TWI522352B - Amids substituted indazole derivatives as poly (adp-ribose) polymerase inhibitors - Google Patents

Description

Indole-substituted carbazole derivative poly(ADP-ribose) polymerase inhibitor

The present invention relates to a guanamine-substituted oxazole and benzotriazole compound which can be used as a poly(ADP-ribose) polymerase (PARP) inhibitor. PARP was previously also known as poly(ADP-ribose) synthase and poly(ADP-ribose) synthase (poly(ADP-ribose) synthase) (poly(ADP-ribosyl)) The compound of the present invention is useful as a monotherapy method for a tumor associated with a specific defect in a DNA repair pathway, as well as a specific DNA damage drug (such as an anticancer drug) and an enhancer for radiation therapy. Further, the present invention The compounds described can also be used to reduce cell necrosis (stroke and myocardial infarction), down-regulate inflammation and tissue damage, treat retroviral infections, and prevent chemotherapy poisoning.

Poly(ADP-ribose) polymerase (PARP) constitutes a superfamily of 18 proteins, all of which contain the PARP catalytic domain (Bioessays (2004) 26: 1148). These proteins include adenosine diphosphate ribose polymerase-1 (PARP-1), PARP-2, PARP-3, tankyrase-1, tankyrase-2, vault PARP, and Ti PARP. PARP member PARP-1 includes three major domains: an amino-N-terminal DNA-binding domain (DBD) containing two zinc finger structures, an automodification domain (AMD), and a Carbonyl C-terminal catalytic domain.

PARP is a cytoplasmic enzyme that degrades Nickel Dinucleotide (NAD ⁺ ) to nicotinamide and ADP-ribose to target proteins (including topoisomerase, tissue protein, and PARP itself) A long chain, branched ADP-ribose polymer is formed ( Biochem. Biophys. Res. Commun. ( 1998 ) 245: 1-10). Poly ADP ribosylation is associated with a variety of biological processes including DNA repair, cell cycle processes, apoptosis, chromatin function, and gene stability ( Biochem. Biophys. Res. Commun. ( 1998 ) 245: 1-10) . The cleavage of the DNA strand can rapidly stimulate the catalytic activity of PARP-1 and PARP-2 ( Pharmacological Research ( 2005 ) 52: 25-33). When DNA is damaged, PARP-1 responds and binds to single-stranded and double-stranded DNA gaps. Under normal physiological conditions, the activity of PARP is the lowest. However, once DNA damage occurs, PARP activity is initiated immediately and up to 500-fold. Both PARP-1 and PARP-2 monitor DNA strand breaks and serve as DNA gap sensors, providing a fast signal for termination of transcription and aggregation of DNA repair enzymes at the site of the lesion. Because radiotherapy for cancer therapy and many chemotherapy methods are achieved by inducing DNA damage, PARP inhibitors are useful as chemosensitizers and radiosensitizers for cancer therapy. Studies have shown that PARP inhibitors can act on radiosensitized anaerobic tumor cells (U.S. Patent Nos. 5,032,617, 5,215,738 and 5,041,653).

The vast majority of biological effects of PARP are related to the poly ADP-ribosylation process, which affects the nature and function of the target protein; PARP oligomers, when cleaved from poly ADP-ribosylated proteins, are given Significant cytological effects; the physical association of PARP and nuclear proteins forms a functional complex; the cellular level of the substrate NAD ⁺ is reduced ( Nature Review ( 2005 ) 4:421-440).

In addition to being involved in DNA repair, PARP can also act as a regulator of apoptosis. Under pathological conditions such as ischemic and reperfusion injury, significant initiation of PARP can result in substantial consumption of NAD ⁺ between cells, causing damage to a variety of NAD ⁺ -dependent metabolic pathways and leading to apoptosis (see Pharmacological Research ( 2005) ) 52:44-59). The NAD ⁺ level is significantly reduced as a result of PARP initiation. A large amount of PARP initiation can result in severe consumption of NAD ⁺ in cells that suffer from large amounts of DNA damage. Due to the short half-life of poly(ADP-ribose), its turnover rate is characterized by rapidity. Because once poly(ADP-ribose) is formed, it is immediately degraded by constitutively activated poly(ADP-ribose) glycohydrolase (PARG). PARP and PARG form a loop that converts a large amount of NAD ⁺ to ADP-ribose, causing NAD ⁺ and ATP to fall below 20% of normal levels. This is quite unfavorable during the ischemic process when hypoxic conditions have completely compromised cellular energy release. Subsequently, during reperfusion, aggregation of free radical products is the main cause of tissue damage. The rapid decline of some ATP is typical during ischemia and reperfusion of many organs. Due to the reversal of poly(ADP-ribose), a rapid drop in some ATP leads to NAD ⁺ consumption. Therefore, the PARP inhibitor needs to preserve the energy level of the cells, thereby enhancing the survival of the ischemic tissue after the injury. Thus, PARP inhibitors are useful in the treatment of PARP-induced diseases that are regulated by apoptosis, including neurological diseases such as stroke, brain trauma, and Parkinson's syndrome.

PARP inhibitors can be used to specifically kill BRCA-1 and BRCA-2 deficient tumors ( Nature ( 2005 ) 434: 913-916 and 917-921; Cancer Biology & Therapy ( 2005 ) 4: 934-936).

PARP inhibitors can also potentiate the effects of anti-tumor drugs ( Pharmacological Research ( 2005 ) 52:25-33), including effects that enhance platinum compounds such as cisplatin and carboplatin ( Cancer Chemother Pharmacol ( 1993) 33:157-162; MoI Cancer Ther ( 2003 ) 2:371-382). PARP inhibitors can also potentiate the antitumor activity of topoisomerase I inhibitors such as irinotecan and topotecan ( MoI Cancer Ther ( 2003 ) 2:371-382; and Clin Cancer Res. ( 2000 ) 6: 2860-2867), and the above enhancements have been further validated in an in vivo model ( J Natl Cancer Inst ( 2004 ) 96: 56-67). PARP inhibitors can also repair the sensitivity of cytotoxic and temozolomide (temozolomide, TMZ) of the anti-proliferative effect of malignant cells (Curr Med Chem (2002) 9 : 1285-1301; Med Chem Rev Online (2004) 1: 144-150) . The results of this study have been in a large number of in vitro models ( Br J Cancer ( 1995 ) 72: 849-856; Br J Cancer ( 1996 ) 74: 1030-1036; MoI Pharmacol ( 1997 ) 52: 249-258; Leukemia ( 1999 ) 13 : 901-909; GHa ( 2002 ) 40:44-54; Clin Cancer Res ( 2000 ) 6:2860-2867 and ( 2004 ) 10:881-889) and in vivo models ( Blood ( 2002 ) 99:2241-2244 Clin Cancer Res ( 2003 ) 9: 5370-5379; J Natl Cancer Inst ( 2004 ) 96: 56-67) was confirmed. PARP inhibitors can also prevent the appearance of tissue necrosis induced by selective N3-adenine methylation drugs such as MeOSO ₂ (CH ₂ )-lexitropsin (Me-Lex) ( Pharmacological Research ( 2005 ) 52: 25-33).

PARP inhibitors are also useful as radiosensitizers. Studies have shown that PARP inhibitors are effective in radiosensitizing (hypoxic) tumor cells; preventing potential lethal damage of DNA from tumor cells after radiation therapy ( Br. J. Cancer ( 1984 ) 49 (Suppl. VI) : 34-42 ; and Int.J.Rαdiαt.Bioi. ( 1999 ) 75:91-100) and sublethal damage ( Clin. Oncol. ( 2004 ) 16(1): 29-39) regeneration, speculated for the cause It is through PARP inhibitors that can prevent recombination of DNA strand breaks and affect multiple DNS damage signaling pathways.

PARP inhibitors are also useful in the treatment of acute and chronic cardiomyopathy ( Pharmacological Research ( 2005 ) 52: 34-43). For example, studies have shown that a single injection of a PARP inhibitor can reduce the infarct size induced by ischemia and reperfusion in rat heart or skeletal muscle. In these studies, a single injection of the PARP inhibitor 3-amino-benzamide (10 mg/kg) one minute prior to occlusion or reperfusion also similarly reduced the infarct size (32% to 42%). In addition, the relative level of myocardial infarct size reduction of another PARP inhibitor 1,5-dihydroxyisoquinoline (1 mg/kg) was 38% to 48%. From the above findings, it can be reasonably concluded that PARP inhibitors can be used to rescue ischemic heart disease or skeletal muscle tissue reperfusion injury in advance ( PNAS ( 1997 ) 94: 679-683). Similar findings are also in pigs ( Eur. J. Pharmacol. ( 1998 ) 359: 143-150; Ann. Thorac. Surg. ( 2002 ) 73: 575-581) and dogs ( Shock. ( 2004 ) 21: 426- 32) The body was found.

PARP inhibitors are also useful in the treatment of certain vascular diseases, septic shock, ischemic injury, and neurotoxicity ( Biochim. Biophys. Acta ( 1989 ) 1014:1-7; J. CHn . Inves t. ( 1997 ) 100: 723-735). Oxygen free radical-induced DNA damage can lead to DNA strand breaks, as revealed by the PARP Inhibitors Institute, which is a major contributor to these disease states, where the DNA strand breaks are subsequently recognized by PARP ( J. Neurosci. Res. ( 1994 ) 39:38-46; PNAS ( 1996 ) 93: 4688-4692). In addition, PARP also plays a role in the pathogenesis of hemorrhagic shock ( PNAS ( 2000 ) 97: 10203-10208). PARP inhibitors are also useful in the treatment of inflammatory diseases ( Pharmacological Research ( 2005 ) 52: 72-82 and 83-92).

Studies have shown that inhibition of PARP activity is effective in blocking efficient retroviral infection of mammalian cells. This inhibition of recombinant retroviral vector infection is also present in a variety of different cell types ( J. Virology , ( 1996 ) 70(6): 3992-4000). Therefore, PARP inhibitors have been developed for antiviral therapy as well as tumor therapy (International Patent Publication No. WO 91/18591).

In vivo and in vitro experiments have shown that PARP inhibitors can be used to treat or prevent autoimmune diseases such as type I diabetes and diabetic complications ( Pharmacological Research ( 2005 ) 52: 60-71).

It is postulated that inhibition of PARP can be used to delay the onset of aging of human fibroblasts ( Biochem. Biophys. Res. Comm. ( 1994 ) 201(2): 665-672; Pharmacological Research ( 2005 ) 52: 93-99). This may be related to the role of PARP in controlling telomere function ( Nature Gen. , ( 1999 ) 23(1): 76-80).

Most of the current PARP inhibitors can interact with the nicotinamide binding domain of the PARP enzyme and can also act as a competitive inhibitor against NAD ⁺ ( Expert Opin. Ther. Patents ( 2004 ) 14: 1531-1551). Structural analogs of the nicotinamide, such as benzamide and its derivatives, belong to the PARP inhibitor compound of the first class of studies. However, these molecules have weak inhibitory activity and have other effects not related to PARP inhibitory activity. Therefore, it is necessary to provide a class of potential PARP enzyme inhibitors.

The present invention provides a class of compounds that inhibit poly(ADP-ribose) polymerase (PARP). In particular, these compounds are useful for inhibiting PARP-1 and/or PARP-2.

The present invention provides a class of compounds of formula (I),

a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein: A and B are each independently selected from CR ₃ or N; and R _{3 is} selected from the group consisting of H, halogen, C _1-6 alkyl, halogenated C _1-6 An alkyl group, a C _2-6 alkenyl group, a halogenated C _2-6 alkenyl group, a C _2-6 alkynyl group, a halogenated C _2-6 alkynyl group or CN; R _{1 is} selected from the group consisting of H, OH, C _1-6 alkane Or NR ₇ R ₈ ; R ₇ and R ₈ are each independently selected from H, C _1-6 alkyl, halo C _1-6 alkyl, C _2-6 alkenyl, halo C _2-6 alkenyl , C _2-6 alkynyl, halogenated C _2-6 alkynyl, C _1-6 alkoxy, halo C _1-6 alkoxy or C _1-6 alkylcarbonyl; R _{2 is} selected from H, halogen , C _1-6 alkyl, halo C _1-6 alkyl, C _1-6 alkoxy, halo C _1-6 alkoxy or CN; X is selected from or Wherein m and n are each independently selected from 1, 2 or 3; and R _{4 is} selected from H, halogen, C _1-6 alkyl, halo C _1-6 alkyl, C _1-6 alkoxy or halo a C _1-6 alkoxy group; R _{5 is} selected from the group consisting of H, -OH, halogen, C _1-6 alkyl, halo C _1-6 alkyl, C _1-6 alkoxy, halo C _1-6 Alkoxy, C _3-6 cycloalkyl, halogenated C _3-6 cycloalkyl, NR ₁₁ R ₁₂ , C _1-6 alkyl (NR ₁₁ R ₁₂ ), C _1-6 alkoxycarbonyl or Wherein R ₁₁ and R ₁₂ are each independently selected from H, C _1-6 alkyl or C _1-6 alkoxy; v is selected from 0, 1 or 2; and R _{9 is} selected from H, C _1-6 alkane , halo C _1-6 alkyl, C _2-6 alkenyl, C _3-6 cycloalkyl or C _3-6 cycloalkyl C _1-4 alkyl; R _{10 is} selected from H, C _1-4 An alkyl group or a C _2-4 alkenyl group; or R ₉ and R ₁₀ , together with the atoms to which they are attached form a dihydrofuran or tetrahydrofuran; Z is selected from substituted or unsubstituted, and contains at least two or more selected from N, O. Or a 5-15 membered non-aromatic monocyclic ring of a hetero atom of S; p is selected from 1 or 2; each R _{6 is} independently selected from H, halo, C _1-6 alkyl or halo C _1-6 alkyl.

With regard to the compounds of formula (I), the invention further provides some preferred technical solutions.

In some embodiments, R ₁ in formula (I) is selected from H, C _1-6 alkyl or OH.

In some embodiments, R ₁ in formula (I) is selected from H, methyl or OH.

In some embodiments, R ₂ in formula (I) is selected from H, halo, C _1-6 alkyl, C _1-6 alkoxy or halo C _1-6 alkoxy.

In some embodiments, R ₂ in formula (I) is selected from H, F, methyl, methoxy or chloro substituted methoxy.

In some embodiments, A in formula (I) is CR ₃ .

In some embodiments, R ₃ in formula (I) is selected from H or C _1-6 alkyl.

In some embodiments, B in formula (I) is N.

In some embodiments, the X in formula (I) is .

In some embodiments, R ₄ in formula (I) is selected from H, C _1-6 alkyl or C _1-6 alkoxy.

In some embodiments, R ₅ in formula (I) is selected from the group consisting of H, OH, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkyl (NR ₁₁ R ₁₂ ) or .

In some embodiments, R ₅ in formula (I) is .

In some embodiments, v in formula (I) is selected from 0 or 1.

In some embodiments, R ₉ in formula (I) is selected from H, C _1-6 alkyl or C _2-6 alkenyl.

In some embodiments, R ₁₀ in formula (I) is selected from H, C _1-6 alkyl or C _2-6 alkenyl.

In some embodiments, R ₉ and R ₁₀ in formula (I) are taken together with the atoms to which they are attached to form dihydrofuran or tetrahydrofuran.

In some embodiments, R ₅ in formula (I) is selected from C _1-6 alkyl (NR ₁₁ R ₁₂ ), and R ₁₁ and R ₁₂ are each independently selected from C _1-3 alkyl.

In some embodiments, R ₁₁ and R ₁₂ in formula (I) are both methyl.

In some embodiments, R ₅ in formula (I) is .

In some embodiments, the X in formula (I) is .

In some embodiments, p in formula (I) is selected from 1 or 2.

In some embodiments, R ₆ in formula (I) are each independently selected from H or C _1-6 alkyl.

In some embodiments, Z in formula (I) is selected from substituted or unsubstituted 5-15 membered non-aromatic monocyclic rings containing at least 2-5 heteroatoms selected from O or S.

In some embodiments, Z in formula (I) is selected from substituted, unsubstituted, 5, 6, 7, 8, 9, 11 having at least 2, 3, 4 or 5 heteroatoms selected from O or S. , 12 or 15 yuan non-aromatic single ring.

In some embodiments, Z in formula (I) is selected from , , , , ,

In some embodiments, the compound of formula (I), ie, the compound of formula (II):

Wherein s is selected from 1, 2 or 3; t is selected from 1 or 2; and R _{5 is} selected from the group consisting of H, OH, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkyl (NR ₁₁ R ₁₂ ) or .

The invention further provides some of the more preferred embodiments of formula (II).

In some embodiments, t in formula (II) is 1 or 2.

In some embodiments, s in formula (II) is 1 or 3.

In some embodiments, R ₅ in formula (II) is selected from H, OH, C _1-3 alkyl or .

In some embodiments, R ₅ in formula (II) is ; v is 0; R _{9 is} selected from H, C _1-4 alkyl or C _2-4 alkenyl; and R _{10 is} selected from H, C _1-3 alkyl or C _2-3 alkenyl.

In some embodiments, the compound of formula (I), ie, the compound of formula (III):

Wherein Z _{1 is} selected from substituted or unsubstituted 5, 6, 7, 8, 9, 11, 12 or 15 non-aromatic singles containing 2, 3, 4 or 5 heteroatoms selected from O or S ring.

In some embodiments, Z ₁ in formula (III) is selected from , , , , ,

The invention further provides some particularly preferred embodiments of the compound of formula (I) selected from the group consisting of Examples 1-45, including: 1) 2-(4-(3-azabicyclo[4.1. 0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 2) 2- (4- (1-methoxymethyl-3-azabicyclo [ 4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 3) 2- (4- (1-methyl-3-azabicyclo [4.1 .0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 4) 2- (4- (1-ethyl-3-azabicyclo [4.1. 0] heptane-6-yl) phenyl) -2 H - indazol-7-amine hydrochloride acyl; 5) 2- (4- (1-isopropyl-3-azabicyclo [4.1. 0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 6) 2- (4- (1-hydroxy-3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 7) 2- (4- (1- (ethoxymethyl) -3-azabicyclo [ 4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 8) 2- (4- (1- (2-methoxyethyl) - 3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 9) 2- (4- (1- (1-hydroxy Allyl)-3-azabicyclo[4.1.0]heptane-6-yl)phenyl)-2H- Oxazole-7-formamide hydrochloride; 10) 2-(4-(1-(hydroxymethyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-2 H -carbazole-7-carbamide hydrochloride; 11) 2-(4-(1-(1-hydroxyethyl)-3-azabicyclo[4.1.0]heptan-6-yl)benzene -2H-carbazole-7-carbamide hydrochloride; 12) 2-(4-(1-(1-hydroxypropyl)-3-azabicyclo[4.1.0]heptane-6- yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 13) 2- (4- (1- (1-hydroxybutyl) -3-azabicyclo [4.1.0] hept- 6-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 14) 2- (4- (1- (1-hydroxy-3-methylbutyl) -3- Azabicyclo[4.1.0]heptane-6-yl)phenyl)-2H-carbazole-7-formamide hydrochloride; 15) 2-(4-(1-(1-hydroxybutan-3) - enyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 16) 2- (4- (1 - (1-methoxyethyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 17) 2 - (4- (1- (1-methoxypropyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl amine salt salt; 18) 2- (4- (1- (1-methoxybutyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole - 7-methanamine hydrochloride; 19) 2-(4 - (1- (1-methoxy-3-methylbutyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-XI Amine hydrochloride; 20) 2-(4-(1-(1-methoxybut-3-enyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)- 2 H - indazole-7-amine hydrochloride acyl; 21) 2- (4- (1- (1-ethoxy-3-enyl) -3-azabicyclo [4.1.0] hept- 6-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 22) 2- (4- (1- (1- (allyloxy) allyl) -3 - azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride, 23) 2- (4- (l- (2,3- dihydro-furan-2-yl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride, 24) 2- ( 4- (1- (tetrahydrofuran-2-yl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 25 2-(4-(1-(methoxymethyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-5-methyl- 2H -carbazole-7 -carbamamine; 26) 5-fluoro-2-(4-(1-(methoxymethyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-2 H -carbazole-7-formamide; 27) 5-methoxy-2-(4-(1-(methoxymethyl)-3-azabicyclo[4.1.0]heptane-6-yl Phenyl)-2 H -carbazole-7-formamide; 28)2- (4-(1-(Methoxymethyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-N-methyl- 2H -carbazole-7-carboxamidine Amine; 29) N-hydroxy-2-(4-(1-(methoxymethyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-2 H -carbazole -7-carbamamine; 30) 2-(4-(1-(methoxymethyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-3-methyl -2 H - indazole-7-Amides; 31) of 3-chloro-2- (4- (1- (methoxymethyl) -3-azabicyclo [4.1.0] heptane-6 yl) phenyl) -2 H - indazole-7-Amides; 32) 2- (4- (1- (2- (dimethylamino) ethyl) -3-azabicyclo [4.1.0 ] heptane-6-yl) phenyl) -2 H - indazole-7-Amides; 33) 2- (4- (3-azabicyclo [3.1.0] hexane-1-yl) benzene yl) -2 H - indazole-7-amine hydrochloride acyl; 34) 2- (4- (octahydro-cyclopenta [c] pyrrol -3a--yl) phenyl) -2 H - indazole - 7-carbamide hydrochloride; 35) 2-(2,3-dihydrobenzo[b][1,4]dioxine-6-yl)-2 H -carbazole-7- Methionine; 36) 2-(3,4-dihydro-2H-benzo[b][1,4]dioxe-7-yl)-2 H -carbazole-7-carboxamide ;37) 2-(benzo[b][1,4]dioxooctyl)-2H-indazole-7-formamide; 38)2-(benzo[d][1,3] Olecyclopentene-5-yl)-2H-carbazole-7-formamide; 39) 2-(2,2- Dimethylbenzo[d][1,3]dioxol-5-yl)-2H-indazole-7-formamide; 40)2-(2,3,5,6,8 ,9-hexahydrobenzo[b][1,4,7,10]tetraoxacyclododecyl)-2H-indazole-7-carboxamide;41)2-(2,3,5 ,6-tetrahydrobenzo[b][1,4,7]trioxacycloalkyl)-2H-carbazole-7-carboxamide;42)2-(2,3,5,6, 8,9,11,12-octahydrobenzo[b][1,4,7,10,13]pentaoxacyclopentadecyl)-2H-carbazole-7-carboxamide;43)2 -(2,3,5,6,8,9-hexahydrobenzo[b][1,4,7,10]tetraoxacyclododecyl)-2H-indazole-7-carboxamide ;44) 2-(2,3,4,6,7,8-hexahydrobenzo[b][1,4,8]-1,4-dioxo-8-thio-11-yl) -2H-carbazole-7-formamide; 45) 2-(2,3,4,6,7,8-hexahydrobenzo[i][1,7,4]-1,4-dioxo -8-8-fluorenyl-11-yl)-2H-carbazole-7-formamide; 46) 2-(4-(3-azabicyclo[4.1.0]heptan-6-yl)phenyl -3a,7a-dihydro-1H-benzo[d]imidazole-4-carboxamide hydrochloride.

The invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound as described above and a pharmaceutically acceptable adjuvant.

Further, the present invention provides the above compound or pharmaceutical composition as a medicament for treating, preventing or delaying a chemotherapeutic agent or radiosensitization in a disease, cancer, cancer treatment, which can be alleviated by inhibiting poly(ADP-ribose) polymerase Application of the agent.

The invention further provides a preferred technical solution for the application: preferably, the compound or pharmaceutical composition is for treating, preventing or delaying cancer, inflammatory diseases, reperfusion injury, ischemic diseases, stroke, acute and chronic Renal failure, cardiovascular disease, other vascular diseases other than cardiovascular disease, diabetes, neurodegenerative diseases, anti-retroviral infection, retinal damage, skin aging or ultraviolet-induced skin damage.

Preferably, the inflammatory disease is selected from the group consisting of organ transplant rejection, inflammatory bowel disease, inflammatory lung disease, inflammatory eye disease, chronic inflammatory gingivitis, inflammatory nephropathy, inflammatory skin disease, inflammatory central nervous system disease , diabetic complications, inflammatory heart disease, and various other diseases with significant inflammatory elements or inflammatory systemic system.

Preferably, the ischemic disease includes diseases caused by anemia and organ transplantation.

Preferably, the diabetes is selected from the group consisting of type I diabetes (insulin dependent diabetes), type II diabetes (non-insulin dependent diabetes), gestational diabetes, autoimmune diabetes, insulin disease, diabetes caused by pancreatic diseases, and other hormonal diseases. Diabetes, type A insulin resistance syndrome, type B insulin resistance syndrome, adipose atrophic diabetes or 3-cytotoxin-induced diabetes.

Preferably, the neurodegenerative disease is selected from the group consisting of a neurodegenerative disease caused by prolonged polyglutamine sequence, Huntington's disease, Kendi disease, cerebellar atrophy, dentate nucleus nucleus pallidus Lewy body atrophy ( DRPLA), neurodegenerative diseases associated with protein aggregation, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, spongiform encephalopathy, prion-related diseases, and multiple sclerosis (MS).

Preferably, the cancer is selected from the group consisting of a solid tumor, a blood-borne cancer, acute or chronic leukemia, lymphoma, central nervous system cancer (CNS), brain cancer, breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, small cell lung cancer. , gastric cancer, neuroepithelial neoplasia, cancer lacking homologous recombination (HR)-dependent DNA double-strand break (DSB) repair activity, BRCA-1 or BRCA-2 deficiency tumor.

Preferably, the cancer lacking homologous recombination (HR)-dependent DNA double-strand break (DSB) repair activity comprises one or more reduced or completely lacking ability of homologous recombinant DNA double-strand break repair compared to normal cells Cancer cells.

Preferably, the cancer cell has a BRCA-1 deficient phenotype or a BRCA-2 deficient phenotype, or the cancer cell lacks BRCA-1 or BRCA-2.

Solvates and salts of the compounds of the formulae (I), (II) and (III), such as hydrates, are also included within the scope of the invention.

The compounds of the present invention may contain asymmetric centers, chiral axes, and chiral surfaces (see Steveochemistry, Carbon Compounds, published by John Wiley & Sons, New York, 1994). Of Carbon Compounds) (pp. 1119-1190 in the book edited by EX. Eliel and SHwilen), which may exist as racemates, racemic mixtures, individual diastereomers, all possible isomers and Mixtures, including optical isomers as well as all such stereoisomers included in the present invention. Furthermore, the compounds described herein may exist as tautomers, and even though only one tautomeric structure is described herein, all tautomers are also included in the scope of the present invention. The compounds of the invention may exist in different isomeric forms, and all isomers are included within the scope of the invention.

The compounds of the invention may exist in a number of different polymorphic forms. In any constituent of a compound, when any variable (such as R ₁ and R _{2 ,} etc.) occurs more than once, the definition of each change in the variable is independent in the case of various other changes. Likewise, combinations of substituents and variables are also permissible as long as such a combination can form a stable compound. A substituted baseline bar drawn within a ring system means that the chemical bond shown can be attached to any substitutable ring atom.

It will be understood that the substituents and substitutions on the compounds described herein can be selected by one of ordinary skill in the art to provide such compounds which are chemically stable and which can be employed by techniques known in the art. The starting materials obtained are relatively easy to prepare. These technical methods will be discussed below. If the substituent itself is substituted by more than one group, it can be understood that as long as the structure of the obtained compound is stable, the above plurality of groups may be bonded to the same or different carbon atoms. The term "optionally containing a substituent" is equivalent to the term "substituent-free or containing one or more substituents". In this case, a preferred embodiment contains 0 to 3 substituents. more Preferably, it contains 0 to 2 substituents. A substituent on a saturated, partially saturated or unsaturated heterocyclic ring can be attached to any substitutable position on the heterocyclic ring.

"C _nm " refers to the number of carbon atoms in a group, where n and m are positive integers. "n-ary" refers to the number of atoms in a cyclic group, for example, a pyridyl group is a 6-membered ring.

As used herein, "alkyl" refers to a branched or straight chain saturated aliphatic hydrocarbon group containing a defined number of carbon atoms. For example, " _C1-6 alkyl" means an alkyl group comprising a straight or branched chain arrangement containing 1, 2, 3, 4, 5 or 6 carbon atoms. For example, "C _1-6 alkyl" specifically includes methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, pentyl and hexyl groups and the like. In some embodiments, preferred alkyl groups are methyl and ethyl.

As used herein, "C _2-6 alkenyl" refers to a straight or branched chain non-aromatic hydrocarbon group containing from 2 to 6 carbon atoms and containing at least one carbon to carbon double bond. It preferably contains one carbon-carbon double bond and may contain up to four non-aromatic carbon-carbon double bonds. The alkenyl group includes a vinyl group, a propenyl group, a butenyl group, and a 2-methylbutenyl group. In some embodiments, preferred alkenyl groups include ethenyl and propenyl.

As used herein, "C _2-6 alkynyl" refers to a straight or branched chain hydrocarbon radical containing from 2 to 6 carbon atoms and containing at least one carbon to carbon triple bond. It can contain up to 3 carbon-carbon triple bonds. The alkynyl group includes an ethynyl group, a propynyl group, a butynyl group, a 3-methylbutynyl group and the like. In some embodiments, preferred alkynyl groups include ethynyl and propynyl.

As used herein, "cycloalkyl" refers to a monocyclic, bicyclic or polycyclic saturated aliphatic hydrocarbon group containing a specified number of carbon atoms. For example, "C _3-6 cycloalkyl" includes cyclopropyl, methylcyclopropyl, 2,2-dimethyl-cyclobutyl, 2-methyl-cyclopentyl, cyclohexyl and the like. Preferred cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

The term "C _3-6 cycloalkyl C _1-4 alkyl" refers to a C _3-6 cycloalkyl group having a defined number of carbon atoms attached through the C _1-4 alkyl side.

"Alkoxy" means an alkyl group having the specified number of carbon atoms attached through an oxygen (O) bridge. Examples of suitable C _1-6 alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy and tert-butoxy. In some embodiments, a preferred alkoxy group is methoxy or ethoxy.

As used herein, "halogen" means fluoro, chloro, bromo and iodo, with fluoro and chloro being preferred.

The terms "halogenated C _1-6 alkyl", "halo C _2-6 alkenyl", "halo C _2-6 alkynyl" and "halo C _1-6 alkoxy" refer to C ₁ respectively. One or more (preferably 1 to 3) hydrogen atoms of the _-6 alkyl group, the C _2-6 alkenyl group, the C _2-6 alkynyl group or the C _1-6 alkoxy group are preferably a halogen atom, and particularly preferably It is a group obtained by substituting a fluorine atom (F) or a chlorine (Cl) atom. In some embodiments, preferred are fluoro C _1-6 alkyl, fluoro C _2-6 alkenyl, fluoro C _2-6 alkynyl and fluoro C _1-6 alkoxy, further preferably fluoro C _1-3 alkyl, such as CF ₃ , CHF ₂ , CH ₂ F, CH ₂ CH ₂ F, CH ₂ CHF ₂ , CH ₂ CF ₃ and fluoro C _1-3 alkoxy, such as OCF ₃ , OCHF ₂ , OCH ₂ F, OCH ₂ CH ₂ F, OCH ₂ CHF ₂ and OCH ₂ CF ₃ , most preferred are CF ₃ , OCF ₃ and OCHF ₂ .

The term "hydroxy C _1-6 alkyl" as used herein means a group obtained by substituting one or more (preferably 1 to 3) hydrogen atoms of an alkyl group with a hydroxyl group. Preferably, the hydroxy C _1-6 alkyl group is selected from the group consisting of CH ₂ OH, CH ₂ CHOH or CHOHCH ₃ .

The terms "C _1-6 alkylcarbonyl" and "C _1-6 alkoxycarbonyl" mean a group which is bonded through a carbonyl group (C=O), respectively. Examples of suitable "C _1-6 alkylcarbonyl" include methylcarbonyl, ethylcarbonyl, propylcarbonyl, isopropylcarbonyl and tert-butylcarbonyl. Examples of the "C _1-6 alkoxycarbonyl group" include a methoxycarbonyl group, an ethoxycarbonyl group, a propoxycarbonyl group, an isopropoxycarbonyl group, and a tert-butylcarbonyl group. Similarly, the term "C _6-10 arylcarbonyl" is explained, an example of which is benzinyl. The ring in the compounds of the invention may be monocyclic or polycyclic, preferably bicyclic. The polycyclic ring can be a fused ring or a spiro ring.

The present invention includes the free base of the compounds of the formulae (I), (II) and (III), and also includes pharmaceutically acceptable salts thereof and stereoisomers thereof. The guanamine-based and/or N-containing heterocyclic moiety-containing N atoms of the compounds of the invention may be protonated to a salt. The term "free base" refers to an amine compound in a non-salt form. The pharmaceutically acceptable salts included in the present invention include not only the salts exemplified herein for the specific compounds, but also all of the typical free forms of the compounds of the formulae (I), (II) and (III). Corresponding pharmaceutically acceptable salts. The free forms of these specific salt compounds can be isolated using techniques known in the art. For example, the free form may be treated with a suitable dilute aqueous base such as NaOH, potassium carbonate, aqueous ammonia, and sodium bicarbonate to treat the salt for regeneration. The physical properties of these free forms, such as the solubility in polar aqueous solutions, will be slightly different from their respective salt forms. However, for the purposes of the present invention, their acid and base salts are pharmaceutically equivalent to their respective free forms.

The pharmaceutically acceptable salts of the compounds of the present invention can be prepared by conventional chemical treatment of the compounds of the invention containing a basic moiety or an acidic moiety. In general, the salt of the basic compound can be prepared by ion exchange chromatography, or can be prepared by reacting the free base with a stoichiometric or excess amount of the corresponding salt-forming inorganic or organic acid in a suitable solvent or various mixed solvents. . Similarly, salts of the acidic compounds can be prepared using suitable inorganic or organic bases. Thus, pharmaceutically acceptable salts of the compounds of the invention include the common non-toxic salts of the basic compounds of the invention formed by mineral, organic or polymeric acids. For example, common non-toxic salts include inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfurous acid, sulfamic acid, phosphoric acid, phosphorous acid, nitric acid, and salts derived from inorganic acids, and organic acids such as acetic acid. Propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, Salicylic acid, sulfamic acid, 2-ethylindenyl-benzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethane disulfonic acid, acetic acid, oxalic acid, isethionic acid, palmitic acid, gluconic acid, Phenylacetic acid, aspartic acid, cinnamic acid, pyruvic acid, ethanesulfonic acid, disulfonic acid, shikimic acid, trifluoroacetic acid, and the like. Examples of suitable high molecular acid salts include polymeric acid-derived salts such as tannic acid and carboxymethyl cellulose. In some embodiments, the pharmaceutically acceptable salts of the present invention comprise one equivalent of a compound of formula (I), (II) and (III) and one, two or three equivalents of a mineral or organic acid. More specifically, the pharmaceutically acceptable salt of the present invention is a trifluoroacetate or hydrochloride. In one embodiment, the salt is a trifluoroacetate salt. In another embodiment, the salt is a hydrochloride salt. When the compound of the present invention is acidic, a suitable "pharmaceutically acceptable salt" means a salt prepared from a pharmaceutically acceptable non-toxic base including inorganic bases and organic bases. These inorganic base-derived salts include aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, manganese (III) salts, manganese (II) salts, potassium salts, sodium salts, A zinc salt or the like similar to an inorganic base derived salt. Particularly preferred are ammonium salts, calcium salts, magnesium salts, potassium salts and sodium salts. Pharmaceutically acceptable organic non-toxic base derived salts include primary, secondary, tertiary amines, amines containing substituents (including natural amines containing substituents), cyclic amines and basic ion exchange resins such as arginine Lysine, betaine, caffeine, choline, N,N-dibenzylethylenediamine, ethylamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, diethanolamine, Ethylenediamine, N-ethylmorpholine, N-ethyl acridine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methyl Glucosamine, morpholine, acridine, pyridazine, polyamine resin, procaine, guanidine, cocoa base, triethylamine, trimethylamine, tripropylamine, tromethamine, two Cyclohexylamine, butylamine, benzylamine, phenylbenzylamine, and the like. A more complete description of the preparation of the above pharmaceutically acceptable salts and other typical pharmaceutically acceptable salts is found in the literature by Berg et al. (1997) J. Pharm. Sci., 'pharmaceutical Salts' , 66: 1-19.

It is also worth noting that the compound of the present invention has a potential internal salt or zwitterion, since the compound deprotonates the acid moiety under physiological conditions, such as a carboxyl group, which may be an anion, and the charge may be resistant to protonation or alkylation. The alkali moiety, such as the quaternary nitrogen atomic cation charge, reaches internal equilibrium.

The present invention provides a compound for treating and/or preventing a disease ameliorated by inhibiting poly(ADP-ribose) polymerase (PARP) (see, for example, Nature Review Drug Discovery ( 2005 ) 4:421-440).

Accordingly, the present invention provides a compound of the formulae (I), (II) and (III) for use in the manufacture of a medicament for the treatment and/or prevention of a disease ameliorated by inhibition of poly(ADP-ribose) polymerase (PARP). The invention also provides a method for treating and/or preventing a disease ameliorated by inhibition of poly(ADP-ribose) polymerase (PARP). The method comprises administering to a patient a therapeutically effective amount of a compound of formula (I), (II) and (III) and/or a compound comprising a compound of formula (I), (II) and (III), according to their needs. combination.

The PARP inhibitors of the present invention are useful in the treatment of the diseases specified in International Publication No. WO2005/082368.

Accordingly, the present invention provides a compound of the formulae (I), (II) and (III) which can be used for the preparation of a medicament for the treatment and/or prevention of diseases including cancer, inflammatory diseases, reperfusion injury, deficiency Blood diseases, stroke, renal failure, cardiovascular disease, non-cardiovascular vascular disease, diabetes, neurodegenerative diseases, retroviral infection, retinal damage, skin aging or skin damage caused by ultraviolet rays.

The compounds of the present invention are useful for the treatment of inflammatory diseases, including those caused by organ transplant rejection, such as chronic inflammatory joint diseases, including arthritis, rheumatoid arthritis, osteoarthritis, and bone resorption associated with increased bone resorption. Disease; inflammatory bowel disease such as ileitis, ulcerative colitis, Barrett's syndrome and Crohn's disease; inflammatory lung diseases such as asthma, adult respiratory distress syndrome and chronic Respiratory obstruction; inflammatory eye diseases, including corneal dystrophy, trachoma, onchocerciasis, uveitis, sympathetic ophthalmia and endophthalmitis; chronic inflammatory diseases of the gums, including gingivitis and periodontitis; tuberculosis; Leprosy; inflammatory nephropathy, including uremic complications, glomerulonephritis and nephrosis; inflammatory skin diseases including sclerosing dermatitis, psoriasis and eczema; inflammatory central nervous system (central nervous system, CNS) diseases, including chronic nervous system demyelinating diseases; multiple sclerosis (MS), AIDS-related neurodegenerative diseases, and Alzheimer's disease, infectious meningitis, encephalomyelitis, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and viral or autoimmune encephalitis; diabetic complications include, but are not limited to, , immune complex vasculitis, systemic lupus erythematosus (SLE); inflammatory heart disease, such as (primary) cardiomyopathy, ischemic heart disease, hypercholesterolemia and atherosclerosis; Various other diseases with significant inflammatory characteristics, including pre-eclampsia, chronic liver failure, brain and spinal cord injury, multiple organ dysfunction syndrome (MODS), and multiple organ failure (MOF) . These inflammatory diseases can also be systemic inflammation of the body, which can be induced by Gram-positive or Gram-negative shock, hemorrhagic or anaphylactic shock, or cancer chemotherapy in response to pro-inflammatory cytokines. An illustration of shock (such as pro-inflammatory cytokine-related shock). These shocks can be triggered by chemotherapeutic agents such as those administered by cancer therapy. Therefore, the present invention provides a Compounds of formula (I), (II) and (III) which are useful in the manufacture of a medicament for the treatment and/or prevention of inflammatory diseases.

The invention also provides a method of treating and/or preventing reperfusion injury comprising administering to a patient a therapeutically effective amount of a compound of formula (I), (II) or (III) or at least one structural formula according to its needs. A composition of the compounds of (I), (II) and (III).

The compounds of the present invention are also useful for the treatment and/or prevention of ischemic diseases, including ischemic diseases caused by organ transplantation, such as stable angina pectoris, unstable angina pectoris, myocardial ischemia, hepatic ischemia, mesenteric artery deficiencies Blood, intestinal ischemia, key limb ischemia, chronic severe limb ischemia, cerebral ischemia, acute cardiac ischemia, ischemic nephropathy, ischemic liver disease, ischemic retinopathy, septic shock, and central nervous system Ischemic disease, such as stroke or cerebral ischemia.

The invention also provides a method of treating and/or preventing stroke comprising administering to a patient a therapeutically effective amount of a compound of formula (I), (II) or (III) or at least one structural formula (I) A combination of compounds of (II) and (III).

The compounds of the invention may also be used to treat and/or prevent chronic or acute renal failure.

The compounds of the invention may also be used to treat and/or prevent vascular diseases other than cardiovascular diseases, such as peripheral vascular occlusion, thromboangiitis obliterans, Reynaud's disease and Raynaud's phenomenon, hand and foot cyanosis, Erythematous limb pain, venous thrombosis, varicose veins, arteriovenous fistula, lymphedema, and fatty edema. Accordingly, the present invention also provides compounds of structural formula (I), (II) and (III) useful in the manufacture of a medicament for the treatment and/or prevention of vascular diseases.

The invention also provides a method of treating and/or preventing a cardiovascular disease comprising administering to a patient a therapeutically effective amount of a compound of formula (I), (II) and (III) or at least one structural formula according to its needs. A composition of the compounds of (I), (II) and (III).

The compounds of the invention may also be used for the treatment and/or prevention of diabetes, including type 1 diabetes (insulin dependent diabetes), type 2 diabetes (non-insulin dependent diabetes), gestational diabetes, autoimmune diabetes, abnormal insulinemia (insulinopathies) Diabetes caused by pancreatic diseases, diabetes associated with other hormonal diseases (such as Cushing's syndrome, acromegaly, pheochromocytoma, glucagonoma, primary aldosteronism) Or somatostatin), type A insulin resistance syndrome, type B insulin resistance syndrome, adipose atrophic diabetes or 3-cytotoxin-induced diabetes. The compounds of the present invention are also useful for the treatment and/or prevention of diabetic complications such as diabetic cataracts, glaucoma, retinopathy, nephropathy (such as microalbuminuria and progressive diabetic nephropathy), multiple neuropathy, foot gangrene, arteries Atherosclerotic coronary artery disease, peripheral vascular disease, non-ketotic hyperglycemia-hyperosmolar coma, mononeuropathy, autonomic neuropathy, foot ulcer, joint problems, and skin or mucosal complications (such as infection, ecchymosis, candida) Infection or diabetes obesity lipid progressive necrosis), hyperlipidemia, hypertension, insulin resistance syndrome, coronary heart disease, retinopathy, diabetic neuropathy, multiple neuropathy, mononeuropathy, autonomic neuropathy, foot ulcer, joint problems, Fungal infections, bacterial infections, and myocardial lesions. Accordingly, the present invention provides a composition of a compound of formula (I), (II) and (III) which can be used in the manufacture of a medicament for the treatment and/or prevention of diabetes.

The compounds of the invention may also be used to treat and/or prevent cancer, including solid tumors, blood-borne cancers, acute and chronic leukemias, lymphomas, central nervous system (CNS), and brain tumors, lacking homology Homologous Recombination (HR)-dependent DNA double-strand breaks (DSB) repair activity of cancer, manifested as BRCA-1 or BRCA-2 deficient tumors.

Solid tumors include fibrosarcoma, mucinous sarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, lymphatic endothelial sarcoma, synovial tumor, mesothelioma, Avon's tumor ( Ewing's tumor), leiomyosarcoma, rhabdomyosarcoma, colon cancer, colorectal cancer, kidney cancer, pancreatic cancer, bone cancer, breast cancer, ovarian cancer, prostate cancer, esophageal cancer, stomach cancer, oral cancer, nasal cancer, throat cancer, scale Cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland cancer, sebaceous gland cancer, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchial carcinoma, renal cell carcinoma, hepatocellular carcinoma, cholangiocarcinoma, villi Membrane cancer, seminoma, embryonic carcinoma, Wilms' tumor, cervical cancer, uterine cancer, testicular cancer, small cell lung cancer, bladder cancer, lung cancer, epithelial cell carcinoma, skin cancer, melanoma, Neuroblastoma and retinoblastoma; blood-borne cancer, such as acute lymphoblastic leukemia (ALL), acute lymphoblastic B-cell leukemia, acute lymphoid Cellular T-cell leukemia, acute myeloblasts leukemia (AML), acute promyelocyte leukemia (APL), acute monocytic leukemia, acute erythrocyte leukemia, acute megakaryoblastic leukemia, acute myeloid Monocytic leukemia, acute non-lymphocytic leukemia, acute undifferentiated leukemia, chronic myelocytic leukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia, and multiple myeloma; Acute and chronic leukemia, such as lymphoblastic, myeloid, lymphocytic, myeloid leukemia; lymphoma, such as Hodgkin's disease, non-Hodgkin's Lymphoma, multiple Myeloma, Waldenstrom's macroglobulinemia, heavy chain disease and polycythemia vera; central Nervous system and brain cancer, such as glioma, hairy cell astrocytoma, astrocytoma, anaplastic astrocytoma, glioblastoma multiforme, medulloblastoma, craniopharyngioma, Ependymoma, pineal tumor, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, vestibular nerve tumor, adenoma, metastatic brain tumor, meningioma, myeloma, and medullary canal Cell tumor.

In some embodiments, the compounds described herein are also useful in the treatment and/or prevention of cancers that lack Homologous Recombination (HR)-dependent DNA double-strand breaks (DBS) repair activity (see International Patent Application Publication No. WO2006/021801).

This HR-dependent DNA DSB repair pathway recombines a continuous DNA helix by repairing a double-strand break of DNA by a homologous mechanism ( Nat. Genet. ( 2001 ) 27(3): 247-254). Factors of the HR-dependent DNA DSB repair pathway include, but are not limited to, ATM (NM-000051), RAD51 (NM-002875), RAD51L1 (NM-002877), RAD51C (NM-002876), RAD51L3 (NM-002878) , DMC1 (NM-007068), XRCC2 (NM7005431), XRCC3 (NM-005432), RAD52 (NM-002879), RAD54L (NM-003579), RAD54B (NM-012415), BRCA-1 (NM-007295), BRCA-2 (NM-000059), RAD5O (NM-005732), MREIIA (NM-005590), NBS1 (NM-002485), ADPRT (PARP-1), ADPRTL2, (PARP02) CTPS, RPA, RPA1, RPA2 RPA3, XPD, ERCC1, XPF, MMS19, RAD51, RAD51p, RAD51C, RAD5 ID ₅ DMC1, XRCCR, XRCC3, BRCA1, BRCA2, RAD52, RAD54, RAD5O, MRE11, NB51, WRN, BLMKU70, RU80, ATM, ATRCHK1, CHK2 FANCA, FANCB, FANCC, FANCD1, FANCD2, FANCE, FANCF, FANCG, FANCC, FANCD1, FANCD2, FANCE, FANCF, FANCG, RAD1 and RAD9. Other HR-dependent DNA DSB repair pathways involve proteins including regulatory factors such as EMSY ( Cell ( 2003 ) 115:523-535).

The invention also provides a method of treating and/or preventing a BRCA-1 or BRCA-2 deficient tumor comprising administering to the patient a therapeutically effective amount of Structural Formula (I), (II) or (III) according to their needs. A compound or a composition comprising at least one compound of the formulae (I), (II) and (III).

In another embodiment, the cancer cells have a BRCA-1 and/or BRCA-2 deficient phenotype. Cancer cells with this phenotype may be deficient in BRCA-1 and/or BRCA-2, for example, the expression and/or activity of BRCA-1 and/or BRCA-2 may be reduced or inhibited in cancer cells, as by coding A manner in which a nucleotide is mutated or polymorphic; or in a manner that amplifies, mutates, or polymorphizes when the gene encodes a regulatory factor, such as an EMSY gene encoding a BRCA-2 regulatory factor (Cell ( 2003 ) 115: 523-535) .

BRCA-1 and BRCA-2 are known tumor suppressors whose wild-type alleles are frequently lost in tumor hybrid vectors ( Oncogene ( 2002 ) 21(58): 8981-93; Trends MoI Med. , ( 2002 ) 8 (12): 571-6). The association of BRCA-1 and/or BRCA-2 mutations with breast cancer has been well characterized ( Exp CHn Cancer Res. ( 2002 ) 21 (3 Suppl): 9-\2). The EMSY gene, which encodes a BRCA-2 binding factor, is also known to be associated with breast and ovarian cancer. Mutation vectors for BRCA-1 and/or BRCA-2 also increase the risk of ovarian, prostate and pancreatic cancer. Mutation detection of BRCA-1 and BRCA-2 is known in the art and is well documented in the literature, for example, European Patent EP 699 754; European Patent EP 705 903; Genet . Test ( 1992 ) 1: 75-83; Cancer Treat Res ( 2002 ) 107 :29-59; Neoplasm ( 2003 ) 50(4): 246-50; described in Ceska Gynekol ( 2003 ) 68(1): 11-16. Measurement of BRCA-2 binding factor EMSY amplification is described in the literature Cell 115:523-535. Studies have shown that PARP inhibitors can be used to specifically kill BRCA-1 and BRCA-2 deficient tumors ( Nature ( 2005 ) 434: 913-916 and 917-920).

The compounds of the present invention are useful for the treatment and/or prevention of neurodegenerative diseases, including neurodegenerative diseases caused by prolonged polyglutamine sequence, Huntington's disease, Kendi disease, cerebellar atrophy, dentate nucleus Nuclear pallidus hypothalamic nuclear atrophy (DRP), protein aggregation-related neurodegenerative diseases, Machado-Joseph disease, Alzheimer's disease, Parkinson's disease, muscle Atrophic lateral sclerosis, spongiform encephalopathy, prion-related diseases, and multiple sclerosis (MS).

The compounds of the invention may also be used to treat and/or prevent retroviral infection (US Patent No. US5652260), retinal damage ( Curr. Eye Res. ( 2004 ), 29: 403), skin aging and UV-induced skin. Injury (U.S. Patent No. 5,589,483; Biochem. Pharmacol ( 2002 ) 63:921).

The compounds of the invention may also be used in the treatment and/or prevention of cancer or tumor cells containing synergist-containing at least one ionizing radiation or chemotherapeutic agent.

The compounds of the present invention may be administered as a medicament, alone or in combination with pharmaceutically acceptable carriers, excipients, diluents, adjuvants, fillers, buffers, stabilizers, preservatives, and lubricants, according to standard pharmaceutical practice. The composition is administered to a mammal, preferably a human.

The compounds of the invention may be administered to the article by any convenient route of administration, whether systemically/peripherally or at a desired point of action, including but not limited to: oral administration (eg, swallowing); topical administration (including, for example, transdermal, intranasal, ocular, buccal, and sublingual); pulmonary administration (eg, by treatment with inhalation or insufflation through the mouth or nose, such as by aerosol); rectal administration Transvaginal administration; parenteral administration (for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, Intra-arterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subepidermal, intraarticular, subarachnoid and sternal); infusion (eg, subcutaneous Or intramuscular).

The invention also provides a pharmaceutical composition comprising one or more compounds of the invention and a pharmaceutically acceptable adjuvant. The term "pharmaceutically acceptable excipient" refers to any diluent, adjuvant, excipient or carrier with which at least one compound disclosed herein is administered.

The pharmaceutical composition containing the active ingredient may be in a form suitable for oral administration, for example, in the form of tablets, troches, lozenges, aqueous or oily suspensions, dispersions or granules, emulsions, hard or soft capsules, A syrup or elixir form. Compositions for oral administration can be prepared according to methods of preparation of pharmaceutical compositions known in the art. These pharmaceutical compositions may contain at least one agent selected from the group consisting of sweeteners, flavoring agents, coloring agents, or preservatives to obtain a pharmaceutically elegant and palatable preparation product. Tablets contain the active ingredient in admixture with a nontoxic pharmaceutically acceptable excipient. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating agents and disintegrating agents such as microcrystalline cellulose, croscarmellose sodium, corn starch Or alginic acid; binders such as starch, gelatin, polyvinyl-pyrrolidone or gum arabic; and lubricants such as magnesium stearate, octadecanoic acid or talc. These tablets may be uncoated or may be coated by prior art techniques to mask the unpleasant taste of the drug or to delay absorption of the gastrointestinal tract thereby providing a sustained action over a longer period of time. For example, a water-soluble taste masking agent such as hydroxypropyl-methylcellulose or hydroxypropylcellulose, or a retarding agent such as ethylcellulose or cellulose acetate butyrate may be employed.

Aqueous suspensions contain the active ingredient in admixture in admixture in the preparation of aqueous dispersions. These excipients may be suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose, sodium alginate, polyvinyl-pyrrolidone, tragacanth and acacia; dispersing agents or wetting agents Agent can be naturally occurring Phospholipids, such as lecithin, or condensates of alkylene oxides and fatty acids, such as polyoxyethylene stearate, or condensates of oxirane groups with long chain fatty alcohols, such as heptadecyl ethoxylates Heptadecaethyleneoxycetanol, or a condensate of a partial ester of ethylene oxide with a fatty acid hexitol anhydride, such as polyoxyethylene sorbitan monooleate. The aqueous suspensions may also contain at least one preservative such as ethyl p-hydroxybenzoate or n-propyl p-hydroxybenzoate, at least one color former, at least one flavoring agent, and at least one sweetener such as Sucrose, saccharin or Abbas.

The oily suspensions may be formulated in the form of a suspension active ingredient in a vegetable oil, such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil, such as paraffin oil. These oily suspensions may contain a thickening agent, such as beeswax, paraffin wax or cetyl alcohol. Sweeteners and flavoring agents such as those described above may be added to provide a palatable oral formulation product. These compositions can be preserved by the addition of an antioxidant such as butylated anisole or alpha-tocopherol.

Dispersible powders and granules are suitable for the preparation of aqueous suspensions, which are prepared by the addition of a water-soluble active ingredient to a mixture comprising a dispersing agent, a wetting agent, a suspending agent and at least one preservative. Examples of suitable dispersing or wetting agents and suspending agents have been mentioned above. Other excipients such as sweeteners, flavoring agents, and coloring agents may also be included. These compositions can be preserved by the addition of an antioxidant such as ascorbic acid. The pharmaceutical composition of the present invention may also be an oil-in-water emulsion. The oil phase may be a vegetable oil such as olive oil or peanut oil, or a mineral oil such as liquid paraffin oil, or a mixture thereof. Suitable emulsifiers may be: naturally occurring phospholipids, such as soy lecithin; and fatty acid and hexitol anhydride derived esters or partial esters, such as sorbitan monooleate; and the partial esters and ethylene oxide a condensate such as polyoxyethylene sorbitan monooleate. These emulsions may also contain sweeteners, flavoring agents, preservatives, and antioxidants. Syrups and elixirs can be formulated with sweeteners, such as glycerin, Propylene glycol, sorbitol or sucrose. These formulations may also contain lubricants, preservatives, flavoring agents, coloring agents, and antioxidants. These pharmaceutical compositions may be in the form of a sterile injectable aqueous solution.

The sterile injectable preparation may also be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oil phase. For example, the active ingredient can be first dissolved in a mixture of soybean oil and lecithin and then the resulting oil solution is poured into a mixture of water and glycerin to form a microemulsion.

These injections or microemulsions can be administered into the patient's bloodstream by topical single injection. As an option, it is advantageous to administer in this manner to maintain a constant loop concentration of the compounds of the invention. In order to maintain this constant concentration, a continuous intravenous infusion device can be employed. An example of such device is the type Deltec CADD-PLUS ^TM 5400 intravenous pump.

The pharmaceutical composition may be in the form of a sterile injectable aqueous suspension or an oily suspension for intramuscular and subcutaneous administration. The suspending agent can be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents as described above. The sterile injectable preparation may also be a sterile injectable solution or suspension, dissolved in a non-toxic parenterally acceptable diluent or solvent, such as a 1,3-butanediol solution. In addition, sterile, non-volatile oils are usually employed as a dissolving or suspending medium. For this purpose, any mild, non-volatile oil, including synthetic mono- or di-glycerides, can be employed. In addition, fatty acids such as oleic acid find use in the preparation of injectables. The compounds of formula (I), (II) and (III) may also be administered rectally in the form of a pharmaceutical suppository. These compositions are prepared by mixing the drug with a suitable non-irritating adjuvant. These excipients are solid at room temperature but liquid at rectal temperatures and will therefore melt in the rectum to release the drug. These excipient materials include cocoa butter, glycerin treated gelatin, hydrogenated vegetable oils, polyethylene glycol mixtures of different molecular weights, and polyvinyl glycerol fatty acid esters.

For topical administration, creams, ointments, gels, solutions or suspensions containing the formulae (I), (II) and (III) may be employed. For the purposes of this application of this application, topical application can include mouthwashes and gargles.

The compounds of the present invention may be administered in the form of intranasal administration, by topical application of a suitable intranasal vehicle and delivery device, or by the administration of transdermal skin patches well known to those of ordinary skill in the art. . Administration is in the form of a transdermal delivery system, of course, the dosage of the entire dosage regimen is continuous rather than intermittent. The compounds of the present invention may also be administered in the form of a suppository, such as a base, such as cocoa butter, glycerin-treated gelatin, hydrogenated vegetable oil, a mixture of polyethylene glycols of various molecular weights, and a polyvinyl glycerol fatty acid ester.

When a compound of the invention is applied to an article, the selected dosage level will depend on a variety of factors including, but not limited to, the activity of the particular compound, the severity of the individual's symptoms, the route of administration, the time of administration, the metabolism of the compound Rate, duration of treatment, other drugs, compounds and/or substances used in combination, age, sex, weight, disease, health status and past medical history of the patient. The dosage and route of administration of the compound will ultimately be judged by the physician, although conventional doses will achieve local concentrations of the site of action. This point of action achieves the desired effect without causing a large number of harmful or toxic side effects.

In vivo administration is effective in a single dose, continuous administration or intermittent administration (e.g., a dispersed dose at appropriate intervals) throughout the course of treatment. Methods for determining the most effective mode of administration and dosage are well known to those skilled in the art. These methods will vary with the therapeutic dosage form, the purpose of the treatment, the target cells being treated, and the subject being treated. The dosage level of the administered drug is the same as the mode of administration, and the single or multiple doses are selected by the treating physician.

In general, a suitable dosage range for the active compound is from about 100 [mu]g/kg (subject body weight) per day to about 50 mg per kilogram (subject body weight) per day. When the active compound is a salt, an ester, a prodrug or the like, the dose administered is calculated based on the parent compound, and thus the weight actually applied is proportionally increased.

The compounds of the invention may also be used in combination with anticancer or chemotherapeutic agents. The compounds of the invention are also useful as chemotherapeutic sensitizers and radiosensitizers for the treatment of cancer. They can be used to treat mammals that have undergone or are undergoing cancer treatment. Such early treatments include early chemotherapy, radiation therapy, surgery or immunotherapy, such as cancer vaccines.

Accordingly, the present invention provides a combination of a compound of the formulae (I), (II) and (III) and an anticancer agent, administered simultaneously, intermittently or sequentially.

The present invention also provides the use of a compound of the formulae (I), (II) and (III) for the preparation of a medicament for assisting cancer treatment or enhancing the effect of ionizing radiation or a chemotherapeutic agent for treating tumor cells.

The invention also provides a method of chemotherapy or radiation therapy comprising administering to a patient a therapeutically effective amount of a compound of formula (I), (II) and (III), or at least one structural formula (I), A combination of a compound of II) and (III) and an ionizing radiation or a radiotherapeutic agent. In combination therapy, the compounds of the invention may be preceded by other anticancer agents according to the needs of the article (eg, 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks), simultaneously or after (for example, 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks Dosing after 6 weeks, 8 weeks, or 12 weeks. In various embodiments, the administration of the compound of the present invention and another anticancer agent are separated by 1 minute, 10 minutes, 30 minutes, less than 1 hour, 1 hour to 2 hours, 2 hours to 3 hours, and 3 hours. ~4 hours, 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 7 hours, 7 hours to 8 hours, 8 hours to 9 hours, 9 hours to 10 hours, 10 hours to 11 hours, 11 hours~ 12 hours, no more than 24 hours or no more than 48 hours.

The compounds of the invention and other anticancer agents may be additive or synergistic. Synergistic combination of a compound of the invention and another anticancer agent may allow for the use of one or both of these agents and/or less frequent doses of one or both of the compounds of the invention and other anticancer agents, and/or Or administration of the agent less frequently can reduce the toxicity associated with the administration of various agents to any article without reducing the effectiveness of the agent in the treatment of cancer. In addition, synergistic effects may enhance the effectiveness of these agents in the treatment of cancer and/or reduce the disadvantages or side effects associated with the use of any of the agents. Examples of cancer or chemotherapeutic agents for use in combination with the compounds described herein can be found in the "Cancer Principles and Practice of Oncology" published by Lippincott Williams & Wilkins, 2010 (VT Devita). And S. Hellman, 6th edition, February 15, 2001). One of ordinary skill in the art will be able to discern which combination of agents will be useful based on the particular characteristics of the drug and the cancer involved. These anticancer agents include, but are not limited to, the following: histone deacetylase (HDAC) inhibitors, estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, Cytotoxin/inhibitory cytotoxic agent, anti-malignant cell proliferation agent, isoprene-protein transferase inhibitor, HMG-CoA reductase inhibitor, HIV protease inhibitor, reverse transcriptase inhibitor or other angiogenesis inhibitor, cell Proliferation inhibitors or survival signal transduction inhibitors, apoptosis inducing agents, and cell cycle checkpoint interfering agents. The compound of the present invention can be preferably applied to a combination of radiotherapy.

Examples of "HDAC inhibitors" include suberoylanilide hydroxamic acid (SAHA), LAQ824, LBH589, PXD101, MS275, FK228, C. Valeric acid, butyric acid and CI-994. "Estrogen receptor modulator" refers to a compound that interferes with or inhibits the binding of estrogen to a receptor, regardless of mechanism. Examples of estrogen receptor modulators include, but are not limited to, tamoxifen, raloxifene, idoxifene, LY353381, LY1 17081, tormifine, fulvestrant, 4-[7-( 2,2-Dimethyl-1-oxopropoxy-4-methyl-2-[4-[2-(1-acridinyl)ethoxy]phenyl-2H-1-benzopyran 3-yl]-phenyl-2,2-dimethylpropionate, 4,4'-dihydroxybenzophenone-2,4-dinitrophenyl-indole and SH646. "Androgen-accepting "Physic modulator" refers to a compound that interferes with or inhibits the binding of androgen to a receptor, regardless of mechanism. Examples of androgen receptor modulators include finasteride and other 5α-reductase inhibitors, nilutamide, Flunar gum, bicalutan, liazodazole and abiraterone acetate.

"Retinoid receptor modulator" refers to a compound that interferes with or inhibits the binding of retinoids to a receptor, regardless of mechanism. Examples of such vitamin A receptor modulators include bexarotene, retinoic acid, 13-cis-retinoic acid, 9-cis-retinoic acid, α-difluoromethylornithine, ILX23-7533, anti-N- (4'-Hydroxyphenyl) retamide (trans-N-(4'-hydroxyphenyl)retinamide) and N-4-carboxyphenyl carbamide.

"Cytotoxin/cytostatic agent" refers to a compound that primarily causes cell death or inhibits cell proliferation by directly interfering with cellular function or inhibiting or interfering with cell mitosis, including alkylating agents, tumor necrosis factors, intercalators, and deficiencies. Oxygen-activated compounds, microtubule inhibitors/microtubule-stabilizers, mitotic kinesin inhibitors, mitotic process-related kinase inhibitors, antimetabolites, biological response modifiers, hormone/antihormone therapeutics, hematopoietic growth factors, monoclonal antibodies Targeted therapeutics, topoisomerase inhibitors, proteasome inhibitors and ubiquitin ligase inhibitors. Examples of cytotoxic agents include, but are not limited to, cyclophosphamide, chlorambucil carmustine (BCNU), lomustine (CCNU), busulfan (busulfan) ), treosulfan, cachectin, ifosfamide, tasonermin, chlordida Lonidamine, carboplatin, altretamine, prednimustine, dibromodulcitol, ranimustine, fotemustine, neda Nedaplatin, Oxaliplatin, Temozolomide, methyl methanesulfonate, procarbazine, dacarbazine, estramustine, and propyl bromide Iprosulfan tosilate, trofosfamide, nimustine, dibrospidium hydrochloride, pumitepa, lobaplatin , satraplatin, profiromycin, cisplatin, irofulven, cis-aminodichloro(2-methyl-pyridine)platinum, benzylguanine, Portuguese Glufosfamide, arsenic trioxide, 1-(1 1-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine, zorubicin, Idabi Star (idarubicin), daunorubicin, bisantrene, mitoxantrone, pirarubicin, pyronubicin (Pinafide), valrubicin, amrubicin, doxorubicin, epirubicin, pirarubicin, anticancer peptide (antineoplaston) , 3'-deamino-3'-morpholino-13-deoxy-10-hydroxyegromycin, galarubicin, elinafide and MEN1 0755.

In some embodiments, the compounds described herein can be used in conjunction with an alkylating agent.

Examples of alkylating agents include, but are not limited to, nitrogen mustard: cyclophosphamide, ifosfamide, trofosfamide, and chlorambucil; nitrosourea: card Carustine (BCNU) and lomustine (CCNU); alkyl sulfonates: white Xiaoan and treosulfan; triazenes: dacarbazine, procarbazine and temozolomide; platinum complexes: cisplatin, carboplatin and oxaliplatin.

In some embodiments, the alkyl reagent is dacarbazine. The dose administered to the target of dacarbazine is from about 150 mg/m ² to about 250 mg/m ² (area of the body surface area). In another embodiment, dacarbazine can be administered intravenously five days of continuous objects, once daily dosage range of about 150mg / m ² ~ about 250mg / m ^2.

In some embodiments, the alkylating agent is procarbazine. The dose of the target administered with procarbazine is from about 50 mg/m ² to about 100 mg/m ² (area of the body surface area). In other embodiments, procarbazine be 5 days continuous intravenous administration object, once daily dosage range of about 50mg / m ² ~ about 100mg / m ^2. In some embodiments, the alkyl reagent is temozolomide. The dose of the subject to be administered with temozolomide ranges from about 150 mg/m ² (area of the body surface area) to about 200 mg/m ² . In other embodiments, the temozolomide may be administered orally five days of continuous objects, once daily dosage range of about 150mg / m ² to about 200mg / m ^2.

Examples of anti-mitotic agents include colchicine, halichonin B, colchicine, colchicine derivatives, maytansine, lysine, thiocolchicine, and tritylcysteine.

An example of an anoxic activating compound is tirapazamine.

Examples of proteasome inhibitors include, but are not limited to, lactacystin, bortezomib, and peptide aldehydes such as MG 132, MG 115, and PSI.

EXAMPLES Examples of microtubule inhibitors/microtubule-stabilizers include paclitaxel, vindesine sulfate, vincristine, vinblastine, vinorelbine, docetaxel, lissoxin, dolastatin, rice volts Mivobulin isethionate, simultane, RPR1 09881, BMS 184476, vinflunine, 2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methyl Oxyphenyl) benzenesulfonamide, anhydrovinblastine, N,N-dimethyl-L-ammonium-L-ammonium-N-methyl-L-amidoxime-L- Amidoxime-L-valine-t-butamidine, TDX258, epothilone (see, for example, U.S. Patent Nos. 6,284,781 and 6,288,237) and BMS 188797. Some examples of topoisomerase inhibitors are topotecan, irinotecan, rubitatecan, esareac, gimetecan, formyl-camptothecin, 9-amino camphor Alkali, camptothecin, cristatol, mitomycin C, 9-methoxy-N,N-dimethyl-5-nitropyrazolo[3,4,5- Kl] acridine-2-(6H)propylamine, 1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de] Pyrano[3',4':b,7]-pyridazino[1,2b]quinoline-10,13(9H,15H)dione, letoticon, 7-[2-(N- Isopropylamino)ethyl]-(20S)camptothecin, BNP1350, BNPI1100, BN80915, BN80942, Etoposide Phosphate, Teniposide, Soproza, 2'-Dimethylamino-2'- Deoxy-etoposide, GL331, N-[2-(dimethylamino)ethyl]-9-hydroxy-5,6-dimethyl-6H-pyrido[4,3-b]carbazole- 1-carbamamine, 6,9-bis[(2-aminoethyl)amino]benzo[g]isoquinoline-5,10-dione, 5-(3-aminopropylamino)-7, 10-dihydroxy-2-(2-hydroxyethylaminomethyl)-6H-pyrazolo[4,5,1-de]acridin-6-one, N-[1-[2(diethyl) Amino)ethylamino]-7-methoxy-9-oxo-9H-thiazepine-4- Methyl]carbamamine, N-(2-(dimethylamino)ethyl)acridine-4-carboxamide, 6-[[2-(dimethylamino)ethyl]amino]-3 -hydroxy-7H-indolo[2,1-c]quinolin-7-one and dimesis; non-camptothecin-type topoisomerase-1 inhibitors, such as carbazoles; and dual topologies Isomerase-1- and II inhibitors, such as benzophenazines, XR20 115761MLN 576, and benzopyridindole. In one embodiment, the topoisomerase inhibitor is irinotecan. The dose range of irinotecan administered is from about 50 mg/m ² to about 150 mg/m ² (area of the body surface area). In another embodiment, irinotecan can be administered intravenously to the drug product, for a continuous dose ranging from about 50 mg/m ² to about 150 mg/m ² for 5 days in 1-5 days, once daily, and then A 5-day continuous intravenous infusion of 28-32, dose ranging from about 50 mg/m ² to about 150 mg/m ² (area of the body surface area), once a day, and finally a continuous intravenous infusion of 5 days at 55-59 For administration, the dosage range is from about 50 mg/m ² to about 150 mg/m ² .

Examples of mitotic kinesin inhibitors, in particular human mitotic kinesin inhibitor KSP are disclosed in International Patent Publication No. WO 01/30768, WO 01/98278, WO 02/056880, WO 03/050,064, WO 03/050, 122, WO 03 /049,527, WO 03/049,679, WO 03/049,678, WO 03/039460, WO 03/079973, WO 03/099211, WO 2004/039774, WO 03/105855, WO 03/106417, WO 2004/087050, WO 2004 It is described in the documents of U.S. Patent Application Nos. US 2004/132830 and US 2004/132719, both of which are incorporated herein by reference. In one embodiment, the mitotic kinesin inhibitors include, but are not limited to, KSP inhibitors, MKLP1 inhibitors, CENP-E inhibitors, MCAK inhibitors, KIFL4 inhibitors, Mphosphl inhibitors, and Rab6-KIFL inhibitors. "Mitosis inhibitors associated with mitosis," including, but not limited to, Aurora kinase inhibitors, Polo-like kinases (PLK) inhibitors, preferably PLK-I inhibitors, Bub-1 inhibitors, and bub- R1 inhibitor.

"Anti-malignant cell proliferation agents" include antisense RNA and antisense DNA oligonucleotides such as G3139, ODN698, RVASKRAS, GEM231 and INX3001, and antimetabolites such as enocitabine, carmofur (carmofur) ), tegafur, pentostatin, doxifluridine, trimetrexate, fludarabine, capecitabine, plus Galocitabine, cytarabine ocfosfate, raltitrexed, emitefur, decitabine, nola Nolatrexed, pemetrexed, 2'-fluoromethylene-2'-deoxycytidine, N-[5-(2,3-dihydro-benzofuranyl)sulfonyl ]-N-(3,4-dichlorophenyl) Urea, ecteinascidin, troxacitabine, aminopterin, 5-flurouracil, alanosine, swainsoniue, lome Lometrexol, dexrazoxane, methioninase and 3-aminopyrimidine-2-carboxamide thiosemicarbazone.

Examples of monoclonal antibody-targeted therapeutic agents include those having a cytotoxic agent or a radioisotope linked to a cancer cell-specific or target cell-specific monoclonal antibody. Examples include Baikesha.

"HMG-CoA reductase inhibitor" means a 3-hydroxy-3-methylpentadienyl-CoA reductase inhibitor. HMG-CoA reductase inhibitors that may be used include, but are not limited to, lovastatin (MEVACOR®; see U.S. Patent Nos. 4,231,938, 4,294,926 and 4,319,039), simvastatin, ZOCOR ®; see U.S. Patent Nos. 4,444,784, 4,820,850 and 4,916,239, pravastatin (pravastatin, PRAVACHOL®; see U.S. Patent Nos. 4,346,227, 4,537,859, 4,410,629, 5,030,447 and 5, 180,589), fluvastatin, LESCOL®; see U.S. Patent Nos. 5,354,772, 4,911,165, 4,929,437, 5,189,164, 5,118,853, 5,290,946 and 5,356,896, and atorvastatin ( Atorvastatin, LIPITOR®; see U.S. Patent Nos. 5,273,995, 4,681,893, 5,489,691 and 5,342,952. These HMG-CoA reductase inhibitors and other HMG-CoA reductase inhibitors useful in the methods of the present invention are disclosed in the literature: Chemical Industry (Chemistry & Industry) Press, published on February 5, 1996, Cholesterol Lowering Drugs (M. Yalpani); U.S. Patent Nos. 4,782,084 and 4,885,314. The term HMG-CoA reductase inhibitor herein includes HMG-CoA reductase inhibiting all pharmaceutically acceptable lactone and ring opening acid forms of active compounds (eg, wherein the lactone ring opens to form a free acid) and a salt And ester form. The use of such salts, esters, ring opening acids and lactone forms is therefore also included within the scope of the invention. "Prenyl-protein transferase inhibitor" means a compound which inhibits any one of isoprenyl-protein transferase or any combination thereof, including farnesyl-protein transferase (FPTase) ), type I geranylgeranyl-protein transferase type I (GGPTase-I) and type II-type geranyl protein transferase (geranylgeranyl-protein transferase type-II, GGPTase-II, also known as Rab GGPTase).

"Angiogenesis inhibitor" refers to a compound that inhibits the formation of new blood vessels, regardless of mechanism. Examples of angiogenesis inhibitors include, but are not limited to, tyrosine kinase inhibitors such as tyrosine kinase receptors FIT-I (VEGFR1) inhibitors and FIK-I/KDR (VEGFR2) inhibitors. Epidermal inhibitor; fibroblast-derived inhibitor or platelet growth inhibitor; matrix metalloprotease (MMP) inhibitor; integrin blocker; interferon-α, interleukin-12; Pentosan polysulfate; epoxidase inhibitors, including nonsteroidal anti-inflammatories (NSAIDs) such as aspirin and ibuprofen, and selective cyclooxygenase-2 inhibitors such as celecoxib And rofecoxib ( PNAS (1992) 89:7384; JNC/ ( 1982 ) 69:475; Arch. Opthalmol. ( 1990 ) 108:573; Anat. Rec. ( 1994 ) 238:68; FEBS Letters ( 1995 ) 372:83; Clin, Orthop. ( 1995 ) 313:76; J. MoI. Endocrinol. ( 1996 ) 16:107; Jpn. J. Pharmacol. ( 1997 ) 75:105; Cancer Res. ( 1997 ) 57:1625 ; Cell (1998) 93: 705 ; Intl.J.MoI.Med (1998) 2:. 715; J.Biol.Chem (1999) 274:. 9116)); steroid Anti-inflammatory agents (such as corticosteroids, mineralocorticoids, dexamethasone, prednisone, prednisolone, methylpred, betamethasone); carboxyl Carboxyamidotriazole; Combretastatin A-4; Squalamine; 6-O-chloroethyl-carbonyl)-tocopherol (6-O-chloroacetyl- Carbonyl)-fumagillol); thalidomide; angiostatin; troponin-1; angiotensin II antagonist ( J. Lab. Clin. Med. ( 1985 ) 105:141-145] and VEGF antibody ( Nature Biotechnology ( 1999 ) 17: 963-968; Kim et al. Nature ( 1993 ) 562: 841-844; International Patent Publication No. WO 00/44777; International Patent Publication No. WO 00/61186).

Other therapeutic agents that modulate or inhibit angiogenesis and which can also be used in combination with the compounds described herein include those that modulate or inhibit coagulation and fibrinolytic systems (see review Clin. Chem. La. Med. ( 2000 ) 38 :679-692). Examples of such drugs that modulate or inhibit the coagulation and fibrinolysis pathway include, but are not limited to, heparin [see Thromb. Haemost. ( 1998 ) 80: 10-23], small molecule heparin and carboxypeptidase U inhibitors ( Also known as inhibitors of active thrombin activatable fibrinolysis inhibitor (TAFIa) [see Thrombosis Res. ( 2001 ) 101: 329-354). TAFIa inhibitors have been published in the international patent publication number. WO 03/013, 526 and U.S. Patent No. US 60/349, 925 (filed on Feb. 18, 2002). "Cell cycle checkpoint interfering agent" refers to a compound that inhibits the signalling of a cell cycle checkpoint signal protein kinase, thereby sensitizing the effects of DNA damage drugs on cancer cells. These include ATR inhibitors, ATM inhibitors, Chk1 kinase inhibitors, Chk2 kinase inhibitors, cdk kinase inhibitors, and cdc kinase inhibitors. Specific examples are 7-hydroxystaurosporin, star Staurosporin, CYC202 (Cyclacel) and BMS-387032. "Inhibitor of cell proliferation and survival signaling pathways" refers to compounds that inhibit cell surface receptors and downstream of the signal transduction pathways of these surface receptors. These drugs include inhibitors of EGFR inhibitors {such as gefitinib and erlotinib}, ERB-2 inhibitors {such as trastuzumab}, IGFR inhibitors ( For example, those disclosed in International Patent Publication No. WO 03/059951), cytokine receptors, MET inhibitors, PI3K inhibitors (eg LY294002), serine/threonine kinases (including, but not limited to, Akt inhibitors, eg Raf inhibition, as disclosed in International Patent Publication No. WO 03/086404, WO 03/086403, WO 03/086394, WO 03/086279, WO 02/083675, WO 02/083139, WO 02/083140, and WO 02/083138 Agents (such as BAY-43-9006), MEK inhibitors (such as CI-1040 and PD-098059) and mTOR inhibitors (such as WCI-779 from Wyeth and AP23573 from Ariad). These drugs include small molecule inhibitor compounds and antibody antagonists.

"Apoptosis-inducing agents" include TNF receptor family member activators (including TRAIL receptors). The invention also includes a combination of NSAID's, wherein the NSAID's are selective COX-2 inhibitors. For purposes of the present invention, a selective COX-2 inhibitor NSAID's is defined as: When the ratio of COX-2 IC ₅₀ values with evaluation test cell or microsomal than COX-I IC ₅₀ values measured, to suppress those COX-2 has at least 100-fold specificity of NS AID's compounds compared to COX-I.

A COX-2 inhibitor specifically for use in the method of treatment of the present invention is 5-chloro-3-(4-methylsulfonyl)phenyl-2-(2-methyl-5-pyridyl)pyridine or A pharmaceutically acceptable salt.

Specific COX-2 inhibitor compound and a compound used in the present invention have been disclosed include, but are not limited to: parecoxib (of parecoxib), acceptable salts CELEBREX ^® and BEXTRA ^® and pharmaceutically acceptable.

Examples of other angiogenesis inhibitors include, but are not limited to, endostatin, 5-amino-1-[[3,5-dichloro-4-(4-chlorobenzylidinyl)phenyl]- -1H-1,2,3-triazole-4-carboxamide, CM101, squalamine, combretastatin, RPI4610, NX31838, sulfated mannose pentasaccharide phosphate 7,7-(Carboxy-bis[imino-N-methyl-4,2-pyrrolocarbonylimino) [N-Methyl-4,2-pyrrole]-carbonylimino]-bis-(1,3-naphthalene disulfonate) and 3-[(2,4-dimethylpyrrole-5-yl) 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone, SU5416).

As used above, "integrin blocker" means a compound that selectively antagonizes, inhibits or blocks the binding of a physiological ligand to the 0Cyβ3 integrin, and selectively antagonizes, inhibits or blocks the physiological ligand and the αvβ5 integrin. A conjugated compound that antagonizes, inhibits, or blocks a physiological ligand that is both a compound of 0Cyβ3 integrin and a 0Cyβ5 integrin, as well as a compound that antagonizes, inhibits, or blocks the expression of a specific integrin on capillary endothelial cells. The term may also refer to antagonists of the integrins OCyβój 0Cvβ8, oqβi, 0C2β1, 0C5βi, 0C6β1 and 0C6β4. The term may also refer to an antagonist of any combination of integrin αvβ3, αvβ5, αvβ6, ocvβ8, oqβi, 0C2β1, βsoq, 0C6β1, and 0C6β4.

Some specific examples of tyrosine kinase inhibition include: N-(trifluoromethylphenyl)-5-methylisoxazole-4-carboxamide, 17-(allylamino)-17-nor Oxygeldanamycin, 4-(3-chloro-4-fluorophenylamino)-7-methoxy-6-[3-(4-morpholinyl)propoxy]quinazoline, N -(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine, BIBX1 382, 2,3,9,10,11,12-hexahydrogen -10-(hydroxymethyl)-10-hydroxy-9-methyl-9,12-epoxy-1H-diindolo[1,2,3-fg:3',2',1'- Kl]pyrrolo[3,4-i][1,6]benzodiazepin-1-one, SH268, genistein, STI571, CEP2563, 4-(3-chlorophenylamino -5,6-Dimethyl-7H-pyrrolo[2,3-d]pyrimidine methanesulfonate, 4-(3-bromo-4-hydroxyphenyl)amino-6,7-dimethoxy Quinazoline, 4-(4'-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, SU6668, STI571A, N-4-chlorophenyl-4-(4-pyridylmethyl) -1-oxazinamide and EMD121974.

In some embodiments, the compounds described herein are useful for treating and or preventing the appearance of gangrene induced by selective N3-adenine methylation agents such as Me-Lex (MeOSO ₂ (CH ₂ )-lexitropsin).

Combinations with compounds other than anti-cancer compounds are also included within the scope of the methods of the invention. For example, a compound claimed herein in combination with a PPAR-[gamma] (eg, PPAR-gamma) agonist and a PPAR-[delta] (eg, PPAR-delta) agonist is used to treat a particular malignancy. PPAR-γ and PPAR-δ are γ and δ of peroxisome proliferator-activated receptors. The expression of PPAR-γ on endothelial cells and its relationship in angiogenesis has been reported in the literature [see J. Cardiovasc. Pharmacol. ( 1998 ) 31: 909-913; J. Biol. Chem. ( 1999 ) 274 : 9116-9121; Invest. Ophthalmol Vis. Sci. ( 2000 ) 41: 2309-2317]. Recently, in vitro studies have shown that PPAR-γ agonists can inhibit VEGF vascular response; troglitazone and rosiglitazone maleic acid can inhibit the progression of retinal neovascularization in mice. [ Arch. Ophthamol. ( 2001 ) 119: 709-717]. Examples of PPAR-gamma agonists and PPAR-gamma/alpha agonists include, but are not limited to, thiazolidinediones {eg DRF2725, CS-OI 1, troglitazone, rosiglitazone, and pioglitazone) , fenotibrate, gemfibrozil, clofibrate, GW2570, SB219994, AR-H039242, JTT-501, MCC-555, GW2331, GW409544, NN2344, KRP297, NPOI 1O, DRF4158, NN622, GI262570, PNU182716, DRF552926, 2-[(5,7-dipropyl-3-trifluoromethyl-1,2-benzisoxazole-6-yl)oxy]-2 -Methylpropionic acid [disclosed in U.S. Patent No. US 09/782,856] and 2(R)-7-(3-(2-chloro-4-(4-fluorophenoxy)phenoxy)propoxy) -2-ethylchroman-2-carboxylic acid [disclosed in U.S. Patent Nos. US 60/235,708 and 60/244,697]. Further embodiments of the invention are the use of the compounds of the invention in combination with antiviral agents {e.g., nucleoside analogs, including ganciclovir} for the treatment of cancer, see International Patent Publication No. WO 98/04290.

Further embodiments of the invention are the use of the compounds of the invention in combination with gene therapy for the treatment of cancer. For genetic strategies for treating cancer, see Hall et al., 1997, Am J Hum Genet , 61: 785-789 and BC Decker, Hamilton, 2000, published in Cancer Medicine (Kufe et al., Fifth Edition). Pages 876-889. Gene therapy can be used to deliver any tumor suppressor gene. Examples of such genes include, but are not limited to, page 53, which can be transferred by recombinant virus-mediated gene transfer [eg, see U.S. Patent No. 6,069,134], uPA/uPAR antagonist ["Adenovirus-Mediated Delivery of a uPA/uPAR" Antagonist Suppresses Angiogenesis-Dependent Tumor Growth and Dissemination in Mice, " Gene Therapy , August ( 1998 ) 5(8): 1105-13] and interferon gamma [ J Immunol ( 2000 ) 164: 217-222] were delivered.

The compounds of the invention may also be administered in conjunction with an intrinsic multidrug resistance (MDR) inhibitor, particularly a MDR inhibitor associated with high level expression of the transporter. The MDR inhibitor includes glycoprotein (p-glycoprotein, P-gp), such as LY335979, XR9576, OC144-093, R101922, VX853, verapamil and vaspod (PSC833, valspodar). Agent.

The compounds of the invention may be used in combination with an antiemetic for the treatment of nausea or vomiting, including acute, delayed, advanced and advanced vomiting, which may be induced by the compounds of the invention, alone or in combination with radiation therapy. For the prevention and/or treatment of vomiting, the compounds of the invention may be used in combination with other antiemetic agents, in particular neurokinin-1 receptor antagonists, 5HT3 receptor antagonists such as ondansetron, glas Granisetron, tropisetron and zatisetron, GABAB receptor antagonists such as baclofen, corticosteroids such as Decadron (dexamethasone) or other such as US patents Nos. 2,789,118, 2,990,401, 3,048,581, 3,126,375, 3,929,768, 3,996,359, 3,928,326, and 3,749,712 Analogous, anti-dopaminergic drugs such as phenothiazines {eg prochlorperazine, fluphenazine, thioridazine and mesoridazine}, metoclopramide (metoclopramide) or dronabinol. In some embodiments, the antiemetic agent is selected from the group consisting of a neurokinin-1 receptor antagonist, a 5HT3 receptor antagonist, and a corticosteroid, and can be administered as an adjuvant to treat and/or prevent the administration of the compound of the present invention. Caused by vomiting.

In some embodiments, the neurokinin-1 receptor antagonist used in combination with a compound of the invention is selected from the group consisting of: 2-(R)-(1-(R)-(3,5-bis(trifluoromethyl) Phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)- Morpholine or a pharmaceutically acceptable salt thereof, which is disclosed in U.S. Patent No. 5,719,147.

The compounds of the invention may also be administered with a medicament for the treatment of anemia. These anemia therapeutic agents are, for example, sustained erythropoietin receptor agonists {e.g., Epoetin alfa}.

The compounds of the invention may also be administered with a medicament for the treatment of neutropenia. These neutropenia therapeutic drugs are, for example, hematopoietic growth factors, which can modulate the production and function of neutrophils, such as granulocyte colony stimulating factor (G-CSF). Examples of G-CSF include filgrastim.

The compounds of the invention may also be administered with immunopotentiating drugs such as levamisole, isoprinosine and Zadaxin. The compounds of the invention may also be combined with bisphosphonates (understood to include bisphosphonates, bisphosphonates, bisphosphonic acids, and diphosphonic acids) for treatment and/or Prevent cancer, including bone cancer. Examples of bisphosphonates include, but are not limited to, etidronate (Didronel), pamidronate (Aredia), alendronate (Fosamax), risedronate (risedronate, Actonel), zoledronate (Zometa), ibandronate (Boniva), incacronate (incadronate) or incaphosphonate (cimadronate), clodronate Salt (clodronate), EB-1053, minodronate, neridronate, piridronate and tiludronate, including any of them Pharmaceutically acceptable salts, derivatives, hydrates and mixtures thereof.

Accordingly, the scope of the invention includes the use of a compound of the invention in combination with ionizing radiation and/or a second compound selected from the group consisting of HD AC inhibitors, estrogen receptor modulators, androgens Receptor Modulators, Retinoid Regulators, Cytotoxins/Cell Inhibitors, Antiproliferatives, Isoprene-Protease Inhibitors, HMG-CoA Reductase Inhibitors, Angiogenesis Inhibitors, PPAR- Gamma agonist, PPAR-delta agonist, antiviral agent, intrinsic multidrug resistance inhibitor, antiemetic, drug for anemia treatment, drug for treating neutropenia, immunopotentiating drug , cell proliferation inhibitors and survival signal inhibitors, cell cycle checkpoint interfering agents, apoptosis inducing agents, and bisphosphonates.

The term "administering" and variations thereof (e.g., "administering" a compound) with respect to a compound described herein refers to introducing the compound or prodrug thereof into an animal system in need of treatment. When a compound of the present invention or a prodrug thereof is provided in combination with one or more other active agents (such as a cytotoxic agent, etc.), the term "administering" and its variables may each be understood to include the simultaneous and sequential introduction of the present invention. a compound or a prodrug thereof and other drugs.

The term "composition" as used herein refers to a product that encompasses a specified ingredient in a defined amount, as well as any product that can be formed directly or indirectly from a defined combination of specified ingredients.

As used herein, the term "therapeutically effective amount" refers to the amount of active compound or pharmaceutical agent that can elicit a biological, medical or medical response to a tissue, system, animal or human, which can be sought by a researcher, veterinarian, physician or other clinician.

The term "treating" refers to treating a mammal suffering from a pathological disease, and may also mean reducing the effect of the disease by killing the cancer cell, but may also cause inhibition of the progression of the disease, and may include slowing the progress of the disease, stopping the progress of the disease, improving the disease, and The effect of treating the disease. It also includes treatment as a preventive method (for example, prevention).

As used herein, the term "pharmaceutically acceptable" means: corresponding to a reasonable probability of benefit/risk, within the scope of sound medical judgment, suitable for contacting an object (eg, human) tissue without causing excessive toxicity, irritation, A compound, substance, combination, and/or dosage form of an allergic reaction or other problem or complication. The various carriers, excipients and the like must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation product.

The term "assisted" refers to the use of a compound in combination with known methods of treatment. Such methods include cytotoxic regimens and/or ionizing radiation used in the treatment of different cancer types. In particular, these active compounds are known compounds which enhance the chemotherapeutic effects of various cancers, including topoisomerase toxicants for the treatment of cancer {eg topotecan, irinotecan, Rubiconcan}, most known alkylating agents {such as DTIC, temozolamide} and platinum-based drugs {such as carboplatin, cisplatin}.

Also included within the scope of the present application is a method of treating cancer comprising administering a therapeutically effective amount of a compound of formula (I), (II), and (III) in combination with radiation therapy and/or the following compounds. These compounds are selected from the group consisting of HDAC inhibitors, estrogen receptor modulators, androgen receptor modulators, and retinoids. Receptor Modulators, Cytotoxins/Cell Inhibitors, Antiproliferatives, Isoprene-Protease Inhibitors, HMG-CoA Reductase Inhibitors, Angiogenesis Inhibitors, PPAR-γ Agonists, PPAR-δ Agonism Agents, antiviral agents, intrinsic multidrug resistance inhibitors, antiemetics, drugs for the treatment of anemia, drugs for the treatment of neutropenia, immunopotentiating drugs, cell proliferation inhibitors and survival signals Inhibitors, cell cycle checkpoint interfering agents, apoptosis inducing agents, and bisphosphonates.

Embodiments of the invention can be explained by the following examples. However, it should be understood that the embodiments of the present invention are not limited to the details of the embodiments, as other variations thereof are known or apparent to those of ordinary skill in the art in light of this disclosure.

All temperatures are in degrees Celsius unless otherwise stated.

Synthetic route 1

Synthetic route 2

detailed description

Example 1

2-(4-(3-Azabicyclo[4.1.0]heptan-6-yl)phenyl)-2H-indazole-7-carboxamide hydrochloride (product 1)

Step 1 Synthesis of Compound 1

Propargylamine hydrochloride (8g), dichloromethane (300ml) and triethylamine (26.4g) were dissolved in a 1L three-necked flask and slowly dissolved in benzyloxycarbonyl hydrazine chloride (19.4g) at 0 °C. A solution of dichloromethane (30 ml) was added for about 30 minutes. The reaction was stirred slowly to room temperature for 3 hours. The reaction solution was washed with saturated sodium chloride solution, layers were separated and the organic phase was dried over anhydrous MgSO _4, concentrated under reduced pressure to give the crude product purified by column chromatography to give compound 1 (3g). MS: 190 (M+H) ⁺ .

Step 2 Synthesis of Compound 2

Compound 1 (3 g) was placed in a 250 ml three-necked flask, and a solution of NaH (60%, 760 mg) dissolved in THF/DMF (4:1, v/v, 100 ml) was slowly added dropwise at 0 ° C for about 20 minutes. The reaction was stirred for 30 minutes, and 3-chloropropene (1.5 g) was added thereto, and the reaction was continued at room temperature for 1 hour. The reaction was quenched by the addition of water and THF was evaporated. The residue was treated with sodium chloride solution and ethyl acetate solution was separated, the organic phase was dried over anhydrous MgSO _4, concentrated under reduced pressure to give the crude product purified by column chromatography (PE: EA = 50: 1 ) to give Compound 2 (2.7g). MS: 230 (M+H) ⁺ .

Step 3 Synthesis of Compound 3

Compound 2 (2.7 g), p-iodonitrobenzene (2.94 g), triethylamine (1.6 g), PdCl ₂ (PPh ₃ ) ₂ (340 mg), CuI (180 mg) and DMF (50 ml) were mixed under nitrogen. Place in a 100 ml sealed tube. The mixture was warmed to 40 ° C and stirred overnight. After the mixture was cooled to room temperature, it was separated by adding an aqueous sodium chloride solution and ethyl acetate. To give compound 3 (3G) crude product purified by column chromatography (:: EA = 30 1 PE ) and washed with saturated aqueous sodium chloride the organic phase three times, the organic phase was dried over anhydrous MgSO _4, and concentrated under reduced pressure to give. MS: 351 (M+H) ⁺ .

Step 4 Synthesis of Compound 4

Under a nitrogen atmosphere, compound 3 (3 g), PtCl ₂ (300 mg) and toluene (50 ml) were placed in a 100 ml sealed tube. The reaction mixture was warmed to 80 ℃ 36 childhood, concentrated under reduced pressure to give the crude product purified by column chromatography to give Compound 4 (700mg). MS: 351 (M+H) ⁺ .

Step 5 Synthesis of Compound 5

Compound 4 (700 mg), iron powder (560 mg), methanol (50 ml) and acetic acid (1 ml) were mixed and placed in a 250 ml single-mouth flask and stirred, and the mixture was heated to reflux for 2 hours and then cooled to room temperature. The residue was separated and concentrated water and ethyl acetate, the organic phase was dried, filtered, and concentrated to give Compound 5 (550mg) decompression. MS: 321 (M+H) ⁺ .

Step 6 Synthesis of Compound 6

Compound 5 (550 mg), methyl-2-nitro-3-methylindenyl-benzoic acid phenyl ester (400 mg) and ethanol (40 ml) were stirred in a 50 ml two-necked flask, and the mixture was heated to 70 ° C and stirred for 5 hours. Concentration under reduced pressure gave compound 6 ( EtOAc, EtOAc) MS: 511 (M+H) ⁺ .

Step 7 Synthesis of Compound 7

Under a nitrogen atmosphere, compound 6 (500 mg), sodium azide (122 mg) and DMF (20 ml) were stirred in a 100 ml single-necked flask, and the mixture was warmed to 95 ° C, stirred overnight and then cooled to room temperature. The residue was separated with a saturated sodium chloride solution and ethyl acetate. Washed with saturated sodium chloride solution, the organic phase was washed four times, dried, filtered, and concentrated under reduced pressure, purified by column chromatography to give compound 7 (400mg). MS: 480 (M+H) ⁺ .

Step 8 Synthesis of Compound 8

Compound 7 (400 mg), EA/MeOH (1:1, v/v, 60 ml), sodium carbonate (150 mg) and Pd/C (200 mg) were placed in a 100 ml single-mouth flask and stirred at room temperature with H ₂ to the mixture. The reaction was overnight. Filtered, concentrated under reduced pressure, the residue was dissolved with ethyl acetate was filtered, the filtrate was concentrated under reduced pressure to give Compound 8 (300mg). MS: 348 (M+H) ⁺ .

Step 9 Synthesis of Compound 9

Compound 8 (300 mg), dichloromethane (30 ml) and triethylamine (110 mg) were stirred in a 100 ml two-necked flask, cooled to 0 ° C, and slowly added Boc ₂ O (210 mg) dissolved in dichloromethane (5 ml) )The solution. Slowly warm to room temperature and stirred for 30 minutes, the reaction mixture was washed with saturated sodium chloride solution, dried, filtered, and concentrated under reduced pressure, by column chromatography to give Compound 9 (120mg). MS: 448 (M+H) ⁺ .

Step 10 Synthesis of Compound 10

Compound 9 (120 mg) was dissolved in a solution of methanol (20 ml) and the mixture was placed in a 100 ml sealed tube. The temperature was lowered to 0 ° C, ammonia gas (2.0 g) was introduced, and the system was sealed, and the temperature was raised to 65 ° C and stirred overnight. The reaction mixture was concentrated under reduced pressure and purified by column chromatography to give Compound 10 (100mg). MS: 433 (M+H) ⁺ .

Step 11 2- (4- (3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-acyl-7-amine hydrochloride (product 1)

Compound 10 (100 mg) was dissolved in dichloromethane (25 ml). The mixture was placed in a 100 ml two-necked flask, cooled to 0 ° C, and dried hydrogen chloride gas was passed for 20 minutes. The mixture was stirred at 0 °C for a further 30 minutes and the filter cake was dried in vacuo to afford product 1 (50mg). ^{1 H NMR (400MHz, DMSO)} : δ 9.30 (1H, s), 8.78 (2H, d), 8.55 (1H, s), 8.10-8.03 (4H, m), 7.89 (1H, s), 7.63-7.62 (2H, m), 7.27 (1H, m), 3.15-2.84 (4H, m), 2.33 - 2.26 (2H, m), 1.51 (1H, m), 1.20 (2H, m). MS: 369 (M+H) ⁺ .

Example 2

2-(4-(1-methoxymethyl-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-2H-indazole-7-carboxamide hydrochloride (product 2)

Step 1 Synthesis of Compound 11

Compound 1 (500 mg) was dissolved in THF/DMF (1:1, v/v, 10 ml) solution, the mixture was placed in a 50 ml two-necked flask, cooled to 0 ° C, NaH (130 mg) was added portionwise, and the mixture was warmed to room. After the temperature was further stirred for 20 minutes, 2-chloromethyl-4-methoxy-butene (400 mg) was added, and stirring was continued for 1 hour at room temperature. TLC (PE: EA = 5:1) was used to complete the reaction. The sodium chloride solution and ethyl acetate were separated. The organic phase was washed with saturated sodium chloride solution, dried, filtered, concentrated under reduced pressure and purified by column chromatography (PE: EA = 20: 1) Compound 11 (500 mg) was obtained. MS: 274 (M+H) ⁺ .

Step 2 Synthesis of Compound 12

Compound 11 (500 mg), p-iodonitrobenzene (455 mg), triethylamine (245 mg), PdCl ₂ (PPh ₃ ) ₂ (52 mg), CuI (30 mg) and DMF (10 ml) were placed in a 100 ml sealed tube and stirred. The temperature was raised to 40 ° C and the reaction was stirred overnight. The mixture was cooled to room temperature, separated with sodium chloride solution and ethyl acetate. The organic phase was washed three times with saturated sodium chloride solution, dried, filtered, concentrated under reduced pressure and purified by column chromatography (PE: EA=15:1) Compound 12 (400 mg) was obtained. MS: 395 (M+H) ⁺ .

Step 3 Synthesis of Compound 13

Under a nitrogen atmosphere, compound 12 (120 mg), PtCl ₂ (30 mg) and toluene (10 ml) were placed in a 100 ml sealed tube, and the mixture was warmed to 80 ° C and stirred overnight. Cooled to room temperature, dried, filtered, and concentrated under reduced pressure, purified by column chromatography to give Compound 13 (90mg). MS: 395 (M+H) ⁺ .

Step 4 Synthesis of Compound 14

Compound 13 (230 mg), iron powder (100 mg), methanol (30 ml), and acetic acid (1 ml) were stirred in a 100 ml single-necked flask, and the mixture was warmed to reflux for 1 hour, cooled to room temperature and concentrated under reduced pressure. The residue was separated water and ethyl acetate, the organic phase was dried, filtered, and concentrated to give Compound 14 (200mg) under reduced pressure. MS: 365 (M+H) ⁺ .

Step 5 Synthesis of Compound 15

Compound 14 (200 mg), methyl-2-nitrophenyl-3-mercaptobenzoic acid (120 mg) and ethanol (10 ml) were placed in a 50 ml single-necked flask and stirred, warmed to 70 ° C for 5 hours, concentrated under reduced pressure. Compound 15 (300 mg) was obtained without further purification. MS: 555 (M+H) ⁺ .

Step 6 Synthesis of Compound 16

Under a nitrogen atmosphere, Compound 15 (300 mg), sodium azide (60 mg) and DMF (10 ml) were placed in a 50 ml single-necked flask and warmed to 95 ° C overnight. After cooling to room temperature, a saturated sodium chloride solution and ethyl acetate were added, and the organic phase was washed four times with saturated sodium chloride solution, dried, filtered, concentrated under reduced pressure and purified by column chromatography to afford compound 16 (150 mg). MS: 524 (M+H) ⁺ .

Step 7 Synthesis of Compound 17

Compound 16 (150mg), EA / MeOH (1: 1,10ml), sodium carbonate (10 mg) and Pd / C (20mg) catalyst is placed in 100ml single-neck flask, into H ₂ at room temperature overnight. Filtered, concentrated under reduced pressure, the residue was washed with ethyl acetate, filtered, the organic phases were combined, to give compound 17 (80mg) and concentrated under reduced pressure. MS: 392 (M+H) ⁺ .

Step 8 Synthesis of Compound 18

Compound 17 (80 mg), dichloromethane (20 ml) and triethylamine (30 mg) were stirred in a 50 ml two-necked flask, cooled to 0 ° C, and slowly added Boc ₂ O (50 mg) dissolved in dichloromethane (5 ml) The solution was added dropwise for about 15 minutes. Warm to room temperature and continue stirring for 30 minutes. The reaction mixture was washed with sodium chloride solution, dried and filtered, to give Compound 18 (100mg) and concentrated under reduced pressure. MS: 492 (M+H) ⁺ .

Step 9 Synthesis of Compound 19

Compound 18 (80 mg) was dissolved in a solution of methanol (15 ml), placed in a 100 ml sealed tube, and ammonia gas (2.0 g) was passed at 0 °C. The sealed reaction system was warmed to 70 ° C and stirred for reaction overnight. The reaction mixture was concentrated to give Compound 19 (EtOAc). MS: 477 (M+H) ⁺ .

Step 10 2- (4- (1-methoxymethyl-3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-acyl-7-amine hydrochloride Salt (product 2)

Compound 19 (80 mg) was dissolved in dichloromethane (20 ml), stirred in a 50 ml two-necked flask, cooled to 0 ° C, dried hydrogen chloride gas for 30 minutes, filtered, and dried under vacuum to give product 2 (15 mg) ). ^{1 H NMR (400MHz, DMSO)} : δ 9.32 (1H, d), 8.12-8.10 (2H, d), 8.10-8.01 (2H, m), 7.73-7.71 (2H, d), 7.28-7.25 (1H, m), 5.67-5.92 (2H, s) 3.51-3.14 (4H, m), 2.59-2.79 (4H, m) 3.07 (3H, s). 1.88-1.26 (2H, m), 0.27-0.52 (2H, m). MS: 413 (M+H) ⁺ .

The product of Example 3-32 was prepared in a similar manner to that of Example 1.

Example 33

2-(4-(3-Azabicyclo[3.1.0]hexane-1-yl)phenyl)-2H-indazole-7-carboxamide hydrochloride (product 33)

Step 1 Synthesis of Compound 20

Under the protection of nitrogen, tert-butyl-1-(4-aminophenyl)-3-azabicyclo[3.1.0]heptane-3-carboxylic acid (211 mg), methyl-2-nitro-3-methyl The mercaptobenzoic acid (178 mg) and methanol (6 ml) were placed in a 50 ml single-necked flask, and the mixture was warmed to 75 ° C for 3 hours, and concentrated under reduced pressure to give compound 20 (350 mg). MS: 466 (M+H) ⁺ .

Step 2 Synthesis of Compound 21

Under a nitrogen atmosphere, compound 20 (350 mg), sodium azide (50 mg) and DMF (10 ml) were placed in a 50 ml sealed tube and warmed to 95 ° C overnight. After being cooled to room temperature, a saturated sodium chloride solution and ethyl acetate were added, and the organic phase was washed three times with a saturated sodium chloride solution, dried, filtered, and concentrated under reduced pressure to afford compound 21 (160 mg). MS: 434 (M+H) ⁺ .

Step 3 Synthesis of Compound 22

Compound 21 (160 mg) and methanol (15 ml) were placed in a 100 ml sealed tube, cooled to 0 ° C, and ammonia gas (3.0 g) was introduced thereto, and then the reaction system was sealed. The reaction was stirred at 75 ° C for 36 hours. After cooling to room temperature, it was concentrated under reduced pressure to give Compound 22 (150 mg). MS: 419 (M+H) ⁺ .

Step 4 2- (4- (3-azabicyclo [3.1.0] hexane-1-yl) phenyl) -2 H - indazole-acyl-7-amine hydrochloride (product 33)

Compound 22 (150 mg) was dissolved in a dichloromethane (35 ml) solution, placed in a 100 ml three-necked flask, cooled to 0 ° C, and allowed to dry with hydrogen chloride gas for 20 minutes and then stirred for 30 minutes. Filtration and drying of the filter cake in vacuo gave product 33 (100 mg). ^{1 H NMR (400MHz, DMSO)} : δ 9.32 (1H, s), 8.10-8.12 (2H, d), 8.06-8.04 (1H, d), 8.02-8.00 (1H, d), 7.49-7.51 (2H, d), 7.28-7.25 (1H, m), 3.77-3.74 (1H, d), 3.55-3.51 (2H, d), 3.15-3.47 (1H, d), 2.25-2.22 (1H, m) 1.50-1.47 (1H, m), 1.21-1.16 (1H, m). MS: 355 (M+H) ⁺ .

Example 34

2-(4-(octahydrocyclopenta[c]pyrrole-3a-yl)phenyl)-2H-indazole-7-carboxamide hydrochloride (product 34)

Step 1 Synthesis of Compound 23

4-(2-Benzyl octahydrocyclopentadienyl[c]pyrrole-3a-yl)aniline (195 mg), methyl-2-nitro-3-aldehydebenzoic acid (153 mg) and ethanol under nitrogen (20 ml) was placed in a 100 ml single-mouth bottle and warmed to reflux overnight. The mixture was then cooled to room temperature, to give compound 23 (320mg), and concentrated under reduced pressure and used without further purification. MS: 484 (M+H) ⁺ .

Step 2 Synthesis of Compound 24

Under a nitrogen atmosphere, compound 23 (300 mg), sodium azide (20 mg) and DMF (10 ml) were placed in a 50 ml single-necked flask and warmed to 90 ° C overnight or TLC (PE: EA = 3:1) to complete the reaction. After cooling to room temperature, sodium chloride solution and ethyl acetate were added, and the organic phase was washed with sodium chloride solution, dried, filtered, and concentrated under reduced pressure to afford compound 24 (130 mg). MS: 452 (M+H) ⁺ .

Step 3 Synthesis of Compound 25

Compound 24 (80 mg) and methanol (15 ml) were placed in a 100 ml sealed tube, and ammonia (2 g) was introduced. The reaction system was sealed and then warmed to 70 ° C overnight. The reaction mixture was concentrated under reduced pressure to give compound 25 (60 mg). . MS: 437 (M+H) ⁺ .

Step 4 2-(4-(octahydrocyclopenta[c]pyrrole-3a-yl)phenyl)-2H-indazole-7-carboxamide hydrochloride (product 34)

The compound 25 (10 mg), methanol (5 ml) and Pd/C (5 mg) catalyst were stirred in a 50 ml single-necked flask, and reacted with H ₂ at room temperature overnight, the reaction was completed, filtered, and concentrated under reduced pressure to give product 34 (6 mg). . MS: 384 (M+H) ⁺ .

Example 35

2-(2,3-Dihydrobenzo[b][1,4]dioxine-6-yl)-2H-indazole-7-carboxamide (product 35)

Step 1 Synthesis of Compound 26

Catechol (2g) and sodium hydroxide (1.8g) were dissolved in DMF (20ml), placed in a 50ml single-mouth bottle, and 1,2-dichloroethane (2.7g) was added, and the temperature was raised to 90 ° C overnight. . The temperature was lowered to room temperature, and the pH was adjusted to about 5-6 by dilute hydrochloric acid, and extracted with ethyl acetate three times. The organic phase was combined, concentrated under reduced pressure and purified by column chromatography to afford compound 26 (2.1 g). MS: 137 (M+H) ⁺ .

Step 2 Synthesis of Compound 27

Concentrated nitric acid (2.5 ml) was placed in a 50 ml three-necked flask, and the temperature was lowered to 0 ° C. Compound 1 (200 mg) was added in portions, the addition time was continued for about 5 minutes, and the stirring was continued for 5 minutes. The reaction mixture was poured into ice water, filtered, and then filtered and dried in vacuo. MS: 182 (M+H) ⁺ .

Step 3 Synthesis of Compound 28

Compound 27 (200 mg) was dissolved in a solution of methanol (20 ml), Pd/C (100 mg) catalyst was added, and the mixture was placed in a 50 ml single-mouth flask, and H _{2 was} reacted overnight at room temperature. The mixture was then filtered and the filtrate concentrated under reduced pressure to give Compound 28 (190mg). MS: 152 (M+H) ⁺ .

Step 4 Synthesis of Compound 29

Under a nitrogen atmosphere, compound 28 (190 mg), methyl-2-nitro-3-carbamidobenzoic acid (20 ml) and ethanol (20 ml) were placed in a 50 ml single-necked flask and warmed to 75 ° C overnight. After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure to give Compound 29 (220 mg). MS: 343 (M+H) ⁺ .

Step 5 Synthesis of Compound 30

Under a nitrogen atmosphere, Compound 29 (220 mg), sodium azide (60 mg) and DMF (20 ml) were placed in a 50 ml single-necked flask and stirred and warmed to 95 ° C overnight. After completion of the reaction, the mixture was poured into ice water, and extracted with ethyl acetate. The organic layer obtained was concentrated under reduced pressure and purified by column chromatography to afford compound 30 (200 mg). MS: 311 (M+H) ⁺ .

Step 6 2-(2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)-2H-indazole-7-carboxamide (product 35)

Compound 30 (200 mg) and methanol (30 ml) were placed in a sealed tube, and after passing ammonia (2 g), the reaction system was sealed and heated to 70 ° C overnight. The mixture was then concentrated under reduced pressure, ethyl acetate / petroleum ether to give product 35 (120mg). ¹ H NMR (400 MHz, DMSO _, ): δ 9.22 (1H, s), 8.54 (1H, s s), 8.05-8.03 (1H, d), 8.00-7.98 (1H, d), 7.85 (1H, br s ), 7.70 (1H, s), 7.64-7.61 (1H, d), 7.27-7.23 (1H, d), 7.10-7.08 (1H, d), 4.34 (4H, s). MS: 296 (M+H) ⁺ .

Example 36

2-(3,4-Dihydro-2H-benzo[b][1,4]dioxe-7-yl)-2H-indazole-7-carboxamide (product 36)

The method and conditions used for the synthesis of product 36 were similar to those of Example 35 , in which 1,3-dichloropropane was used in place of 1,2-dichloroethane. _{^{1 H NMR (400MHz, DMSO,}} ): δ 9.26 (1H, s), 8.54 (1H, s), 8.06-8.04 (1H, d), 8.00-7.98 (1H, d), 7.86 (1H, s), 7.80-7.79 (1H, d), 7.76-7.73 (1H, d), 7.28-7.26 (1H, d), 7.20-7.18 (1H, d), 4.27-4.20 (4H, m), 2.20-2.15 (2H , m). MS: 310 (M+H) ⁺ .

The products of Example 37 - Example 45 are shown in the table below.

Pharmacodynamic experiment

Kinase level test

A Trevigen colorimetric assay kit was used to screen for PARP-1 inhibitors and to detect PARP-1 activity.

experiment method:

A. Ribosylation reaction

1. Remove the strips, add 50 μL of 1xPARP buffer hydrated histone to each well, incubate for 30 min at room temperature, discard the liquid in the wells, and beat on a paper towel to remove residual buffer.

2. Add serial dilutions of the inhibitor to the appropriate wells (10 μL/well).

3. Add serially diluted PARP enzyme (0.15 U/well) to the wells and incubate for 10 min at room temperature. (Note: i. Positive control group: including 0.15 U of PARP enzyme, but no inhibitor, as a 100% active control. ii. Negative control group: a negative control without PARP enzyme to determine the background absorption value.)

4. Add 25 μL of 1xPARP cocktail to each well with a lance and incubate for 60 min at room temperature.

B. Testing

1. Wash the plate twice with 1xPBS + 0.1% Triton X-100 (1x PBST, 200 μL/well), wash the plate twice with 1x PBS, and beat on a paper towel to wash off the residual liquid each time.

2. Add 50 μL of Strep-HRP to each well and incubate for 60 min at room temperature.

3. Wash the plate twice with 1x PBST (200 μL/well), wash the plate twice with 1×PBS, and beat it on a paper towel to wash off the residual liquid each time.

4. Add TACS-Sapphire chromogenic substrate solution (50 μL/well) pre-warmed to room temperature, and react at room temperature for 15 min in the dark. The reaction was stopped by adding 50 μl of 0.2 M HCl per well, and the absorbance value was measured at 450 nm.

The results of the study are shown in Table 1 .

Cell proliferation assay

experiment method:

Cell proliferation was measured by the MTS method. Hcc1937, T47D, NCI-H69, HGC27 and MKN45 cells were cultured in RPMI1640 medium, MDA-MB-436, SK-OV-3, MGC803 cells were cultured in DMEM medium, SK-N-MC cells were cultured in MEM medium, and all cells were grown. In a culture medium containing 10% fetal calf serum, in a culture environment of 37 ° C, 5% CO 2 . The cells in the logarithmic growth phase were used for testing and counting. The viability of trypan blue staining cells was above 98%, and the cell density was adjusted to 1.5×10 ⁴ or 2×10 ⁴ cells/mL with the corresponding culture medium in 96-well plates. 100 μL of the cell suspension was added to each well (3 replicate wells per concentration), and the final seeding density of the cells was 1.5 × 10 ³ or 2 × 10 ³ /well. The next day, the test drug and the positive drug were dissolved in DMSO as a stock solution, and 2 μL of the drug stock solution was added to 1 mL of the culture solution. Discard the original culture solution in the well, add 200 μL of the drug-containing culture solution to each well, and the final concentration of the drug is 0, 0.03, 0.1, 0.3, 1, 3, 10, 30, and 100 μM, and continue to incubate in the incubator. 10 days, then tested by MTS method. Prepare MTS/PMS mixture before use, add 20μLMTS/PMS mixture to 100μL culture medium, continue to incubate for 1-4 hours in 37°C, 5% CO2 culture environment, and read absorbance value at 490nm using Victor X5 microplate reader. .

The results of the study are shown in Table 2 .

Intracellular ADP ribose (PAR) detection

DNA damage, PARP-1 as a major member of the PARP family, was quickly initiated. Once PARP-1 is initiated, NAD ⁺ is consumed to synthesize highly negatively charged polymer poly ADP ribose (PAR). The PAR content in the cells was measured using a high-throughput chemiluminescence ELISA kit from Trevigen.

experiment method:

A. Prepare cell extracts

1. adherent or suspension cells at 2-10x10 ⁶ cells were inoculated in 6-well plates. The next day, the original culture solution was discarded, and 2 mL of the drug-containing culture solution was added to each well.

2. Suspension washing: Transfer the cell suspension to a 5 mL pre-cooled centrifuge tube, centrifuge at 1500 rpm for 5 min at 4 ° C, discard the supernatant, wash at least once with 4 mL of ice PBS, and suspend the cell pellet in 1 mL PBS and transfer. Transfer to a 1.5 mL centrifuge tube, centrifuge at 2000 rpm for 5 min at 4 ° C, and discard the supernatant.

Adherent cells: discard the original culture solution, add 5 ml of PBS, discard the PBS, repeat at least once, place the plate on ice, scrape the cells with a cell scraper, transfer to a 1.5 mL centrifuge tube, centrifuge at 2000 rpm, and centrifuge at 5 °C for 5 min. Discard the supernatant.

3. Resuspend the cells in 50-100 μL of cell lysate and incubate for 15 min on ice.

4. Add 20% SDS to a final concentration of 1%.

5. Incubate at 100 ° C for 5 min and cool to room temperature.

6. Add 0.01 volumes of 100x magnesium cations and 2 μL of DNase I, mix and incubate at 37 °C for 90 min in order to remove the viscosity of the DNA and extracts in the cells.

7. Centrifuge at 12000 rpm for 10 min at 4 ° C to remove cell debris and store the supernatant.

8. Determine the protein concentration in the extract by the BCA method, measure the PAR content immediately or store the extract at -80 °C.

B. Detection steps:

1. Remove the coated slats from the foil pouch.

2. Dilute the PAR standard and the sample to be tested with a sample buffer series.

3. Add 50 μL of PAR standard, sample to be tested and sample buffer (background control) to each well and set 2 replicate wells.

4. To maximize binding, cover each well with a parafilm and incubate overnight (16 ± 1 hour) at 4 °C.

5. Dilute the PAR polyclonal detection antibody 250-fold with the antibody dilution and incubate for 1 hour at room temperature before use. The strip was removed at 4 ° C, the strip was carefully torn off, and the plate was washed 4 times with PBST (200 μL/well), and the paper towel was beaten to remove the residual liquid.

6. Add 50 μL of PAR polyclonal detection antibody to each well, cover the strip with a parafilm, and incubate at room temperature for 2 hours at 25 °C.

7. Dilute the IgG-HRP conjugate 250-fold with the antibody dilution and incubate for 1 hour at room temperature before use. The strips were removed, and the strips were carefully torn off, and the plate was washed 4 times with PBST (200 μL/well), and the paper towel was beaten to remove the residual liquid.

8. Add 50 μL of goat anti-rabbit IgG-HRP conjugate to each well, cover the strip with a parafilm, and incubate for 1 hour at room temperature at 25 °C. The PeroxyGlow ^TM A and B preheated to room temperature.

9. Carefully tear the seal strip, wash the plate 4 times (200 μL/well) with PBST, and beat it on a paper towel to remove the remaining liquid.

10. Before use, the PeroxyGlow ^TM A and B are mixed in the same volume, 100 L added per well, luminosity value detected immediately.

The results of the study are shown in Table 3 .

Transplanted tumor model

PARP-1 plays an important role in DNA single-strand repair by scissor excision repair. DNA alkylating agents, such as temozolomide (TMZ) and carboplatin, cause DNA single-strand damage, and it is necessary to initiate base excision repair. If the activity of PARP-1 is inhibited, base excision repair cannot be initiated, DNA single Chain breaks cannot be repaired, eventually leading to DNA double-strand breaks. The present invention is a potent PARP-1 inhibitor, and the compound as described in Example 33 in combination with a chemotherapeutic agent can improve the antitumor effect of the chemotherapeutic drug in a tumor xenograft model in vivo.

Human breast cancer cells MX-1 and mouse melanoma cells B16F10 were expanded and collected, and inoculated subcutaneously into the right flank of BALB/c nude mice and C57 mice. Tumor growth was measured by measuring the tumor volume twice a week, and the tumor volume inhibition rate (tumor growth inhibition rate) was calculated in comparison with the control group, and the change in body weight of the animal was measured to reflect the toxicity of the drug.

In the MX-1 xenograft model, Example 33 (30, 60, 120 mg/kg, po, qd) was treated with carboplatin (10 mg/kg, ip), and Example 33 (120 mg/kg) was administered alone with a TGI of 40.98. %, carboplatin (10mg/kg) TGI was 13.22%, and Example 33 (120mg/kg) combined with carboplatin TGI was 68.64%. The combination therapy was significantly better than carboplatin alone. In the B16F10 model, Example 33 (30, 60, 120 mg/kg, po) was administered from day 1 to day 11 of the group, and TMZ (50 mg/kg, po, qd) was administered from day 1 to day 5, The TGI of TMZ (50 mg/kg) alone was 27.8%, and the TGI of Example 33 (30, 60, 120 mg/kg) combined with TMZ (50 mg/kg) were 61.0, 69.8 and 73.1%, respectively, Example 33 (30, 60, The combination of 120mg/kg) and TMZ was significantly better than TMZ alone. The mice of Example 33 and carboplatin/TMZ combination were well tolerated, and the animals in each group were stable during the test, and no toxicity of the drug was found.

The above embodiments are merely illustrative of the specific embodiments of the present invention. The scope of the present invention is defined by the scope of the claims, and is not limited to the specific embodiments described above. It is also within the scope of the present invention to make equivalent substitutions or alterations of the present invention without departing from the scope of the invention.

Claims (42)

a compound of the formula (I), a pharmaceutically acceptable salt thereof, or a mixture of the two, Wherein A and B are each independently selected from CR ₃ or N; R _{3 is} selected from H, halogen, C _1-6 alkyl or halo C _1-6 alkyl; R _{1 is} selected from H, OH or C _{1- 6} alkyl; R _{2 is} selected from H, halogen, C _1-6 alkyl, halo C _1-6 alkyl, C _1-6 alkoxy or halo C _1-6 alkoxy; X is selected from or Wherein m and n are each independently selected from 1, 2 or 3; R _{4 is} selected from H or halogen; R _{5 is} selected from H, -OH, halogen, C _1-6 alkyl, halo C _1-6 alkane a group, a C _1-6 alkoxy group, a halogenated C _1-6 alkoxy group, a C _1-6 alkyl group (NR ₁₁ R ₁₂ ) or Wherein R ₁₁ and R ₁₂ are each independently selected from C _1-6 alkyl; v is selected from 0, 1 or 2; and R _{9 is} selected from H, C _1-6 alkyl, halo C _1-6 alkyl Or C _2-6 alkenyl; R _{10 is} selected from H, C _1-4 alkyl or C _2-4 alkenyl; or R ₉ and R ₁₀ together with the atoms to which they are attached form dihydrofuran or tetrahydrofuran; a 5-15 membered non-aromatic monocyclic ring selected from methyl substituted or unsubstituted, containing at least 2 or more heteroatoms selected from O or S; p is selected from 1 or 2; each R _{6 is} independently selected from H , halogen, C _1-6 alkyl or halogenated C _1-6 alkyl. A compound, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R _{1 is} selected from the group consisting of H, methyl or OH. The compound of claim 1, the pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R _{2 is} selected from the group consisting of H, halogen, C _1-6 alkyl, C _1-6 alkoxy Or a halogenated C _1-6 alkoxy group. The compound of claim 3, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R _{2 is} selected from the group consisting of H, F, methyl, methoxy or chloro substituted methoxy. A compound, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein A is CR ₃ . The compound of claim 5, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R _{3 is} selected from H or C _1-6 alkyl. The compound according to claim 1, the pharmaceutically acceptable salt thereof, or a mixture of the two, wherein B is N. The compound of claim 1, the pharmaceutically acceptable salt thereof, or a mixture of the two, wherein X is . The compound of claim 8, the pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R _{4 is} selected from H. The compound of claim 1, the pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R _{5 is} selected from the group consisting of H, OH, C _1-6 alkyl, C _1-6 alkoxy , C _1-6 alkyl (NR ₁₁ R ₁₂ ) or The compound of claim 10, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R ₅ is . The compound of claim 10, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein v is selected from 0 or 1. The compound of claim 10, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R _{9 is} selected from the group consisting of H, C _1-6 alkyl or C _2-6 alkenyl. The compound of claim 10, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R _{10 is} selected from the group consisting of H, C _1-3 alkyl or C _2-3 alkenyl. The compound of claim 10, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R ₉ and R ₁₀ together with the atoms to which they are attached form dihydrofuran or tetrahydrofuran. The compound of claim 10, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R _{5 is} selected from C _1-6 alkyl (NR ₁₁ R ₁₂ ), and R ₁₁ and R ₁₂ are each independently selected from a C _1-3 alkyl group. The compound of claim 16, the pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R ₁₁ and R ₁₂ are each a methyl group. The compound of claim 16, the pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R ₅ is . The compound of claim 1, the pharmaceutically acceptable salt thereof, or a mixture of the two, wherein X is . The compound of claim 19, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein p is selected from 1 or 2. The compound of claim 20, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R _{6 is} independently selected from H or C _1-6 alkyl. The compound of claim 20, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein Z is selected from methyl substituted or unsubstituted, at least 2-5 selected from O or A 5-15 member non-aromatic monocyclic ring of a hetero atom of S. The compound of claim 20, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein Z is selected from methyl substituted or unsubstituted, and contains at least 2, 3, 4 or 5 A 5, 6, 7, 8, 9, 11, 12 or 15 membered non-aromatic monocyclic ring selected from heteroatoms of O or S. The compound of claim 20, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein Z is selected from the group consisting of , , , , , , , The compound of claim 1, the pharmaceutically acceptable salt thereof, or a mixture of the two, wherein the compound is as shown in formula (II): Wherein s is selected from 1, 2 or 3; t is selected from 1 or 2; and R _{5 is} selected from the group consisting of H, OH, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkyl (NR ₁₁ R ₁₂ ) or . The compound of claim 25, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein t is 2. The compound of claim 25, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein s is 1 or 3. The compound of claim 25, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R _{5 is} selected from the group consisting of H, OH, C _1-3 alkyl or . The compound of claim 25, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein R ₅ is ; v is 0; R _{9 is} selected from H, C _1-4 alkyl or C _2-4 alkenyl; and R _{10 is} selected from H, C _1-3 alkyl or C _2-3 alkenyl. The compound of claim 1, the pharmaceutically acceptable salt thereof, or a mixture of the two, wherein the compound is as shown in formula (III): Wherein Z _{1 is} selected from 5, 6, 7, 8, 9, 11, 12 or 15 membered non-substituted or unsubstituted, 2, 3, 4 or 5 heteroatoms selected from O or S Aromatic single ring. The compound of claim 30, a pharmaceutically acceptable salt thereof, or a mixture of the two, wherein Z _{1 is} selected from the group consisting of , , , , , , The compound according to claim 1, the pharmaceutically acceptable salt thereof, or a mixture of the two, wherein the compound is: 1) 2-(4-(3-azabicyclo[4.1.0] ] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 2) 2- (4- (1-methoxymethyl-3-azabicyclo [4.1 .0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 3) 2- (4- (1-methyl-3-azabicyclo [4.1. 0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 4) 2- (4- (1-ethyl-3-azabicyclo [4.1.0 ] heptane-one -6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 5) 2- (4- (1-isopropyl-3-azabicyclo [4.1.0 ] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 6) 2- (4- (1-hydroxy-3-azabicyclo [4.1.0] hept- 6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 7) 2- (4- (1- (ethoxymethyl) -3-azabicyclo [4.1 .0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 8) 2- (4- (1- (2-methoxyethyl) -3 - azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-acyl-amine hydrochloride; 9) 2- (4- (1- (1-hydroxy-ene Propyl)-3-azabicyclo[4.1.0]heptane-6-yl)phenyl)-2 H -carbazole-7 -carbamidine hydrochloride; 10) 2-(4-(1-(hydroxymethyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-2 H -carbazole -7-carbamidine hydrochloride; 11) 2-(4-(1-(1-hydroxyethyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-2 H -carbazole-7-formamide hydrochloride; 12) 2-(4-(1-(1-hydroxypropyl)-3-azabicyclo[4.1.0]heptan-6-yl)benzene yl) -2 H - indazole-7-amine hydrochloride acyl; 13) 2- (4- (1- (1-hydroxybutyl) -3-azabicyclo [4.1.0] heptane -6 - yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 14) 2- (4- (1- (1-hydroxy-3-methylbutyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 15) 2- (4- (1- (1-hydroxy-3-en yl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 16) 2- (4- (1- ( 1-methoxyethyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 17) 2- ( 4- (1- (1-methoxypropyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-acyl-7-amine hydrochloride ; 18) 2- (4- (1- (1-methoxybutyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7 Formamide hydrochloride; 19) 2-(4-(1- (1-methoxy-3-methylbutyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-acyl-7-amine hydrochloride ;20) 2-(4-(1-(1-methoxybut-3-enyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-2 H -indole Zylo-7-carbamide hydrochloride; 21) 2-(4-(1-(1-ethoxybut-3-enyl)-3-azabicyclo[4.1.0]heptane-6- yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 22) 2- (4- (1- (1- (allyloxy) allyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 23) 2- (4- (1- (2,3-dihydrofuran - 2- yl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 24) 2- (4- (1 - (tetrahydrofuran-2-yl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 25) 2- ( 4-(1-(Methoxymethyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-5-methyl- 2H -carbazole-7-carboxamide ; 26) 5-fluoro-2- (4- (1- (methoxymethyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole - 7-carbamamine; 27) 5-methoxy-2-(4-(1-(methoxymethyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl) -2 H -carbazole-7-formamide; 28) 2-(4-( 1-(methoxymethyl)-3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-N-methyl- 2H -indazole-7-carboxamide;29) N- hydroxy-2- (4- (1- (methoxymethyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -2 H - indazole-7-carboxylic Amides; 30) 2- (4- (1- (methoxymethyl) -3-azabicyclo [4.1.0] heptane-6-yl) phenyl) -3-methyl -2 H - Oxazole-7-formamide; 31) 3-chloro-2-(4-(1-(methoxymethyl)-3-azabicyclo[4.1.0]heptane-6-yl)phenyl ) -2 H - indazole-7-Amides; 32) 2- (4- (1- (2- (dimethylamino) ethyl) -3-azabicyclo [4.1.0] heptane - 6-yl) phenyl) -2 H - indazole-7-Amides; 33) 2- (4- (3-azabicyclo [3.1.0] hexane-1-yl) phenyl) -2 H - indazole-7-amine hydrochloride acyl; 34) 2- (4- (octahydro-cyclopenta [c] pyrrol -3a--yl) phenyl) -2 H - indazole-7-XI hydrochloride; 35) 2- (2,3-dihydrobenzo [b] [1,4] dioxin-6-yl) -2 H - indazole-7-Amides; 36) 2- (3,4-dihydro -2H- benzo [b] [1,4] dioxepin-7-yl) -2 H - indazole-7-Amides; 37) 2 -(Benzo[b][1,4]dioxooctyl)-2H-indazole-7-formamide; 38)2-(benzo[d][1,3]dioxole Alkene-5-yl)-2H-carbazole-7-formamide; 39) 2-(2,2-di Benzo[d][1,3]dioxol-5-yl)-2H-carbazole-7-formamide; 40)2-(2,3,5,6,8,9 - hexahydrobenzo[b][1,4,7,10]tetraoxacyclododecyl)-2H-indazole-7-carboxamide; 41)2-(2,3,5,6 -tetrahydrobenzo[b][1,4,7]trioxinyl)-2H-carbazole-7-formamide 42)2-(2,3,5,6,8,9 ,11,12-octahydrobenzo[b][1,4,7,10,13]pentaoxacyclopentadecyl)-2H-indazole-7-carboxamide;43)2-(2 ,3,5,6,8,9-hexahydrobenzo[b][1,4,7,10]tetraoxacyclododecyl)-2H-indazole-7-carboxamide; 44) 2-(2,3,4,6,7,8-hexahydrobenzo[b][1,4,8]-1,4-dioxo-8-thio-11-yl)-2H- Carbazole-7-formamide; 45) 2-(2,3,4,6,7,8-hexahydrobenzo[i][1,7,4]-1,4-dioxo-8 -mercapto-11-yl)-2H-carbazole-7-formamide; 46) 2-(4-(3-azabicyclo[4.1.0]heptan-6-yl)phenyl)-3a , 7a-dihydro-1H-benzo[d]imidazole-4-carboxamide hydrochloride. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 32, a pharmaceutically acceptable salt thereof, or a mixture of both and a pharmaceutically acceptable excipient. A compound according to any one of claims 1 to 32, a pharmaceutically acceptable salt thereof, or a mixture thereof, or a pharmaceutical composition according to claim 33, For the preparation of a medicament for treating, preventing or delaying a disease which can be alleviated by inhibition of poly(ADP-ribose) polymerase, or in the preparation of a medicament for treating, preventing or delaying cancer, or when preparing for use as a cancer treatment a chemotherapeutic agent or a radiosensitizer. The application according to claim 34, wherein the medicament is for treating, preventing or delaying cancer, inflammatory diseases, reperfusion injury, ischemic diseases, stroke, acute and chronic renal failure, cardiovascular diseases Other vascular diseases other than cardiovascular diseases, diabetes, neurodegenerative diseases, anti-retroviral infections, retinal damage, skin aging or ultraviolet-induced skin damage. The application according to claim 35, wherein the inflammatory disease is selected from the group consisting of organ transplant rejection, inflammatory bowel disease, inflammatory lung disease, inflammatory eye disease, chronic inflammatory gingivitis, inflammatory nephropathy, Inflammatory skin disease, inflammatory central nervous system disease, diabetic complications, inflammatory heart disease. The application of claim 35, wherein the ischemic disease comprises anemia and an organ transplant. The application according to claim 35, wherein the diabetes is selected from the group consisting of type I diabetes (insulin dependent diabetes), type II diabetes (non-insulin dependent diabetes), gestational diabetes, autoimmune diabetes, insulin disease, pancreas Disease-induced diabetes, type A insulin resistance syndrome, type B insulin resistance syndrome, adipose atrophic diabetes, or 3-cytotoxin-induced diabetes. The application according to claim 35, wherein the neurodegenerative disease is selected from the group consisting of a neurodegenerative disease caused by prolonged polyglutamine sequence, Huntington's disease, Kendi disease, cerebellar atrophy, teeth Nucleus red nucleus globus pallidus hypothalamic nuclear atrophy (DRPLA), protein aggregation-related neurodegenerative diseases, Machado-Joseph disease, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Spongiform encephalopathy, prion-related diseases, and multiple sclerosis (MS). The application of claim 35, wherein the cancer is selected from the group consisting of a solid tumor, a blood-borne cancer, acute or chronic leukemia, lymphoma, central nervous system cancer (CNS), brain cancer, breast cancer, ovary Cancer, pancreatic cancer, prostate cancer, small cell lung cancer, gastric cancer, neuroepithelial neoplasia, cancer lacking homologous recombination (HR)-dependent DNA double-strand break (DSB) repair activity, BRCA-1 or BRCA-2 deficiency tumor. The use according to claim 40, wherein the cancer lacking homologous recombination (HR)-dependent DNA double-strand break (DSB) repair activity comprises one or more homologs compared to normal cells. Recombinant DNA double-strand break repair ability is reduced or completely absent in cancer cells. The use according to claim 41, wherein the cancer cell has a BRCA-1 deficient phenotype or a BRCA-2 deficient phenotype, or the cancer cell lacks BRCA-1 or BRCA-2.