Compounds used as PARP inhibitors

Abstract

The present invention provides poly(ADP-ribose) polymerase (PARP) inhibitors, methods for preparing them, pharmaceutical compositions comprising them, and methods of using them for treating, preventing, and / or ameliorating PARP-mediated diseases or disorders. Specifically, the compounds described herein can be used to treat breast cancer, ovarian cancer, liver cancer, lung cancer, small cell lung cancer, esophageal cancer, gallbladder cancer, pancreatic cancer, and gastric cancer.

Description

[0001] This invention claims the benefit of Indian Patent Application No. 202041018149, filed on April 28, 2020, and Indian Patent Application No. 202041047713, filed on November 2, 2020, the entire contents of each of which are incorporated herein by reference. Technical Field

[0002] The present invention relates to compounds used as poly(ADP-ribose) polymerase (PARP) inhibitors, methods for preparing them, pharmaceutical compositions containing them, and methods for treating, preventing, and / or ameliorating diseases or disorders involving PARP. Background Art

[0003] Poly(ADP-ribose) polymerase (PARP) defines a family of 17 enzymes that cleave NAD+ into nicotinamide and ADP-ribose to form long and branched (ADP-ribose) polymers on the glutamate residues of many target proteins, including PARP itself. The addition of the negatively charged polymer greatly alters the properties and functions of the receptor proteins. Poly(ADP-ribosyl)ation is involved in the regulation of many cellular processes, such as DNA repair, gene transcription, cell cycle progression, cell death, chromatin function, and genomic stability. These functions are mainly attributed to PARP-1, which is considered the most well-characterized member of the PARP family. However, the identification of new genes encoding PARP, as well as the characterization of their structures and subcellular localizations, has revealed different roles of poly(ADP-ribosyl)ation in cells, including telomere replication and cell trafficking.

[0004] Recently, poly(ADP-ribose) binding sites have been found in many DNA damage checkpoint proteins, such as the tumor suppressor p53, the cyclin-dependent kinase inhibitor p21 Cip1 / waf1, DNA damage recognition factors (i.e., nucleotide excision repair xeroderma pigmentosum complementation group A protein and mismatch repair protein MSH6), base excision repair (BER) proteins (i.e., DNA ligase III, X-ray repair cross-complementing protein 1, and XRCC1), DNA-dependent protein kinase (DNA-PK), and regulators of cell death and survival (i.e., NF-κB, inducible nitric oxide synthase, and telomerase). These findings suggest that different components of the PARP family may be involved in the DNA damage signaling network, thereby regulating protein-protein and protein-DNA interactions and, consequently, different types of cellular responses to genotoxic stress. In addition to its involvement in BER and single-strand break (SSB) repair, PARP-1 also appears to contribute to the non-homologous end joining (NHEJ) and homologous recombination (HR) pathways of double-strand break (DSB) repair. See Lucio Tentori et al., Pharmacological Research, Vol. 45, No. 2, 2002, pp. 73-85.

[0005] PARP inhibition may be a useful therapeutic strategy not only for treating BRCA mutations but also for treating a broader range of tumors with multiple defects in the HR pathway. In addition, existing clinical data (e.g., Csaba Szabo et al., British Journal of Pharmacology (2018) 175:192-222) also indicate that stroke, traumatic brain injury, circulatory shock, and acute myocardial infarction are some of the indications in which PARP activation has been shown to contribute to tissue necrosis and inflammatory responses.

[0006] To date, regulatory agencies around the world have approved four PARP inhibitors for human use, namely olaparib, talazoparib, niraparib, and rucaparib.

[0007] Patent documents related to PARP inhibitors include International Publication Nos. WO 2000 / 42040, WO 2001 / 016136, WO 2002 / 036576, WO 2002 / 090334, WO2003 / 093261, WO 2003 / 106430, WO 2004 / 080976, WO2004 / 087713, WO 2005 / 012305, WO 2005 / 012524, WO 2005 / 012305, WO 2005 / 012524, WO2005 / 053662, W02006 / 033003, W02006 / 033007, WO 2006 / 033006, WO 2006 / 021801, WO2006 / 067472, WO 2007 / 144637, WO 2007 / 144639, WO 2007 / 144652, WO 2008 / 047082, WO2008 / 114114, WO 2009 / 050469, WO 2011 / 098971, WO 2015 / 108986, WO 2016 / 028689, WO2016 / 165650, WO 2017 / 153958, WO 2017 / 191562, WO 2017 / 123156, WO 2017 / 140283, WO2018 / 197463, WO 2018 / 038680 and WO 2018 / 108152, and for all purposes, each document is incorporated herein by reference in its entirety.

[0008] There is still an unmet need for new PARP inhibitors for the treatment of various diseases and disorders related to cell proliferation, such as cancer. SUMMARY OF THE INVENTION

[0009] The present invention relates to compounds of formula (I), their pharmaceutically acceptable salts, pharmaceutical compositions containing them, methods for their preparation and methods for treating with them. Specifically, the compounds of formula (I) and their pharmaceutically acceptable salts can be used for treating, preventing and / or ameliorating diseases or disorders involving PARP.

[0010] In one aspect, the present invention relates to a compound of formula (I):

[0011]

[0012] or a tautomer, a prodrug, an N-oxide, a stereoisomer, a pharmaceutically acceptable ester or a pharmaceutically acceptable salt thereof,

[0013] wherein

[0014] Ra 、R b 、R c and R d are each independently selected from hydrogen, halogen, and substituted or unsubstituted C 1-3 alkyl (e.g., C 1-3 haloalkyl);

[0015] X is CR x or N;

[0016] Y is CR y or N;

[0017] Z is CR Z or N;

[0018] R x 、R y and R z may be the same or different and are each independently selected from hydrogen, halogen, and substituted or unsubstituted alkyl;

[0019] G is selected from

[0020]

[0021] wherein

[0022] R e and R f are the same or different and are each independently selected from hydrogen, halogen, hydroxy, substituted or unsubstituted alkyl (e.g., haloalkyl), substituted or unsubstituted alkoxy, and -OR g , or R e and R f both directly bonded to a common atom are capable of connecting to form a C 3-6 cycloalkyl or heterocycle (e.g., 3- to 7-membered heterocycle);

[0023] R g is selected from hydrogen, substituted or unsubstituted C 1-3 alkyl, and -(CO)R h ;

[0024] R h is selected from hydrogen and substituted or unsubstituted alkyl; and

[0025] R 1 、R 2 、R 3 and R 4 are each independently selected from hydrogen, halogen, and substituted or unsubstituted alkyl (e.g., haloalkyl).

[0026] In another aspect, the present invention relates to a compound of formula (I):

[0027]

[0028] or a tautomer thereof, a prodrug thereof, an N-oxide thereof, a stereoisomer thereof, a pharmaceutically acceptable ester thereof or a pharmaceutically acceptable salt thereof,

[0029] wherein

[0030] R a 、R b 、R c and R d are each independently selected from hydrogen, halogen and substituted or unsubstituted C 1-3 alkyl;

[0031] X is CR x or N;

[0032] Y is CR y or N;

[0033] Z is CR Z or N;

[0034] R x 、R y and R z may be the same or different and are each independently selected from hydrogen, halogen and substituted or unsubstituted C 1-3 alkyl;

[0035] G is selected from

[0036]

[0037] wherein

[0038] R e and R f may be the same or different and are each independently selected from hydrogen, halogen, hydroxy, substituted or unsubstituted C 1-3 alkyl (e.g., C 1-3 haloalkyl), substituted or unsubstituted C 1-3 alkoxy and -O(CO)R h ;

[0039] R g is selected from hydrogen, substituted or unsubstituted C 1-3 alkyl and -(CO)R h ;

[0040] R h is selected from hydrogen and substituted or unsubstituted alkyl; and

[0041] R 1 、R 2 、R 3 and R 4Each independently selected from hydrogen, halogen, and substituted or unsubstituted C 1-3 alkyl (e.g., C 1-3 haloalkyl).

[0042] In another aspect, the present invention relates to a compound of formula (IA) or (IB):

[0043]

[0044] or a tautomer thereof, a prodrug thereof, an N-oxide thereof, a stereoisomer thereof, a pharmaceutically acceptable ester thereof, or a pharmaceutically acceptable salt thereof,

[0045] wherein the variables R a 、R b 、R c 、R d 、X、Y、Z、R e 、R f 、R 1 、R 2 、R 3 and R 4 are as defined above for the compounds of formula (I).

[0046] One specific embodiment is a compound of formula (I), (IA), or (IB), wherein any one or more of R a 、R b 、R c and R d is halogen.

[0047] Another embodiment is a compound of formula (I), (IA), or (IB), wherein any one or more of R a 、R b 、R c and R d is hydrogen.

[0048] Another embodiment is a compound of formula (I), (IA), or (IB), wherein R a 、R c and R d are hydrogen and R b is halogen.

[0049] Another embodiment is a compound of formula (I), (IA), or (IB), wherein R b is fluorine or chlorine.

[0050] Another embodiment is a compound of formula (I), (IA), or (IB), wherein R a 、R b and R d are hydrogen and R cis a halogen.

[0051] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R c is fluorine or chlorine.

[0052] Another embodiment is a compound of formula (I), (IA) or (IB), wherein X, Y and Z are each independently CH or N.

[0053] Another embodiment is a compound of formula (I), (IA) or (IB), wherein X is CH or N.

[0054] Another embodiment is a compound of formula (I), (IA) or (IB), wherein Y is CH or N.

[0055] Another embodiment is a compound of formula (I), (IA) or (IB), wherein Z is CH or N.

[0056] Another embodiment is a compound of formula (I), (IA) or (IB), wherein X is N.

[0057] Another embodiment is a compound of formula (I), (IA) or (IB), wherein Y is N.

[0058] Another embodiment is a compound of formula (I), (IA) or (IB), wherein Z is N.

[0059] Another embodiment is a compound of formula (I), (IA) or (IB), wherein Y and Z are CH, and X is N.

[0060] Another embodiment is a compound of formula (I), (IA) or (IB), wherein X and Z are CH, and Y is N.

[0061] Another embodiment is a compound of formula (I), (IA) or (IB), wherein X and Y are CH, and Z is N.

[0062] Another embodiment is a compound of formula (I), (IA) or (IB), wherein Y is CH, and Z is CR Z (wherein R z is as defined above).

[0063] Another embodiment is a compound of formula (I), (IA) or (IB), wherein X and Y are CH, and Z is CR Z , wherein R z is a halogen.

[0064] Another embodiment is a compound of formula (I), (IA) or (IB), wherein X and Y are CH, and Z is CR Z , wherein R z is fluorine.

[0065] Another embodiment is a compound of formula (I), (IA) or (IB), wherein Z is CH.

[0066] Another embodiment is a compound of formula (I), (IA) or (IB), wherein X, Y and Z are CH.

[0067] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R e and R f are each independently selected from hydrogen, hydroxy, substituted or unsubstituted C 1-3 alkyl (e.g., C 1-3 haloalkyl), substituted or unsubstituted C 1-3 alkoxy and acetoxy.

[0068] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R e is hydroxy, substituted or unsubstituted C 1-3 alkoxy or acetoxy.

[0069] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R e is hydroxy.

[0070] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R f is substituted or unsubstituted C 1-3 alkyl (e.g., C 1-3 haloalkyl).

[0071] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R f is substituted or unsubstituted C 1-3 alkyl.

[0072] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R e is hydroxy and R f is substituted or unsubstituted C 1-3 alkyl.

[0073] Another embodiment is a compound of formula (I), (IA) or (IB), wherein

[0074] (i) R e and Rf All are hydrogen;

[0075] (ii) R e and R f are each independently selected from substituted or unsubstituted C 1-3 alkyl; or

[0076] (iii) R e and R f both connected to form a C 3-6 cycloalkyl or heterocycle

[0077] (e.g., 3- to 7-membered heterocycle).

[0078] Another embodiment is a compound of formula (I), (IA) or (IB), wherein

[0079] (i) R 1 , R 2 , R 3 and R 4 are independently selected from hydrogen or halogen;

[0080] (ii) R 1 , R 2 , R 3 and R 4 are hydrogen;

[0081] (iii) R 1 , R 2 , R 3 and R 4 are halogen;

[0082] (iv) R 1 , R 3 and R 4 are hydrogen, and R 2 is halogen; or

[0083] (v) R 1 , R 2 and R 4 are hydrogen, and R 3 is halogen.

[0084] Another embodiment is a compound of formula (I), (IA) or (IB), wherein

[0085] (i) Any one or more of R 1 , R 2 , R 3 and R 4 are fluorine or chlorine;

[0086] (ii) R 1 , R 3 and R4 is hydrogen and R 2 is fluorine; or

[0087] (iii) R 1 , R 2 and R 4 are hydrogen and R 3 is fluorine.

[0088] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R e and R f are hydrogen.

[0089] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R e and R f are methyl.

[0090] Another embodiment is a compound of formula (I), (IA) or (IB), wherein Re and R directly bonded to the common atom f are both connected to form a C 3-6 cycloalkyl ring or a heterocycle (e.g., a 3- to 7-membered heterocycle).

[0091] Another embodiment is a compound of formula (I), (IA) or (IB), wherein any one or more of R 1 , R 2 , R 3 and R 4 are selected from hydrogen and halogen.

[0092] Another embodiment is a compound of formula (I), (IA) or (IB), wherein any one or more of R 1 , R 2 , R 3 and R 4 are hydrogen.

[0093] Another embodiment is a compound of formula (I), (IA) or (IB), wherein any one or more of R 1 , R 2 , R 3 and R 4 are halogen.

[0094] Another embodiment is a compound of formula (I), (IA) or (IB), wherein any one or more of R 1 , R 2 , R 3 and R 4 are fluorine or chlorine.

[0095] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R 1 , R 3 and R 4 are hydrogen and R 2 is halogen.

[0096] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R 1 , R 3 and R 4 are hydrogen and R 2 is fluorine.

[0097] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R 1 , R 2 and R 4 are hydrogen and R 3 is halogen.

[0098] Another embodiment is a compound of formula (I), (IA) or (IB), wherein R 1 , R 2 and R 4 are hydrogen and R 3 is fluorine.

[0099] Another embodiment is a compound of formula (I), wherein G is selected from

[0100]

[0101] Another embodiment is a compound of formula (I), wherein G is selected from

[0102]

[0103] Representative compounds of the invention include those specified below and their pharmaceutically acceptable salts. The invention should not be construed as limited to these specific compounds.

[0104] In one embodiment, the invention relates to a compound selected from the following:

[0105] 1. 4 - ((5 - (3 - hydroxy - 3 - methyl - 2 - oxodihydroindol - 1 - yl)pyridin - 3 - yl)methyl)phthalazin - 1(2H) - one;

[0106] 2. (R)-(+)-4 - ((5 - (3 - hydroxy - 3 - methyl - 2 - oxodihydroindol - 1 - yl)pyridin - 3 - yl)methyl)phthalazin - 1(2H) - one;

[0107] 3. (S)-(-)-4-((5-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0108] 4. 4-(4-Fluoro-3-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one;

[0109] 5. 4-((5-(3-Ethyl-3-hydroxy-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0110] 6. 7-Fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0111] 7. (+)-7-Fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0112] 8. (-)-7-Fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0113] 9. 6-Fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0114] 10. 7-Fluoro-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0115] 11. (+)-7-Fluoro-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0116] 12. (-)-7-Fluoro-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0117] 13. 4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0118] 14. (+)-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0119] 15. (-)-4-((5-(5-Fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0120] 16. 7-Fluoro-4-((5-(6-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0121] 17. 7-Fluoro-4-((5-(6-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0122] 18. 4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one;

[0123] 19. (-)-4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one;

[0124] 20. (+)-4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one;

[0125] 21. 7-Fluoro-4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one;

[0126] 22. (-)-7-Fluoro-4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one;

[0127] 23. (+)-7-Fluoro-4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one;

[0128] 24. 7-Fluoro-4-((2-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one;

[0129] 25. 4-((5-(3-Hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0130] 26. (+)-4-((5-(3-Hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0131] 27. (-)-4-((5-(3-Hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0132] 28. 4-(3-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one;

[0133] 29. 7-Fluoro-4-(4-fluoro-3-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one;

[0134] 30. 4-((5-(3-Methoxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0135] 31. 4-(3-(3-Hydroxy-3-methyl-2-oxo-2,3-dihydro-1H-pyrrolo[2,3-c]pyridin-1-yl)benzyl)phthalazin-1(2H)-one;

[0136] 32. 3-Methyl-2-oxo-1-(5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)pyridin-3-yl)dihydroindol-3-yl acetate;

[0137] 33. 4-(4-Fluoro-3-(2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one;

[0138] 34. 4-(3-(3,3-Dimethyl-2-oxodihydroindol-1-yl)-4-fluorobenzyl)phthalazin-1(2H)-one;

[0139] 35. 4-((5-(3,3-Dimethyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0140] 36. 4-((5-(3,3-Dimethyl-2-oxo-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one;

[0141] 37. 4-(3-(3,3-Dimethyl-2-oxo-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-1-yl)-4-fluorobenzyl)phthalazin-1(2H)-one;

[0142] 38.1'-(5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)pyridin-3-yl)spiro[cyclopropane-1,3'-dihydroindol]-2'-one;

[0143] and its pharmaceutically acceptable salts.

[0144] Table 1

[0145]

[0146]

[0147] Another embodiment of the present invention is a method of inhibiting PARP in a patient (such as a patient in need) by administering an effective amount of at least one compound of the present invention (e.g., a compound of formula (I) as defined above or its pharmaceutically acceptable salt).

[0148] Another embodiment of the present invention is a method of treating an inflammatory, autoimmune or proliferative disease (e.g., by inhibiting PARP) in a patient (such as a patient in need) by administering an effective amount of at least one compound of the present invention.

[0149] In one embodiment, the compound of the present invention inhibits PARP (i.e., an effective amount of the compound is administered to inhibit PARP). In one embodiment, the compound of the present invention inhibits PARP1 and / or PARP2 (i.e., an effective amount of the compound is administered to inhibit PARP1 and / or PARP2).

[0150] Another embodiment of the present invention is a method of treating an inflammatory, autoimmune or proliferative disease (e.g., by inhibiting PARP) in a patient (such as a patient in need) by administering an effective amount of a combination of at least one compound of the present invention and at least one other anti-inflammatory agent, immunomodulator and / or anti-cancer agent (simultaneously or sequentially).

[0151] The compounds of formula (I) and their pharmaceutically acceptable esters and salts can be used for treating, preventing and / or ameliorating diseases or disorders associated with PARP, particularly ameliorating PARP-mediated diseases or disorders, including but not limited to inflammatory diseases or disorders, autoimmune diseases or disorders, and cancer and other proliferative diseases or disorders.

[0152] The compounds of the present invention can be used for treating various cancers, including but not limited to:

[0153] · Malignant tumors, including bladder cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer (including small cell lung cancer), esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, gastric cancer, cervical cancer, thyroid cancer, prostate cancer and skin cancer (including squamous cell carcinoma);

[0154] · Lymphoid hematopoietic tumors, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma, and Burkitt lymphoma;

[0155] · Myeloid hematopoietic tumors, including acute and chronic myelogenous leukemia, myelodysplastic syndrome, and promyelocytic leukemia;

[0156] · Mesenchymal-derived tumors, including fibrosarcoma and rhabdomyosarcoma;

[0157] · Central and peripheral nervous system tumors, including astrocytoma, neuroblastoma, glioma, and schwannoma; and

[0158] · Other tumors, including melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, follicular thyroid carcinoma, and Kaposi's sarcoma.

[0159] The compounds of the present invention as apoptosis regulators can be used for the treatment of cancer (including but not limited to those types mentioned above), viral infections (including but not limited to herpes virus, poxvirus, Epstein-Barr virus, Sindbis virus, and adenovirus), prevention of the development of AIDS in HIV-infected individuals, autoimmune diseases (including but not limited to systemic lupus, lupus erythematosus, autoimmune-mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease, and autoimmune diabetes), neurodegenerative diseases (including but not limited to Alzheimer's disease, AIDS-related dementia, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentosa, spinal muscular atrophy, and cerebellar degeneration), myelodysplastic syndrome, aplastic anemia, ischemic injury associated with myocardial infarction, stroke, and reperfusion injury, arrhythmia, atherosclerosis, toxin-induced or alcohol-related liver diseases, blood diseases (including but not limited to chronic anemia and aplastic anemia), degenerative diseases of the musculoskeletal system (including but not limited to osteoporosis and arthritis), aspirin-sensitive sinusitis, cystic fibrosis, multiple sclerosis, kidney diseases, and cancer pain.

[0160] The compounds of the present invention can regulate the levels of cellular RNA and DNA synthesis. Accordingly, the compounds described herein can be used for the treatment of viral infections (including but not limited to HIV, human papillomavirus, herpes virus, poxvirus, Epstein-Barr virus, Sindbis virus, and adenovirus).

[0161] The compounds of the present invention can be used for the chemoprevention of cancer. Chemoprevention is defined as the inhibition of the development of invasive cancer by blocking initiating mutagenic events or by blocking the progression of premalignant cells that have suffered damage or by inhibiting tumor recurrence. The compounds described herein can also be used to inhibit tumor angiogenesis and metastasis. One embodiment of the present invention is a method for inhibiting tumor angiogenesis or metastasis in a patient (such as a patient in need) by administering an effective amount of one or more compounds of the present invention.

[0162] Another embodiment of the present invention is a method for treating immune system-related diseases (e.g., autoimmune diseases), diseases or disorders involving inflammation (e.g., asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, inflammatory bowel disease, glomerulonephritis, neurodegenerative diseases, multiple sclerosis, uveitis, and immune system disorders), cancer or other proliferative diseases, liver diseases or disorders, or kidney diseases or disorders. The method comprises administering to a patient (such as a patient in need) an effective amount of one or more compounds of the present invention.

[0163] Examples of immune disorders include, but are not limited to, psoriasis, rheumatoid arthritis, vasculitis, inflammatory bowel disease, dermatitis, osteoarthritis, asthma, inflammatory muscle diseases, allergic diseases (e.g., allergic rhinitis), vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, allograft or xenograft (organ, bone marrow, stem cell, and other cell and tissue) transplant rejection, graft-versus-host disease, lupus erythematosus, inflammatory diseases, type I diabetes, pulmonary fibrosis, dermatomyositis, Sjogren's syndrome, thyroiditis (e.g., Hashimoto's and autoimmune thyroiditis), myasthenia gravis, autoimmune hemolytic anemia, multiple sclerosis, cystic fibrosis, chronic recurrent hepatitis, primary biliary cirrhosis, allergic conjunctivitis, and atopic dermatitis.

[0164] In one embodiment, the compounds described herein are used as immunosuppressants to prevent transplant rejection, allograft or xenograft rejection (organ, bone marrow, stem cell, other cell and tissue), and graft-versus-host disease. In other embodiments, transplant rejection is caused by tissue or organ transplantation. In further embodiments, graft-versus-host disease is caused by bone marrow or stem cell transplantation. One embodiment is a method for preventing or reducing the risk of transplant rejection, allograft or xenograft rejection (organ, bone marrow, stem cell, other cell and tissue), or graft-versus-host disease by administering to a patient (such as a patient in need) an effective amount of one or more compounds of the present invention.

[0165] The compounds of the present invention can also be used in combination with the following (administered together or sequentially): known anti-cancer treatments such as, but not limited to, radiotherapy; or cytostatic agents, cytotoxic agents or anti-cancer agents such as, but not limited to, DNA-interacting agents such as cisplatin or doxorubicin; topoisomerase II inhibitors such as etoposide; topoisomerase I inhibitors such as CPT-11 or topotecan; naturally occurring or synthetic tubulin-interacting agents such as paclitaxel, docetaxel or epothilones (e.g., ixabepilone); hormonal agents such as tamoxifen; thymidylate synthase inhibitors such as 5-fluorouracil; and antimetabolites such as methotrexate, other tyrosine kinase inhibitors such as Iressa and OSI-774; angiogenesis inhibitors; EGF inhibitors; VEGF inhibitors; CDK inhibitors; HDAC inhibitors, SRC inhibitors; c-Kit inhibitors; Her1 / 2 inhibitors and monoclonal antibodies against growth factor receptors such as Erbitux (EGF) and Herceptin (Her2) and other protein kinase modulators, or any combination of the foregoing.

[0166] The compounds of the present invention can also be used in combination with one or more steroidal anti-inflammatory drugs, non-steroidal anti-inflammatory drugs (NSAIDs) or immunoselective anti-inflammatory derivatives (ImSAIDs) (administered together or sequentially).

[0167] In another aspect, the present invention also provides a pharmaceutical composition comprising one or more compounds of the present invention (such as a compound of formula (I) or a pharmaceutically acceptable salt thereof) and one or more pharmaceutically acceptable carriers. The pharmaceutical composition may also comprise one or more other active ingredients identified herein, such as other anti-cancer agents.

[0168] In one embodiment, the pharmaceutical composition comprises a therapeutically effective amount of one or more compounds of formula (I) or a pharmaceutically acceptable salt thereof.

[0169] Another embodiment is a method of treating cancer in a patient, such as a patient in need thereof, by administering a therapeutically effective amount of a compound of the present invention. For example, the compounds of the present invention can be effective in treating lymphoid hematopoietic tumors, leukemia, acute lymphoblastic leukemia, acute lymphocytic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma, and Burkitt lymphoma; myeloid hematopoietic tumors, acute myeloid leukemia, chronic myeloid leukemia, myelodysplastic syndrome, and promyelocytic leukemia. The compounds of the present invention can also be effective in treating bladder cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer, small cell lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, gastric cancer, cervical cancer, thyroid cancer, prostate cancer, skin cancer, squamous cell carcinoma, tumors of mesenchymal origin, fibrosarcoma, rhabdomyosarcoma, central and peripheral nervous system tumors, astrocytoma, neuroblastoma, glioma, schwannoma, melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, follicular thyroid cancer, and Kaposi's sarcoma.

[0170] Another embodiment is a method of treating leukemia in a patient, such as a patient in need thereof, by administering a therapeutically effective amount of a compound of the present invention. For example, the compounds of the present invention can be effective in treating breast cancer, ovarian cancer, liver cancer, lung cancer, small cell lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, or gastric cancer. Detailed Description

[0171] As used herein, unless otherwise specified, the following definitions shall apply. In addition, many of the groups defined herein can be optionally substituted. The substituents listed in the definitions are exemplary and should not be construed as limiting the substituents defined elsewhere in this specification.

[0172] Unless otherwise indicated, the term "alkyl" refers to a straight-chain or branched-chain hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing no unsaturation, having from one to eight carbon atoms, and attached to the remainder of the molecule by a single bond such as methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, and 1,1-dimethylethyl (tert-butyl). The term "C 1-6 alkyl" refers to an alkyl as defined above having up to 6 carbon atoms. The term "C 1-4 alkyl" refers to an alkyl as defined above having up to 4 carbon atoms. Where appropriate, the term "alkyl" refers to a divalent hydrocarbon chain group as described above.

[0173] Unless otherwise specified, the term "alkenyl" refers to an aliphatic hydrocarbon group containing a carbon-carbon double bond and can be a straight-chain, branched-chain or branched-chain group having from about 2 to about 10 carbon atoms, such as vinyl, 1-propenyl, 2-propenyl (allyl), isopropenyl, 2-methyl-1-propenyl, 1-butenyl and 2-butenyl. The term "(C 2-6 ) alkenyl" refers to an alkenyl as defined above having at most 6 carbon atoms.

[0174] Unless otherwise specified, the term "alkynyl" refers to a straight-chain or branched-chain hydrocarbon group having at least one carbon-carbon triple bond and having from 2 to a maximum of 12 carbon atoms (currently preferably a group having from 2 to a maximum of 10 carbon atoms), such as ethynyl, propynyl and butynyl. The term "(C 2-6 ) alkynyl" refers to an alkynyl as defined above having at most 6 carbon atoms.

[0175] Unless otherwise specified, the term "alkoxy" means an alkyl, cycloalkyl or cycloalkylalkyl as defined above connected to the remainder of the molecule by an oxygen bond. The term "substituted alkoxy" refers to an alkoxy in which the alkyl moiety is substituted (i.e., -O-(substituted alkyl), where the term "substituted alkyl" is the same as defined above for "alkyl"). For example, "alkoxy" refers to the group -O-alkyl, including straight-chain, branched-chain, cyclic configurations of 1 to 8 carbon atoms connected to the parent structure by oxygen and combinations thereof. Examples include methoxy, ethoxy, propoxy, isopropoxy, cyclopropoxy and cyclohexyloxy.

[0176] Unless otherwise specified, the term "cycloalkyl" refers to a non-aromatic monocyclic or polycyclic ring system of about 3 to 12 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. Examples of polycyclic cycloalkyls include perhydronaphthyl, adamantyl and norbornyl, bridged ring groups and spirobicyclic groups, such as spiro(4,4)non-2-yl. The term "(C 3-8 ) cycloalkyl" refers to a cycloalkyl as defined above having at most 8 carbon atoms.

[0177] Unless otherwise specified, the term "cycloalkylalkyl" refers to a ring-containing group having from about 3 to 8 carbon atoms, these carbon atoms being directly connected to an alkyl group and then connected to the main structure at any carbon from the alkyl group that results in a stable structure, such as cyclopropylmethyl, cyclobutylethyl and cyclopentylethyl.

[0178] Unless otherwise specified, the term "cycloalkenyl" refers to a ring-containing group having from about 3 to a maximum of 8 carbon atoms and having at least one carbon-carbon double bond, such as cyclopropenyl, cyclobutenyl and cyclopentenyl. The term "cycloalkenylalkyl" means that the cycloalkenyl is directly connected to an alkyl group and then connected to the main structure at any carbon of the alkyl group, resulting in a stable structure.

[0179] Unless otherwise specified, the term "aryl" refers to an aromatic group having 6 to 20 carbon atoms, such as phenyl, naphthyl, tetrahydronaphthyl, indanyl, and biphenyl.

[0180] Unless otherwise specified, the term "arylalkyl" refers to an aryl as defined above directly bonded to an alkyl as defined above, such as -CH2C6H5 and -C2H5C6H5.

[0181] Unless otherwise specified, the term "heterocycle" refers to a non-aromatic 3- to 15-membered ring group composed of carbon atoms and at least one heteroatom selected from nitrogen, phosphorus, oxygen, and sulfur. For the purposes of this invention, the heterocyclic group can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which can include fused rings, bridged rings, or spiro ring systems, and the nitrogen, phosphorus, carbon, oxygen, or sulfur atoms in the heterocyclic group can optionally be oxidized to various oxidation states. In addition, the nitrogen atom can optionally be quaternized. The heterocyclic ring group can be attached to the main structure at any heteroatom or carbon atom, thus producing a stable structure.

[0182] Unless otherwise specified, the term "heterocyclic group" refers to a heterocyclic group as defined above. The heterocyclic ring group can be attached to the main structure at any heteroatom or carbon atom, thus producing a stable structure.

[0183] Unless otherwise specified, the term "heterocyclic alkyl" refers to a heterocyclic ring group as defined above directly bonded to an alkyl. The heterocyclic alkyl can be attached to the main structure at a carbon atom in the alkyl, thus producing a stable structure. Examples of such heterocyclic alkyls include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidinyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thioxomorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl.

[0184] Unless otherwise indicated, the term "heteroaryl" refers to an optionally substituted 5- to 14-membered aromatic ring having one or more heteroatoms selected from N, O, and S as ring atoms. The heteroaryl can be a monocyclic, bicyclic, or tricyclic ring system. Examples of such "heterocyclic" or "heteroaryl" groups include, but are not limited to, oxazolyl, thiazolyl, imidazolyl, pyrrolyl, furyl, pyridyl, pyrimidinyl, pyrazinyl, benzofuryl, indolyl, benzothiazolyl, benzoxazolyl, carbazolyl, quinolinyl, isoquinolinyl, azetidinyl, acridinyl, benzodioxolyl, benzodioxanyl, benzofuryl, carbazolyl, cinnolinyl, dioxolanyl, indazolyl, naphthyridinyl, perhydroazepinyl groups, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, quinazolinyl, quinoxalinyl, tetrazolyl, tetrahydroisoquinolinyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl group, azepinyl group, 4-piperidone, pyrrolidinyl, pyridazinyl, oxazolinyl, oxazolidinyl, triazolyl, indanyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiazolinyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, isoindolyl, dihydroindolyl, isoindolinyl, octahydroindolyl, octahydroisoindolyl, decahydroisoquinolinyl, benzimidazolyl, thiadiazolyl, benzopyranyl, tetrahydrofuranyl, tetrahydropyranyl, thienyl, benzothienyl, thiomorpholinyl, thiomorpholine sulfoxide, thiomorpholine sulfone, dioxaphospholanyl, oxadiazolyl, chromanyl and isochromanyl. The heteroaryl ring group may be attached to the main structure at any heteroatom or carbon atom to give a stable structure. The term "substituted heteroaryl" also includes ring systems substituted with one or more oxide (-O-) substituents such as pyridyl N-oxide.

[0185] Unless otherwise stated, the term "heteroarylalkyl" refers to a heteroaryl ring group as defined above directly bonded to an alkyl group. The heteroarylalkyl may be attached to the main structure at any carbon atom from the alkyl group to give a stable structure.

[0186] The term "ring" refers to a ring containing 3 to 10 carbon atoms.

[0187] Unless otherwise specified, the term "substituted" means substituted with any one or any combination of the following substituents, which may be the same or different and are independently selected from hydrogen, hydroxy, halogen, carboxy, cyano, nitro, oxo(=O), thioxo(=S), substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted cycloalkenylalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted heterocycle, substituted heterocyclic alkyl ring, substituted or unsubstituted guanidine, -COOR t , -C(O)R v , -C(S)R V , -C(O)NR t R u , -C(O)ONR t R u , -NR t R u , -NRt CONR u R v 、 -N(R t )SOR u 、 -N(R t )SO2R u 、 -(=N - N(R t )R u )、 -NR t C(O)OR u 、 -NR t R u 、 -NR t C(O)R u -、 -NR t C(S)R u -NR t C(S)NR t R u 、 -SONR t R u -、 -SO2NR t R u -、 -OR t 、 -OR t C(O)NR u R v 、 -OR t C(O)OR u -、 -OC(O)R t 、 -OC(O)NR t R u 、 -R t NR u C(O)R v 、 -R t OR u 、 -R t C(O)OR u 、 -R t C(O)NR u R v 、 -R t C(O)R u 、 -R t OC(O)R u 、 -SR t 、 -SOR t 、 -SO2R t and -ONO2, wherein R in each of the above groups t 、 R u and R vmay be hydrogen, a substituted or unsubstituted alkyl, a substituted or unsubstituted alkoxy, a substituted or unsubstituted alkenyl, a substituted or unsubstituted alkynyl, a substituted or unsubstituted aryl, a substituted or unsubstituted arylalkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted cycloalkylalkyl, a substituted or unsubstituted cycloalkenyl, a substituted or unsubstituted amino, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted heteroarylalkyl, a substituted or unsubstituted heterocycle, a substituted heterocyclic alkyl ring, or R t 、R u and R v any two of and may be joined to form a substituted or unsubstituted saturated or unsaturated 3- to 10-membered ring, which may optionally contain heteroatoms which may be the same or different and are selected from O, NR q (wherein R q may be hydrogen or C 1-6 alkyl) or S. The combinations of substituents envisaged by the present invention are preferably those which result in the formation of stable or chemically viable compounds. As used herein, the term "stable" refers to a compound or structure which does not substantially change when subjected to conditions which allow its generation, detection and preferably its recovery, purification and incorporation into a pharmaceutical composition. The substituents on the above "substituted" groups cannot be further substituted. For example, when the substituent on "substituted alkyl" is "substituted aryl", the substituent on "substituted aryl" cannot be "substituted alkenyl".

[0188] The terms "halo", "halide" or "halogen" refer to fluorine, chlorine, bromine or iodine. The terms "haloalkyl", "haloalkenyl", "haloalkynyl" and "haloalkoxy" include alkyl, alkenyl, alkynyl and alkoxy structures substituted with one or more halogen groups or combinations thereof. For example, the terms "fluoroalkyl" and "fluoroalkoxy" include haloalkyl and haloalkoxy, respectively, wherein the halogen is fluorine.

[0189] The term "protecting group" or "PG" refers to a substituent used to block or protect a specific functional group. Other functional groups on the compound can remain reactive. For example, an "amino protecting group" is a substituent attached to an amino group that blocks or protects the amino functional group in a compound. Suitable amino protecting groups include, but are not limited to, acetyl, trifluoroacetyl, tert-butoxycarbonyl (BOC), benzyloxycarbonyl (CBz), and 9-fluorenylmethyloxycarbonyl (Fmoc). Similarly, a "hydroxy protecting group" refers to a substituent of a hydroxyl group that blocks or protects the hydroxyl functional group. Suitable hydroxy protecting groups include, but are not limited to, acetyl and silyl. A "carboxyl protecting group" refers to a substituent of a carboxyl group that blocks or protects the carboxyl functional group. Suitable carboxyl protecting groups include, but are not limited to, -CH2CH2SO2Ph, cyanoethyl, 2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl, 2-(p-nitrophenylsulfinyl)ethyl, 2-(diphenylphosphino)-ethyl, and nitroethyl. For a general description of protecting groups and their use, see T.W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

[0190] Certain compounds described herein can contain one or more asymmetric centers and can thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined as (R)- or (S)- according to absolute stereochemistry. The chemical entities, pharmaceutical compositions, and methods of the present invention are intended to include all such possible isomers, including racemic mixtures, optically pure forms, and mixtures of intermediates. Non-limiting examples of mixtures of intermediates include, for example, isomer mixtures in a ratio of 10:90, 13:87, 17:83, 20:80, or 22:78. Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When the compounds described herein contain an olefinic double bond or other geometrically asymmetric center, unless otherwise specified, the compounds are intended to include E and Z geometric isomers.

[0191] As used herein, the term "tautomer" refers to a compound characterized by relatively facile interconversion of isomeric forms under equilibrium conditions. These isomers are intended to be encompassed by the present invention. "Tautomers" are structurally distinct isomers that interconvert by tautomerization. "Tautomerization" is a form of isomerization and includes prototropic or prototropic tautomerization, which is considered a subset of acid-base chemistry. "Prototropic tautomerization" or "prototropic tautomerization" involves proton migration accompanied by a change in bond order, typically the interchange of a single bond with an adjacent double bond. Where tautomerization is possible (e.g., in solution), a chemical equilibrium of tautomers can be achieved. An example of tautomerization is keto-enol tautomerization. A specific example of keto-enol tautomerization is the interconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerization is phenol-keto tautomerization. A specific example of phenol-keto tautomerization is the interconversion of pyridin-4-ol and pyridin-4(1H)-one tautomers.

[0192] A "leaving group or atom" is any group or atom that will cleave from the starting material under the reaction conditions, thereby facilitating the reaction at the designated site. Suitable examples of these groups are halogen atoms and mesyloxy, p-nitrobenzenesulfonyloxy, and tosyloxy, unless otherwise specified.

[0193] The term "prodrug" refers to a compound that is an inactive precursor of a compound and is converted in vivo to its active form through normal metabolic processes. Prodrug design is discussed generally in Hardman et al. (eds.), Goodman and Gilman, The Pharmacological Basis of Therapeutics, 9th ed., pp. 11-16 (1996). In-depth discussions are provided in Higuchi et al., Prodrugs as Novel Delivery Systems, Vol. 14, ASCD Symposium Series and Roche (ed.), Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press (1987). By way of illustration, a prodrug can be converted to a pharmacologically active form by hydrolysis of, for example, an ester or amide bond, thereby introducing or exposing a functional group on the resulting product. Prodrugs can be designed to react with endogenous compounds to form water-soluble conjugates that further enhance the pharmacological properties of the compound, such as increasing the circulating half-life. Alternatively, a prodrug can be designed to covalently modify a functional group with, for example, glucuronic acid, sulfate, glutathione, an amino acid, or acetate. The resulting conjugate can be inactivated and excreted in the urine, or made more effective than the parent compound. High molecular weight conjugates can also be excreted into the bile, undergo enzymatic cleavage, and be released back into the circulation, thereby effectively increasing the biological half-life of the initially administered compound.

[0194] The term "ester" refers to a compound formed by the reaction between an acid and an alcohol with the removal of water. Esters can be represented by the general formula RCOOR'.

[0195] These prodrugs and esters are intended to be encompassed within the scope of the present invention.

[0196] In addition, the present invention also includes compounds that differ only in the presence of one or more isotopically enriched atoms, such as the replacement of hydrogen with deuterium or tritium, or the replacement of carbon with enriched 13C or 14C carbon.

[0197] The compounds of the present invention can also contain unnatural proportions of atomic isotopes at one or more atoms that make up such compounds. For example, the compounds can be radiolabeled with a radioactive isotope such as tritium ( 3 3H), iodine-125 ( 125 125I), or carbon-14 ( 14 14C). All isotopic variants of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

[0198] Pharmaceutically acceptable salts forming part of the present invention include, for example, salts derived from inorganic bases such as Li, Na, K, Ca, Mg, Fe, Cu, Zn, and Mn; salts of organic bases such as N,N'-diacetyldiethylenetriamine, glucosamine, triethylamine, choline, hydroxide, dicyclohexylamine, metformin, benzylamine, trialkylamine, and thiamine; chiral bases such as alkyl aniline, glycinol, and phenylglycinol; salts of natural amino acids such as glycine, alanine, valine, leucine, isoleucine, norleucine, tyrosine, cystine, cysteine, methionine, proline, hydroxyproline, histidine, ornithine, lysine, arginine, and serine; quaternary ammonium salts of the compounds of the present invention with alkyl halides, alkyl sulfates such as MeI and (Me)2SO4; unnatural amino acids such as D-isomers or substituted amino acids; guanidine; and substituted guanidines, wherein the substituents are selected from nitro, amino, alkyl, alkenyl, alkynyl, ammonium, or substituted ammonium salts, and aluminum salts. The salts may include, where appropriate, acid addition salts, which are sulfate, nitrate, phosphate, perchlorate, borate, hydrohalide (e.g., hydrochloride), acetate, tartrate, maleate, citrate, fumarate, succinate, palmitate, mesylate, benzoate, salicylate, benzenesulfonate, ascorbate, glycerophosphate, and ketoglutarate.

[0199] When ranges are used herein for physical properties (such as molecular weight) or chemical properties (such as chemical formula), all combinations and subcombinations of the ranges and specific embodiments therein are intended to be included. When referring to a numerical value or a range of numerical values, the term "about" means that the numerical value or range of numerical values is an approximation within experimental variability (or within statistical experimental error), so that the numerical value or range of numerical values can vary, for example, from 1% to 15% of the stated numerical value or range. The term "comprising" (and related terms such as "containing" or "having" or "including") includes those embodiments that "consist of the recited features" or "consist essentially of the recited features", for example, embodiments of any composition of matter, composition, method, or process, etc.

[0200] The following abbreviations and terms have the indicated meanings throughout: AIDS = acquired immunodeficiency syndrome; HIV = human immunodeficiency virus; the abbreviations used herein have their conventional meanings in the fields of chemistry and biology.

[0201] The term "cell proliferation" refers to the phenomenon of a change in the number of cells due to division. The term also includes cell growth, by which the cell morphology changes (e.g., increases in size) in accordance with the proliferation signal.

[0202] As used herein, the terms "co-administer", "administer in combination with", and their grammatical equivalents encompass the administration of two or more agents to an animal such that the two agents and / or their metabolites are present in the animal simultaneously. Co-administration includes simultaneous administration in separate compositions, administration in separate compositions at different times, or administration in a composition in which both agents are present.

[0203] The term "effective amount" or "therapeutically effective amount" means an amount of a compound as described herein sufficient to represent the intended application, including but not limited to the treatment of diseases as defined below. The therapeutically effective amount can vary according to the intended application (in vitro or in vivo) or the subject and disease condition being treated, such as the weight and age of the subject, the severity of the disease condition, the mode of administration, etc., which can be readily determined by one of ordinary skill in the art. The term also applies to doses that will elicit a specific response in a target cell, such as reducing platelet adhesion and / or cell migration. The specific dose will vary according to the selected compound, the dosing regimen to be followed, whether administered in combination with other compounds, the time of administration, the tissue to which it is administered, and the physical delivery system carrying it. In one embodiment, the amount of the compound administered is in the range of about 0.1 mg to 5 g, about 1 mg to 2.0 g, about 100 mg to 1.5 g, about 200 mg to 1.5 g, about 400 mg to 1.5 g, and about 400 mg to 1.0 g.

[0204] As used herein, the terms "treat" or "improve" are used interchangeably. These terms refer to methods of obtaining a beneficial or desired result, including but not limited to therapeutic benefits and / or prophylactic benefits. A so-called therapeutic benefit means eradicating or ameliorating the underlying disease being treated. Additionally, a therapeutic benefit is achieved by eradicating or ameliorating one or more of the physiological symptoms associated with the underlying disease such that an improvement is observed in the patient, even though the patient may still be suffering from the underlying disease. For a prophylactic benefit, the composition can be administered to a patient at risk of developing a particular disease or to a patient who has reported one or more physiological symptoms of a disease, even if a diagnosis of such disease has not yet been made.

[0205] The term "therapeutic effect" as used herein includes the therapeutic and / or prophylactic benefits as described above. Prophylactic effects include delaying or eliminating the onset of a disease or disorder, delaying or eliminating the onset of symptoms of a disease or disorder, slowing, halting, or reversing the progression of a disease or disorder, or any combination thereof.

[0206] The term "subject" or "patient" refers to an animal, such as a mammal, for example a human. The methods described herein can be used for human therapy and veterinary applications (e.g., dogs, cats, cows, sheep, pigs, horses, goats, chickens, turkeys, ducks, and geese).

[0207] In some embodiments, the patient is a mammal, and in some embodiments, the patient is a human.

[0208] As used herein, "radiotherapy" refers to exposing a patient to a radiation emitter using conventional methods and compositions known to a practitioner, such as radionuclides that emit alpha particles (e.g., actinium and thorium radionuclides), low linear energy transfer (LET) radiation emitters (i.e., beta emitters), conversion electron emitters (e.g., strontium-89 and samarium-153-EDTMP), or high-energy radiation, including but not limited to x-rays, gamma rays, and neutrons.

[0209] As used herein, the term "pharmaceutically acceptable excipient" includes, but is not limited to, any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, one or more suitable diluents, fillers, salts, disintegrants, binders, lubricants, glidants, wetting agents, controlled release matrices, coloring / flavoring agents, carriers, buffers, stabilizers, solubilizers, and combinations thereof. Unless any conventional medium or agent is incompatible with the active ingredient, its use in the therapeutic compositions of the present invention is contemplated. Supplementary active ingredients can also be incorporated into the compositions.

[0210] Any method of the present invention described herein can be applied to cell populations in vivo or ex vivo. "In vivo" refers to within a living individual, such as within an animal or a human or within a subject. In this regard, the methods of the present invention can be used for the treatment or prevention of an individual. "Ex vivo" or "in vitro" refers to outside of a living individual. Examples of ex vivo cell populations include in vitro cell cultures and biological samples, including but not limited to fluid or tissue samples obtained from an individual. Such samples can be obtained by methods known in the art. Exemplary biological fluid samples include blood, cerebrospinal fluid, urine, and saliva. Exemplary tissue samples include tumors and their biopsies. In this regard, the present invention can be used for a variety of purposes, including therapeutic and experimental purposes. For example, the present invention can be used ex vivo or in vitro to determine the optimal administration regimen and / or dosage of a PARP inhibitor for a given indication, cell type, individual, and other parameters. Information collected from such uses can be used for experimental or diagnostic purposes, or for clinical use to set a regimen for in vivo treatment. Other ex vivo uses to which the present invention may be applicable are described below or will be apparent to those skilled in the art.

[0211] Drug composition

[0212] The present invention provides a pharmaceutical composition comprising one or more compounds of the present invention. The pharmaceutical composition can comprise one or more additional active ingredients, as described herein. The pharmaceutical composition can be administered for any disorder described herein.

[0213] The subject pharmaceutical compositions are generally formulated to provide a therapeutically effective amount of the compounds of the invention as the active ingredient. If desired, the pharmaceutical compositions contain the compounds of the invention as the active ingredient and one or more pharmaceutically acceptable carriers or excipients, such as inert solid diluents and fillers, diluents (including sterile aqueous solutions and various organic solvents), penetration enhancers, solubilizers, and adjuvants.

[0214] The pharmaceutical compositions can be administered alone or in combination with one or more other agents, which are also generally administered in the form of pharmaceutical compositions. When needed, the subject compounds and the other agents can be combined into one formulation or the two components can be formulated into separate formulations for using them alone or in combination simultaneously.

[0215] The methods include administering the compounds of the invention alone or in combination as described herein, and in each case optionally including one or more suitable diluents, fillers, salts, disintegrants, binders, lubricants, glidants, wetting agents, controlled release matrices, coloring / flavoring agents, carriers, excipients, buffers, stabilizers, solubilizers, and combinations thereof.

[0216] Formulations of various pharmaceutical compositions are known in the art. See, for example, Anderson, Philip O.; Knoben, James E.; Troutman, William G editors, Handbook of Clinical Drug Data, 10th Edition, McGraw-Hill, 2002; Pratt and Taylor editors, Principles of Drug Action, 3rd Edition, Churchill Livingston, New York, 1990; Katzung editor, Basic and Clinical Pharmacology, 9th Edition, McGraw Hill, 2003; Goodman and Gilman editors, The Pharmacological Basis of Therapeutics, 10th Edition, McGraw Hill, 2001; Remingtons Pharmaceutical Sciences, 20th Edition, Lippincott Williams & Wilkins., 2000; Martindale, The Extra Pharmacopoeia, 32nd Edition (The Pharmaceutical Press, London, 1999), all of which are hereby incorporated by reference in their entireties.

[0217] The compounds or pharmaceutical compositions of the present invention can be administered by any route capable of delivering the compound to the site of action, such as oral route, intraduodenal route, parenteral injection (including intravenous, intraarterial, subcutaneous, intramuscular, intracascular, intraperitoneal or infusion), topical administration (e.g., transdermal administration), rectal administration, local delivery through a catheter or stent or by inhalation. The compound can also be administered intralipally or intrathecally.

[0218] The composition can be administered in solid, semi-solid, liquid or gaseous form, or can be in dry powder form, such as lyophilized form. The pharmaceutical composition can be packaged in a manner convenient for delivery, including for example solid dosage forms such as capsules, sachets, cachets, gelatin, paper, tablets, capsules, suppositories, pills, boluses, lozenges and troches. The type of packaging will generally depend on the desired route of administration. Implantable sustained release formulations are also contemplated, as are transdermal formulations.

[0219] Therapeutic method

[0220] The present invention also provides methods of using the compounds or pharmaceutical compositions of the present invention to treat disease conditions, including but not limited to diseases associated with and / or caused by PARP overexpression.

[0221] The treatment methods provided herein include administering a therapeutically effective amount of the compound of the present invention to a subject (such as a subject in need). In one embodiment, the present invention provides a method of treating an inflammatory disease, including an autoimmune disease, in a mammal. The method includes administering a therapeutically effective amount of the compound of the present invention to the mammal.

[0222] It should be understood that the treatment methods of the present invention described herein can be used in the fields of human medicine and veterinary medicine. Thus, the individual to be treated can be a mammal, preferably a human or other animal. For veterinary purposes, the individuals include but are not limited to farm animals, including cows, sheep, pigs, horses and goats; companion animals, such as dogs and cats; wild animals and / or zoo animals; laboratory animals, including mice, rats, rabbits, guinea pigs and hamsters; and poultry, such as chickens, turkeys, ducks and geese.

[0223] The present invention also relates to a method for treating a hyperproliferative disorder in a mammal, the method comprising administering to the mammal a therapeutically effective amount of a compound of the present invention. In some embodiments, the method relates to treating cancer, such as acute myeloid leukemia, thymic carcinoma, brain cancer, lung cancer, squamous cell carcinoma, skin cancer, eye cancer, retinoblastoma, intraocular melanoma, oral and oropharyngeal cancer, bladder cancer, gastric cancer, gastric cancer, pancreatic cancer, bladder cancer, breast cancer, cervical cancer, head cancer, neck cancer, kidney cancer, kidney cancer, liver cancer, ovarian cancer, prostate cancer, colorectal cancer, esophageal cancer, testicular cancer, gynecological cancer, thyroid cancer, CNS, PNS, AIDS-related cancers (such as lymphoma and Kaposi's sarcoma) or virus-induced cancers. In some embodiments, the method relates to treating non-cancerous hyperproliferative disorders, such as benign skin hyperplasia (e.g., psoriasis), restenosis or benign hyperplasia of the prostate (e.g., benign prostatic hyperplasia (BPH)).

[0224] The present invention also relates to a method for treating a disease in a mammal that is related to angiogenesis or vasculogenesis, the method comprising administering to the mammal a therapeutically effective amount of a compound of the present invention. In some embodiments, the method is used to treat a disease selected from the group consisting of: tumor angiogenesis, chronic inflammatory diseases such as rheumatoid arthritis, atherosclerosis, inflammatory bowel disease, skin diseases such as psoriasis, eczema and scleroderma, diabetes, diabetic retinopathy, retinopathy of prematurity, age-related macular degeneration, hemangioma, glioma, melanoma, Kaposi's sarcoma and ovarian cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, colon cancer and epidermoid carcinoma.

[0225] Patients who can be treated with the compounds of the invention according to the methods of the invention include, for example, patients who have been diagnosed with the following diseases: psoriasis; restenosis; atherosclerosis; BPH; breast cancer, such as ductal carcinoma, medullary carcinoma, colloid carcinoma, tubular carcinoma, and inflammatory breast cancer in breast duct tissue; ovarian cancer, including epithelial ovarian tumors, such as ovarian adenocarcinoma and adenocarcinoma that has migrated from the ovary to the abdominal cavity; uterine cancer; cervical cancer, such as adenocarcinoma in the epithelium of the cervix, including squamous cell carcinoma and adenocarcinoma; prostate cancer, such as prostate cancer selected from the following: adenocarcinoma or adenocarcinoma that has migrated to the bone; pancreatic cancer, such as epithelial carcinoma in pancreatic duct tissue and adenocarcinoma in the pancreatic duct; bladder cancer, such as transitional cell carcinoma, urothelial carcinoma ( leukemias, such as acute myeloid leukemia (AML), acute lymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, myelodysplasia, myeloproliferative disorders, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), mastocytosis, chronic lymphocytic leukemia (CLL), multiple myeloma (MM) and myelodysplastic syndrome (MDS); bone cancer; lung cancer, such as non-small cell lung cancer (NSCLC), which is divided into squamous cell carcinoma, adenocarcinoma and large cell undifferentiated carcinoma and small cell lung cancer; skin cancer, such as basal cell carcinoma, melanoma, squamous cell carcinoma, and actinic keratosis, a skin condition that sometimes develops into squamous cell carcinoma; retinoblastoma of the eye; skin or intraocular (eye) melanoma; primary liver cancer (cancer that starts in the liver); kidney cancer; thyroid cancers, such as papillary, follicular, medullary, and anaplastic thyroid cancers; AIDS-related lymphomas, such as diffuse large B-cell lymphoma, B-cell immunoblastic lymphoma, and small non-lytic cell lymphoma; Kaposi's sarcoma; virus-induced cancers, including hepatitis B virus (HBV), hepatitis C virus (HCV), and hepatocellular carcinoma; human lymphotropic virus type 1 (HTLV-I) and adult T-cell leukemia / lymphoma; and human papillary HPV and cervical cancer; central nervous system cancers (CNS), such as primary brain tumors, including gliomas (astrocytomas, anaplastic astrocytomas, or glioblastoma multiforme), oligodendrogliomas, ependymomas, meningiomas, lymphomas, schwannomas, and medulloblastomas; peripheral nervous system (PNS) cancers, such as acoustic neuromas and malignant peripheral nerve sheath tumors (MPNSTs), including neurofibromas and schwannomas, malignant fibrous cell tumors, malignant fibrous histiocytomas, malignant meningiomas, malignant mesotheliomas, and malignant mixed Müllerian tumors; oral and oropharyngeal cancers, such as hypopharyngeal, laryngeal, nasopharyngeal, and oropharyngeal cancers; gastric cancers, such as lymphomas, gastric stromal tumors, and carcinoid tumors;Testicular cancers, such as germ cell tumors (GCTs), including seminomas and non-seminomas, and gonadal stromal tumors, including Leydig cell tumors and Sertoli cell tumors; thymic cancers, such as thymomas, thymic carcinomas, Hodgkin's disease, non-Hodgkin's lymphoma, carcinoids or carcinomata; rectal cancer; and colon cancer.

[0226] In another aspect of the invention, there is provided a method of treating an ophthalmic disease by administering to the eye of a subject one or more of the compounds or pharmaceutical compositions described herein.

[0227] The invention also provides a method of inhibiting PARP by contacting PARP with an amount of a compound of the invention sufficient to inhibit PARP enzyme activity. In some embodiments, the invention provides a method of inhibiting PARP enzyme activity by contacting a PARP enzyme with an amount of a compound of the invention sufficient to inhibit PARP enzyme activity. In some embodiments, the invention provides a method of inhibiting PARP enzyme activity. Such inhibition can occur in solution, in a cell expressing one or more PARP enzymes, in a tissue comprising cells expressing PARP, or in an organism expressing PARP. In some embodiments, the invention provides a method of inhibiting PARP activity in an animal (including a mammal, such as a human) by contacting the animal with an amount of a compound of the invention sufficient to inhibit PARP enzyme activity in the animal.

[0228] The following general methods described herein provide ways and means of preparing and using the compounds of the invention and are illustrative rather than limiting. Further modifications and additional new methods of the provided methods can also be designed in order to achieve and be used for the purposes of the invention. Thus, it should be understood that there may be other embodiments that fall within the spirit and scope of the invention as defined by the appended specification.

[0229] General preparation method

[0230] The compounds of the invention can be prepared by the following methods. Unless otherwise specified, variables (e.g., X, Y, Z, G, R a 、R b 、R c 、R d 、R e 、R f 、R 1 、R 2 、R 3 and R 4 ) are to be understood as representing those groups described above for the compounds of formulas (I), (IA) and (IB) when used in the following formula. These methods can be similarly applied to other compounds of the formulas with or without modifications as provided above herein.

[0231] Scheme 1

[0232]

[0233] The compound of formula (1) as an aldehyde can be protected with a protecting group (PG), such as by reaction with 1,2-ethanediol, to give the acetal (2). The compound of formula (3) (G-H) can be N-arylated with the acetal (2) using Buchwald-type reaction conditions to give the compound of formula (4). The acetal of formula (4) can be deprotected using a suitable acid such as hydrochloric acid to give the aldehyde of formula (5). The aldehyde of formula (5) can be reacted with the Wittig salt of formula (6) to give the olefin of formula (7). The olefin of formula (7) can be reacted with hydrazine hydrate to form the compound of formula (I). The compound of formula (6) can be prepared by brominating a compound (Int-1) such as with N-bromosuccinimide to form a compound (Int-2) and reacting it with triphenylphosphine. The compound of formula (I) can be converted to a salt by methods known in the art. This scheme is illustrated below in Scheme 1A, Example 1, and Example 2.

[0234] Scheme 1A

[0235]

[0236] In Scheme 1A, the compound of formula (1) as an aldehyde (wherein X, Y, Z are independently selected from CH or N) can be protected, such as by reaction with 1,2-ethanediol, to give the acetal (2). The compound of formula (a) (wherein R e is selected from hydrogen, methyl, ethyl or trifluoromethyl, R f is selected from hydroxy, acetoxy or methoxy, and R 1 -R 4 is as defined herein) can be N-arylated with the acetal (2) using Buchwald-type reaction conditions to give the compound of formula (4a). The acetal of formula (4a) can be deprotected using a suitable acid such as hydrochloric acid to give the aldehyde of formula (5a). The aldehyde of formula (5a) can be reacted with the Wittig salt of formula (6) (wherein R a 、R b 、R c and R d are independently selected from hydrogen or halogen) to give the olefin of formula (7a). The olefin of formula (7a) can be reacted with hydrazine hydrate to form the compound of formula (I).

[0237] Similarly, the compound of formula (I) can be prepared using a compound of formula (b), (c), (d) or (e) in place of the compound of formula (a) in Scheme 1A. This scheme is illustrated below in Example 1.

[0238] Example 1

[0239]

[0240] Similar methods with certain modifications known to those skilled in the art can be used to synthesize the compounds of formula (I) using suitable intermediates and reagents, wherein all variables should be understood to represent the groups described above.

[0241] Another method for preparing the compounds of formula (I) is provided in Scheme 2 below.

[0242] Scheme 2

[0243]

[0244] The compound of formula (1) can be reacted with the Wittig salt of formula (6) to give the olefin of formula (8). The olefin of formula (8) can be reacted with hydrazine hydrate to form the compound of formula (9), which can be N-arylated with the compound of formula (3) using Buchwald-type reaction conditions to give the compound of formula (I). The compound of formula (I) can be converted to a salt by methods known in the art.

[0245] Experimental data

[0246] General procedure for Buchwald coupling reaction - 1: Dissolve the aryl halide (1 equivalent), oxindole (2-hydroxyindole) and related derivatives or NH-containing heterocycles (1 equivalent), trans-4-hydroxy-L-proline (0.4 equivalent) and potassium carbonate (1 equivalent) in DMSO (6 volumes) and degas with nitrogen for 15 minutes. Add copper(I) iodide (0.2 equivalent) to the above mixture and degas again for 15 minutes. After degassing, heat the reaction mixture to 130 °C and stir at the same temperature for 4 hours. After completion of the reaction, dilute the reaction mixture with water and extract with MeOH:DCM (1:9) to give the crude product. Purify the crude product by column chromatography to give the N-arylated product.

[0247] General procedure for Wittig reaction - 2: Dissolve the aldehyde (1 equivalent) and the Wittig salt (1 equivalent) in dichloromethane (100 volumes). Add triethylamine (2 equivalents) to the mixture. Stir the reaction mixture at room temperature for 1 hour. After completion of the reaction, dilute the reaction mixture with water and separate the organic layer. Evaporate the organic layer on a rotary evaporator to give the olefin, which is used in the next step without further purification.

[0248] General procedure for phthalazinone formation - 3: Dissolve the alkene (1 equivalent) and hydrazine hydrate (1.2 equivalents) in THF (15 volumes). Stir the mixture at room temperature for 1 hour. After 1 hour, add acetic acid (0.5 equivalent) and reflux the reaction mixture at 80 °C. Monitor the progress of the reaction by TLC. After completion of the reaction, dilute the reaction mixture with water and extract with a mixture of MeOH and DCM (1:9). Dry the organic layer over anhydrous Na2SO4 and distill to obtain the crude product. Purify the crude product by combinatorial flash or column chromatography using a suitable mixture of MeOH and DCM.

[0249] General procedure for chiral separation of racemic intermediates and examples - 4: Chiral intermediates and examples obtained synthetically in racemic form can be separated into pure enantiomers by appropriate HPLC methods using the following preparative chiral separations. By using the following methods, Examples 2, 3, 7, 8, 11, 12, 14, 15, 19, 20, 22 and 23 can be resolved into pure enantiomers.

[0250] Preparative method - 1:

[0251] Column: CHIRALCEL OJ-H, (250×30) mm, 5 Mobile phase: Hexane / EtOH / MeOH / DEA / 80 / 10 / 10 / 0.1 v / v / v / v

[0252] Flow rate: 40 mL / min

[0253] Detection: UV 210 nm

[0254] Temperature: 25 °C

[0255] Feed concentration: 10 mg / mL

[0256] Injection volume: 5 mL (on column: 50 mg)

[0257] Run time: 30 min

[0258] Cycle time: 12 min

[0259] Preparative method 2:

[0260] Column: CHIRALCEL OX-H, (250×30) mm, 5 Mobile phase: CO2 / cosolvent 65 / 35

[0261] Cosolvent: MeOH / ACN / DEA 50 / 50 / 03 v / v / v

[0262] Flow rate: 120 mL / min

[0263] Detection: UV 260 nm

[0264] Temperature: 25 °C

[0265] Feed concentration: 20 mg / mL

[0266] Injection volume: 5 mL (on column: 100 mg)

[0267] Run time: 20 min

[0268] Cycle time: 15 min

[0269] Preparative method 3:

[0270] Chromatographic column: CHIRALCEL OX-H, (250×30) mm, 5 Mobile phase: CO2 / cosolvent 65 / 35

[0271] Cosolvent: MeOH / ACN 50 / 50

[0272] Flow rate: 120 mL / min

[0273] Detection: UV 260 nm

[0274] Temperature: 25 °C

[0275] Feed concentration: 20 mg / mL

[0276] Injection volume: 5 mL (on column: 100 mg)

[0277] Run time: 20 min

[0278] Cycle time: 15 min

[0279] Preparative method 4:

[0280] Chromatographic column: CHIRALPAK AS-H, (250×21) mm, 5 Mobile phase: MeOH / ACN 10 / 90

[0281] Cosolvent: MeOH / ACN 50 / 50

[0282] Flow rate: 20 mL / min

[0283] Detection: UV 300 nm

[0284] Temperature: 25 °C

[0285] Feed concentration: 40 mg / mL

[0286] Injection volume: 4 mL (on column: 160 mg)

[0287] Run time: 10 min

[0288] Cycle time: 7 min

[0289] Preparative method 5:

[0290] Column: CHIRALPAK IG, (250×30) mm, 5 Mobile phase: ACN / MeOH / DEA (70 / 30 / 0.1 v / v / v)

[0291] Flow rate: 40 mL / min

[0292] Detection: UV 245 nm

[0293] Temperature: 25 °C

[0294] Feed concentration: 10 mg / mL

[0295] Injection volume: 5 mL (on column: 50 mg)

[0296] Run time: 15 min

[0297] Concentration: 30 °C

[0298] Analytical method 1:

[0299] Column: CHIRALCEL OX-3R (150x4.6) mm, 3.0 μm

[0300] Mobile phase: Solution of 30 mM ammonium acetate in [water / ACN / MeOH (40 / 10 / 50, v / v / v)] Flow rate: 1.0 mL / min

[0301] Detection: UV 210 nm

[0302] Temperature: 40 °C

[0303] Analytical method 2:

[0304] Column: CHIRALCEL AS-H (250x4.6) mm, 3.0 μm

[0305] Mobile phase: ACN / MeOH (90 / 10 v / v)

[0306] Flow rate: 1.0 mL / min

[0307] Detection: UV 247 nm

[0308] Temperature: 25 °C

[0309] Analytical method 3:

[0310] Chromatographic column: CHIRALCEL IG (250 x 4.6) mm, 5.0 μm

[0311] Mobile phase: ACN / MeOH / DEA (70 / 30 / 0.1 v / v / v)

[0312] Flow rate: 1.0 mL / min

[0313] Detection: UV 254 nm

[0314] Temperature: 25 °C

[0315] Intermediate 1: 2-(4-Fluoro-3-iodophenyl)-1,3-dioxolane :

[0316] Suspend 4-fluoro-3-iodobenzaldehyde (5 g, 19.99 mmol) in toluene (22 ml). Add camphorsulfonic acid (23 mg, 0.1 mmol) and ethylene glycol (1.61 ml, 29.99 mmol) to the mixture and reflux for 4 hours under a dean-stark condenser. After completion of the reaction, dilute the reaction mixture with ethyl acetate (200 ml) and wash with saturated aqueous sodium bicarbonate. Dry the organic layer with anhydrous Na2SO4. Distill off the organic layer in vacuo to obtain the title compound (5.88 g) as a brown liquid. 1 1H-NMR (δ ppm, CDCl3, 400 MHz): 7.88 (d, J 4.2, 1H), 7.42 (t, J 6, 1H), 7.05 (t, J 8.1, 1H), 5.74 (s, 1H), 4.17 - 3.98 (m, 4H).

[0317] Intermediate 2: 3-Bromoisobenzofuran-1(3H)-one :

[0318] Phthalide (100 g, 0.745 mol) was suspended in carbon tetrachloride (500 ml), and N-bromosuccinimide (146 g, 0.82 mol) was added to the mixture. The reaction mixture was heated to 85 °C, and azobisisobutyronitrile AIBN (6.12 g, 37.2 mmol) was added to the reaction mixture in portions (10 portions). After 4 hours, the reaction mixture was cooled to room temperature. The reaction mixture was quenched with water, and the organic layer was separated. The aqueous layer was extracted with DCM, and the combined organic layers were dried over anhydrous Na2SO4. The organic layer was evaporated on a rotary evaporator to give a crude solid. The crude solid was suspended in petroleum ether (300 ml) and stirred for 15 minutes to give a solid. The solid was filtered and washed with petroleum ether (100 ml). The solid was dried in vacuo for 1 hour to give the title compound as a brown solid (145 g). Yield: 91.39%. 1 1H-NMR (δ ppm, CDCl3, 400 MHz): 7.94 (d, J 8, 1H), 7.79 (t, J 7.6, 1H), 7.64 (d, J 7.6, 2H), 7.40 (s, 1H).

[0319] Intermediate 3: (3-oxo-1,3-dihydroisobenzofuran-1-yl)triphenylphosphonium bromide:

[0320] Intermediate 2 (50 g, 0.234 mol) was suspended in acetonitrile (180 ml), and triphenylphosphine (61.54 g, 0.234 mol) was added. The mixture was heated to 90 °C and stirred for 2.5 hours. The reaction mixture was cooled to room temperature to give a solid. The solid was filtered and washed with ether (125 ml). The solid was dried in vacuo for 30 minutes to give the title compound as a white solid (94 g). Yield: 84%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.61 (s, 1H), 8.00 - 7.93 (m, 3H), 7.86 - 7.70 (m, 15H), 6.95 (d, J 7.6, 1H).

[0321] Intermediate 4: 3-Bromo-5-(1,3-dioxolan-2-yl)pyridine :

[0322] 5-Bromonicotinaldehyde (35 g, 0.19 mol) was suspended in toluene (500 ml), and camphorsulfonic acid (350 mg, 0.15 mmol) and ethylene glycol (15.23 ml, 0.28 mol) were added. The mixture was refluxed under a dean-stark condenser for 4 hours. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (200 ml) and washed with saturated aqueous sodium bicarbonate. The organic layer was dried over anhydrous Na2SO4. The organic layer was distilled in vacuo to give the title compound (5.88 g) as a brown liquid. Yield: 100%.1 H-NMR (δ ppm, CDCl3, 400 MHz): 7.88 (d, J 4.2, 1H), 7.42 (t, J 6, 1H), 7.05 (t, J 8.1, 1H), 5.74 (s, 1H), 4.17 - 3.98 (m, 4H).

[0323] Intermediate 5: 3-Hydroxy-3-methyldihydroindol-2-one :

[0324] Indigo red (3 g, 20.39 mmol) was dissolved in THF (50 ml) under a nitrogen atmosphere and cooled to -10 °C. A THF solution of 3 M methylmagnesium chloride (20.39 ml, 61.1 mmol) was added dropwise to the above mixture, and the reaction mixture was stirred at -10 °C for 2 hours. After 2 hours, the reaction mixture was quenched with an aqueous ammonium chloride solution and extracted with MeOH and DCM (1:9) (3 * 150 ml). The combined organic layers were washed with water (100 ml) and dried over anhydrous Na2SO4. The organic layer was distilled to obtain the crude product. The crude product was suspended in ether (50 ml) and stirred for 15 minutes to obtain a solid. The solid was filtered and washed with ether (10 ml). The solid was dried in vacuo for 30 minutes to obtain the title compound (2.35 g) as a yellow solid. Yield: 71%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.18 (s, 1H), 7.26 (d, J 7.6, 1H), 7.17 (t, J 7.6, 1H), 6.94 (t, J 7.6, 1H), 6.78 (d, J 7.6, 1H), 5.82 (s, 1H), 1.33 (s, 3H). MS (m / z): 162.2 ([M-H] - ). The racemic 3-hydroxy-3-methylindolin-2-one was resolved into (+) and (-) enantiomers by one of the preparation methods shown in General Procedure 4 and used as such when needed.

[0325] ( (R)-(+)-3-Hydroxy-3-methyldihydroindol-2-one :

[0326] 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.19 (s, 1H), 7.26 (d, J 7.6, 1H), 7.17 (t, J 7.6, 1H), 6.94 (t, J 7.6, 1H), 6.78 (d, J 7.6, 1H), 5.82 (s, 1H), 1.33 (s, 3H). [α] D 25 : +45.90° (MeOH; c 1.0)

[0327] (S)-(-)-3-Hydroxy-3-methyldihydroindol-2-one:

[0328] 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.19 (s, 1H), 7.26 (d, J 7.6, 1H), 7.17 (t, J 7.6, 1H), 6.94 (t, J 7.6, 1H), 6.78 (d, J 7.6, 1H), 5.83 (s, 1H), 1.33 (s, 3H). [α] D 25 : -45.94° (MeOH; c 1.0).

[0329] Intermediate 6: 1-(5-(1,3-Dioxolan-2-yl)-2-fluorophenyl)-3-hydroxy-3-methyl- dihydroindol-2-one :

[0330] According to the general procedure 1, the title compound was synthesized from intermediate 1 (1.63 g, 5.54 mmol) and intermediate 5 (0.9 g, 5.54 mmol). The crude product obtained was purified by the combination flash method using MeOH and DCM (2.1:97.9) as the eluent. The combined pure fractions from the combination flash method were distilled to give the title compound as a brown gel (740 mg). Yield: 40.88%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 7.66 - 7.58 (m, 1.5H), 7.56 - 7.41 (m, 2.5H) 7.28 - 7.21 (m, 1H), 7.16 - 7.08 (m, 1H), 6.55 (t, J 8.2, 1H) 6.28 (s, 0.45H), 6.16 (s, 0.55H) 5.81 (s, 0.55H), 5.78 (s, 0.45H), 4.10 - 4.03 (m, 2H), 3.98 - 3.93 (m, 2H), 1.53 (s, 1.35H), 1.49 (s, 1.65H).

[0331] Intermediate 7: 4-Fluoro-3-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)benzaldehyde :

[0332] Intermediate 6 (700 mg, 2.13 mmol) was dissolved in THF (10 ml) and 6N hydrochloric acid (3 ml) was added. The mixture was stirred at room temperature for 1 hour. After 1 hour, the reaction material was cooled to 0 °C and the pH of the reaction mixture was adjusted to about 7 using saturated aqueous sodium bicarbonate. The aqueous solution of the reaction mixture was extracted with MeOH and DCM (1:9) (2 * 100 ml). The combined organic layers were dried over anhydrous Na2SO4 and evaporated to give the title compound as a brown gel (605 mg). Yield: 100%. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.04 (s, 0.55H), 10.01 (s, 0.45H), 8.19 - 8.02 (m, 2H), 7.74 (t, J 9.5, 1H), 7.50 - 7.43 (m, 1H), 7.30 - 7.22 (m, 1H), 7.18 - 7.10 (m, 1H), 6.70 - 6.62 (m, 1H), 6.32 (s, 0.45H), 6.21 (s, 0.55H), 1.55 (s, 1.35H), 1.50 (s, 1.65H).

[0333] Intermediate 8: 1-(2-Fluoro-5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)phenyl)-3-hydroxy-3- methyldihydroindol-2-one :

[0334] According to General Procedure 2, the title compound was synthesized from Intermediate 7 (600 mg, 2.1 mmol) and Intermediate 3 (1.79 g, 3.8 mmol) as a pale yellow gel (840 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0335] Intermediate 9: 1-(5-(1,3-Dioxolan-2-yl)pyridin-3-yl)-3-hydroxy-3-methyldihydroindol-2- one :

[0336] According to General Procedure 1, the title compound was synthesized from Intermediate 4 (2 g, 8.68 mmol) and Intermediate 5 (1.4 g, 8.68 mmol). The crude product obtained was purified by the combined flash method using MeOH and DCM (3.1:96.9) as the eluent. The combined pure fractions from the combined flash method were distilled to give the title compound as a pale yellow gel (865 mg). Yield: 31.9%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.71 (bs, 2H), 7.92 (s, 1H), 7.46 (d, J 7.3, 1H), 7.27 (t, J 7.6, 1H), 7.13 (t, J 7.4, 1H), 6.75 (d, J 7.8, 1H), 6.15 (s, 1H), 5.93 (s, 1H), 4.15 - 4.09 (m, 2H), 4.02 - 3.95 (m, 2H), 1.52 (s, 3H). MS (m / z): 312.8 ([M + H] + )

[0337] Intermediate 10: 5-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)nicotinaldehyde :

[0338] Intermediate 9 (850 mg, 2.72 mmol) was dissolved in THF (10 ml) and concentrated hydrochloric acid (2.5 ml) was added. The mixture was refluxed at 70 °C for 1 h. After 1 h, the reaction mixture was cooled to 0 °C and the pH of the reaction mixture was adjusted to about 7 using saturated aqueous sodium bicarbonate. The aqueous solution of the reaction mixture was extracted with MeOH and DCM (1:9) (2 * 100 ml). The combined organic layers were dried over anhydrous Na2SO4 and evaporated to give the title compound as a pale yellow gel (729 mg). Yield: 100%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.18 (s, 1H), 9.13 (d, J 1.2, 1H), 8.96 (d, J 2.4, 1H), 8.34 (t, J 2, 1H), 7.48 (d, J 7.2, 1H), 7.28 (td, J 8, 1.2, 1H), 7.16 (t, J 7.2, 1H), 6.89 (d, J 8, 1H), 6.19 (s, 1H), 1.53 (s, 3H). MS (m / z): 268.8 ([M+H] + )。

[0339] Intermediate 11: 3-Hydroxy-3-methyl-1-(5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin- 3-yl)dihydroindol-2-one :

[0340] According to General Procedure 2, the title compound was synthesized from Intermediate 10 (720 mg, 2.7 mmol) and Intermediate 3 (2.29 g, 4.8 mmol) as a yellow gel (1.03 g). Yield: 100%. MS (m / z): 384.8 ([M+H] + )。

[0341] Intermediate 12: 2-Bromo-4-(1,3-dioxolan-2-yl)pyridine :

[0342] 2-Bromoisonicotinaldehyde (5 g, 26.9 mmol) was suspended in toluene (40 ml). Camphorsulfonic acid (20 mg, 0.09 mmol) and ethylene glycol (2.25 ml, 0.28 mol) were added to the mixture and the mixture was refluxed for 15 h under a dean-stark condenser. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (200 ml) and washed with saturated aqueous sodium bicarbonate. The organic layer was dried over anhydrous Na2SO4 and distilled to give the title compound (6 g) as a brown liquid. Yield: 97% 1 1H-NMR (δ ppm, CDCl3, 400 MHz): 8.39 (d, J 4.9, 1H), 7.60 (s, 1H), 7.35 (d, J 4.9, 1H), 5.80 (s, 1H), 4.06 (s, 4H).

[0343] Intermediate 13: 1-(4-(1,3-Dioxolan-2-yl)pyridin-2-yl)-3-hydroxy-3-methyldihydroindol-2- one :

[0344] According to general procedure 1, the title compound was synthesized from intermediate 12 (4.3 g, 18.69 mmol) and intermediate 5 (3 g, 18.69 mmol). The crude product obtained after workup was used in the next step without further purification (900 mg). Yield: 17%. The compound was used in the next step without any characterization.

[0345] Intermediate 14: 2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)isonicotinaldehyde :

[0346] Intermediate 13 (900 mg, 2.88 mmol) was dissolved in THF (10 ml) and concentrated hydrochloric acid (0.9 ml) was added. The mixture was refluxed at 80 °C for 1 h. After 1 h, the reaction mass was cooled to 0 °C and the pH of the reaction mixture was adjusted to about 7 using saturated aqueous sodium bicarbonate. The aqueous solution of the reaction mixture was extracted with MeOH and DCM (1:9) (2 * 100 ml). The organic layer was dried over anhydrous Na2SO4 and evaporated on a rotary evaporator to give the title compound as a pale yellow gel (700 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0347] Intermediate 15: 3-Hydroxy-3-methyl-1-(4-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin- 2-yl)dihydroindol-2-one :

[0348] According to general procedure 2, the title compound was synthesized from intermediate 14 (350 mg, 1.30 mmol) and intermediate 3 (1.1 g, 2.35 mmol) as a yellow gel (500 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0349] Intermediate 16: 3-Ethyl-3-hydroxydihydroindol-2-one :

[0350] The compound was prepared by the following procedure. Magnesium turnings (2.48 g, 0.101 mol) and iodine (2 mg) were placed in diethyl ether (50 ml) under a nitrogen atmosphere. Ethyl bromide (7.57 ml, 0.101 mol) was added dropwise to the mixture over 20 min and the reaction mixture was stirred at room temperature for 2 h to give ethylmagnesium bromide.

[0351] Indigo (5 g, 0.033 mol) was dissolved in THF (50 ml) under a nitrogen atmosphere and cooled to -15 °C. An ethereal solution of ethylmagnesium bromide from the above reaction was added dropwise to the reaction mixture at -15 °C and stirred at the same temperature for 2 hours. After 2 hours, the reaction mixture was quenched with an aqueous ammonium chloride solution (200 ml) and extracted with MeOH and DCM (1:9) (3 * 100 ml). The combined organic layers were dried over anhydrous Na2SO4 and distilled to obtain the crude product. The crude product was purified by the combinatorial flash method using MeOH and DCM (3:97) as the eluent. The pure fractions from the combinatorial flash method were distilled to obtain the title compound as a brown solid (1.24 g). Yield: 21%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.19 (s, 1H), 7.22 (d, J 7.31, 1H), 7.18 (t, J 7.7, 1H), 6.95 (t, J 7.4, 1H), 6.78 (d, J 7.7, 1H), 5.79 (s, 1H), 1.75 (m, 2H), 0.59 (t, J 7.4, 3H). MS (m / z): 176.1 ([M-H]-).

[0352] Intermediate 17: 1-(5-(1,3-Dioxolan-2-yl)pyridin-3-yl)-3-ethyl-3-hydroxydihydroindol-2- one :

[0353] According to General Procedure 1, the title compound was synthesized from Intermediate 4 (1.43 g, 6.2 mmol) and Intermediate 16 (1.1 g, 6.2 mmol). The crude product obtained was purified by the combinatorial flash method using MeOH and DCM (2.5:97.5) as the eluent. The combined pure fractions from the combinatorial flash method were distilled to obtain the title compound as a light yellow gel (800 mg). Yield: 39%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.71 (s, 1H), 8.67 (s, 1H), 7.88 (d, J 1.9, 1H), 7.429 (d, J 7.3, 1H), 7.28 (td, J 7.1, 1, 1H), 7.15 (t, J 7.4, 1H), 6.76 (d, J 7.8, 1H), 6.15 (s, 1H), 5.93 (s, 1H), 4.12 - 4.06 (m, 2H), 4.01 - 3.96 (m, 2H), 1.97 - 1.88 (m, 2H), 0.72 (t, J 7.4, 3H).

[0354] Intermediate 18: 5-(3-Ethyl-3-hydroxy-2-oxodihydroindol-1-yl)nicotinaldehyde :

[0355] Intermediate 17 (800 mg, 2.45 mmol) was dissolved in THF (20 ml) and concentrated hydrochloric acid (2 ml) was added. The mixture was refluxed at 80 °C for 2 h. After 2 h, the reaction material was cooled to 0 °C and the pH of the reaction mixture was adjusted to about 7 using saturated aqueous sodium bicarbonate. The aqueous solution of the reaction mixture was extracted with MeOH and DCM (1:9) (3 * 50 ml). The combined organic layers were dried over anhydrous Na2SO4 and evaporated to give the title compound as a brown liquid (700 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0356] Intermediate 19: 3-Ethyl-3-hydroxy-1-(5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin- 3-yl)dihydroindol-2-one :

[0357] According to General Procedure 2, the title compound was synthesized from Intermediate 18 (700 mg, 2.48 mmol) and Intermediate 3 (2.12 g, 4.46 mmol) as a pale yellow gel (987 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0358] Intermediate 20: (2-Bromo-4-fluorophenyl)methanol :

[0359] The compound was prepared by the following procedure. 2-Bromo-4-fluorobenzaldehyde (12 g, 59.11 mmol) was suspended in MeOH (120 ml) and cooled to 0 °C. Sodium borohydride (4.47 g, 118.2 mmol) was added to the above mixture in portions and stirred at 0 °C for 1 h. After 1 h, the reaction mixture was quenched with saturated aqueous ammonium chloride (150 ml). The reaction mixture was distilled to remove MeOH, and the aqueous layer was extracted with ethyl acetate (2 * 200 ml). The combined ethyl acetate layers were dried over anhydrous Na2SO4 and evaporated to give the title compound as an off-white solid (11.6 g). Yield:

[0360] 95.71%. 1 1H-NMR (δ ppm, CDCl3, 400 MHz): 7.46 (dd, J 8.4, 6.1, 1H), 7.31 (dd, J 8.2, 2.5, 1H), 7.05 (dd, J 8.3, 5.9, 1H), 4.72 (d, J 5.1, 2H), 1.96 (t, J 5.7, 1H).

[0361] Intermediate 21: 6-Fluorisobenzofuran-1(3H)-one :

[0362] The compound was prepared by the following procedure. Intermediate 20 (11.5 g, 56.1 mmol) was dissolved in DMF (50 ml) under a nitrogen atmosphere. Cuprous cyanide (10.04 g, 112.2 mmol) was added thereto and the reaction mixture was heated to 180 °C for 2.5 hours. After 2.5 hours, the reaction mixture was cooled to 100 °C and water was added to the reaction mixture. The reaction was continued at 100 °C for 18 hours. After 18 hours, the reaction mixture was cooled to room temperature and diluted with ethyl acetate (250 ml). The reaction mixture was filtered through a celite plug and the celite bed was washed with ethyl acetate (100 ml). The combined ethyl acetate filtrate was washed with water (200 ml), brine solution (200 ml) and saturated lithium chloride aqueous solution (200 ml). The organic layer was dried over anhydrous Na2SO4 and evaporated to give the crude product. The crude product was purified by the combined flash method using ethyl acetate and petroleum ether (16:84) as the eluent. The combined pure fractions from the column were evaporated to give the title compound as a brown solid (3 g). Yield: 36%. 1 1H-NMR (δ ppm, CDCl3, 400 MHz): 7.59 (dd, J 7.2, 2, 1H), 7.48 (dd, J 8.3, 4.3, 1H), 7.41 (td, J 8.5, 2.2, 1H), 5.31 (s, 2H).

[0363] Intermediate 22: 3-Bromo-6-fluoroisobenzofuran-1(3H)-one :

[0364] Intermediate 21 (1.5 g, 9.86 mmol) was suspended in carbon tetrachloride (20 ml) and N-bromosuccinimide (1.93 g, 10.84 mmol) was added. The reaction mixture was heated to 85 °C and azobisisobutyronitrile AIBN (80 mg, 0.49 mmol) was added to the reaction mixture in two portions. After 4 hours, the reaction mixture was cooled to room temperature. The reaction mixture was quenched with water and the organic layer was separated. The aqueous layer was extracted with DCM and the combined organic layers were dried over anhydrous Na2SO4. The organic layer was evaporated to give the title compound as a brown gel (2.27 g). Yield: 100%. 1 1H-NMR (δ ppm, CDCl3, 400 MHz): 7.62 (dd, J 8.4, 4.2, 1H), 7.57 (dd, J 6.8, 2.2, 1H), 7.50 (td, J 8.5, 2.3, 1H), 7.38 (s, 1H).

[0365] Intermediate 23: (5-Fluoro-3-oxo-1,3-dihydroisobenzofuran-1-yl)triphenylphosphonium bromide :

[0366] Intermediate 22 (5 g, 21.6 mmol) was suspended in acetonitrile (20 ml), and triphenylphosphine (5.67 g, 21.6 mmol) was added. The mixture was heated to 90 °C and stirred at the same temperature for 2.5 hours. The reaction mixture was cooled to room temperature to give a solid. The solid was filtered and washed with ether (20 ml). The solid was dried under vacuum for 1 hour to give the crude product (8.2 g). The crude compound was mixed with ethanol (25 ml) and refluxed at 90 °C for 1 hour. After one hour, the heterogeneous mixture was stirred at room temperature for 15 hours. The solid was filtered and the solid was washed with ethanol (5 ml). The solid was dried under vacuum to give the title compound (6 g) as an off-white solid. Yield: 56%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.57 (s, 1H), 8.02 - 7.94 (m, 3H), 7.84 - 7.67 (m, 14H), 7.00 - 6.94 (m, 1H).

[0367] Intermediate 24: 3-Hydroxy-3-methyl-1-(5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin- 3-yl)indoline-2-one :

[0368] The title compound was synthesized from Intermediate 10 (450 mg, 1.68 mmol) and Intermediate 23 (1.48 g, 3.01 mmol) according to General Procedure 2 as a pale yellow gel (674 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0369] Intermediate 25: 5-Fluoro-1(3H)-isobenzofuranone :

[0370] The compound was prepared by the following procedure. Under a nitrogen atmosphere, (2-bromo-5-fluorophenyl)methanol (10 g, 48.8 mmol) was dissolved in DMF (50 ml). Cuprous cyanide (8.74 g, 97.54 mmol) was added thereto and the reaction mixture was heated to 180 °C for 3 hours. After 3 hours, the reaction mixture was cooled to 100 °C and water was added to the reaction mixture. The reaction was continued at 100 °C for 18 hours. After 18 hours, the reaction mixture was cooled to room temperature and diluted with ethyl acetate (250 ml). The reaction mixture was filtered through a Celite plug and the Celite bed was washed with ethyl acetate (100 ml). The combined ethyl acetate filtrate was washed with water (200 ml), brine solution (200 ml), and saturated lithium chloride aqueous solution (200 ml). The organic layer was dried over anhydrous Na2SO4 and evaporated to give the crude product. The crude product was purified by the combined flash method using ethyl acetate and petroleum ether (16:84) as the eluent. The combined pure fractions from the combined flash method were evaporated to give the title compound as a brown solid (4.2 g). Yield: 57%. 1H-NMR (δ ppm, CDCl3, 400 MHz): 7.93 (dd, J 8.5, 5, 1H), 7.24 (td, J 8.4, 1.8, 1H), 7.18 (dd, J 7.8, 1.4, 1H), 5.30 (s, 2H).

[0371] Intermediate 26: 3-Bromo-5-fluoro-1(3H)-isobenzofuranone :

[0372] The intermediate 25 (3 g, 19.72 mmol) was suspended in carbon tetrachloride (40 ml) and N-bromosuccinimide (3.86 g, 21.69 mmol) was added. The reaction mixture was heated to 85 °C and azobisisobutyronitrile AIBN (161 mg, 0.98 mmol) was added to the reaction mixture in portions (3 portions). After 4 h, the reaction mixture was cooled to room temperature. The reaction mixture was quenched with water and the organic layer was separated. The aqueous layer was extracted with DCM and the combined organic layers were dried over anhydrous Na2SO4. The organic layer was evaporated on a rotary evaporator to give the crude product. The crude product was purified by combinatorial flash method using ethyl acetate and petroleum ether (11:89) as the eluent. The combined pure fractions from the combinatorial flash method were distilled to give the title compound as a brown liquid (3.96 g). Yield: 87%. 1 H-NMR (δ ppm, CDCl3, 400 MHz): 7.94 (dd, J 8.3, 4.7, 1H), 7.38 - 7.29 (m, 3H).

[0373] Intermediate 27: (6-Fluoro-3-oxo-1,3-dihydroisobenzofuran-1-yl)triphenylphosphonium bromide :

[0374] The intermediate 26 (3.46 g, 14.98 mmol) was suspended in acetonitrile (10 ml) and triphenylphosphine (3.92 g, 14.98 mmol) was added. The mixture was heated to 90 °C and stirred at the same temperature for 2.5 h. The reaction mixture was cooled to room temperature to give a solid. The solid was filtered and washed with ether (50 ml). The solid was dried in vacuo for 1 h to give the crude product (6.2 g). The crude compound was suspended in ethanol (30 ml) and refluxed at 90 °C for 1 h. After 1 h, the heterogeneous mixture was stirred at room temperature for 18 h. The solid was filtered and the solid was washed with ethanol (5 ml). The solid was dried in vacuo to give the title compound (2.7 g) as an off-white solid. Yield: 37%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.58 (s, 1H), 8.02 - 7.90 (m, 4H), 7.77 - 7.65 (m, 12H), 7.61 (t, J 8.8, 1H), 6.65 (d, J 7.9, 1H).

[0375] Intermediate 28: 1-(5-((6-Fluoro-3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin-3-yl)-3-hydroxy-3-methylandoline-2-one :

[0376] According to general procedure 2, the title compound was synthesized from intermediate 10 (450 mg, 1.68 mmol) and intermediate 27 (1.48 g, 3.01 mmol) as a pale yellow gel (674 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0377] Intermediate 29: 5-Fluoro-3-hydroxy-3-methylandoline-2-one :

[0378] 5-Fluoroindole-2,3-dione (10 g, 60.56 mmol) was suspended in THF (150 ml) under a nitrogen atmosphere and cooled to -10 °C. A THF solution of 3 M methylmagnesium chloride (60.56 ml, 181.69 mmol) was added dropwise to the above mixture and the reaction mixture was stirred at -10 °C for 3 hours. After 3 hours, the reaction mixture was quenched with an aqueous ammonium chloride solution (250 ml) and extracted with ethyl acetate (2 × 300 ml). The combined organic layers were washed with water (300 ml) and dried over anhydrous Na2SO4. The organic layer was distilled to give the crude product (10 g). The crude product was suspended in diethyl ether (50 ml) and stirred for 15 minutes to give a solid. The solid was filtered and washed with diethyl ether (2 × 25 ml). The solid was dried in vacuo for 30 minutes to give the title compound (8 g) as a brown solid. Yield: 72%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.22 (s, 1H), 7.15 (dd, J 8.1, 2.6, 1H), 7.04 - 6.98 (m, 1H), 6.78 (dd, J 8.4, 4.3, 1H), 5.95 (s, 1H), 1.34 (s, 3H).

[0379] Intermediate 30: 1-(5-(1,3-Dioxolan-2-yl)pyridin-3-yl)-5-fluoro-3-hydroxy-3-methylandoline -2-one :

[0380] According to general procedure 1, the title compound was synthesized from intermediate 4 (1.5 g, 6.51 mmol) and intermediate 29 (1.18 g, 6.51 mmol). The crude product obtained was purified by the combined flash method using MeOH and DCM (2.5:97.5) as the eluent. The combined pure fractions from the combined flash method were distilled to give the title compound as a pale yellow solid (650 mg). Yield: 30%. 11H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.76 (bs, 2H), 7.92 (s, 1H), 7.38 (dd, J 7.9, 2.5, 1H), 7.10 (td, J 9.2, 2.5, 1H), 6.77 (dd, J 8.6, 4.1, 1H), 6.27 (s, 1H), 5.93 (s, 1H), 4.12 - 4.05 (m, 2H), 4.01 - 3.95 (m, 2H), 1.53 (s, 3H).

[0381] Intermediate 31: 5-(5-Fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)nicotinaldehyde :

[0382] The intermediate 30 (650 mg, 1.97 mmol) was dissolved in THF (20 ml) and concentrated hydrochloric acid (2 ml) was added. The mixture was refluxed at 80 °C for 2.5 h. After 2.5 h, the reaction material was cooled to room temperature and the pH of the reaction mixture was adjusted to about 7 using an aqueous solution of saturated sodium bicarbonate. The aqueous solution of the reaction mixture was extracted with MeOH and DCM (1:9) (3 * 50 ml). The combined organic layers were dried over anhydrous Na2SO4 and evaporated to give the title compound as a brown liquid (700 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0383] Intermediate 32: 5-Fluoro-3-hydroxy-3-methyl-1-(5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl) pyridin-3-yl)indoline-2-one :

[0384] According to General Procedure 2, the title compound was synthesized from intermediate 31 (250 mg, 0.87 mmol) and intermediate 3 (746 mg, 1.57 mmol) as a light yellow gel (351 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0385] Intermediate 33: 5-Fluoro-1-(5-((5-fluoro-3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin-3- yl)-3-hydroxy-3-methylandoline-2-one :

[0386] According to General Procedure 2, the title compound was synthesized from intermediate 31 (250 mg, 0.87 mmol) and intermediate 23 (0.77 g, 1.57 mmol) as a light yellow gel (366 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0387] Intermediate 34: 6-Fluoro-3-hydroxy-3-methylandoline-2-one :

[0388] 6-Fluoroindole-2,3-dione (5 g, 30.28 mmol) was suspended in THF (75 ml) under a nitrogen atmosphere and cooled to -10 °C. A THF solution of 3 M methylmagnesium chloride (60.56 ml, 181.69 mmol) was added dropwise to the above mixture, and the reaction mixture was stirred at -10 °C for 2 hours. After 2 hours, the reaction mixture was quenched with an aqueous ammonium chloride solution (200 ml) and extracted with ethyl acetate (3 × 100 ml). The combined organic layers were dried over anhydrous Na2SO4 and the organic layer was distilled to obtain the crude product (6.2 g). The crude product was suspended in ether (100 ml) and stirred for 15 minutes to obtain a solid. The solid was filtered and washed with ether (50 ml). The solid was dried in vacuo for 3 hours to obtain the title compound (4 g) as a brown solid. Yield: 73%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.34 (s, 1H), 7.28 (dd, J 8.1, 5.7, 1H), 6.73 (td, J 8.2, 2.4, 1H), 6.60 (dd, J 9.3, 2.3, 1H), 5.87 (s, 1H), 1.33 (s, 3H).

[0389] Intermediate 35: 1-(5-(1,3-Dioxolan-2-yl)pyridin-3-yl)-6-fluoro-3-hydroxy-3-methylandoline -2-one :

[0390] According to General Procedure 1, the title compound was synthesized from Intermediate 4 (1.5 g, 6.51 mmol) and Intermediate 34 (1.18 g, 6.51 mmol). The crude product obtained was purified by the combined flash method using MeOH and DCM (2.5:97.5) as the eluent. The combined pure fractions from the combined flash method were distilled to obtain the title compound as a light yellow solid (650 mg). Yield: 30%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.72 (s, 1H), 8.69 (s, 1H), 7.92 (s, 1H), 7.48 (dd, J 8.2, 5.6, 1H), 6.94 (td, J 8.4, 2.2, 1H), 6.63 (dd, J 9.4, 2.2, 1H), 6.18 (s, 1H), 5.94 (s, 1H), 4.11 - 4.05 (m, 2H), 4.01 - 3.95 (m, 2H), 1.52 (s, 3H).

[0391] Intermediate 36: 5-(6-Fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)nicotinaldehyde :

[0392] Intermediate 35 (730 mg, 2.21 mmol) was dissolved in THF (20 ml) and concentrated hydrochloric acid (3.5 ml) was added. The mixture was refluxed at 80 °C for 3 h. After 3 h, the reaction mixture was cooled to 0 °C and the pH of the reaction mixture was adjusted to about 7 using saturated aqueous sodium bicarbonate. The aqueous solution of the reaction mixture was extracted with MeOH and DCM (1:9) (2 * 200 ml). The combined organic layers were dried over anhydrous Na2SO4 and evaporated to give the title compound as a brown liquid (632 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0393] Intermediate 37: 6-Fluoro-3-hydroxy-3-methyl-1-(5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl) pyridin-3-yl)indoline-2-one :

[0394] According to General Procedure 2, the title compound was synthesized from Intermediate 36 (300 mg, 1.04 mmol) and Intermediate 3 (895 mg, 1.88 mmol) as a yellow solid (420 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0395] Intermediate 38: 6-Fluoro-1-(5-((5-fluoro-3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin-3- yl)-3-hydroxy-3-methylandoline-2-one :

[0396] According to General Procedure 2, the title compound was synthesized from Intermediate 36 (300 mg, 1.04 mmol) and Intermediate 23 (929 mg, 1.88 mmol) as a yellow gel (440 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0397] Intermediate 39: 1-(4-((5-Fluoro-3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin-2-yl)-3-hydroxy -3-methylandoline-2-one :

[0398] According to General Procedure 2, the title compound was synthesized from Intermediate 14 (350 mg, 1.30 mmol) and Intermediate 23 (1.15 g, 1.8 mmol) as a dark brown gel (800 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0399] Intermediate 40: 1-(4-(1,3-Dioxolan-2-yl)pyridin-2-yl)-5-fluoro-3-hydroxy-3-methylandoline -2-one :

[0400] According to General Procedure 1, the title compound was synthesized from Intermediate 12 (2 g, 8.7 mmol) and Intermediate 29 (1.57 g, 8.7 mmol). The crude product obtained was purified by the combined flash method using MeOH and DCM (5:95) as the eluent. The combined pure fractions from the combined flash method were distilled to give the title compound as a brown solid (900 mg). Yield: 31%. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.61 (d, J 4.8, 1H), 7.81 (s, 1H), 7.64 (dd, J 8.8, 4.4, 1H), 7.42 (dd, J 5.2, 1.2, 1H), 7.35 (dd, J 8, 2.8, 1H), 7.14 (td, J 9.2, 2.8, 1H), 6.33 (s, 1H), 5.89 (s, 1H), 4.05 - 3.98 (m, 4H), 1.51 (s, 3H).

[0401] Intermediate 41: 2-(5-Fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)isonicotinaldehyde :

[0402] The intermediate 40 (765 mg, 2.31 mmol) was dissolved in THF (8 ml) and concentrated hydrochloric acid (3 ml) was added. The mixture was refluxed at 80 °C for 3 h. After 3 h, the reaction mass was cooled to 0 °C and the pH of the reaction mixture was adjusted to about 7 using saturated aqueous sodium bicarbonate. The aqueous solution of the reaction mixture was extracted with MeOH and DCM (1:9) (2 * 50 ml). The combined organic layers were dried over anhydrous Na2SO4 and evaporated on a rotary evaporator to give the title compound as a brown liquid (662 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0403] Intermediate 42: 5-Fluoro-1-(4-((5-fluoro-3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin-2- yl)-3-hydroxy-3-methylandoline-2-one :

[0404] According to General Procedure 2, the title compound was synthesized from intermediate 41 (662 mg, 2.31 mmol) and intermediate 23 (2.05 g, 4.16 mmol) as a dark brown gel (930 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0405] Intermediate 43: 3-Hydroxy-3-(trifluoromethyl)indoline-2-one :

[0406] This compound was prepared by the following reported procedure (Chen Zang et al., Organic & Biomolecular Chemistry, 2013, 11, 5621 - 5633). Aniline (6 g, 64.4 mmol) and ethyl 3,3,3-trifluoro-2-oxopropionate (13.1 g, 77.3) were placed in a microwave vial. 1,2-Dichlorobenzene (17.16 ml) was added to the vial and heated in a microwave oven to 150 °C for 20 min. The reaction mass was diluted with water (150 ml) and brine (150 ml). The aqueous layer was extracted with EtOAc to give the crude product. The crude product was purified by column chromatography to give the title compound as an off-white solid (4.9 g). Yield: 35%. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.80 (s, 1H), 7.59 (s, 1H), 7.42 - 7.33 (m, 2H), 7.05 (t, J 7.6, 1H), 6.89 (d, J 8, 1H). MS (m / z): 216.21 ([M-H] + )。

[0407] Intermediate 44: 1-(5-(1,3-dioxolan-2-yl)pyridin-3-yl)-3-hydroxy-3-(trifluoromethyl)dihydroind ole-2-one :

[0408] According to General Procedure 1, the title compound was synthesized from Intermediate 4 (2.64 g, 11.5 mmol) and Intermediate 43 (2.49 g, 11.5 mmol). The crude product obtained was purified by the combinatorial flash method using MeOH and DCM (2.4:97.6) as the eluent. The combined pure fractions from the combinatorial flash method were distilled to give the title compound as a brown solid (700 mg). Yield: 17%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.77 (s, 1H), 8.70 (d, J 1.6, 1H), 7.94 (s, 1H), 7.91 (s, 1H), 7.59 (d, J 7.2, 1H), 7.45 (t, J 7.6, 1H), 7.25 (t, J 7.6, 1H), 6.85 (d, J 8, 1H), 5.95 (s, 1H), 4.12 - 4.07 (m, 2H), 4.04 - 3.95 (m, 2H). MS (m / z): 367.34 ([M+H] + )。

[0409] Intermediate 45: 5-(3-hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)nicotinaldehyde :

[0410] Intermediate 44 (680 mg, 1.9 mmol) was dissolved in a mixture of water (17 ml) and acetone (17 ml). Oxalic acid hydrate (2.34 g, 18.6 mmol) was added to the mixture and stirred at 70 °C for 16 h. Acetone was distilled off from the reaction mixture and the mixture was basified with 10% aqueous sodium bicarbonate. The aqueous layer was extracted with DCM and the combined DCM layers were distilled to give the crude product. The crude product was triturated with ether to give the title compound as a brown solid (490 mg). Yield: 82%. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.19 (s, 1H), 9.18 (d, J 1.7, 1H), 8.96 (d, J 2.4, 1H), 8.36 (t, J 2, 1H), 7.60 (t, J 7.5, 1H), 7.47 (td, J 7.8, 1.1, 1H), 7.28 (t, J 7.5, 1H), 7.00 (d, J 7.9, 1H). MS (m / z): 323.0 ([M+H] + )。

[0411] Intermediate 46: 3-hydroxy-1-(5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin-3-yl)-3- (trifluoromethyl)dihydroindole-2-one :

[0412] According to General Procedure 2, the title compound was synthesized from Intermediate 45 (490 mg, 1.5 mmol) and Intermediate 3 (800 mg, 1.7 mmol) as a yellow solid (660 mg). MS (m / z): 437.36 ([M-H]-).

[0413] Intermediate 47: 2-(3-iodophenyl)-1,3-dioxolane :

[0414] 3-Iodobenzaldehyde (14.5 g, 62.5 mmol) was suspended in toluene (145 ml). p-Toluenesulfonic acid monohydrate (594 mg, 3.12 mmol) and ethylene glycol (4.65 g, 75 mmol) were added to the mixture and refluxed for 7 hours under a Dean-Stark condenser. After 7 hours, the reaction mixture was cooled to room temperature, diluted with toluene (29 ml), and washed with saturated aqueous sodium bicarbonate (75 ml). The organic layer was washed with water (2 * 75 ml) and dried over anhydrous Na2SO4. The organic layer was distilled off in vacuo to give the title compound (15 g) as a yellow liquid. Yield: 87%. 1 H-NMR (δ ppm, CDCl3, 400 MHz): 7.86 (d, J 1.6, 1H), 7.73 (dd, J 7.6, 1.2, 1H), 7.45 (d, J 7.2, 1H), 7.14 (td, J 7.6, 1H), 5.78 (s, 1H), 4.17 - 4.10 (m, 2H), 4.08 - 4.01 (m, 2H). MS (m / z): 276.98 ([M+H] + )。

[0415] Intermediate 48: 1-(3-(1,3-dioxolan-2-yl)phenyl)-3-hydroxy-3-methyldihydroindole-2-one :

[0416] According to General Procedure 1, the title compound was synthesized from Intermediate 47 (12 g, 52.39 mmol) and Intermediate 5 (8.55 g, 52.39). The crude product was purified by combinatorial flash method using ethyl acetate and petroleum ether (1:1) as the eluent. The combined pure fractions from the combinatorial flash method were distilled to give the title compound as a black gel (7.8 g). Yield: 48%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 7.59 (t, J 7.2, 1H), 7.51 (d, J 7.6, 1H), 7.48 - 7.40 (m, 3H), 7.24 (t, J 7.2, 1H), 7.10 (t, J 7.2, 1H), 6.68 (d, J 7.6, 1H), 6.11 (s, 1H), 5.80 (s, 1H), 4.09 - 4.03 (m, 2H), 3.99 - 3.92 (m, 2H), 1.49 (s, 3H).

[0417] Intermediate 49: 3-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)benzaldehyde :

[0418] Intermediate 48 (7.7 g, 25 mmol) was dissolved in a mixture of water (60 ml) and acetone (60 ml). Hydrated oxalic acid (8 g, 125 mmol) was added to the mixture and stirred at 55 °C for 3 hours. Acetone was distilled off from the reaction mixture and the residue was basified with 10% aqueous sodium bicarbonate. The aqueous layer was extracted with DCM (3 * 100 ml) and the combined DCM layers were distilled to give the title compound as a brown solid (6 g). It was used in the next step without further purification. Yield: 91%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.07 (s, 1H), 8.01 - 7.94 (m, 2H), 7.85 - 7.75 (m, 2H), 7.46 (d, J 6.8, 1H), 7.26 (t, J 7.2, 1H), 7.13 (t, J 7.2, 1H), 6.80 (d, J 7.6, 1H), 6.16 (s, 1H), 1.52 (s, 3H).

[0419] Intermediate 50: 3-hydroxy-3-methyl-1-(3-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)phenyl) dihydroindole-2-one :

[0420] According to General Procedure 2, the title compound was synthesized from Intermediate 49 (5.5 g, 20.6 mmol) and Intermediate 3 (10.8 g, 22.6 mmol) as a black gel (7.9 g). Yield: 100%. The compound was used in the next step without any characterization.

[0421] Intermediate 51: 1-(2-fluoro-5-((5-fluoro-3-oxoisobenzofuran-1(3H)-ylidene)methyl)phenyl)-3-hydroxy -3-methyldihydroindole-2-one :

[0422] According to General Procedure 2, the title compound was synthesized from intermediate 7 (2 g, 7 mmol) and intermediate 23 (4 g, 8 mmol) as a black gel (3.1 g). Yield: 100%. The compound was used in the next step without any characterization.

[0423] Intermediate 52: 4-((5-(3-chloro-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1 (2H)-one :

[0424] 4-((5-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one (Example 1) (8 g, 20.1 mmol) and thionyl chloride (33.9 g, 285 mmol) were mixed at 0 °C and refluxed at 90 °C for 1 h. Thionyl chloride was distilled off from the reaction mixture to give a solid. The solid was suspended in water and basified with aqueous sodium bicarbonate. The solid was filtered and washed with a mixture of EtOAc and diethyl ether (1:1) to give the title compound (7.7 g) as a white solid. Yield: 92%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.58 (s, 1H), 8.68 (d, J 1.2, 1H), 8.59 (d, J 3.2, 1H), 8.26 (d, J 7.2, 1H), 8.06 (d, J 8.4, 1H), 7.97 - 7.91 (m, 2H), 7.85 (t, J 7.2, 1H), 7.66 (d, J 6.4, 1H), 7.32 (td, J 8.0, 1.2, 1H), 7.20 (t, J 7.2, 1H), 6.76 (d, J 7.6, 1H) 4.47 (s, 2H), 1.96 (s, 3H). MS (m / z): 417.29 ([M + H] + )

[0425] Intermediate 53: tert-butyl pyridin-3-ylcarbamate :

[0426] 3-Aminopyridine (45 g, 0.478 mol) was dissolved in 2-propanol (150 ml) and water (29 ml) and cooled to 0 °C. Di-tert-butyl dicarbonate (119 g, 0.545 mol) was dissolved in 2-propanol (75 ml) and added to the above mixture at 0 °C. The mixture was stirred at room temperature for 18 h. After 18 h, 2-propanol was distilled off from the reaction mixture to give a residue. Water (100 ml) was added to the residue and the mixture was extracted with MTBE (2 × 200 ml). The combined organic layers were washed with brine (100 ml) and dried over anhydrous Na2SO4. The organic layer was distilled in vacuo to give a crude product. The crude product was purified by column chromatography using EtOAc and petroleum ether (3:7) as the eluent to give the title compound as a white solid (74 g). Yield: 80%.1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 7.77 (s, 1H), 7.36 (d, J 4, 1H), 7.17 (d, J 7.6, 1H), 6.55 (dd, J 7.6, 4, 1H), 0.74 (s, 9H). MS (m / z): 194.93 ([M+H] + )。

[0427] Intermediate 54: ethyl 2-(3-((tert-butoxycarbonyl)amino)pyridin-4-yl)-2-oxoacetate :

[0428] Dissolve intermediate 53 (17 g, 87.5 mmol) in THF (400 ml). Add N,N,N,N-tetramethylethylenediamine (25.33 g, 218 mmol) to the mixture and cool to -78 °C. Add t-BuLi (14.02 g, 219 mmol) dropwise to the above mixture over 45 minutes. Warm the reaction mixture to -10 °C to -20 °C and stir for 2 hours. After 2 hours, cool the reaction mixture to -60 °C again and add diethyl oxalate (38.4 g, 262.6 mmol). Warm the reaction mixture to 0 °C and stir at 0 °C for 3 hours. After 3 hours, quench the reaction mixture with aqueous NH4Cl solution at 0 °C. Extract the aqueous layer with EtOAc (2 * 200 ml). Wash the combined organic layers with water (200 ml) and dry over anhydrous Na2SO4. Distill the organic layer under vacuum to obtain the crude product. Purify the crude product by column chromatography on silica gel (60 - 120 mesh) using EtOAc and petroleum ether (2:8) as the eluent to obtain the title compound as a brown oil (25 g). Yield: 25%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.34 (s, 1H), 8.51 (d, J 2.4, 1H), 8.41 (dd, J 4.8, 2.4, 1H), 7.46 (dd, J 4.8, 2.4, 1H), 4.24 (q, J 7.2, 2H), 1.41 (s, 9H), 1.26 (t, J 7.2, 3H).

[0429] Intermediate 55: 1H-pyrrolo[2,3-c]pyridine-2,3-dione :

[0430] Heat intermediate 54 (1.7 g, 5.77 mmol) to 180 °C under high vacuum for 7 minutes. After 7 minutes, cool the reaction mixture to room temperature. Repeat the reaction on the above scale 6 more times. Purify the combined residue by combinatorial flash method using ethyl acetate as the eluent to obtain the title compound as a yellow solid (2.23 g). Yield: 37%. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 11.20 (s, 1H), 8.44 (t, J 5.2, 1H), 8.33 (d, J 5.2, 1H), 7.41 (t, J 5.2, 1H). MS (m / z): 147.20 ([M-H] - )。

[0431] Intermediate 56: 3-hydroxy-3-methyl-1,3-dihydro-2H-pyrrolo[2,3-c]pyridin-2-one :

[0432] Under a nitrogen atmosphere, intermediate 55 (2.1 g, 14 mmol) was dissolved in THF (5 ml) and cooled to -5 °C. A THF solution of 3M methylmagnesium chloride (14 ml, 43 mmol) was added dropwise to the above mixture. The reaction mixture was stirred at 0 °C to 5 °C for 2 hours. After 2 hours, the reaction mixture was quenched with an aqueous ammonium chloride solution (50 ml) and extracted with MeOH and DCM (1:9) (6 * 50 ml). The combined organic layers were dried over anhydrous Na2SO4. The organic layer was distilled to obtain a crude product. The crude product was purified by the combinatorial flash method using MeOH and DCM (8:92) as the eluent to obtain the title compound (1.2 g) as a brown solid. Yield: 52%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.45 (s, 1H), 8.30 (d, J 4.8, 1H), 8.15 (d, J 4.8, 1H), 7.36 (d, J 4.8, 1H), 6.16 (s, 1H), 1.38 (s, 3H). MS (m / z): 165.05 ([M+H] + )。

[0433] Intermediate 56: 1-(3-(1,3-dioxolan-2-yl)phenyl)-3-hydroxy-3-methyl-1,3-dihydro-2H-pyrro lo[2,3-c]pyridin-2-one :

[0434] According to General Procedure 1, the title compound was synthesized from intermediate 47 (680 mg, 2.5 mmol) and intermediate 56 (400 mg, 2.5 mmol). The crude product was purified by the combinatorial flash method using MeOH and DCM (4:96) as the eluent. The combined pure fractions from the combinatorial flash method were distilled to obtain the title compound as a black gel (300 mg). Yield: 32%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.44 (bs, 1H), 8.06 (bs, 1H), 7.60 (t, J 7.9, 1H), 7.55 - 7.49 (m, 4H), 6.36 (s, 1H), 5.82 (s, 1H), 4.09 - 4.00 (m, 2H), 3.99 - 3.91 (m, 2H), 1.52 (s, 3H). MS (m / z): 313.29 ([M+H]+ )。

[0435] Intermediate 57: 3-(3-hydroxy-3-methyl-2-oxo-2,3-dihydro-1H-pyrrolo[2,3-c]pyridin-1-yl) benzaldehyde :

[0436] Intermediate 56 (280 mg, 0.5 mmol) was dissolved in a mixture of water (4 ml) and acetone (4 ml). Oxalic acid hydrate (570 mg, 4.5 mmol) was added to the mixture and stirred at 55 °C for 4 h. Acetone was distilled off from the reaction mixture and the mixture was basified with 10% aqueous sodium bicarbonate. The aqueous layer was extracted with DCM (3 * 100 ml) and the combined DCM layers were distilled to give the title compound as a brown solid (160 g). It was used in the next step without further purification. Yield: 67%. MS (m / z): 269.22 ([M+H] + )。

[0437] Intermediate 58: 3-hydroxy-3-methyl-1-(3-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)phenyl)- 1,3-dihydro-2H-pyrrolo[2,3-c]pyridin-2-one :

[0438] According to General Procedure 2, the title compound was synthesized from Intermediate 57 (150 mg, 0.56 mmol) and Intermediate 3 (290 mg, 0.62 mmol) as a black gel (210 mg). Yield: 100%. The compound was used in the next step without any characterization. MS (m / z): 383.61 ([M-H]-).

[0439] Intermediate 59: 1-(naphthalen-1-ylmethyl)dihydroindole-2,3-dione :

[0440] Indoline-2,3-dione (37 g, 0.25 mol) was dissolved in DMF (740 ml) and cooled to 0 °C. Sodium hydride was added portionwise to the above mixture at 0 °C and stirred at the same temperature for 10 min. 1-(Chloromethyl)naphthalene was added to the above reaction mixture at 0 °C, stirred at 0 °C for 30 min and stirred at room temperature for 12 h. After 12 h, the reaction mixture was quenched with saturated NH4Cl (1 L) to give a solid. The solid was filtered and washed with water (500 ml). The solid was dissolved in a mixture of MeOH and DCM (1:9) (2 L) and dried over anhydrous Na2SO4. The solution was distilled to give the crude product. The crude product was purified by column chromatography on silica gel (60 - 120 mesh) using DCM as the eluent. The combined pure fractions from the column were distilled to give the title compound (43.8 g) as a brown solid. Yield: 61%. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.20 (d, J 8.4, 1H), 7.99 (d, J 8.4, 1H), 7.88 (d, J 8.4, 1H), 7.68 - 7.50 (m, 5H), 7.42 (t, J 8, 1H), 7.12 (t, J 7.6, 1H), 6.90 (d, J 8, 1H), 5.39 (s, 2H).

[0441] Intermediate 60: (R)-3-methyl-1-(naphthalen-1-ylmethyl)-2-oxodihydroindol-3-yl acetate :

[0442] Dissolve (R)-N-(4-chlorobenzyl)-2-hydroxy-2-phenylacetamide (3.148 g, 12.03 mmol) in DCM (1200 ml) and stir under a nitrogen atmosphere. Add 2M dimethylzinc (60.14 ml, 120.3 mmol) to the mixture and stir at room temperature for 30 minutes. Dissolve intermediate 59 (17.28 g, 60.14 mmol) in DCM (600 ml) and add it dropwise to the above reaction mixture over 1 hour. Stir the reaction mixture at room temperature for another 1 hour.

[0443] Cool the reaction mixture to 0 °C and add acetic anhydride (22.7 ml, 240 mmol) dropwise to the reaction mixture. After adding acetic anhydride, warm the reaction mixture to room temperature and stir for 1 hour. At this stage, add pyridine (19.03 ml, 240 mmol) and DMAP (733 mg, 6 mmol) to the reaction mixture and stir at room temperature for 1 hour. After 1 hour, add pyridine (9.52 ml, 120 mmol), DMAP (733 mg, 6 mmol) and acetic anhydride (11.37 ml, 120 mmol) again, and stir the reactant at room temperature for 1 hour. Quench the reaction mixture with an aqueous ammonium chloride solution (75 g in 750 ml of water). Separate the organic layer and the aqueous layer. Extract the aqueous layer with DCM (2 * 350 ml). Wash the combined DCM layers with water (700 ml) and brine (700 ml). Distill the organic layer to obtain a crude solid (22 g). Suspend the crude solid (22 g) in a mixture of EtOAc (12 ml) and hexane (108 ml) and stir for 2 hours. Filter the solid and wash it with hexane (35 ml). Dry the solid under vacuum to obtain the title compound (17 g) as an off-white solid. Yield: 82%. Chiral HPLC purity: 90.45%, retention time: 22.92 min. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.18 (d, J 8, 1H), 7.97 (d, J 8, 1H), 7.85 (d, J 8, 1H), 7.58 (quintet, J 7.2, 2H), 7.45 (d, J 6.8, 1H), 7.41 (d, J 8, 1H), 7.37 (d, J 7.2, 1H), 7.18 (t, J 8, 1H), 7.02 (t, J 7.6, 1H), 6.75 (d, J 8, 1H), 5.44 (d, J 16.8, 1H), 5.32 (d, J 16.8, 1H), 2.07 (s, 3H), 1.59 (s, 3H).

[0444] Intermediate 61: (R)-3-methyl-2-oxodihydroindol-3-yl acetate :

[0445] Intermediate 60 (1.25 g, 3.62 mmol) was suspended in chlorobenzene (70 ml), and N-bromosuccinimide (772 mg, 4.34 mmol) and AIBN (118 mg, 0.723 mmol) were added. The mixture was refluxed at 130 °C for 3 hours. After 3 hours, chlorobenzene was distilled from the reaction mixture to obtain the crude product. The crude product was diluted with water and extracted with EtOAc. The organic layer was dried over anhydrous Na2SO4 and distilled to obtain the crude product. The crude product was purified by a combined flash method using EtOAc and petroleum ether (2:8) as the eluent to obtain the title compound as a light brown solid (360 mg). Yield: 48%. Chiral HPLC purity: 89.85%, retention time: 5.18 min. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.52 (s, 1H), 7.24 (d, J 7.2, 1H), 7.22 (td, J 7.6, 1.2, 1H), 6.95 (td, J 7.6, 1.2, 1H), 6.82 (d, J 8, 1H), 2.00 (s, 3H), 1.47 (s, 3H).

[0446] Intermediate 62: (R)-(+)-3-hydroxy-3-methyldihydroindole-2-one :

[0447] Intermediate 61 (300 mg, 1.46 mmol) and LiOH (135 mg, 2.2 mmol) were suspended in MeOH (1.5 ml) and THF (1.5 ml). The mixture was stirred at room temperature for 3 hours. After 3 hours, the reaction mixture was diluted with water (10 ml) and extracted with EtOAc (2 * 10 ml). The combined organic layers were dried over anhydrous Na2SO4 and distilled to obtain the title compound as a light brown solid (150 mg). Yield: 48%. Chiral HPLC purity: 82.81%, retention time: 3.01 min. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.19 (s, 1H), 7.26 (d, J 7.2, 1H), 7.17 (td, J 7.6, 1.2, 1H), 6.94 (td, J 7.6, 1.2, 1H), 6.78 (d, J 7.6, 1H), 5.82 (s, 1H), 1.33 (s, 3H).

[0448] Intermediate 63: 1-(5-(1,3-dioxolan-2-yl)-2-fluorophenyl)dihydroindole-2-one :

[0449] According to General Procedure-1, the title compound was synthesized from Intermediate 1 (10 g, 34.00 mmol) and oxindole (4.53 g, 34.00 mmol) as a brown liquid (700 mg). Yield: 6%. The compound was used in the next step without any characterization.

[0450] Intermediate 64: 4-fluoro-3-(2-oxodihydroindol-1-yl)benzaldehyde :

[0451] Intermediate 63 (350 mg, 1.17 mmol) was dissolved in THF (5 ml) and 6N hydrochloric acid (4 ml) was added. The mixture was stirred at room temperature for 1 hour. After 1 hour, the reaction mixture was cooled to 0 °C and the pH was adjusted to about 7 using saturated aqueous sodium bicarbonate (50 ml). The aqueous solution of the reaction mixture was extracted with MeOH:DCM (1:9) and the organic layer was evaporated on a rotary evaporator to give the crude product. The crude product was purified by combinatorial flash method using MeOH and DCM (1:9) as the eluent. The combined pure fractions from the combinatorial flash method were distilled on a rotary evaporator to give the title compound as a brown gel (160 mg). Yield: 53.69%. The compound was used in the next step without any characterization.

[0452] Intermediate 65: 1-(2-Fluoro-5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)phenyl)indolin- -2-one :

[0453] According to General Procedure 2, the title compound (240 mg) was synthesized from Intermediate 64 (150 mg, 0.58 mmol) and Intermediate 3 (500 mg, 1.06) as a yellow gel. Yield: 100%. The compound was used in the next step without any characterization.

[0454] Intermediate 66: tert-Butyl 2-oxoindoline-1-carboxylate :

[0455] Dissolve oxindole (16 g, 0.12 mol) in THF (382 ml) and cool to 0 °C. Add sodium carbonate (101.89 g, 0.96 mol) and boc-anhydride (41.4 ml, 0.18 mol) to the mixture at 0 °C. Heat the reaction mixture to 70 °C and stir at the same temperature for 24 hours. Cool the reaction mixture to room temperature and filter the insoluble solid from the reaction mixture. Stir the filtered solid with ethyl acetate (200 ml) and filter. Distill the combined filtrate to obtain the crude product. Purify the crude product by flash chromatography using ethyl acetate and petroleum ether (2:98) as the eluent. Distill the combined pure fractions from flash chromatography to obtain the title compound as a white solid (5.5 g). Yield: 19.6%. 1 H-NMR (δ ppm, CDCl3, 400 MHz): 7.78 (d, J 8.2, 1H), 7.30 (t, J 7.8, 1H), 7.24 (d, J 7.8, 1H), 7.13 (t, J 7.4, 1H), 3.65 (s, 2H), 1.64 (s, 9H).

[0456] Intermediate 67: tert-Butyl 3,3-dimethyl-2-oxoindoline-1-carboxylate :

[0457] Dissolve intermediate 66 (5.5 g, 23.6 mmol) and methyl iodide (4.42 ml, 70.77 mmol) in THF (45 ml) and cool to 0 °C. Add sodium hydride (60%) (2.06 g, 51.9 mmol) to the mixture portionwise at the same temperature. Warm the reaction mixture to room temperature and stir for 3 hours. Cool the reaction mixture to 0 °C and quench with water (100 ml). Extract the aqueous solution of the reaction mixture with ethyl acetate and distill the ethyl acetate layer to obtain the crude product. Purify the crude product by flash chromatography using ethyl acetate and petroleum ether (15:85) as the eluent. Distill the combined pure fractions from flash chromatography to obtain the title compound as an off-white solid (2.4 g). Yield: 39%. 1 H-NMR (δ ppm, CDCl3, 400 MHz): 7.84 (d, J 8.1, 1H), 7.29 (d, J 7.2, 1H), 7.24 - 7.13 (m, 2H), 1.65 (s, 9H), 1.42 (s, 6H).

[0458] Intermediate 68: 3,3-Dimethylindolin-2-one :

[0459] The intermediate was prepared according to the report in the literature (Ref: Li, Honghe et al., Angewandte Chemie, International Edition (2019), 58(20), 6732 - 6736). The intermediate 67 (2 g, 7.65 mmol) was dissolved in dichloromethane (10 ml) and cooled to 0 °C. Trifluoroacetic acid (5.9 ml, 76.5 mmol) was added to the above reaction mixture and stirred at 0 °C for 30 minutes. The pH of the reaction mixture was adjusted to about 7 using saturated aqueous sodium bicarbonate. The aqueous layer was extracted with MeOH:DCM (1:9) and the organic layer was evaporated to obtain the crude product. The crude product was suspended in diethyl ether (20 ml) and stirred to obtain a solid. The solid was filtered and dried in vacuo to obtain the title compound (1.25 g) as a brown solid. Yield: 100%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.29 (s, 1H), 7.25 (d, J 7.2, 1H), 7.14 (t, J 7.6, 1H), 6.94 (t, J 7.5, 1H), 6.82 (d, J 7.6, 1H), 1.22 (s, 6H).

[0460] Intermediate 69: 1-(5-(1,3-Dioxolan-2-yl)-2-fluorophenyl)-3,3-dimethylindolin-2-one :

[0461] According to General Procedure 1, the title compound was synthesized from Intermediate 1 (1.64 g, 5.58 mmol) and Intermediate 68 (0.9 g, 5.58). The crude product was purified by combinatorial flash method using ethyl acetate and petroleum ether (20:80) as the eluent. The combined pure fractions from the combinatorial flash method were distilled to obtain the title compound as a brown solid. Yield: 32%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 7.63 - 7.60 (m, 2H), 7.50 (t, J 9.5, 1H), 7.45 (d, J 7.1, 1H), 7.21 (t, J7.7, 1H), 7.10 (t, J 7.2, 1H), 6.56 (d, J 7.7, 1H), 5.79 (s, 1H), 4.10 - 4.03 (m, 2H), 3.97 - 3.90 (m, 2H), 1.41 (s, 3H), 1.37 (s, 3H).

[0462] Intermediate 70: 3-(3,3-Dimethyl-2-oxoindolin-1-yl)-4-fluorobenzaldehyde :

[0463] The intermediate 69 (600 mg, 1.84 mmol) was dissolved in THF (6 ml) and 6N hydrochloric acid (2 ml) was added. The mixture was stirred at room temperature for 30 minutes. After 30 minutes, the reaction mixture was cooled to 0 °C and the pH was adjusted to about 7 using saturated aqueous sodium bicarbonate (50 ml). The aqueous solution of the reaction mixture was extracted with a dichloromethane and MeOH mixture (1:9). The organic layer was dried over anhydrous Na2SO4 and evaporated to give the crude product. The crude product was stirred with petroleum ether (3 ml) to give a solid and the solid was filtered. The solid was dried in vacuo to give the title compound as a brown solid (450 mg). Yield: 86%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.02 (s, 1H), 8.17 (d, J 7.2, 1H), 8.14 - 8.11 (m, 1H), 7.73 (t, J 9.4, 1H), 7.48 (d, J 7, 1H), 7.22 (t, J 6.8, 1H), 7.13 (t, J 7.4, 1H), 6.67 (d, J 7.8, 1H), 1.43 (s, 3H), 1.39 (s, 3H).

[0464] Intermediate 71: 1-(2-Fluoro-5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)phenyl)-3,3-dimeth ylindolin-2-one :

[0465] According to General Procedure 2, the title compound was synthesized from intermediate 70 (450 mg, 1.58 mmol) and intermediate 3 (1.36 g, 2.86 mmol) as a yellow solid. Yield: 100%.

[0466] Intermediate 72: 1-(5-(1,3-Dioxolan-2-yl)pyridin-3-yl)-3,3-dimethylindolin-2-one :

[0467] According to General Procedure 1, the title compound was synthesized from intermediate 4 (1.28 g, 5.58 mmol) and intermediate 68 (899 mg, 5.58). The crude product was purified by combinatorial flash method using ethyl acetate and petroleum ether (20:80) as the eluent. The combined pure fractions from the combinatorial flash method were distilled to give the title compound as a brown solid (800 mg). Yield: 46%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.70 (d, J 6.5, 2H), 7.96 (s, 1H), 7.46 (d, J 7.7, 1H), 7.22 (t, J7.7, 1H), 7.12 (t, J 7.4, 1H), 6.75 (d, J 7.7, 1H), 5.92 (s, 1H), 4.12 - 4.07 (m, 2H), 4.01 - 3.95 (m, 2H), 1.40 (s, 6H).

[0468] Intermediate 73: 5-(3,3-Dimethyl-2-oxoindolin-1-yl)nicotinaldehyde :

[0469] The intermediate 72 (800 mg, 2.6 mmol) was dissolved in THF (15 ml) and concentrated hydrochloric acid (2.5 ml) was added. The mixture was stirred at 80 °C for 2 h. After 2 h, the reaction mass was cooled to room temperature and water (50 ml) was added. The pH of the reaction mixture aqueous solution was adjusted to about 7 using saturated aqueous sodium bicarbonate. The reaction mixture aqueous solution was extracted with dichloromethane. The organic layer was dried over anhydrous Na2SO4 and evaporated to give the title compound as a brown solid (600 mg). Yield: 87%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.17 (s, 1H), 9.12 (s, 1H), 8.99 (s, 1H), 8.40 (s, 1H), 7.48 (d, J 7.2, 1H), 7.24 (t, J 7.4, 1H), 7.14 (t, J 7.3, 1H), 6.89 (d, J 7.7, 1H), 1.42 (s, 6H).

[0470] Intermediate 74: 3,3-Dimethyl-1-(5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin-3- yl)indolin-2-one :

[0471] According to General Procedure 2, the title compound was synthesized from intermediate 73 (550 mg, 2.7 mmol) and intermediate 3 (1.76 g, 3.71 mmol) as a yellow solid (620 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0472] Intermediate 75: Diethyl 2-(3-nitropyridin-2-yl)malonate :

[0473] Sodium hydride (60%) (18.22 g, 0.46 mol) was suspended in DMSO (360 ml) under a nitrogen atmosphere and cooled to 0 °C. Diethyl malonate (69.18 ml, 0.46 mol) was added to the above mixture, warmed to room temperature and stirred for 30 min. 2-Chloro-3-nitropyridine (31 g, 0.195 mol) was added to the above reaction mixture and heated to 100 °C for 15 min. After 15 min, the reaction mixture was cooled to 0 °C and quenched with aqueous ammonium chloride solution. The reaction mixture aqueous solution was extracted with ethyl acetate (2 * 250 ml) and the ethyl acetate layer was washed with water (2 * 500 ml). The organic layer was distilled in vacuo to give the title compound as a brown gel (55.2 g). Yield: 100%. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.88 (dd, J 4.6, 1.3, 1H), 8.61 (dd, J 8.4, 1.4, 1H), 7.75 (dd, J 8.3, 4.7, 1H), 5.59 (s, 1H), 4.20 - 4.14 (m, 4H), 1.17 (t, J 7.1, 6H).

[0474] Intermediate 76: Ethyl 2-(3-nitropyridin-2-yl)acetate :

[0475] A mixture of intermediate 75 (55 g, 0.282 mol), lithium chloride (20.65 g, 0.487 mol), DMSO (550 ml) and water (3.5 ml, 0.194 mol) was heated to 100 °C for 16 h. The reaction mixture was cooled to room temperature and quenched with brine solution (500 ml). The aqueous solution of the reaction mixture was extracted with ethyl acetate (2 × 200 ml). The combined organic layers were washed with water (5 × 200 ml) and dried over anhydrous Na2SO4. The organic layer was distilled to obtain the crude product. The crude product was purified by combinatorial flash method using ethyl acetate and petroleum ether (15:85) as the eluent. The pure fractions from the combinatorial flash method were distilled to obtain the title compound as a brown gel (35.9 g). Yield: 88%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.84 (d, J 3.5, 1H), 8.54 (d, J 8.3, 1H), 7.68 (dd, J 8.2, 4.8, 1H), 4.23 (s, 2H), 4.08 (q, J 7.1, 2H), 1.15 (t, J 7.1, 3H).

[0476] Intermediate 77: Ethyl 2-methyl-2-(3-nitropyridin-2-yl)propionate :

[0477] Intermediate 76 (3.8 g, 18 mmol) was dissolved in DMF (30 ml) under a nitrogen atmosphere. Methyl iodide (3.3 ml, 54 mmol) and 18-crown-6 (0.48 g, 1.8 mmol) were added to the solution and cooled to 0 °C. Sodium hydride (60%) (1.59 g, 39.8 mmol) was added portionwise to the reaction mixture and stirred at 0 °C for 1 h. The reaction mixture was quenched with water at 0 °C and extracted with ethyl acetate (2 × 100 ml). The combined organic layers were washed with water (5 × 100 ml) and evaporated on a rotary evaporator to obtain the crude product. The crude product was purified by combinatorial flash method using ethyl acetate and petroleum ether (8:92) as the eluent. The pure fractions from the combinatorial flash method were combined and distilled to obtain the title compound as a brown gel (3 g). Yield: 70%. 1H-NMR (δ ppm, CDCl3, 400 MHz): 8.79 (d, J 4.5, 1H), 8.24 (d, J 8.1, 1H), 7.40 (dd, J 8.1, 4.6, 1H), 4.12 (q, J 7.1, 2H), 1.71 (s, 6H), 1.20 (t, J 7.1, 3H).

[0478] Intermediate 78: 3,3-Dimethyl-1,3-dihydro-2H-pyrrolo[3,2-b]pyridin-2-one :

[0479] The intermediate 77 (3 g, 12 mmol) was suspended in ethanol (25 ml) and ammonium formate (3.17 g, 50.37 mmol) and 10% Pd / C (300 mg) were added. The mixture was stirred at 100 °C for 2 h. The reaction mixture was filtered through a bed of diatomaceous earth and the diatomaceous earth bed was washed with MeOH and DCM (1:9) (250 ml). The combined filtrate was distilled on a rotary evaporator to give a residue. The residue was dissolved in a mixture of MeOH and DCM (1:9) (250 ml) and washed with water (2 * 100 ml). The organic layer was dried over anhydrous Na2SO4 and distilled to give the title compound as a brown solid (1.45 g). Yield: 71%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.51 (s, 1H), 8.07 (d, J 4.4, 1H), 7.20 - 7.11 (m, 2H), 1.23 (s, 6H).

[0480] Intermediate 79: 1-(5-(1,3-Dioxolan-2-yl)pyridin-3-yl)-3,3-dimethyl-1,3-dihydro-2H-pyrro lo[3,2-b]pyridin-2-one :

[0481] According to General Procedure 1, the title compound was synthesized from intermediate 4 (1.5 g, 6.5 mmol) and intermediate 78 (1.06 g, 6.5 mmol). The crude product obtained was purified by the combination flash method using MeOH and DCM (3:97) as the eluent. The combined pure fractions from the combination flash method were distilled to give the title compound as a pale yellow gel (1.2 g). Yield: 59%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.75 (s, 1H), 8.70 (s, 1H), 8.24 (d, J 4.8, 1H), 8.01 (s, 1H), 7.27 - 7.21 (m, 1H), 7.18 (d, J 8, 1H), 5.92 (s, 1H), 4.11 - 4.05 (m, 2H), 4.01 - 3.96 (m, 2H), 1.41 (s, 6H).

[0482] Intermediate 80: 5-(3,3-Dimethyl-2-oxo-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-1-yl)nicotina ldehyde :

[0483] The intermediate 79 (620 mg, 2 mmol) was dissolved in THF (5 ml) and concentrated hydrochloric acid (1.5 ml) was added. The mixture was stirred at 80 °C for 3 hours. After 3 hours, the reaction mass was cooled to 0 °C and the pH of the reaction mixture was adjusted to about 7 using saturated aqueous sodium bicarbonate. The aqueous solution of the reaction mixture was extracted with MeOH and DCM (1:9) (2 * 100 ml). The combined organic layers were dried over anhydrous Na2SO4 and evaporated to give the title compound as a pale yellow gel (532 mg). Yield: 100%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.18 (s, 1H), 9.13 (s, 1H), 9.02 (s, 1H), 8.43 (s, 1H), 8.26 (d, J 4.2, 1H), 7.32 (d, J 8, 1H), 7.29 - 7.22 (m, 1H), 1.43 (s, 6H).

[0484] Intermediate 81: 3,3-Dimethyl-1-(5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin-3- yl)-1,3-dihydro-2H-pyrrolo[3,2-b]pyridin-2-one :

[0485] According to General Procedure 2, the title compound was synthesized from intermediate 80 (532 mg, 2 mmol) and intermediate 3 (1.70 g, 4 mmol) as a yellow solid (762 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0486] Intermediate 82: 1-(5-(1,3-Dioxolan-2-yl)-2-fluorophenyl)-3,3-dimethyl-1,3-dihydro-2H-pyrro lo[3,2-b]pyridin-2-one :

[0487] According to General Procedure 1, the title compound was synthesized from intermediate 1 (730 mg, 2.48 mmol) and intermediate 78 (402 mg, 2.48 mmol). The crude product obtained was purified by the combined flash method using MeOH and DCM (2.5:97.5) as the eluent. The combined pure fractions from the combined flash method were distilled to give the title compound as a brown gel (460 mg). Yield: 57%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.23 (d, J 5, 1H), 7.67 (dd, J 7.2, 2, 1H), 7.65 - 7.60 (m, 1H), 7.51 (t, J 9.9, 1H), 7.23 (dd, J 8, 5, 1H), 6.99 (d, J 8, 1H), 5.79 (s, 1H), 4.10 - 4.03 (m, 2H), 3.97 - 3.91 (m, 2H), 1.43 (s, 3H), 1.38 (s, 3H).

[0488] Intermediate 83: 3-(3,3-Dimethyl-2-oxo-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-1-yl)-4- fluorobenzaldehyde :

[0489] The intermediate 82 (440 mg, 1.34 mmol) was dissolved in THF (10 ml) and concentrated hydrochloric acid (2 ml) was added. The mixture was stirred at room temperature for 30 minutes. After 30 minutes, the reaction material was cooled to 0 °C and the pH of the reaction mixture was adjusted to about 7 using saturated aqueous sodium bicarbonate. The aqueous solution of the reaction mixture was extracted with MeOH and DCM (1:9). The organic layer was dried over anhydrous Na2SO4 and evaporated to give the title compound as a pale yellow gel (380 mg). Yield: 100%. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.02 (s, 1H), 8.25 (dd, J 5, 1.2, 1H), 8.23 (dd, J 6.8, 4.8, 1H), 8.16 - 8.10 (m, 1H), 7.74 (t, J 9.6, 1H), 7.25 (dd, J 8, 5, 1H), 7.13 (d, J 8, 1H), 1.45 (s, 3H), 1.40 (s, 3H).

[0490] Intermediate 84: 1-(2-Fluoro-5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)phenyl)-3,3-dimeth yl-1,3-dihydro-2H-pyrrolo[3,2-b]pyridin-2-one :

[0491] According to General Procedure 2, the title compound was synthesized from intermediate 83 (380 mg, 1.33 mmol) and intermediate 3 (1.70 g, 4 mmol) as a yellow gel (404 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0492] Intermediate 85: tert-Butyl 2'-oxospiro[cyclopropane-1,3'-dihydroindole]-1'-carboxylate :

[0493] Under a nitrogen atmosphere, intermediate 66 (2.5 g, 10.72 mmol) was dissolved in DMF (25 ml). 1,2-Dibromoethane (2.77 ml, 32.16 mmol) and 18-crown-6 (0.28 g, 1.072 mmol) were added to the solution and cooled to 0 °C. Sodium hydride (60%) (943 mg, 23.6 mmol) was added portionwise to the mixture at 0 °C. The reaction mixture was stirred at room temperature for 3 hours and at 70 °C for 3 hours. The reaction mixture was quenched with water at 0 °C and extracted with MeOH and DCM (1:9) (3 * 200 ml). The combined organic layers were washed with water (3 * 250 ml) and evaporated to give the crude product. The crude product was purified by combinatorial flash method using ethyl acetate and petroleum ether (4:96) as the eluent. The pure fractions from the combinatorial flash method were combined and distilled to give the title compound as a white solid (1.1 g). Yield: 40%. 1H-NMR (δ ppm, CDCl3, 400 MHz): 7.90 (d, J 8.2, 1H), 7.30 - 7.22 (m, 1H), 7.12 (t, J 7.5, 1H), 6.81, (d, J 7.4, 1H), 1.82 (q, J 4.2, 2H), 1.65 (s, 9H), 1.54 (q, J 3.9, 2H).

[0494] Intermediate 86: Spiro[cyclopropane-1,3-dihydroindole]-2'-one :

[0495] The intermediate 85 (1.1 g, 4.24 mmol) was dissolved in dichloromethane (40 ml) and cooled to 0 °C. Trifluoroacetic acid (3.27 ml, 42.4 mmol) was added to the mixture and stirred at 0 °C for 30 minutes. The pH of the reaction mixture was adjusted to approximately 7 using saturated aqueous sodium bicarbonate. The aqueous layer was extracted with MeOH:DCM (1:9) (4 * 100 ml) and the organic layer was evaporated to give the title compound as a brown solid (650 mg). Yield: 96%.

[0496] 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 10.52 (s, 1H), 7.13 (td, J 7.6, 1.7, 1H), 6.97 - 6.85 (m, 3H), 1.56 - 1.50 (m, 2H), 1.46 - 1.41 (m, 2H).

[0497] Intermediate 87: 1'-(5-(1,3-dioxolan-2-yl)pyridin-3-yl)spiro[cyclopropane-1,3'-dihydroindole]-2'-one Intermediate 88: 5-(2'-oxospiro[cyclopropane-1,3'-dihydroindole]-1'-yl)nicotinaldehyde :

[0498] According to General Procedure 1, the title compound was synthesized from intermediate 4 (940 mg, 4.08 mmol) and intermediate 86 (649 mg, 4.08 mmol). The crude product obtained was purified by combinatorial flash method using ethyl acetate and petroleum ether (35:65) as the eluent. The combined pure fractions from the combinatorial flash method were distilled to give the title compound as a brown gel (501 mg). Yield: 40%. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 8.74 (d, J 2.3, 1H), 8.70 (d, J 1.7, 1H), 7.97 (d, J2.1, 1H), 7.25 - 7.19 (m, 1H), 7.15 - 7.06 (m, 2H), 6.84 (d, J 7.8, 1H), 5.92 (s, 1H), 4.11 - 4.03 (m, 2H), 4.01 - 3.93 (m, 2H), 1.78 - 1.72 (m, 2H), 1.67 - 1.62 (m, 2H).

[0499] Intermediate 89: 1'-(5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin-3-yl)spiro[cyclopropane-1,3'-dihydroindole]-2'-one :

[0500] The intermediate 87 (480 mg, 1.56 mmol) was dissolved in THF (15 ml) and concentrated hydrochloric acid (2 ml) was added. The mixture was refluxed at 80 °C for 3.5 h. After 3.5 h, the reaction material was cooled to 0 °C and the pH of the reaction mixture was adjusted to about 7 using saturated aqueous sodium bicarbonate. The reaction mixture aqueous solution was extracted with MeOH and DCM (1:9) (2 * 150 ml). The organic layer was dried over anhydrous Na2SO4 and evaporated to give the title compound as a yellow gel (411 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0501] Example 1 4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one :

[0502] According to general procedure 2, the title compound was synthesized from intermediate 88 (411 mg, 1.56 mmol) and intermediate 3 (1.33 g, 2.8 mmol) as a yellow gel (411 mg). Yield: 100%. The compound was used in the next step without any characterization.

[0503] Example 2

[0504] (R)-(+)-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin- 1(2H)-one :

[0505] According to general procedure 3, the title compound was synthesized from intermediate 11 (1 g, 2.6 mmol), hydrazine hydrate (156 mg, 3.12 mmol) and acetic acid (78 mg, 1.3 mmol). Purification: combined rapid method. Eluent: MeOH and DCM: 5.3:94.7. Appearance: off-white solid. Yield: 236 mg. % Yield: 23%. M.P.: 110 °C - 113 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.59 (s, 1H), 8.65 (s, 1H), 8.52 (s, 1H), 8.26 (d, J 7.8, 1H), 8.06 (d, J8.1, 1H), 7.93 (t, J 8, 1H), 7.85 (t, J 8, 1H), 7.79 (s, 1H), 7.43 (d, J 7.2, 1H), 7.21 (t, J 7.6, 1H), 7.11 (t, J 7.2, 1H), 6.69 (d, J 7.7, 1H), 6.12 (s, 1H), 4.46 (s, 2H), 1.48 (s, 3H) MS (m / z): 399.29 ([M+H] + )

[0506] Method-1

[0507] Method-2 Method-3 :

[0508] Example 3 :

[0509] Using the preparation method reported in General Procedure 4, the title compound was resolved into pure enantiomers from the racemic mixture of 4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one (Example 1). Chiral HPLC purity: 99.16, retention time: 11.48 min. M.P.: 97 °C - 100 °C. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.58 (s, 1H), 8.65 (d, J 1.7, 1H), 8.52 (d, J 2.2, 1H), 8.26 (d, J 7.4, 1H), 8.06 (d, J 8.0, 1H), 7.93 (t, J 7.8, 1H), 7.85 (t, J 7.8, 1H), 7.79 (t, J 2.0, 1H), 7.43 (d, J 6.9, 1H), 7.22 (t, J 7.8, 1H), 7.11 (t, J 7.6, 1H), 6.69 (d, J 7.6, 1H), 6.12 (s, 1H), 4.47 (s, 2H), 1.48 (s, 3H). MS (m / z): 399.39 ([M+H] + )。[α] D 25 : +30.36° [MeOH: chloroform (1:9); c 1.0]

[0510] (S)-(-)-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin- :

[0511] 5-Bromonicotinaldehyde was converted to the acetal by treatment with ethylene glycol. The above acetal was reacted with (R)-(+)-3-methyl-2-oxoindolin-3-yl acetate under Buchwald conditions to give (R)-(+)-1-(5-(1,3-dioxolan-2-yl)pyridin-3-yl)-3-methyl-2-oxoindolin-3-yl acetate. (R)-(+)-1-(5-(1,3-Dioxolan-2-yl)pyridin-3-yl)-3-methyl-2-oxoindolin-3-yl was deprotected under acidic conditions to give (R)-(+)-1-(5-formylpyridin-3-yl)-3-methyl-2-oxoindolin-3-yl acetate. The resulting (R)-(+)-1-(5-formylpyridin-3-yl)-3-methyl-2-oxoindolin-3-yl acetate was reacted with (3-oxo-1,3-dihydroisobenzofuran-1-yl)triphenylphosphonium bromide under Wittig reaction conditions to give (R)-(+)-3-methyl-2-oxo-1-(5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin-3-yl)indolin-3-yl acetate. (R)-(+)-3-methyl-2-oxo-1-(5-((3-oxoisobenzofuran-1(3H)-ylidene)methyl)pyridin-3-yl)indolin-3-yl acetate was reacted with hydrazine hydrate to give (R)-(+)-4-((5-(3-hydroxy-3-methyl-2-oxoindolin-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one.

[0512] 1(2H)-one The title compound was also synthesized according to the procedure in Example-1 and using the chiral intermediate-5 (R)-(+)-3-hydroxy-3-methylindolin-2-one.

[0513] Example 4

[0514] 4-(4-fluoro-3-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one Example 5 :

[0515] Using the preparation method reported in General Procedure 4, the title compound was resolved into pure enantiomers from the racemic mixture of 4-((5-(3-hydroxy-3-methyl-2-oxoindolin-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one (Example 1). Chiral HPLC purity: 99.82, retention time: 13.61 min. M.P.: 94 °C - 97 °C. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.59 (s, 1H), 8.65 (d, J 1.7, 1H), 8.52 (d, J 2.2, 1H), 8.26 (d, J 7.8, 1H), 8.06 (d, J 7.9, 1H), 7.93 (t, J 7.6, 1H), 7.85 (t, J 8.0, 1H), 7.79 (t, J 2, 1H), 7.43 (d, J 7.2, 1H), 7.21 (t, J 7.8, 1H), 7.11 (t, J 7.8, 1H), 6.69 (d, J 7.8, 1H), 6.12 (s, 1H), 4.46 (s, 2H), 1.48 (s, 3H). MS (m / z): 399.41 ([M+H] + )。[α] D 25 : -26.44° [MeOH: chloroform (1:9); c 1.0]

[0516] 4-((5-(3-ethyl-3-hydroxy-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-

[0517] one :

[0518] According to the general procedure 3, the title compound was synthesized from intermediate 8 (800 mg, 1.99 mmol), hydrazine hydrate (119 mg, 2.4 mmol) and acetic acid (59 mg, 1 mmol). Purification: combined rapid method. Eluent: MeOH and DCM: 3:97. Appearance: off-white solid. Yield: 240 mg. % Yield: 29%. M.P.: 131 °C - 134 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.60 (s, 1H), 8.25 (d, J 7.7, 1H), 7.99 (d, J 7.9, 1H), 7.90 (t, J 7.3, 1H), 7.81 (t, 7.3, 1H), 7.54 - 7.49 (m, 1H), 7.47 - 7.35 (m, 3H), 7.24 - 7.17 (m, 1H), 7.09 (t, J 7.3, 1H), 6.46 (d, J7.7, 1H), 6.25 (s, 0.4H), 6.13 (s, 0.6H), 4.39 (s, 1.2H), 4.36 (s, 0.8H), 1.50 (s, 1.8H), 1.45 (s, 1.2H). MS (m / z): 416.1 ([M+H] + )。

[0519] Example 6

[0520] 7-fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1 (2H)-one :

[0521] According to General Procedure 3, the title compound was synthesized from Intermediate 19 (900 mg, 2.26 mmol), hydrazine hydrate (135 mg, 2.71 mmol) and acetic acid (67 mg, 1.13 mmol). Purification: combined rapid method. Eluent: MeOH and DCM: 5:95. Appearance: off-white solid. Yield: 200 mg. % Yield: 21%. M.P.: 117 °C - 120 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.58 (s, 1H), 8.65 (d, J 1.4, H), 8.50 (d, J 2.1, 1H), 8.26 (d, J 7.7, 1H), 8.07 (d, J 8, 1H), 7.94 (t, J 7, 1H), 7.85 (t, J 7.2, 1H), 7.79 (s, 1H), 7.40 (d, J7.2, 1H), 7.23 (t, J 7.1, 1H), 7.12 (t, J 7.5, 1H), 6.70 (d, J 7.8, 1H), 6.12 (s, 1H), 4.47 (s, 2H), 1.94 - 1.83 (m, 2H), 0.68 (t, J 7.4, 3H). MS (m / z): 413.43 ([M+H] + )。

[0522] Example 7

[0523] (+)-7-fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthal azin-1(2H)-one :

[0524] According to General Procedure 3, the title compound was synthesized from Intermediate 24 (650 mg, 1.61 mmol), hydrazine hydrate (97 mg, 1.94 mmol) and acetic acid (48 mg, 0.81 mmol). Purification: combined rapid method. Eluent: MeOH and DCM: 4:96. Appearance: off-white solid. Yield: 110 mg. % Yield: 16%. M.P.: 133 °C - 136 °C. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.69 (s, 1H), 8.65 (d, J 1.5, 1H), 8.53 (d, J 2.1, 1H), 8.19 (dd, J 8.9, 5.0, 1H), 7.94 (dd, J 8.8, 2.7, 1H), 7.84 (td, J 8.8, 2.7, 1H), 7.81 (t, J 2.7, 1H), 7.44 (d, J 7, 1H), 7.23 (t, J 7.7, 1H), 7.11 (t, J 7.4, 1H), 6.71 (d, J 7.8, 1H), 6.13 (s, 1H), 4.47 (s, 2H), 1.48 (s, 3H). MS (m / z): 417.47 ([M-H]-).

[0525] Example 8

[0526] (-)-7-fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthal azin-1(2H)-one :

[0527] Using the preparation method reported in General Procedure 4, the title compound was resolved from the racemic mixture of 7-fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one (Example 6) into pure enantiomers. Chiral HPLC purity: 99.54, retention time: 5.86 min. M.P.: 148 °C - 151 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.69 (s, 1H), 8.65 (d, J 1.6, 1H), 8.53 (d, J 2.0, 1H), 8.19 (dd, J 9.2, 5.2, 1H), 7.94 (dd, J 8.8, 2.7, 1H), 7.84 (td, J 8.8, 2.7, 1H), 7.80 (t, J 2.7, 1H), 7.44 (d, J 7, 1H), 7.23 (t, J 7.6, 1H), 7.12 (t, J 7.6, 1H), 6.71 (d, J 7.6, 1H), 6.13 (s, 1H), 4.47 (s, 2H), 1.48 (s, 3H). MS (m / z): 417.32 ([M+H] + ). [α] D 25 : +23.51° [MeOH: chloroform (1:9); c 0.5]

[0528] Example 9

[0529] 6-fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1 (2H)-one :

[0530] Using the preparation method reported in General Procedure 4, the title compound was resolved from the racemic mixture of 7-fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one (Example 6) into pure enantiomers. Chiral HPLC purity: 99.64%, retention time: 6.75 min. M.P.: 138 °C - 141 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.69 (s, 1H), 8.65 (d, J 1.5, 1H), 8.53 (d, J 2.1, 1H), 8.19 (dd, J 8.9, 5, 1H), 7.94 (dd, J 8.8, 2.7, 1H), 7.84 (t, J 8.8, 2.7, 1H), 7.80 (t, J 2.7, 1H), 7.44 (d, J 7.6, 1H), 7.23 (t, J 7.6, 1H), 7.12 (t, J 7.6, 1H), 6.71 (d, J 7.6, 1H), 6.13 (s, 1H), 4.47 (s, 2H), 1.48 (s, 3H). MS (m / z): 417.28 ([M+H] + )。[α] D 25 : -27.74° [MeOH: chloroform (1:9); c 0.5]

[0531] Example 10

[0532] 7-fluoro-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthal azin-1(2H)-one :

[0533] According to General Procedure 3, the title compound was synthesized from Intermediate 28 (650 mg, 1.61 mmol), hydrazine hydrate (97 mg, 1.94 mmol) and acetic acid (48 mg, 0.80 mmol). Purification: combined flash method. Eluent: MeOH and DCM: 4.1:95.9. Appearance: light yellow solid. Yield: 210 mg. % Yield: 31%. M.P.: 132 °C - 135 °C. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.66 (s, 1H), 8.65 (d, J 1.6, 1H), 8.53 (d, J 2.2, 1H), 8.33 (dd, J 8.8, 5.7, 1H), 7.93 (dd, J 9.8, 2.2, 1H), 7.83 (s, 1H), 7.72 (t, J 8.8, 1H), 7.44 (d, J 7, 1H), 7.23 (t, J 7.7, 1H), 7.12 (t, J 7.1, 1H), 6.73 (d, J 7.8, 1H), 6.14 (s, 1H), 4.44 (s, 2H), 1.49 (s, 3H). MS (m / z): 417.43 ([M+H] + )。

[0534] Example 11

[0535] (+)-7-fluoro-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthal azin-1(2H)-one :

[0536] According to general procedure 3, the title compound was synthesized from intermediate 33 (350 mg, 0.83 mmol), hydrazine hydrate (49 mg, 1 mmol) and acetic acid (24 mg, 0.42 mmol). Purification: combinatorial flash method. Eluent: MeOH and DCM: 4.2:95.8. Appearance: off-white solid. Yield: 140 mg. % Yield: 38%. M.P.: 157 °C - 160 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.68 (s, 1H), 8.64 (d, J 2, 1H), 8.52 (d, J 2, 1H), 8.19 (dd, J 8.8, 5, 1H), 7.94 (dd, J 8.8, 2.6, 1H), 7.84 (td, J 8.8, 2.6, 1H), 7.81 (t, J 2, 1H), 7.35 (dd, J 7.8, 2.4, 1H), 7.07 (td, J 8.8, 2.5, 1H), 6.75 (dd, J 8.8, 4.0, 1H), 6.25 (s, 1H), 4.46 (s, 2H), 1.50 (s, 3H). MS (m / z): 435.32 ([M+H] + )。

[0537] Example 12

[0538] (-)-7-fluoro-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthal azin-1(2H)-one

[0539] Using the preparation method reported in General Procedure 4, the title compound was resolved from the racemic mixture of 7-fluoro-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one (Example 10) into pure enantiomers. Chiral HPLC purity: 99.90%, retention time: 5.88 min. M.P.: 148 °C - 151 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.69 (s, 1H), 8.64 (d, J 2, 1H), 8.52 (d, J 2, 1H), 8.19 (dd, J 8.8,, 5, 1H), 7.94 (dd, J 8.4, 2.4, 1H), 7.84 (td, J 8.8, 2.4, 1H), 7.81 (t, J 2, 1H), 7.35 (dd, J 8, 2.8, 1H), 7.08 (td, J 8.8, 2.4, 1H), 6.75 (dd, J 8.4, 4.0, 1H), 6.26 (s, 1H), 4.46 (s, 2H), 1.50 (s, 3H). MS (m / z): 435.39 ([M+H] + )。[α] D 25 :+17.79° [MeOH: chloroform (1:9); c 0.5]

[0540] Example 13

[0541] 4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1 (2H)-one

[0542] Using the preparation method reported in General Procedure 4, the title compound was resolved from the racemic mixture of 7-fluoro-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one (Example 10) into pure enantiomers. Chiral HPLC purity: 99.28%, retention time: 6.92 min. M.P.: 154 °C - 157 °C. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.69 (s, 1H), 8.64 (d, J 2, 1H), 8.52 (d, 1H), 8.19 (dd, J 8.8, 5, 1H), 7.94 (dd, J 8.4, 2.4, 1H), 7.84 (td, J 8.4, 2.4, 1H), 7.81 (t, J 2, 1H), 7.36 (dd, J 7.6, 2.4, 1H), 7.08 (td, J 9.2, 2.8, 1H), 6.75 (dd, J 8.4, 4.0, 1H), 6.26 (s, 1H), 4.46 (s, 2H), 1.50 (s, 3H). MS (m / z): 435.33 ([M+H] + )。[α] D 25 : -25.52° [MeOH: chloroform (1:9); c 0.5]

[0543] Example 14

[0544] ​ ​ :

[0545] According to the general procedure 3, the title compound was synthesized from intermediate 32 (351 mg, 0.87 mmol), hydrazine hydrate (52 mg, 1.05 mmol) and acetic acid (26 mg, 0.44 mmol). Purification: combined rapid method. Eluent: MeOH and DCM: 5.1:94.9. Appearance: brown-yellow solid. Yield: 90 mg. % Yield: 25%. M.P.: 116 °C - 120 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.58 (s, 1H), 8.65 (d, J 2, 1H), 8.52 (d, J 2, 1H), 8.26 (d, J 7.4 1H), 8.06 (d, J 7.9, 1H), 7.93 (t, J 8.4, 1H), 7.85 (t, J 7.4, 1H), 7.80 (t, J 2, 1H), 7.35 (dd, J 7.9, 2.6, 1H), 7.06 (td, J 8.8, 2.7, 1H), 6.72 (dd, J 8.6, 4.1, 1H), 6.24 (s, 1H), 4.46 (s, 2H), 1.49 (s, 3H). MS (m / z): 417.35 ([M+H] + )。

[0546] ​

[0547] (+)-4-((5-(5-Fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthal azin-1(2H)-one

[0548] Using the preparation method reported in General Procedure 4, the title compound was resolved from the racemic mixture of 4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one (Example 13) into the pure enantiomers. Chiral HPLC purity: 99.65%, retention time: 6.05 min. M.P.: 198 °C - 201 °C. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.59 (s, 1H), 8.65 (d, J 2.5, 1H), 8.52 (d, J 1.2, 1H), 8.26 (d, J 8, 1H), 8.06 (d, J 8, 1H), 7.94 (t, J 7.6, 1H), 7.85 (t, J 7.4, 1H), 7.80 (t, J 2, 1H), 7.35 (dd, J 8, 2.4, 1H), 7.06 (td, J 8.8, 2.8, 1H), 6.72 (dd, J 8.8, 4, 1H), 6.25 (s, 1H), 4.46 (s, 2H), 1.49 (s, 3H). MS (m / z): 417.33 ([M+H] + )。[α] D 25 : +23.63° [MeOH: chloroform (1:9); c 0.5]

[0549] Example 15

[0550] (-)-4-((5-(5-Fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthal azin-1(2H)-one

[0551] Using the preparation method reported in General Procedure 4, the title compound was resolved from the racemic mixture of 4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one (Example 13) into the pure enantiomers. Chiral HPLC purity: 98.81%, retention time: 7.61 min. M.P.: 197 °C - 200 °C. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.59 (s, 1H), 8.65 (d, J 2.5, 1H), 8.52 (d, J 2, 1H), 8.26 (d, J8, 1H), 8.06 (d, J 8, 1H), 7.94 (t, J 7.6, 1H), 7.85 (t, J 7.4, 1H), 7.80 (t, J 2, 1H), 7.35 (dd, J 8, 2.6, 1H), 7.06 (td, J 8.8, 2.8, 1H), 6.72 (dd, J 8.8, 4.2, 1H), 6.25 (s, 1H), 4.46 (s, 2H), 1.49 (s, 3H). MS (m / z): 417.34 ([M+H] + )。[α] D 25 : -28.17° [MeOH: chloroform (1:9); c 0.5]

[0552] Example 16

[0553] 7-Fluoro-4-((5-(6-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthal azin-1(2H)-one

[0554] According to the general procedure 3, the title compound was synthesized from intermediate 37 (400 mg, 1 mmol), hydrazine hydrate (59 mg, 1.19 mmol) and acetic acid (29 mg, 0.5 mmol). Purification: combinatorial flash method. Eluent: MeOH and DCM: 4.7:95.3. Appearance: off-white solid. Yield: 150 mg. % Yield: 36%. M.P.: 125 °C - 128 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.57 (s, 1H), 8.66 (s, 1H), 8.52 (s, 1H), 8.26 (d, J 7.3, 1H), 8.06 (d, J8, 1H), 7.92 (t, J 7, 1H), 7.84 (t, J 7.2, 1H), 7.80 (s, 1H), 7.46 (dd, J 8.1, 5.6, 1H), 6.93 - 6.88 (m, 1H), 6.54 (dd, J 9.4, 2.1, 1H), 6.15 (s, 1H), 4.47 (s, 2H), 1.48 (s, 3H). MS (m / z): 417.33 ([M+H] + )。

[0555] Example 17

[0556] 7-Fluoro-4-((5-(6-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthal azin-1(2H)-one :

[0557] According to general procedure 3, the title compound was synthesized from intermediate 38 (400 mg, 0.95 mmol), hydrazine hydrate (72 mg, 1.56 mmol) and acetic acid (39 mg, 0.47 mmol). Purification: combined rapid method. Eluent: MeOH and DCM: 3.8:96.2. Appearance: light yellow solid. Yield: 90 mg. % Yield: 22%. M.P.: 219 °C - 221 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.68 (s, 1H), 8.66 (d, J 7.1, 1H), 8.529 d, J 2.2, 1H), 8.17 (dd, J 9, 5.04, 1H), 7.93 (dd, J8.8, 2.7, 1H), 7.87 - 7.76 (m, 2H), 7.45 (dd, J 8.2, 5.7, 1H), 6.91 (t, J 10.1, 1H), 6.15 (s, 1H), 4.47 (s, 2H), 1.48 (s, 3H). MS (m / z): 435.32 ([M + H] + )。

[0558] Example 18

[0559] 4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)- one :

[0560] According to general procedure 3, the title compound was synthesized from intermediate 15 (500 mg, 1.3 mmol), hydrazine hydrate (78 mg, 1.56 mmol) and acetic acid (39 mg, 0.65 mmol). Purification: column chromatography on silica gel of 100 - 200 mesh. Eluent: MeOH and DCM: 1.5:98.5. Appearance: off-white solid. Yield: 35 mg. % Yield: 6.7%. M.P.: 203 °C - 205 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.63 (s, 1H), 8.51 (d, J 5.2, 1H), 8.28 (d, J 6.8, 1H), 7.99 (d, J8, 1H), 7.91 (td, J 7.2, 1.2, 1H), 7.84 (t, J 7.2, 1H), 7.69 (s, 1H), 7.49 (d, J 8, 1H)7.44 (d, J 6.8, 1H), 7.39 (d, J 5.2, 1H), 7.26 (td, J 7.6, 1.2, 1H), 7.14 (t, J7.6, 1H), 6.15 (s, 1H), 4.46 (s, 2H), 1.48 (s, 3H). MS (m / z): 399.11 ([M + H] + )。

[0561] Example 19

[0562] (-)-4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1 (2H)-one :

[0563] Using the preparation method reported in General Procedure 4, the title compound was resolved into pure enantiomers from the racemic mixture of 4-((2-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one (Example 18). Chiral HPLC purity: 99.54%, retention time: 9.19 min. M.P.: 207 °C - 210 °C. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.63 (s, 1H), 8.49 (d, J 5.2, 1H), 8.26 (d, J 7.6, 1H), 7.98 (d, J 8, 1H), 7.90 (td, J 7.2, 1.2, 1H), 7.83 (t, J 7.2, 1H), 7.69 (s, 1H), 7.48 (d, J 8, 1H) 7.42 (d, J 7.2, 1H), 7.37 (d, J 5.0, 1H), 7.26 (td, J 7.6, 1.6, 1H), 7.12 (t, J 7.6, 1H), 6.15 (s, 1H), 4.45 (s, 2H), 1.48 (s, 3H). MS (m / z): 399.29 ([M+H] + )。[α] D 25 :-33.01° [MeOH: chloroform (1:9); c 0.5]

[0564] Example 20

[0565] (+)-4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1 (2H)-one

[0566] Using the preparation method reported in General Procedure 4, the title compound was resolved into pure enantiomers from the racemic mixture of 4-((2-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one (Example 18). Chiral HPLC purity: 99.11, retention time: 11.22 min. M.P.: 205 °C - 208 °C. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.63 (s, 1H), 8.49 (d, J 5.2, 1H), 8.26 (d, J 7.6, 1H), 7.98 (d, J 8, 1H), 7.90 (td, J 7.2, 1.2, 1H), 7.83 (t, J 7.2, 1H), 7.68 (s, 1H), 7.48 (d, J 8, 1H) 7.44 (d, J 6.8, 1H), 7.37 (d, J 5.0, 1H), 7.26 (td, J 7.6, 1.2, 1H), 7.12 (t, J 7.6, 1H), 6.15 (s, 1H), 4.45 (s, 2H), 1.47 (s, 3H). MS (m / z): 399.29 ([M+H] + )。[α] D 25 : +29.62° [MeOH: chloroform (1:9); c 0.5]

[0567] Example 21

[0568] 7-Fluoro-4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1 (2H)-one :

[0569] According to the general procedure 3, the title compound was synthesized from intermediate 39 (640 mg, 1.3 mmol), hydrazine hydrate (32 mg, 0.65 mmol) and acetic acid (16 mg, 0.27 mmol). Purification: Column chromatography was performed on silica gel of 60 - 120 mesh: 3.8:96.2. Appearance: off-white solid. Yield: 23 mg. % Yield: 5.2%. M.P.: 187 °C - 189 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.74 (s, 1H), 8.50 (d, J 5.2, 1H), 8.10 (dd, J 9.0, 5, 1H), 7.94 (dd, J 8.8, 2.4 1H), 7.80 (td, J 8.8, 2.8, 1H), 7.67 (s, 1H), 7.48 (d, J 8, 1H), 7.42 (d, J 7.6, 1H), 7.36 (d, J 5.2, 1H), 7.26 (td, J 8, 1, 1H), 7.13 (t, J 7.2, 1H), 6.16 (s, 1H), 4.46 (s, 2H), 1.48 (s, 3H). MS (m / z): 417.16 ([M+H] + )。

[0570] Example 22

[0571] (-)-7-Fluoro-4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthal azin-1(2H)-one :

[0572] Using the preparation method reported in General Procedure 4, the title compound was resolved from the racemic mixture of 7-fluoro-4-((2-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one (Example 21) into pure enantiomers. Chiral HPLC purity: 99.95%, retention time: 8.23 min. M.P.: 165 °C - 168 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.74 (s, 1H), 8.50 (d, J 5.2, 1H), 8.10 (dd, J 9.0, 5.2, 1H), 7.94 (dd, J 8.8, 2.4 1H), 7.80 (td, J 8.8, 2.8, 1H), 7.67 (s, 1H), 7.48 (d, J 8, 1H), 7.42 (d, J 7.6, 1H), 7.36 (d, J 5.2, 1H), 7.26 (td, J 8, 1, 1H), 7.13 (t, J 7.2, 1H), 6.16 (s, 1H), 4.46 (s, 2H), 1.47 (s, 3H). MS (m / z): 417.28 ([M+H] + )。[α] D 25 : -29.17° [MeOH: chloroform (1:9); c 0.5]

[0573] Example 23

[0574] (+)-7-Fluoro-4-((2-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthal azin-1(2H)-one :

[0575] Using the preparation method reported in General Procedure 4, the title compound was resolved from the racemic mixture of 7-fluoro-4-((2-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one (Example 21) into pure enantiomers. Chiral HPLC purity: 99.23, retention time: 10.79 min. M.P.: 167 °C - 170 °C. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.74 (s, 1H), 8.50 (d, J 5.2, 1H), 8.10 (dd, J 8.8, 4.8, 1H), 7.95 (dd, J 8.8, 2.4, 1H), 7.80 (td, J 8.8, 2.8, 1H), 7.67 (s, 1H), 7.48 (d, J 8, 1H), 7.42 (d, J 7.2, 1H), 7.36 (d, J 5.2, 1H), 7.26 (d, J 7.6, 1H), 7.13 (t, J 7.2, 1H), 6.15 (s, 1H), 4.46 (s, 2H), 1.47 (s, 3H). MS (m / z): 417.30 ([M+H] + )。[α] D 25 : +24.29° [MeOH: chloroform (1:9); c 0.5]

[0576] Example 24

[0577] 7-Fluoro-4-((2-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthal azin-1(2H)-one :

[0578] According to General Procedure 3, the title compound was synthesized from Intermediate 42 (508 mg, 1.2 mmol), hydrazine hydrate (72 mg, 1.45 mmol) and acetic acid (36 mg, 0.6 mmol). Purification: Column chromatography on silica gel 100 - 200 mesh: 2:98. Appearance: off-white solid. Yield: 80 mg. % Yield: 15.2%. M.P.: 225 °C - 228 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.73 (s, 1H), 8.49 (d, J 4.8, 1H), 8.10 (dd, J 8.8, 4.8, 1H), 7.95 (dd, J 8.8, 2.8, 1H), 7.81 (td, J 8.4, 2.8, 1H), 7.72 (s, 1H), 7.61 (dd, J 9.2, 4.8, 1H), 7.37 (dd, J 4.8, 1.5, 1H), 7.33 (dd, J 8.0 2.8 1H), 7.12 (td, J 9.2, 2.8, 1H), 6.28 (s, 1H), 4.46 (s, 2H), 1.49 (s, 3H). MS (m / z): 435.14 ([M+H] + )。

[0579] Example 25

[0580] 4-((5-(3-Hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1 (2H)-one

[0581] According to General Procedure 3, the title compound was synthesized from intermediate 45 (670 mg, 1.5 mmol), hydrazine hydrate (100 mg, 2 mmol) and acetic acid (41 mg, 0.69 mmol). Purification: combinatorial rapid method. Eluent: MeOH and DCM: 2:98. Appearance: off-white solid. Yield: 360 mg. % Yield: 52%. M.P.: 258 °C - 260 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.57 (s, 1H), 8.70 (d, J 2, 1H), 8.52 (d, J 2, 1H), 8.26 (d, J 7.6, 1H), 8.08 (d, J 8, 1H), 7.94 (t, J 8.4, 1H), 7.93 (s, 1H), 7.89 - 7.82 (m, 2H), 7.57 (d, J7.6, 1H), 7.42 (t, J 7.6, 1H), 7.23 (t, J 7.6, 1H), 6.81 (d, J 8, 1H), 4.49 (s, 2H). MS (m / z): 453.1 ([M+H] + )。

[0582] Example 26

[0583] (+)-4-((5-(3-Hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)pyridin-3-yl)methyl)phthal azin-1(2H)-one

[0584] Using the preparation method reported in General Procedure 4, the title compound was resolved from the racemic mixture of 4-((5-(3-hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one (Example 25) into pure enantiomers. Chiral HPLC purity: 99.5, retention time: 4.2 min. M.P.: 170 °C - 174 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.60 (s, 1H), 8.71 (d, J 1.6, 1H), 8.54 (d, J 2.4, 1H), 8.29 (dd, J8.0, 1.2, 1H), 8.10 (d, J 8.0, 1H), 7.98 (dt, J 7.2, 1.2, 1H), 7.94 (s, 1H), 7.89 - 7.83 (m, 2H), 7.59 (d, J 7.6, 1H), 7.46 (dt, J 7.6, 1.2, 1H), 7.27 (dt, J 8.0, 0.8, 1H), 6.83 (d, J8.0, 1H), 4.50 (s, 2H). MS (m / z): 453.33 ([M+H] + )。[α] D25 :+46.33° [chloroform; c 0.15],

[0585] Example 27

[0586] (-)-4-((5-(3-Hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)pyridin-3-yl)methyl)phthal azin-1(2H)-one

[0587] Using the preparation method reported in General Procedure 4, the title compound was resolved from the racemic mixture of 4-((5-(3-hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one (Example 25) into pure enantiomers. Chiral HPLC purity: 99.9, retention time: 7.3 min. M.P.: 210 °C - 214 °C. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.60 (s, 1H), 8.71 (d, J 2.0, 1H), 8.54 (d, J 2.4, 1H), 8.29 (dd, J 8.0, 0.8, 1H), 8.11 (d, J 8.0, 1H), 7.98 (dt, J 7.6, 1.6, 1H), 7.94 (s, 1H), 7.89 - 7.83 (m, 2H), 7.59 (d, J 7.2, 1H), 7.46 (dt, J 8.0, 1.2, 1H), 7.27 (dt, J 7.6, 0.8, 1H), 6.83 (d, J 8.4, 1H), 4.50 (s, 2H). MS (m / z): 453.33 ([M+H] + )。[α] D 25 :-41.50° [chloroform; c 0.15],

[0588] Example 28

[0589] 4-(3-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one

[0590] Intermediate 49 (7.5 g, 19.6 mmol), hydrazine hydrate (1.26 g, 25.2 mmol) and 2-propanol (150 ml) were mixed and refluxed at 100 °C for 3 hours. After 3 hours, the reaction mixture was cooled to room temperature and stirred at room temperature for 1 hour. Water (100 ml) was added to the reaction mixture. The aqueous layer was extracted with MeOH and DCM (1:9) (3 * 150 ml). The combined organic layers were dried over anhydrous Na2SO4 and distilled to obtain a solid. Purification: combined flash method. Eluent: MeOH and DCM: 4:96. Appearance: off-white solid. Yield: 4.7 g. % Yield: 60%. M.P.: 240 °C - 243 °C. 11H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.61 (s, 1H), 8.26 (d, J 8, 1H), 8.00 (d, J 8, 1H), 7.90 (t, J 7.6, 1H), 7.83 (t, J 7.6, 1H), 7.49 (t, J 7.6, 1H), 7.41 (t, J 6.2, 2H), 7.35 (s, 1H), 7.26 (d, J 7.6, 1H), 7.18 (t, J 8, 1H), 7.08 (t, J 7.6, 1H), 6.60 (d, J 7.6, 1H), 6.09 (s, 1H), 4.40 (, 2H), 1.47 (s, 3H). MS (m / z): 398.27 ([M+H] + )。

[0591] Example 29

[0592] 7-Fluoro-4-(4-fluoro-3-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one

[0593] Intermediate 51 (3 g, 7 mmol), hydrazine hydrate (500 mg, 9 mmol) and 2-propanol (60 ml) were mixed and refluxed at 100 °C for 3 hours. After 3 hours, the reaction mixture was cooled to room temperature and stirred at room temperature for 1 hour. Water (50 ml) was added to the reaction mixture. The aqueous layer was extracted with MeOH and DCM (1:9) (2 * 25 ml). The combined organic layers were dried over anhydrous Na2SO4 and distilled to obtain a solid. Purification: combinatorial flash method. Eluent: MeOH and DCM: 3:97. Appearance: light brown solid. Yield: 1.3 g. % Yield: 40%. M.P.: 207 °C - 210 °C. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.72 (s, 1H), 8.16 - 8.09 (m, 1H), 7.94 (d, J 7.6, 1H), 7.81 (t, J 8, 1H), 7.51 (d, J 5.2, 1H), 7.44 (d, J 6.8, 2H), 7.39 (d, J 6.4, 1H), 7.22 (t, J 7.6, 1H), 7.11 (t, J 7.2, 1H), 6.50 (d, J 5.6, 1H), 6.27 (s, 0.4H), 6.15 (s, 0.6H), 4.41 (s, 1.2H), 4.38 (s, 0.8H), 1.51 (s, 1.2H), 1.47 (s, 1.8H). MS (m / z): 434.26 ([M+H] + )。

[0594] Example 30

[0595] 4-((5-(3-Methoxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one

[0596] Suspend intermediate 52 (1.25 g, 3 mmol) in MeOH (5 ml) and cool to 0 °C. Add sodium methoxide (972 mg, 4.5 mmol) to the mixture. Warm the reaction mixture to room temperature and stir at room temperature for 2 hours. After 2 hours, quench the reaction mixture with water (250 ml) and extract the aqueous layer with a mixture of DCM and MeOH (9:1) (2 * 50 ml). Wash the combined organic layers with water (50 ml) and 5% aqueous citric acid solution (50 ml). Dry the organic layer over anhydrous Na2SO4 and distill in vacuo to obtain the crude product. Purify the crude product by column chromatography using EtOAc as the eluent to obtain the title compound as a white solid. Yield: 65 mg. % Yield: 5.3%. M.P.: 207 °C - 210 °C. 1 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.59 (s, 1H), 8.65 (s, 1H), 8.57 (d, J 2, 1H), 8.28 (d, J 8, 1H), 8.08 (d, J 7.6, 1H), 7.95 (t, J 7.6, 1H), 7.90 (s, 1H), 7.86 (t, J 7.6, 1H), 7.45 (d, J 7.6, 1H), 7.31 (t, J 7.6, 1H), 7.19 (t, J 7.6, 1H), 6.75 (d, J 7.6, 1H) 4.47 (s, 2H), 2.97 (s, 3H), 1.54 (s, 3H). MS (m / z): 413.34 ([M+H] + )。

[0597] Example 31

[0598] 4-(3-(3-Hydroxy-3-methyl-2-oxo-2,3-dihydro-1H-pyrrolo[2,3-c]pyridin-1-yl)benzyl)phthal azin-1(2H)-one

[0599] Mix intermediate 58 (210 mg, 0.55 mmol), hydrazine hydrate (35 mg, 0.7 mmol) and 2-propanol (5.2 ml) and reflux at 100 °C for 3 hours. After 3 hours, cool the reaction mixture to room temperature and stir at room temperature for 1 hour. Add water (50 ml) to the reaction mixture. Extract the aqueous layer with MeOH and DCM (1:9) (2 * 50 ml). Dry the combined organic layers over anhydrous Na2SO4 and distill to obtain a solid. Purification: combinatorial flash method. Eluent: MeOH and DCM: 5:95. Appearance: light yellow solid. Yield: 30 mg. % Yield: 14%. M.P.: 133 °C - 136 °C. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.60 (s, 1H), 8.39 (d, J 4.4, 1H), 8.27 (dd, J 8, 1.2, 1H), 8.02 (d, J 8, 1H), 7.99 (s, 1H), 7.90 (td, J 8, 1.2, 1H), 7.83 (td, J 8, 1.2, 1H), 7.53 - 7.48 (m, 2H), 7.47 - 7.40 (m, 2H), 7.32 (d, J 8, 1H), 6.34 (s, 1H), 4.41 (s, 2H), 1.50 (s, 3H). MS (m / z): 399.40 ([M+H] + )。

[0600] Example 32

[0601] 3-Methyl-2-oxo-1-(5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)pyridin-3-yl)dihydroind ole-3-yl acetate

[0602] Dissolve intermediate 52 (5 g, 12 mmol) in DMF (60 ml), add 18-crown-6 (3.17 g, 12 mmol) and cesium acetate (9.21 g, 48 mmol). Stir the mixture at 80 °C for 1.5 h. After completion of the reaction, dilute the reaction mixture with water and extract with DCM (3 * 100 ml). Distill the combined organic layers to obtain the crude product. Purification: Perform column chromatography on silica gel of 60 - 120 mesh. Eluent: MeOH and DCM: 4:96. Appearance: off-white solid. Yield: 1.2 g. % Yield: 22.7%. M.P.: 237 °C - 239 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.58 (s, 1H), 8.67 (s, 1H), 8.49 (d, J 2, 1H), 8.27 (d, J 8, 1H), 8.07 (d, J 8, 1H), 7.94 (t, J 7.2, 1H), 7.85 (t, J 8, 1H), 7.79 (s, 1H), 7.43 (d, J 7.6, 1H), 7.26 (t, J 7.6, 1H), 7.12 (t, J 7.6, 1H), 6.69 (d, J 7.6, 1H), 4.48 (s, 2H), 2.03 (s, 3H), 1.63 (s, 3H). MS (m / z): 441.3 ([M+H] + )。

[0603] Example 33

[0604] 4-(4-Fluoro-3-(2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one

[0605] According to general procedure 3, the title compound was synthesized from intermediate 65 (200 mg, 0.54 mmol), hydrazine hydrate (32 mg, 0.65 mmol) and acetic acid (16 mg, 0.27 mmol). Purification: combined rapid method. Eluent: MeOH and DCM: 2.7:97.3. Appearance: off-white solid. Yield: 12 mg. % Yield: 5.8%. M.P.: 232 °C - 235 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.59 (s, 1H), 8.25 (d, J 7.8, 1H), 7.99 (d, J 7.8, 1H), 7.94 - 7.88 (m, 1H), 7.87 - 7.80 (m, 1H), 7.50 - 7.37 (m, 3H), 7.32 (d, J 7.2, 1H), 7.14 (t, J 7.6, 1H), 7.06 (t, J 7.2, 1H), 6.44 (d, J 7.6, 1H), 4.37 (s, 2H), 3.76 (d, J 5, 2H). MS (m / z): 386.0 ([M+H] + )。

[0606] Example 34

[0607] 4-(3-(3,3-Dimethyl-2-oxodihydroindol-1-yl)-4-fluorobenzyl)phthalazin-1(2H)-one

[0608] According to general procedure 3, the title compound was synthesized from intermediate 71 (600 mg, 1.5 mmol), hydrazine hydrate (90 mg, 1.8 mmol) and acetic acid (45 mg, 0.75 mmol). Purification: combined rapid method. Eluent: MeOH and DCM: 2:98. Appearance: off-white solid. Yield: 340 mg. % Yield: 54%. M.P.: 129 °C - 132 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.59 (s, 1H), 8.25 (d, J 7.7, 1H), 8.00 (d, J 7.8, 1H), 7.90 (t, J 7, 1H), 7.87 - 7.80 (m, 1H), 7.52 (d, J 6.2, 1H), 7.48 - 7.35 (m, 3H), 7.15 (t, J 7.5, 1H), 7.09 (t, J 7.2, 1H), 6.48 (d, J7.7, 1H), 4.37 (s, 2H), 1.38 (s, 3H), 1.35 (s, 3H). MS (m / z): 414.45 ([M+H]

[0609] Example 35

[0610] 4-((5-(3,3-Dimethyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-oneAccording to General Procedure 3, the title compound was synthesized from intermediate 74 (600 mg, 1.57 mmol), hydrazine hydrate (94 mg, 1.88 mmol) and acetic acid (47 mg, 0.78 mmol). Purification: combined rapid method. Eluent: MeOH and DCM: 3:97. Appearance: off-white solid. Yield: 260 mg. % Yield: 41%. M.P.: 221 °C - 224 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.58 (s, 1H), 8.63 (s, 1H), 8.54 (s, 1H), 8.26 (d, J 7.8, 1H), 8.06 (d, J 7.8, 1H), 7.93 (t, J 7.8, 1H), 7.88 - 7.80 (m, 2H), 7.43 (d, J 7.2, 1H), 7.18 (t, J 7.6, 1H), 7.10 (t, J 7.3, 1H), 6.70 (d, J 7.7, 1H), 4.45 (s, 2H), 1.37 (s, 6H). MS (m / z): 397.47 ([M+H] + )。

[0611] Example 36

[0612] 4-((5-(3,3-Dimethyl-2-oxo-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-1-yl)pyridin-3- yl)methyl)phthalazin-1(2H)-one

[0613] According to General Procedure 3, the title compound was synthesized from intermediate 81 (762 mg, 1.99 mmol), hydrazine hydrate (119 mg, 2.38 mmol) and acetic acid (59 mg, 0.99 mmol). Purification: combined rapid method. Eluent: MeOH and DCM: 5.2:94.8. Appearance: off-white solid. Yield: 300 mg. % Yield: 38%. M.P.: 242 °C - 245 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.58 (s, 1H), 8.64 (s, 1H), 8.58 (s, 1H), 8.26 (d, J 7.8, 1H), 8.22 (d, J 4.6, 1H), 8.05 (d, J 8, 1H), 7.93 (t, J 7.6, 1H), 7.89 (s, 1H), 7.84 (t, J 7.4, 1H), 7.24 - 7.18 (m, 1H), 7.12 (d, J 7.8, 1H), 4.45 (s, 2H), 1.38 (s, 6H). MS (m / z): 398.1 ([M+H] + )。

[0614] Example 37

[0615] 4-(3-(3,3-Dimethyl-2-oxo-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-1-yl)-4-fluorobenzyl) phthalazin-1(2H)-one

[0616] According to general procedure 3, the title compound was synthesized from intermediate 84 (390 mg, 0.97 mmol), hydrazine hydrate (49 mg, 1.17 mmol), and acetic acid (29 mg, 0.49 mmol). Purification: combinatorial flash method. Eluent: MeOH and DCM: 3.2:96.8. Appearance: off-white solid. Yield: 168 mg. % Yield: 42%. M.P.: 114 °C - 117 °C. 1 H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.59 (S, 1H), 8.25 (d, J 7.8, 1H), 8.23 - 8.21 (m, 1H), 7.99 (d, J 8, 1H), 7.90 (t, J7, 1H), 7.85 - 7.81 (m, 1H), 7.60 - 7.53 (m, 1H), 7.50 - 7.45 (m, 1H), 7.44 - 7.38 (m, 1H), 7.20 (dd, J 8, 5.04, 1H), 6.91 (d, J 7.9, 1H), 4.37 (s, 2H), 1.40 (s, 3H), 1.36 (s, 3H) MS (m / z): 415.2 ([M+H] + )。

[0617] Example 38

[0618] 1'-(5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)pyridin-3-yl)spiro[cyclopropane-1,3'-dihydroind ole]-2'-one

[0619] According to general procedure 3, the title compound was synthesized from intermediate 89 (591 mg, 1.55 mmol), hydrazine hydrate (93 mg, 1.86 mmol), and acetic acid (46 mg, 0.78 mmol). Purification: combinatorial flash method. Eluent: MeOH and DCM: 3.1:96.9. Appearance: light yellow solid. Yield: 160 mg. % Yield: 26%. M.P.: 219 °C - 222 °C. 1H-NMR (δ ppm, DMSO-d6, 400 MHz): 12.57 (s, 1H), 8.63 (d, J 1.7, 1H), 8.57 (d, J 2.3, 1H), 8.26 (d, J 7.8, 1H), 8.07 (d, J 8, 1H), 7.93 (t, J 7, 1H), 7.90 - 7.81 (m, 2H), 7.20 - 7.16 (m, 1H), 7.12 - 7.04 (m, 2H), 6.79 (d, J 7.8, 1H), 4.46 (s, 2H), 1.76 - 1.70 (m, 2H), 1.65 - 1.60 (m, 2H). MS (m / z): 395.35 ([M+H] + )。

[0620] Biological assay

[0621] The pharmacological properties of the compounds of the present invention can be confirmed by a variety of pharmacological assays. Suitable pharmacological assays that can be performed to evaluate the compounds described herein are given below.

[0622] Example -1

[0623] Determine the GI of the compound of formula (I) in the HCT-116 and UWB1.289 cell lines 50 Cell proliferation assay (MTT assay) assay)

[0624] Assay protocol :

[0625] On day "0", the test cells are plated in triplicate in complete medium in a 96-well plate at 100 μL / well and the plate is incubated at 37 °C and 5% CO2. On day 1, 10 μL of MTT (5 mg / ml) is added to the columns designated for day "0". The columns are mixed well and incubated at 37 °C and 5% CO2 for 3.5 hours. The cells are pelleted at 4000 rpm for 10 minutes. The medium is aspirated, 150 μL of DMSO is added to the cells and mixed by pipetting to dissolve the crystals. The plate is read at A560 nm and A640 nm. The DMSO dilution of the inhibitor is diluted in growth medium to 3 times the desired concentration. The cells in each well are treated with 50 μL of complete medium containing the inhibitor. The DMSO concentration in the wells is 0.1%. As needed, the plate is incubated at 37 °C and 5% CO2 for 144 hours. 15 μL of MTT (5 mg / mL) is added to the wells. The plate is incubated at 37 °C and 5% CO2 for 3.5 hours. After incubation, the cells are pelleted at 4000 rpm for 10 minutes. The medium is aspirated and 150 μL of DMSO is added to each well to dissolve the formazan crystals. The plate is read at A560 nm and A640 nm.

[0626] Results: The GI of the compounds of the present invention was determined 50Value. GI 50 The values are given in Table - 2 below:

[0627] Example 2

[0628] PARP enzyme assay for determining the IC of the compound of formula (1) 50 ​

[0629] Assay protocol :

[0630] Step 1 - Coating :

[0631] 1) Coat a 384 - well plate with 25 μL of histone solution as described below:

[0632] · Dilute the 5x histone mixture 1:5 with PBS.

[0633] · Add 25 μL of the diluted histone solution to each well and incubate overnight at 4 °C.

[0634] · Wash the plate three times with 100 μL of PBST buffer (1x PBS containing 0.05% Tween 20) / well.

[0635] · Tap the plate on a clean paper towel to remove the liquid.

[0636] · Block the wells by adding 100 μL of blocking buffer 3 to each well.

[0637] Incubate for 1 hour and 15 minutes at room temperature.

[0638] · Wash the plate three times with 100 μL of PBST buffer as described above.

[0639] · Tap the plate on a clean paper towel to remove the liquid.

[0640] Step 2: Ribosylation reaction :

[0641] 1) Preparation of 1X PARP buffer:

[0642] · Prepare 1x PARP buffer by adding 1 part of 10x PARP buffer to 9 parts of distilled water (v / v). To prepare 500 μL of 1X PARP buffer, add 50 μL of 10X PARP assay buffer to 450 μL of H2O and mix well.

[0643] 2) Preparation of ribosylation master mixture:

[0644] · N wells x (1.25 μL of 10x PARP buffer + 1.25 μL of 10X PARP assay mixture + 2.5 μL of activated DNA + 7.5 μL of distilled water). Add 12.5 μL to each well.

[0645] 3) Preparation of inhibitor solution:

[0646] · Prepare a 10 mM inhibitor stock solution in DMSO. From the 10 mM stock solution, prepare a 250X concentration of the inhibitor in DMSO.

[0647] · Further dilute the inhibitor at 250X DMSO concentration to 10X concentration in 1X kinase buffer.

[0648] · Add 2.5 μL (10X concentration) of the kinase buffer containing the inhibitor to each well.

[0649] · For "positive control" and "blank", add 2.5 μL of the kinase buffer containing DMSO

[0650] (4%).

[0651] · The final DMSO concentration in the wells is 0.4%.

[0652] 4) Preparation of enzyme solution:

[0653] · Thaw the PARP1 enzyme on ice. After the first thaw, briefly spin the tube containing the enzyme to recover all the contents of the tube. Calculate the amount of PARP1 required for the assay,

[0654] and dilute the enzyme to 2.0 ng / μL with 1x PARP buffer.

[0655] · Start the reaction by adding 10 μL of the diluted PARP1 enzyme to the wells designated as "positive control" and "test inhibitor".

[0656] · Add 10 μL of 1X PARP buffer to the well designated as "blank".

[0657] · Centrifuge the plate at 1000 rpm for 30 seconds and incubate at room temperature for 60 minutes.

[0658] · After 1 hour, discard the reaction mixture, wash the plate three times with 100 μL of PBST buffer, and tap the plate on a clean paper towel as described above.

[0659] Step 3: Detection :

[0660] 1) Preparation of streptavidin-HRP:

[0661] · Dilute streptavidin-HRP 1:50 in blocking buffer.

[0662] · Add 25 μL of the diluted streptavidin-HRP to each well. Incubate at room temperature for 30 minutes.

[0663] · Wash three times with 100 μL of PBST buffer and tap the plate on a clean paper towel as described above.

[0664] 2) Preparation of ECL substrate:

[0665] · Immediately before use, mix equal volumes of ELISA ECL substrate A and ELISA ECL substrate B on ice and add 50 μL per well.

[0666] · Immediately read the plate in the microtiter plate in the chemiluminescence setting. Subtract the "blank" value from all other values.

[0667] Results: Inhibition % values and IC 50 values are given in Table - 2 below:

[0668] Example 3

[0669] Seed UWB 1.289 cells at a predetermined density in their respective media in 6 - well plates and incubate the plates overnight at 37 °C and 5% CO₂. After overnight incubation, treat the plates with the desired concentration of the compound or DMSO and incubate the plates at 37 °C and 5% CO₂ for 24 hours as needed.

[0670] After 24 - hour incubation is complete, prepare total cell lysates from the treated samples.

[0671] Analyze the lysates by Western blot analysis and probe with a human PAR - specific monoclonal antibody or a human - specific β - actin monoclonal antibody (loading control).

[0672] Develop the Western blot image after exposing it to the chemiluminescent substrate and capture the image on an imaging system.

[0673] Measure the target - specific band and the loading - specific band using densitometric analysis. Normalize the densitometric value of the target - specific band to the loading control value. Calculate the % inhibition of PAR relative to the DMSO - treated samples.

[0674] Results: Inhibition % values are given in Table - 2 below:

[0675] Table - 2

[0676]

[0677] Inhibition %: A represents > 75% to 100%; B represents > 50% to ≤ 75%; C represents > 25% to ≤ 50% and D represents ≤ 25%, and IC50: + represents ≤ 10 nM and ++ represents ≥ 10 nM; and GI50: A1 represents ≤ 2500 nM; B1 represents > 2500 nM to ≤ -5000 nM; C1 represents > 5000 nM to ≤ 7500 nM; D1 represents > 7500 nM to ≤ 10000 nM, and E1 represents > 10000 nM

[0678] Although the invention herein has been described with reference to specific embodiments, it should be understood that these embodiments are merely illustrative of the principles and applications of the invention. Accordingly, it should be understood that numerous modifications may be made to the exemplary embodiments, and other arrangements may be designed without departing from the spirit and scope of the invention as set forth above. It is intended that the appended claims define the scope of the invention and thereby cover methods and structures within the scope of these claims and their equivalents.

[0679] All publications, patents and / or patent applications cited in this application are hereby incorporated by reference as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

Claims

1. A compound of formula (I): or a tautomer thereof or a pharmaceutically acceptable salt thereof, wherein R a , R b , R c and R d are each independently selected from hydrogen and halogen; X is CR x or N; Y is CR y or N; Z is CR Z or N; R x 、R y and R z each independently selected from hydrogen and halogen; G is selected from R e and R f each independently selected from hydrogen, hydroxy, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy and acetoxy, or R e and R f are both capable of joining to form cyclopropyl; R g selected from hydrogen, C 1-3 alkyl, and -(CO)R h ; R h is C 1-3 alkyl; and R 1 、R 2 、R 3 and R 4 are each independently selected from hydrogen and halogen.

2. The compound according to claim 1, wherein R a 、R b 、R c and R d are each independently selected from hydrogen and halogen; X is CR x or N; Y is CR y or N; Z is CR Z or N; R x 、R y and R z each independently selected from hydrogen and halogen; G is selected from R e and R f each independently selected from hydrogen, halogen, C 1-3 alkyl, haloalkyl or -O(CO)R h ; R g selected from hydrogen, C 1-3 alkyl, and -(CO)R h ; R h selected from hydrogen and C 1-3 alkyl; and R 1 、R 2 、R 3 and R 4 are each independently selected from hydrogen and halogen.

3. The compound according to claim 1, wherein the compound has the formula (IA) or (IB): or a tautomer thereof or a pharmaceutically acceptable salt thereof.

4. The compound according to any one of claims 1-3, wherein (i)R a 、R b 、R c and R d each independently selected from the group consisting of hydrogen and halogen; (ii) R a , R c and R d are hydrogen, and R b is a halogen; or (iii) R a 、R b and R d are hydrogen and R c is a halogen.

5. The compound according to claim 4, wherein R b is fluorine or chlorine.

6. The compound according to claim 4, wherein R c is fluorine or chlorine.

7. The compound according to any one of claims 1-3, wherein X, Y and Z are each independently selected from CH and N.

8. The compound according to claim 7, wherein (i) Y and Z are CH, and X is N; (ii) X and Z are CH, and Y is N; or (iii) X and Y are CH, and Z is N.

9. A compound according to any one of claims 1 - 3, wherein X and Y are CH, and Z is CR Z .

10. The compound according to claim 9, wherein R z is a halogen.

11. The compound according to claim 10, wherein R z is fluorine.

12. The compound according to claim 1, wherein R e is hydroxy, C 1-3 alkoxy or acetoxy.

13. The compound according to claim 12, wherein R e is a hydroxyl group.

14. The compound according to claim 1, wherein R f is C 1-3 alkyl or C 1-3 haloalkyl.

15. The compound according to claim 14, wherein R f is methyl, ethyl or CF3.

16. The compound according to claim 1, wherein R e is a hydroxyl group, and R f is C 1-3 alkyl or C 1-3 haloalkyl.

17. The compound according to claim 1, wherein (i)R e and R f are both hydrogen; (ii) R e and R f are each independently selected from C 1-3 alkyl; or (iii) R e and R f are both joined to form C 3-6 cycloalkyl group.

18. The compound according to claim 17, wherein R e and R f are both hydrogen.

19. The compound according to claim 17, wherein R e and R f are both methyl groups.

20. The compound according to claim 17, wherein R e and R f are both joined to form a C 3-6 cycloalkyl ring.

21. The compound according to any one of claims 1-3, wherein (i)R 1 、R 2 、R 3 and R 4 are each independently selected from hydrogen or halogen; (ii) R 1 , R 2 , R 3 and R 4 are hydrogen; (iii) R 1 、R 2 、R 3 and R 4 each independently selected from halogen; (iv) R 1 , R 3 and R 4 is hydrogen, and R 2 is a halogen; or (v)R 1 、R 2 and R 4 are hydrogen, and R 3 is a halogen.

22. The compound according to claim 1, wherein (i)R 1 、R 2 、R 3 and R 4 each independently is fluorine or chlorine; (ii) R 1 , R 3 and R 4 is hydrogen, and R 2 is fluorine; or (iii) R 1 , R 2 and R 4 are hydrogen, and R 3 is fluorine.

23. The compound according to any one of claims 1-3, wherein G is selected from 24. The compound according to claim 23, wherein G is selected from 25. A compound selected from: 4-((5-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; (R)-(+)-4-((5-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; (S)-(-)-4-((5-(3-Hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 4-(4-Fluoro-3-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one; 4-((5-(3-Ethyl-3-hydroxy-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 7-Fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; (+)-7-Fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; (-)-7-Fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 6-Fluoro-4-((5-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 7-Fluoro-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; (+)-7-Fluoro-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; (-)-7-Fluoro-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; (+)-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; (-)-4-((5-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 7-Fluoro-4-((5-(6-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 7-Fluoro-4-((5-(6-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 4-((2-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one; (-)-4-((2-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one; (+)-4-((2-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one; 7-Fluoro-4-((2-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one; (-)-7-Fluoro-4-((2-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one; (+)-7-Fluoro-4-((2-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one; 7-Fluoro-4-((2-(5-fluoro-3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-4-yl)methyl)phthalazin-1(2H)-one; 4-((5-(3-hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; (+)-4-((5-(3-hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; (-)-4-((5-(3-hydroxy-2-oxo-3-(trifluoromethyl)dihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 4-(3-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one; 7-Fluoro-4-(4-fluoro-3-(3-hydroxy-3-methyl-2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one; 4-((5-(3-methoxy-3-methyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 4-(3-(3-Hydroxy-3-methyl-2-oxo-2,3-dihydro-1H-pyrrolo[2,3-c]pyridin-1-yl)benzyl)phthalazin-1(2H)-one; 3-Methyl-2-oxo-1-(5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)pyridin-3-yl)dihydroindol-3-yl acetate; 4-(4-Fluoro-3-(2-oxodihydroindol-1-yl)benzyl)phthalazin-1(2H)-one; 4-(3-(3,3-Dimethyl-2-oxodihydroindol-1-yl)-4-fluorobenzyl)phthalazin-1(2H)-one; 4-((5-(3,3-Dimethyl-2-oxodihydroindol-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 4-((5-(3,3-Dimethyl-2-oxo-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-1-yl)pyridin-3-yl)methyl)phthalazin-1(2H)-one; 4-(3-(3,3-Dimethyl-2-oxo-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-1-yl)-4-fluorobenzyl)phthalazin-1(2H)-one; and 1'-(5-((4-Oxo-3,4-dihydrophthalazin-1-yl)methyl)pyridin-3-yl)spiro[cyclopropane-1,3'-dihydroindole]-2'-one; and pharmaceutically acceptable salts thereof.

26. A pharmaceutical composition comprising a compound according to any one of claims 1-25 and a pharmaceutically acceptable carrier.

27. The pharmaceutical composition according to claim 26, further comprising one or more additional therapeutic agents.

28. The pharmaceutical composition according to claim 27, wherein the one or more additional therapeutic agents are anti-cancer agents, anti-inflammatory agents, immunosuppressive agents, antihistamines, analgesics or any combination of any of the foregoing drugs.

29. The pharmaceutical composition according to claim 28, wherein the anti-inflammatory agent is a steroid or a non-steroidal anti-inflammatory agent.

30. Use of a compound according to any one of claims 1-25 in the preparation of a drug for inhibiting the catalytic activity of PARP enzyme present in cells.

31. The use according to claim 30, wherein the inhibition occurs in a subject suffering from cancer, bone disease, inflammatory disease, immune disease, neurological disease, metabolic disease, respiratory disease, thrombosis or heart disease.

32. Use of a compound according to any one of claims 1-25 in the preparation of a drug for treating a disease that would benefit from inhibition of the catalytic activity of PARP enzyme.

33. Use of a compound according to any one of claims 1-25 in the manufacture of a medicament for use in a method for treating a PARP-related disease, said method comprising administering to a subject in need thereof an effective amount of said compound.

34. Use according to claim 33, said method further comprising the step of administering to said subject simultaneously or sequentially at least one other anti-cancer agent, anti-inflammatory agent, immunosuppressive agent, antihistamine, analgesic or any combination of any of the foregoing agents.

35. Use according to claim 33 or claim 34, wherein said PARP-related disease is an immune system-related disease, a disease involving inflammation, cancer or other proliferative diseases, liver diseases, or kidney diseases.

36. Use according to claim 33 or claim 34, wherein said PARP-related disease is selected from inflammation, liver diseases, kidney diseases, chronic obstructive pulmonary disease, osteoporosis, eczema, autoimmune hemolytic anemia, cystic fibrosis, multiple sclerosis, Guillain-Barré disease, pernicious anemia, autoimmune thrombocytopenia, antiphospholipid syndrome, Wegener's granulomatosis, Behçet's disease, psoriasis, pemphigus vulgaris, vitiligo, Crohn's disease, primary biliary cirrhosis, Parkinson's disease, Huntington's disease, and cancer.

37. Use according to claim 33 or claim 34, wherein said PARP-related disease is selected from glomerulonephritis, uveitis, rheumatoid arthritis, inflammatory bowel disease, dermatitis, osteoarthritis, autoimmune uveitis, temporal arteritis, vasculitis, dermatitis herpetiformis, and colitis.

38. Use according to claim 37, wherein said colitis is ulcerative colitis.

39. Use according to claim 33 or claim 34, wherein said PARP-related disease is selected from lymphoid hematopoietic tumors, leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma, and Burkitt lymphoma, myeloid hematopoietic tumors, myelodysplastic syndromes, bladder cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer, esophageal cancer, cervical cancer, thyroid cancer, prostate cancer, and skin cancer.

40. Use according to claim 39, wherein said leukemia is acute lymphoblastic leukemia, acute lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, or promyelocytic leukemia.

41. Use according to claim 33 or claim 34, wherein said PARP-related disease is selected from breast cancer, ovarian cancer, liver cancer, lung cancer, esophageal cancer, gallbladder cancer, pancreatic cancer, and gastric cancer.

42. Use according to claim 33 or claim 34, wherein said PARP-related disease is small cell lung cancer, breast cancer, or ovarian cancer.

43. Use according to claim 34, wherein said anti-inflammatory agent is a steroid or a non-steroidal anti-inflammatory agent.

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