrile to prepare a 500 μM compound solution. Liver microsomes (Sekisui XenoTech, LLC., human liver microsomes, cat. #H0610; Beagle dog liver microsomes, cat. #D1000; cynomolgus monkey liver microsomes, cat. #P2073; rat liver microsomes, cat. #R1073; mouse Liver microsomes, cat. #M1000) were taken and slowly melted on ice, then 18.75 μL of the liver microsome solution (20 mg/mL) and 1.5 μL of 500 μM compound solution were added to 479.75 μL of 0.1M potassium phosphate buffer solution to prepare a 0.75 mg/mL liver microsome working solution with a concentration of 1.5 μM compound. 6 mM NADPH (SIGMA, cat. #V900362) aqueous solution was prepared. 0.2 mL 2 mg/mL internal standard stock solution (imipramine solution in acetonitrile) was added to 1000 mL acetonitrile (SIGMA, cat. #271004) to prepare the reaction stop solution and stored at 4° C.

[0512] Six deep-well plates were labeled as T0, T5, T15, T30, T45 and NCF60 (negative control, without NADPH). 30 μL of 1.5 mM control compound or test compound solution in 0.75 mg/mL liver microsome working solution was added to each well of the corresponding plates. After being sealed with film, all the microplates were placed in a 37° C. water bath to preheat for 5 minutes, and 6 mM NADPH solution was placed in a 37° C. water bath to preheat for 5 minutes. Then 15 μL of NADPH solution was added to each cell successively in reverse time order, that is, first adding the T45 plate, 15 minutes later the T30 plate, 15 minutes later again the T15 plate, and 10 minutes later gain the T5 plate. NCF60 plate was not added with NADPH but 15 μL of deionized water. At this time, to the T0 plate were added 135 μL of ice-cooled reaction stop solution and 15 μL of NADPH solution. By the end of the reaction, 135 μL of ice-precooled reaction stop solution was added successively to each well of the T5, T15, T30, and T45 plates. All samples were centrifuged at 4° C. at 600 rpm for 10 minutes, and then at 5594 g for 15 minutes (Thermo Multifugex 3R). Six 96-well plates were respectively labeled and added 50 μL of ultrapure water (Millipore, ZMQS50F01) to each well, then 50 μL of the centrifuged supernatant was added to the corresponding well of the corresponding plate, for LC-MS/MS-014 (API4000), UPLC-MS/MS-11 (API4000), LC-MS/MS-034 (API-6500+) analysis. The concentration (C0) of the test compound at T0 time point was taken as 100%, and the concentrations at other incubation time points were converted into residue percentages. The natural logarithm of the residue percentage at each time point was subjected to linearly regression as a factor of incubation time to obtain Slope k, and then the liver microsome clearance rate (CL.sub.int.sup.mic) and in vitro half-life (T½) were calculated according to the following formula:

[00001]$C_t=\frac{1}{2}{C}_0$$T_{1/2}=\frac{\ln 2}{K}=\frac{0.693}{K}$${\mathrm{CL}}_{\mathrm{int}}^{\mathrm{mic}}=\frac{K}{\mathrm{mg}\text{/}\mathrm{ml}\mathrm{Incubated}\mathrm{microsomal}\mathrm{protein}}$

TABLE-US-00019 TABLE 11 Comparison of liver microsomal metabolism parameters between Example 12 of the present disclosure and Merestinib human mouse Rat dog monkey T½ (minute) Merestinib 78.95 27.52 52.01 89.91 25.04 Example 12 288.48 39.21 73.39 74.28 55.99 Cl.sub.int (mL/min/kg) Merestinib 22.02 90.26 104.93 38.43 80.96 Example 12 6.03 63.34 74.36 46.51 36.20

[0513] The study of the present disclosure shows that, the compounds of the present disclosure such as Example 12 showed significantly improved metabolic stability in liver microsomes, a longer metabolic half-life, and a lower intrinsic clearance rate as compared with the prior art inhibitor Merestinib. Therefore, it is expected to provide a longer-lasting effect in clinical use, and correspondingly can reduce the drug dose required to achieve the same therapeutic strength.

Example 14: Inhibition of the Compounds of the Present Disclosure on Cytochrome P450 Enzyme System

[0514] According to the standard methods conventional in the art for cytochrome P450 enzyme system study, for example those described in Kerns, Edward H. and Di Li (2008). Drug-like Properties: Concepts, Structure Design and Methods: from ADME to Toxicity Optimization. San Diego: Academic Press; Lin, Tong et al., In Vitro Assessment of Cytochrome P450 Inhibition: Strategies for Increasing LC/MS-Based Assay Throughput Using a One-Point IC50 Method and Multiplexing High-Performance Liquid Chromatography; J. Pharm Sci. 2007, 96(9), 2485), the inhibitory effect of the compounds of the present disclosure on the cytochrome P450 enzyme system was investigated analogously as follows.

[0515] The test compounds were evaluated by detecting the metabolism of human liver microsomes for the substrates of different subtypes of Cyp450. The metabolites, substrates and specific inhibitors of each Cyp450 enzyme are shown in Table 10. The liver microsomes (Sekisui XenoTech, LLC., human liver microsomes, cat. #H0610) were slowly melted on ice, and 0.482 mL of the fiver microsome solution (20 mg/mL) was added to 30 mL of 100 mM potassium phosphate solution to prepare a liver microsome working solution with a concentration of 0.316 mg/mL. The test compound and specific inhibitors of different Cyp450 subtypes (as a control) were prepared into 10 mM DMSO solutions, and then 20 μL of the 10 mM DMSO compound solution was added to 180 μL of acetonitrile to obtain a 1 mM compound solution. 2 μL of such 1 mM test compound or inhibitor solution was added to 150 μL of 0.316 mg/mL fiver microsome working solution to obtain a 13.16 μM compound working solution. 45.3 mg of NADPH (SIGMA, cat. #V900362) was weighted and added into 13.59 mL of deionized water to prepare a 4 mM NADPH aqueous solution. 0.2 mL of 2 mg/mL internal standard stock solution (dexamethasone in acetonitrile) was added to 1000 mL of acetonitrile (SIGMA, cat. #271004) to prepare a reaction stop solution and stored at 4° C.

TABLE-US-00020 TABLE 12 Sources of P450 metabolites P450 subtype Metabolites Vendor cat# 1A2 Acetaminophen Sigma PHR1531 2C9 4′-Hydroxydiclofenac Sigma 32412 2C19 4′-Hydroxymephenytoin Sigma H146 2D6 Dextrorphan Sigma PHR1974 3A4 1′-Hydroxymidazolam Caymen 10385 3A4 6′-Hydroxytestosterone Sigma H2898

TABLE-US-00021 TABLE 13 Sources of P450 specific inhibitors P450 subtype specific inhibitors Vendor cat# 1A2 7-ethoxycoumarin Sigma E1379 2C9 Sulfaphenazole Sigma S0758 2C19 Omeprazole Sigma PHR1059 2D6 Promethazine Sigma PHR1467 3A4 Fluconazole Sigma PHRU60 3A4 Ketoconazole Sigma PHR1385

TABLE-US-00022 TABLE 14 P450 substrate sources and substrate solutions (diluted in potassium phosphate solution) Working sol. Final P450 Methanol concentration concentration Subtype Substrate Vendor cat # Storage sol. (at 4mM NADPH) (mg/ml) 1A2 Phenacetin Sigma V900730 10 mM 40 uM 10 uM 2C9 Diclofenac Sigma PHR1144 10 mM 40 uM 10 uM 2C19 S-Mephenytoin Sigma UC175 40 mM 160 uM 40 uM 2D6 Dextromethorphan Sigma D0740000 10 mM 40 uM 10 uM 3A4 Midazolam Tocris 2832 3 mM 20 uM 5 uM 3A4 Testosterone Tocris 2822 50 mM 500 uM 125 uM

TABLE-US-00023 TABLE 15 Human liver microsome solution prepared according to the following table (diluted with potassium phosphate solution) Protein con. of working Final protein con. P450 subtype solution (mg/ml) (mg/ml) 1A2 0.133 0.10 2C9 0.0668 0.05 2C19 0.266 0.20 2D6 0.133 010 3A4 0.0668 0.05 3A4 0.133 0.10

[0516] 150 μL 0.316 mg/mL human liver microsome working solution was added to each well of a 96-well deep plate, and 2 μL of 1 mM specific inhibitor or test compound solution was added to the corresponding wells, and then added to each well 50 μL preheated working solution of each substrate (in 4 mM NADPH solution). After incubating for 5-40 minutes respectively (different enzymes have different incubation times), 200 μL of pre-cooled reaction stop solution was added. All samples were centrifuged at 4000 rpm for 20 minutes. 150 μL of the centrifugal supernatant was then added to a new 96-well deep plate, and added to each well 150 μL of 0.1% formic acid aqueous solution, for analysis by LC/MS/MS (SCI AP4000). The single point IC50 values were calculated according to the following formula.

[00002]${\mathrm{IC}}_{50}=C_0*\frac{100\%-\%\mathrm{inhibition}\mathrm{at}{C}_0}{\%\mathrm{inhibition}\mathrm{at}{C}_0}$

(Assuming Hill slope=1)
C.sub.0=concentration of test compound and inhibitor solution

TABLE-US-00024 TABLE 16 Comparison of the inhibitory activity of Example 12 of the present disclosure and Merestinib on several CYP450 isoenzymes IC50 (μM) P450 Enzyme 1A2 2C9 2C19 2D6 3A4 Control 6.3 0.7 8.9 3.2 7.3 Merestinib >10 4.2 >10 >10 >10 Example 12 >10 9.8 >10 >10 >10 (Control 10 μM, test compound 10 μM)

[0517] Among the tested CYP450 isoenzymes, the protein encoded by the P450 2C9 gene is abundant in human liver microsomes, accounting for about 20% of the total P450. P450 2C9 can metabolize many drugs of different properties, and plays a role in the activation of pre-carcinogen/protoxins and mutagenic agents. The results of this experiment show that, the inhibitory activity of the compounds of the present disclosure, such as Example 12, on 2C9 is significantly lower than that of the prior inhibitor Merestinib. This indicates that the compound of the present disclosure has a smaller effect on P450 2C9 than Merestinib, and thus is expected to have a smaller impact on the metabolism of other co-adminstered drugs, that is, the potential of drug interaction is expected to be significantly reduced.

Example 15: Investigation of Solubility of the Compounds of the Present Disclosure

[0518] According to the well-known standard methods conventional in the art for solubility determination, for example those described in Kerns, Edward H. and Di Li (2008). Drug-like Properties: Concepts, Structure Design and Methods: from ADME to Toxicity Optimization. San Diego: Academic Press), the solubility properties of the compounds of the present disclosure were investigated as follows, in testing systems of FaSSGF (pH 1.6) (Biorelevant, Fasted State Simulated gastric fluids), FeSSIF (pH 5.8) (Biorelevant, Fed State Simulated Intestinal Fluid), FaSSIF (pH 6.5) (Biorelevant, Fasted State Simulated Intestinal Fluid) and PBS buffer (Hyclone, Cat No. SH30256.01B).

[0519] Accurately weighed ˜1 mg of test compound dry powder was charged into a glass bottle, and added an appropriate volume of each of the above test system solutions, to prepare a 4 mg/mL working solution. The sample bottle was placed on a shaker at room temperature (25° C.) and shook at 1000 rpm for 1 hour, then stood overnight for equilibrium. All samples were centrifuged at 12000 rpm at 25° C. for 10 minutes. Solutions of different concentrations for each compound were prepared in DMSO as the standard solutions. 580 μL of 20% (v/v) acetonitrile aqueous solution (containing 40 ng/mL tolbutamide as internal standard) was added into each well of three microplates, and mixed uniformly. 20 μL of 4 mg/mL working solution or standard solution was added to the wells of the first microplate and mixed uniformly, from which 20 μL was taken and added to the corresponding well of the second microplate, and mixed uniformly, from which again 20 μL was taken from each well and added to the corresponding wells of the third microplate, and mixed uniformly. Mass spectrometry was adopted to analyze the samples (LC-MS/MS-014, API4000), a standard curve was developed based on the concentrations of the standard solutions, and the solubility of the test compound was calculated.

TABLE-US-00025 TABLE 17 Comparison of the solubility of Example 12 of the present disclosure and Merestinib Solubility (μg/mL) FaSSGF FeSSEF FaSSIF PBS (pH 1.6) (pH 5.8) (pH 6.5) (pH 7.4) Merestinib 19.28 33.70 3.07 0.02 Example 12 27.66 53.30 5.47 0.01

[0520] The above data show that, the compounds of the present disclosure such as Example 12 show a significantly improved solubility as compared with Merestinib, which could provide better pharmaceutical properties and is expected to have improved bioavailability.